text:
|Home | About | Journals | Submit | Contact Us | Français
Whole-cell patch-clamp recordings and high resolution morphometry were used to assess functional and structural properties of layer 3 pyramidal neurons in early (< 4 months) and advanced (> 8 months) stages of tauopathy in frontal cortical slices prepared from rTg4510 tau mutant (P301L) mice. In early tauopathy, dendritic architecture is preserved. In advanced tauopathy, neurons can be categorized as either "atrophic" (58%)- exhibiting marked atrophy of the apical tuft, or "intact" (42%)- with normal apical tufts and, in some instances, proliferative sprouting of oblique branches of the apical trunk. Approximately equal numbers of atrophic and intact neurons contain neurofibrillary tangles (NFTs) or are tangle-free, lending further support to the idea that NFTs per se are not toxic. Spine density is decreased due to a specific reduction in mushroom spines, but filopodia are increased in both atrophic and intact neurons. By contrast to these morphological changes, which are robust only in the advanced stage, significant electrophysiological changes are present in the early stage and persist in the advanced stage in both atrophic and intact neurons. The most marked of these changes are: a depolarized resting membrane potential, an increased depolarizing sag potential and increased action potential firing rates- all indicative of hyperexcitability. Spontaneous excitatory postsynaptic currents are not reduced in frequency or amplitude in either stage. The difference in the time course of functionally important electrophysiological changes versus regressive morphological changes implies differences in pathogenic mechanisms underlying functional and structural changes to neurons during progressive tauopathy.
During the pathogenesis of neurodegenerative tauopathies, the microtubule-associated protein tau becomes hyperphosphorylated, disengages from microtubules and missorts from the axon to the somatodendritic compartment where it ultimately aggregates into a mature neurofibrillary tangle (NFT). NFTs were long believed to contribute to neuronal dysfunction and ultimately lead to neuron death in tauopathies such as Alzheimer's disease (AD), in which the number of NFTs correlates with the degree of neuron loss and the severity of cognitive impairment [16,4,17]. However, there is now strong evidence that NFTs per se are not toxic, but rather that soluble hyperphosphorylated tau species, present before and during NFT aggregation, are the key pathogenic entities (review [50]). Evidence that soluble hyperphosphorylated tau species drive neuronal dysfunction and degeneration in tauopathy has been generated primarily from studies of the regulatable rTg4510 mouse model, which expresses mutant (P301L) human tau and closely recapitulates key characteristics of human tauopathy [15,47,49,19,25,5]. Early in the progression of tauopathy in these mice (< 4 months of age) many different soluble hyperphosphorylated tau species are present in the neocortex, but NFTs and neurodegeneration are largely absent. Soluble hyperphosphorylated tau species persist in advanced tauopathy (> 8 months of age), when NFTs are also prevalent and neuron loss in the frontal cortex is extensive [41,47,49]. We have recently shown that individual layer 3 frontal cortical pyramidal neurons from 9-month-old rTg4510 mice exhibit major structural and functional changes independent of the presence or absence of a mature NFT [9,43]. These changes include: loss of the apical dendritic tuft; ~30% reduction in dendritic spine density; increased density of axonal boutons; depolarized resting membrane potential; markedly increased depolarizing sag potential; increased action potential firing rates, and; increased frequency of low amplitude excitatory synaptic currents [43,9].
In the present study, we extended our analyses of layer 3 frontal cortical pyramidal neurons in rTg4510 mice to a younger "early stage tauopathy" cohort (1 to 3 months of age). In addition, by expanding both the number of neurons and the age of mice assessed in the "advanced stage tauopathy" cohort (9 to 13 months of age), we were able to differentiate morphologically distinct types of neurons -atrophic and intact- and to demonstrate that this distinction occurs independent of the presence or absence of a mature NFT. Neurons from rTg4510 mice treated with lifelong doxycycline to suppress expression of the mutant tau transgene exhibited functional and structural properties that did not differ from those of neurons from non-transgenic littermates. These studies revealed that significant electrophysiological changes occur in early tauopathy and persist in the advanced stage, while morphological changes are prominent only in the advanced stage. Thus, we provide evidence for a dissociation between functional and structural changes to individual neurons during progressive neurodegenerative tauopathy in this widely used mouse model.
A total of 19 rTg4510 (TG) and 17 age-matched non-transgenic (NT) littermate controls (strain FVB/129) were used in this study. Two age cohorts were used: an "early" 1- to 3-month-old cohort (n: TG- 8, NT- 8 mice) and an "advanced" 9- to 13-month-old cohort (TG: 11 mice, NT: 9 mice). Mice treated to suppress the tau transgene were aged to 9 months (n: TG- 5, NT- 3 mice) and received doxycycline (dox) from conception and then first in drinking water and subsequently in chow containing 200mg/kg dox (Harlan Teklad, South Easton, MA) to the time of sacrifice. Mice were group housed in a pathogen-free barrier facility on a 12/12-hour light/dark cycle with unrestricted access to food and water. DNA extracted from tail samples was screened for both activator and responder transgenes as described using standard PCR methodology [41]. All experiments were approved by the Boston University Institutional Animal Care and Use Committee (IACUC) and were conducted in strict accordance with the animal care guidelines outlined in the NIH Guide for the Care and Use of Laboratory Animals and the U.S. Public Health Service Policy on Humane Care and Use of Laboratory Animals.
Mice were sacrificed by decapitation and their brains were extracted and placed in oxygenated (95% O2 and 5% CO2) ice-cold Ringer's solution, concentrations in mM: 25 NaHCO3, 124 NaCl, 1 KCl, 2 KH2PO4, 10 Glucose, 2.5 CaCl2, and 1.3 MgCl2, (pH 7.4; Sigma-Aldrich, St. Louis, MO). After blocking, frontal cortical hemispheres were sectioned into 8 to 10 coronal sections (300 µm thick) with a vibrating microtome. Slices were maintained in oxygenated room temperature (RT) Ringer's solution for equilibration for at least 1 hour. Following equilibration, individual slices were situated in submersion-type recording chambers (Harvard Apparatus, Holliston, MA) affixed to the stages of Nikon E600 infrared-differential interference contrast (IR-DIC) microscopes (Micro Video Instruments, Avon, MA) for whole-cell patch-clamp recordings. During recordings (~15 min in duration), slices were superfused with oxygenated RT Ringer's solution at a rate of 2.5 ml/min.
Whole-cell patch-clamp recordings were made using patch electrode pipettes fabricated from non-heparinized microhematocrit capillary tubes (Thermo Fisher Scientific, Pittsburg, PA) on a Flaming and Brown horizontal pipette puller (Model P-87, Sutter Instrument, Novato, CA). All neurons from which recordings were obtained were layer 3 frontal (dorsal premotor) cortical pyramidal cells identified under IR-DIC optics as previously described [43,9]. Electrodes were filled with potassium methanesulfonate internal solution, concentrations in mM: 122 KCH3SO3, 2 MgCl2, 5 EGTA, and 10 NaHEPES containing 1% biocytin (pH 7.4; Sigma-Aldrich, St. Louis, MO) and had resistances from 4 to 6 MΩ when immersed in Ringer's solution. EPC-9 and EPC-10 patch-clamp amplifiers and PatchMaster software were used for data acquisition (HEKA Elektronik, Lambrecht, Germany). Access resistance was monitored throughout the experiment and all recordings were low-pass filtered at 10 kHz.
Passive membrane and action potential (AP) firing properties were assessed in current-clamp mode as previously described [43]. Passive membrane properties were determined using voltage responses to a series of 200 ms hyperpolarizing and depolarizing current steps (10 pA steps, −40 to +40 pA; 12.5 kHz sampling frequency). Resting membrane potential (Vr) was measured as the membrane voltage in the absence of current injection. Responses to current steps were measured at steady state and plotted on a voltage-current plot, and input resistance (Rn) was calculated as the slope of the line of best fit through the linear portion of the graph. Membrane time constant was determined by fitting a single-exponential function to the membrane voltage response to the 10 pA hyperpolarizing current step. Rheobase (amount of current required to evoke the first AP) was assessed with a 10 s depolarizing current ramp (0 to 200 pA; 3.03 kHz sampling frequency). Single AP properties were measured on the first AP elicited in a 200 ms current-clamp series using an expanded timescale and the linear measurement tool in FitMaster analysis software (HEKA Elektronik). Single AP properties assessed included threshold and amplitude (voltage difference from threshold to peak). The event detection tool in FitMaster analysis software was used to evaluate the number of APs generated during a series of 2 s hyperpolarizing and depolarizing current steps (50 pA steps, −170 to +380 pA; 12.5 kHz sampling frequency) and AP firing rates were subsequently calculated. In most neurons, a slow depolarizing voltage sag potential (an H-current-mediated response) was evoked by strong hyperpolarizing current steps. This sag potential was eliminated by the H-current (HCN channel) blocker ZD-7288 (10 µM; Tocris Bioscience, Ellisville, MO) [60]. Depolarizing sag potential amplitudes were measured using the 2 s, −170 pA hyperpolarizing current step with the FitMaster linear measurement tool.
AMPA receptor-mediated spontaneous excitatory postsynaptic currents (sEPSCs) were recorded from a holding potential of −80 mV for a period of 2 min (6.67 kHz sampling frequency). sEPSCs were unaffected by the NMDA-receptor antagonist APV or by the GABAA receptor antagonist BMI, but were fully blocked by application of the non-NMDA receptor antagonist CNQX. MiniAnalysis software was used to quantify synaptic current properties including frequency, amplitude, and kinetics (rise time constant and decay time constant). For assessment of kinetics, the rise and decay of averaged traces were each fit to a single exponential function. The MiniAnalysis detection threshold was set at the maximum of the RMS noise level (5 pA) for all synaptic current analyses.
All neurons from which recordings were obtained were simultaneously filled with 1% biocytin in the internal solution (pH 7.4; Sigma-Aldrich, St. Louis, MO). Following recording, slices containing biocytin-filled neurons were fixed in 4% paraformaldehyde in 0.1M phosphate buffered saline (PBS) solution (pH 7.4) for 4 days at 4°C. After three rinses in PBS (10 min each), slices were bathed in 0.1% Triton X-100/PBS for a period of 2 h at RT. For visualization and subsequent confocal laser scanning of neurons, slices were incubated in Streptavidin-Alexa 546 (1:500; Vector Labs, Burlingame, CA) for 2 days at 4°C. For detection of mature neurofibrillary tangles (NFTs), slices from both TG and NT mice were incubated in filtered 0.1% Thioflavin-S dissolved in PBS/10% ethanol solution (30 min). Prolong Gold mounting medium (InVitrogen, Eugene, OR) was used to mount stained slices on slides, which were then cover-slipped.
A Zeiss LSM-510 confocal laser-scanning microscope outfitted with a 40× Plan Apochromatic 1.3 NA oil-immersion objective lens (210 µm working distance) was used to obtain confocal images for subsequent 3D neuron reconstructions and for assessment of Thioflavin-S staining. Completely filled neurons were imaged in their entirety at a resolution of 0.4 × 0.4 × 0.41 µm per voxel (0.7× digital zoom) and required acquisition of 2 to 4 z-stacks per neuron. To determine whether neurons contained a Thioflavin-S positive NFT, one z-stack was captured through each soma at a resolution of 0.05 × 0.05 × 0.5 µm per voxel (3.0× digital zoom). For spine density and subtype assessments, 100× (2.0× digital zoom) image stacks were acquired using a UPlan-FL 100×/1.3 NA oil-immersion objective lens (100 µm working distance) from one distal apical dendritic branch and from one basal dendritic branch at a resolution of 0.022 × 0.022 × 0.1 µm per voxel; 2 to 4 z-stacks were acquired per dendritic branch. Emissions from Streptavidin-Alexa 546 (Helium/Neon laser excitation) and Thioflavin-S (Argon laser excitation) were detected using two separate channels with filters set at 560 nm and 480 to 520 nm, respectively. To reduce z-plane signal blurring, all confocal z-stacks were subject to deconvolution using AutoQuant software (Media Cybernetics, Bethesda, MD). Following deconvolution, z-stacks obtained for each individual neuron were aligned in 3D and integrated into a single volumetric dataset using Volume Integration and Alignment System (VIAS) software [44]. The distance from the soma to the pial surface was measured for each neuron using the VIAS measure tool.
For 3D reconstruction of individual neurons, integrated volumetric datasets obtained following VIAS z-stack stitching of 40× confocal images were imported into Neurolucida neuron reconstruction software (MBF Bioscience, Williston, VT). First, Neurolucida automatically traced the entire dendritic structure and soma of each neuron and then reconstructions were manually corrected. Reconstruction data were subsequently imported into NeuroExplorer software (MBF Bioscience). Total dendritic length and complexity (number of bifurcation nodes) were assessed for the apical and basal dendritic arbor of each neuron. To determine regional measures of dendritic length and complexity, a Sholl analysis [48] was also used to divide apical and basal dendritic arbors separately into proximal, middle and distal thirds (based on the total 3D extent of the arbor) using concentric rings originating from the center of each soma. The enclosed volume of each soma was also determined using NeuroExplorer software.
Integrated volumetric datasets of 100× confocal images were imported into the 64-bit version of NeuronStudio [57,46; available at: http://www.mssm.edu/cnic]. First, using a Rayburst-based algorithm analysis method, NeuronStudio automatically traced each dendritic segment to generate a dendritic .swc file. Then, spines were automatically detected and then manually corrected on dendritic .swc files [45]. To calculate spine density, the dendritic length and the number of spines for a given dendritic segment were extracted from .swc and .txt files respectively. Spine densities were determined for one complete distal apical dendritic branch and one complete basal dendritic branch per neuron. For spine subtyping analyses, spines on distal apical segments were classified as mushroom-, stubby-, thin- or filopodia-type spines as previously described [9].
Neurons included in this study were required to meet both electrophysiological and morphological inclusion criteria. Electrophysiological criteria were: a resting membrane potential of ≤ −55 mV, stable access resistance (< 10% change over the course of the recording), and the ability to fire a train of APs during sustained depolarizing current injection. Morphological criteria were: a well-filled dendritic arbor with no cut dendrites in the distal apical dendritic arbor, an intact soma, a soma-to-pial surface distance between 220 and 550 µm, and, for TG neurons from 9- to 13-month-old mice, unequivocal Thioflavin-S labeling for classification of neurons as NFT+ or NFT−.
It is important to note that we cannot rule out that brains of transgenic mice are inherently more susceptible to damage during slice preparation. However, TG neurons from the early tauopathy cohort were assessed at an age prior to neurodegeneration and widespread tau pathology in the frontal cortices of TG mice. Thus, we believe that it is unlikely that neurons from TG mice are less resistant to insult than neurons from NT mice at this age. While it is possible that TG neurons may become increasingly vulnerable with increasing age, the membrane potential is depolarized to the same level in TG neurons from younger relative to older mice, suggesting that this finding is indeed a real phenomenon that occurs independent of tissue damage. In addition, basic electrophysiological responses of TG neurons examined in the present study met criteria for "healthy neurons" and in most respects did not differ from NT neurons, exhibiting a regular spiking pattern, high amplitude APs, and an AP threshold close to −30 mV. Lastly, under IR-DIC visualization, somata of all neurons from which recordings were obtained appeared healthy, lacking signs of neuron death such as pyknotic nuclei.
Statistical analyses were performed using SPSS Statistics software version 19 (SPSS Inc., Chicago, IL). A k-means cluster analysis of distal apical dendritic length (followed by a one-way ANOVA) was used to verify morphologically distinct populations of neurons (atrophic vs. intact) from advanced-stage TG mice. A Multivariate ANOVA with group as the fixed factor was used to compare all morphological and electrophysiological parameters. Fisher's Least Significant Difference (LSD) post hoc tests were performed to assess between-group differences. Student's t-tests (two-tailed, equal variance not assumed) were used to compare means between groups where indicated. A Pearson Product-Moment correlation was used to determine relationships between variables. Cumulative histograms were compared using a Kolmogorov-Smirnov (K-S) test. The significance level was p ≤ 0.05 and all data are presented as the mean ± the standard error of the mean.
A total of 18 transgenic (TG) and 23 non-transgenic (NT) neurons in early tauopathy (1- to 3-month-old mice) and 36 TG and 34 NT neurons in advanced tauopathy (9- to 13-month-old mice) met both electrophysiological and morphological criteria for inclusion in the present study. Qualitative and quantitative approaches were combined to classify neurons from early and advanced-stage tauopathy. First, qualitative visual assessment of neuron reconstruction data revealed that while dendritic architecture was no different in TG compared to NT neurons in early tauopathy (Fig. 1a, top and middle rows), many TG (58%) neurons in advanced tauopathy exhibited marked regression of the apical dendritic tuft (Fig. 1a, bottom row, arrowheads). Then, to quantitatively distinguish distinct populations of TG neurons in advanced tauopathy, a cluster analysis was performed on total dendritic length within the distal third of the apical arbor (ANOVA, F(2,32) = 160.9, p < 0.001). Using this analysis, a 500 µm cutoff value was determined and hence the two populations of TG neurons in advanced tauopathy were: 1) neurons with intact apical dendritic tufts and distal apical lengths of > 500 µm, and; 2) neurons with dramatic regression of apical dendritic tufts and distal apical lengths of < 500 µm. These two populations are hereafter referred to as TGintact and TGatrophic respectively. A scatter plot of the distal apical dendritic lengths of neurons is shown in Figure 1b. The 500 µm cutoff value for distal apical length revealed discrete subpopulations of neurons within the advanced TG neuron group only (intact and atrophic); such subpopulations were not evident in the early TG, early NT, or advanced NT groups. When advanced NT and TG neurons were compared without clustering, significant differences between groups were found for each of the same parameters revealed by the cluster analysis (not shown).
Of TGintact neurons, 47% were Thioflavin-S positive (NFT+) and 53% were Thioflavin-S negative (NFT−), and of TGatrophic neurons, 43% were NFT+ and 57% were NFT−. Thus, approximately equal proportions of TGintact and TGatrophic neurons were NFT+ or NFT− indicating that atrophy of the apical tuft occurs in TG neurons regardless of the presence or absence of a mature NFT. A pie chart showing relative proportions of TG neurons in the advanced group that were NFT+ or NFT− and had either an intact or an atrophic apical dendritic tuft is shown in Figure 1c. Confocal images of the somata of representative neurons with and without NFTs are shown in Figure 1d.
Total and regional lengths and complexities of dendritic arbors were determined for each electrophysiologically characterized neuron. A multivariate ANOVA revealed significant differences between groups for all but four measures of dendritic morphology (the complexity of the middle third of the apical arbor; the length of the proximal third of the basal arbor, and; the length and complexity of distal third of the basal arbor). Significant differences in the apical dendritic arbor were: total and regional lengths (total: F(4,105) = 14.8, p < 0.001; proximal: F(4,105) = 7.0, p < 0.001; middle: F(4,105) = 3.2, p = 0.016; distal: F(4,105) = 18.4, p < 0.001) and, total complexity and complexity of the proximal and distal divisions (total: F(4,105) = 8.8, p < 0.001; proximal: F(4,105) = 4.7, p = 0.002; distal: F(4,105) = 9.9, p < 0.001). Significant differences in the basal dendritic arbor were: total length and length of the middle division (total: F(4,106) = 3.4, p = 0.012; middle: F(4,106) = 5.3, p = 0.001) and, total complexity and complexity of the proximal and middle divisions (total: F(4,106) = 8.3, p < 0.001; proximal: F(4,106) = 4.9, p = 0.001; middle: F(4,106) = 5.0, p = 0.001). Lastly, the volume of the soma did not differ significantly between groups (Table 1).
Post hoc comparisons (Fisher's LSD test) showed that in early tauopathy, the total lengths and complexities of the apical dendritic arbor were not significantly different between TG and NT neurons (Table 1). Similarly regional lengths (Fig. 2a, left) and complexities (not shown) did not differ. The total and regional lengths of the basal arbor were not significantly different between the two groups (Table 1; Fig. 2b, left). However, the total complexity of the basal arbor was significantly reduced (p = 0.002; Table 1) due to a decreased number of nodes in the proximal (p = 0.049) and middle (p = 0.006) divisions, but not the distal division in TG neurons (not shown). Thus, changes to dendritic architecture in early tauopathy were restricted to the basal dendritic arbor, which was less complex in TG than in NT neurons.
In advanced tauopathy, the total length of the apical dendritic arbor was not significantly different between TGintact and NT neurons (Table 1). TGintact neurons showed an increase in length in proximal (p = 0.019) but not middle or distal divisions of the apical dendritic arbor compared to NT neurons (Fig. 2a, right). An unusually high number of oblique dendritic branches extending from the main apical trunk were apparent in a number of TGintact neurons (7/15 neurons). These branches are clearly demonstrated by the dendrograms shown in Figure 2a (bottom). Consistent with this observation, there was a significantly greater number of bifurcation nodes in the proximal division of the apical dendritic arbor (p = 0.039; not shown). However, the total complexity (Table 1) and the complexity of the middle and distal divisions of the apical dendritic arbor (not shown) were the same in TGintact and NT neurons. Basal dendritic length of TGintact neurons was significantly decreased in the middle division (p = 0.040; Fig. 2b, right); there were no additional differences in basal length between TGintact and NT neurons. However, the total complexity of the basal dendritic arbors of TGintact neurons was significantly reduced (p = 0.002; Table 1) due to fewer bifurcation nodes in the proximal (p = 0.044) and middle divisions (p = 0.007), but not the distal division (not shown). In summary, changes to dendritic arbors of TGintact neurons in advanced tauopathy occurred within relatively close proximity to the soma, with the most notable changes being increased length and complexity of the apical arbor and decreased complexity of the basal arbor.
Both apical and basal dendritic arbors showed marked regression in TGatrophic neurons. TGatrophic neurons exhibited a significant reduction in total apical length (p < 0.001; Table 1) as well as in all three divisions of the apical dendritic arbor (proximal: p = 0.020; middle: p = 0.019; distal: p < 0.001; Fig. 2a, right). Moreover, the overall complexity of the apical dendritic arbor was reduced in TGatrophic neurons (p < 0.001; Table 1) due to a decreased number of bifurcation nodes in the distal division (p < 0.001), but not the proximal or the middle division (not shown). Total basal dendritic length was significantly reduced in TGatrophic compared to NT neurons (p = 0.001; Table 1) due to a specific reduction in length in the middle division of the arbor (p < 0.001; Fig. 2b, right). TGatrophic neurons also had fewer bifurcation nodes in the proximal division of the basal dendritic arbor (p < 0.001; not shown) that contributed to a significant reduction in total basal dendritic complexity (p < 0.001; Table 1).
The total and regional lengths (total: p < 0.001; proximal: p < 0.001; middle: p = 0.001; distal: p < 0.001) and complexities (total: p < 0.001; proximal: p = 0.001; distal: p < 0.001) of apical dendritic arbors were significantly greater in TGintact versus TGatrophic neurons. Moreover, the total length of the basal arbor was greater in TGintact versus TGatrophic neurons (p = 0.008). Both total and regional complexity of basal dendritic arbors was the same in TGintact and TGatrophic neurons. Thus, while the length of the basal arbor was relatively more preserved in TGintact neurons, both groups of TG neurons in advanced tauopathy exhibit similar reductions in basal complexity.
Representative examples of apical dendritic segments imaged at 100× are shown in Figure 3a. Spine density was the same in apical and basal branches (data not shown) and thus these data were combined for each neuron. There was an overall difference in spine density between groups (F(4,77) = 57.7, p < 0.001). In early tauopathy, spine density was not significantly different in TG compared to NT neurons (Fig. 3a,b). In advanced tauopathy, spine density was significantly reduced in TGatrophic relative to NT neurons (p = 0.016; Fig. 3a,b). While spine density was lower in TGintact compared to NT neurons, this did not reach the significance level (TGintact vs. NT, p = 0.119); TGintact versus TGatrophic did not differ with regard to spine density (Fig. 3a,b). There was an overall difference in the relative proportion (percentage of total spines) of mushroom- (F(4,27) = 9.2, p < 0.001) and filopodia- (F(4,27) = 13.7, p < 0.001) type spines, but not of stubby- or thin-type spines along distal apical dendrites between groups. The relative proportion of mushroom- and filopodia-type spines was no different in TG versus NT neurons in early-stage tauopathy (Fig. 3c, left). However, in the advanced stage, the relative proportion of mushroom-type spines was significantly reduced (TGintact vs. NT, p = 0.001; TGatrophic vs. NT, p = 0.003), while the relative proportion of filopodia was significantly increased in TGintact and TGatrophic neurons relative to NT neurons (TGintact vs. NT, p = 0.011; TGatrophic vs. NT, p = 0.010); TGintact and TGatrophic neurons did not significantly differ with regard to relative proportions of spine subtypes (Fig. 3c, right).
Mean electrophysiological data are presented in Table 2. A multivariate ANOVA demonstrated a significant difference in resting membrane potential (Vr; F(4,97) = 14.0, p < 0.001) and input resistance (Rn; F(4,97) = 5.9, p < 0.001), but not in membrane time constant between groups. Importantly, TG neurons in both early and advanced tauopathy had a significantly depolarized mean resting membrane potential compared to NT neurons (early: TG vs. NT, p = 0.003; advanced: TGintact vs. NT, p < 0.001; TGatrophic vs. NT: p < 0.001). In early tauopathy, TG neurons had significantly higher Rn than did NT neurons (p = 0.018). Similarly in advanced tauopathy, TGatrophic neurons had a significantly higher Rn compared to both NT (p < 0.001) and TGintact neurons (p = 0.002); Rn was not different between TGintact and NT neurons in this stage.
A multivariate ANOVA demonstrated that action potential (AP) amplitude (F(4, 83) = 2.5, p < 0.048) was significantly different between groups while AP threshold was the same. While AP amplitude did not differ between groups in early tauopathy, AP amplitude was significantly lower in TGatrophic compared to both NT (p = 0.020) and TGintact neurons (p = 0.013, Table 2). TGintact and NT neurons did not significantly differ with regard to AP amplitude. Exemplar traces of repetitive AP firing elicited by a 2 s depolarizing current step and a current ramp protocol are shown in Figure 4a (middle and bottom respectively). The significant positive linear relationship between AP firing rate (+130 pA current step) and Vr across all neurons (Pearson Product-Moment correlation; r (109) = 0.564, p < 0.001) is shown in Figure 4b (left). Graphs of mean AP firing rates at the +130 pA step and at each depolarizing current step are shown in Figure 4b (middle and right respectively). Firing rates differed between groups at depolarizing current steps greater than +80 pA (+130 to +380 pA; F(4,68) = 5.7, 6.4, 5.2, 5.5, 5.6, 5.1, p < 0.001). In early tauopathy, TG neurons had a significantly higher AP firing rate relative to NT neurons in response to the +130 pA depolarizing current step (p = 0.032). In advanced tauopathy, TGintact neurons exhibited increased firing rates at every step from +130 to +230 pA (p = 0.045, 0.033, 0.032) and TGatrophic neurons had an increased firing rate every step from +130 to +380 pA (p < 0.001 for each step).
Rheobase, the amount of current required to elicit an AP, was significantly different between groups (F(4, 83) = 6.2, p < 0.001). In early tauopathy, rheobase was significantly lower in TG relative to NT neurons (p = 0.010; Fig. 4a, bottom; Table 2). In advanced tauopathy, rheobase was significantly lower in TGatrophic versus NT neurons (p = 0.002; Fig. 4a, bottom; Table 2). While rheobase was also lower in TGatrophic compared to NT neurons, this did not reach statistical significance (p = 0.067; Fig. 4a, bottom; Table 2); TGintact and TGatrophic neurons did not significantly differ from one another with regard to rheobase.
Depolarizing sag potential amplitude was significantly different between groups (F(4, 97) = 11.4, p < 0.001). This sag potential was significantly larger in amplitude in TG neurons in both early (TG vs. NT, p = 0.005) and advanced tauopathy groups (TGintact vs. NT, p < 0.001; TGatrophic vs. NT, p < 0.001; Fig. 5a; Fig. 5c, left) and was eliminated by HCN channel blocker ZD-7288 (Fig. 5b). There was a significant positive linear relationship between sag potential amplitude and Rn across all neurons (Pearson Product-Moment correlation; r (99) = 0.418, p < 0.001; Fig. 5c, right). There was also a significant positive linear relationship between sag potential amplitude and Vr across all neurons (Pearson Product-Moment correlation; r (101) = 0.669, p < 0.001; Fig. 5d, left). To determine if the greater sag potential amplitude was dependent upon the significantly depolarized Vr exhibited by TG neurons, sag potential amplitude was compared between TG and NT neurons that had the same Vr. When TG and NT neurons were matched by Vr,, the sag potential amplitude was still significantly greater in TG neurons (Student's t-test, p = 0.016; Fig. 5d, right). Thus the difference in sag potential amplitude was not dependent on the difference in Vr.
Electrophysiological traces of sEPSCs from representative neurons in early and advanced tauopathy are shown in Figure 6a1,2. The mean frequency of sEPSCs did not differ between groups at either the early or the advanced stage (Fig. 6b, Table 2). The rise and decay time constants of sEPSCs also did not significantly differ as a function of group (Fig. 6a1,2, Table 2). However, the mean sEPSC amplitude was significantly different between groups (F(4,93) = 3.8, p = 0.007). In early tauopathy, the mean sEPSC amplitude was the same in TG and NT neurons (Fig. 6b, Table 2). In advanced tauopathy, the mean amplitude of sEPSCs was significantly higher in TGintact neurons relative to both NT (p = 0.043) and TGatrophic neurons (p = 0.003; Fig. 6b, Table 2). On the other hand, TGatrophic and NT neurons did not significantly differ from one another with regard to mean sEPSC amplitude. Cumulative percentile histograms revealed an increased proportion of small amplitude events in TG relative to NT neurons in early tauopathy (K-S test, p < 0.001; Fig. 6c) and in TGatrophic relative to NT neurons in advanced tauopathy (p = 0.027; Fig. 6c). Conversely, TGintact neurons had an increased number of large amplitude events compared to NT (p = 0.013) and TGatrophic neurons (p < 0.001; Fig. 6c).
Within genotype cross-age comparisons of structural and functional parameters are shown in Supplementary Tables 1 and 2. Notably, since differences in morphological and electrophysiological properties between early and advanced NT neurons were minor, most differences between early and advanced TG neurons were dependent upon the effects of tauopathy rather than development. However, spine density was significantly higher in early compared advanced NT neurons. When spine density of each TG group was expressed as a percentage of the corresponding NT mean value (thereby normalizing for differences due to development), early TG neurons still had a significantly greater spine density than advanced TG neurons (Student's t-test, p < 0.001; data not shown) indicating that spine density did not decrease as a function of development alone, but more likely decreased as a function of tauopathy.
To determine if structural and functional changes to neurons were dependent on expression of high levels of human mutant tau, electrophysiological and morphological properties of neurons from TG and NT mice that received lifelong dox treatment were examined (data not shown). These two groups of neurons were designated TGdox and NTdox respectively. Apical dendritic tufts of each well-filled TGdox neuron that met morphological (but not electrophysiological) inclusion criteria (n: 21 neurons) were qualitatively the same as those of normal, healthy NT neurons. A total of 9 TGdox and 6 NTdox neurons met both morphological and electrophysiological inclusion criteria and were further assessed with regard to detailed properties. Neuron reconstruction data confirmed that each TGdox neuron had an intact distal apical dendritic arbor (> 500 µm in length). Moreover, assessment of spine density in distal apical dendritic branches of TGdox and NTdox neurons revealed that spine density was not significantly different between the two groups. Finally, functional electrophysiological properties were indistinguishable between TGdox and NTdox neurons. In summary, the major structural and functional changes to TG neurons we observed were dependent on high levels of human mutant tau expression.
In this study, we assessed structural and functional changes to layer 3 frontal cortical pyramidal neurons from rTg4510 mice in early and advanced stages of tauopathy. The key findings are that: 1) apical dendritic tufts are unchanged in early tauopathy, but are either preserved (TGintact) or severely atrophic (TGatrophic) in advanced tauopathy; 2) Thioflavin-S positive NFTs are present (or are absent) in approximately equal proportions in TGintact and TGatrophic neurons; 3) some TGintact neurons exhibit proliferative sprouting of oblique branches of the apical trunk; 4) spine density is decreased due to a specific loss of mushroom spines, but filopodia are increased in advanced tauopathy; 5) TG neurons exhibit stage-independent depolarized resting membrane potentials, increased depolarizing sag potentials, and increased action potential firing rates; 6) the mean amplitude and the proportion of high amplitude sEPSCs is increased in TGintact neurons, while the proportion of low amplitude sEPSCs is greater in both early TG and TGatrophic neurons, and; 7) suppression of the human mutant tau transgene with lifelong dox treatment prevents structural and functional changes to TG neurons.
No major changes in dendritic morphology were observed at an early stage of tauopathy. This is consistent with observations in hippocampal neurons from early-stage tau-P301L mice [6] and in frontal cortical neurons from early-stage htau mice [10]. However, AAV-tauP301L infection in the hippocampus of mice [23] and overexpression of human tau in the lamprey neurons [29] result in early dendritic regression. Further, in early AD, dendritic regression precedes substantial tangle formation and neurodegeneration in the hippocampus [7]. Differences in experimental models, types and/or levels of tau species present or, in the case of AD, the potentially confounding interactions between tau- and amyloid-beta-mediated pathology may account for the apparent discrepancies between these findings.
By comparison to early tauopathy, changes to the dendritic morphology of many TG neurons in advanced tauopathy were dramatic. At this stage, TGatrophic neurons exhibited significant regression of both basal and apical dendritic arbors; the apical dendritic tuft was reduced to only two or three branches or was completely lost. These degenerative changes are reminiscent of pronounced regression and dystrophy of distal apical dendrites and loss of basal dendrites of hippocampal neurons in AD [33,7]. Jaworski and coworkers [23] reported dystrophy and loss of apical dendrites of hippocampal neurons from AAV-tauP301L-infected mice that were particularly pronounced distally. Similarly, successive tau-mediated loss of microtubules and subsequent dendritic degeneration of first distal and then more proximal dendrites was observed in lamprey neurons [29]. In both of these models of tauopathy, as with rTg4510 mice, particularly high levels of tau may substantially contribute to the observed dendritic degeneration.
What causes dendritic degeneration during tau pathology? Since in healthy neurons tau binds to and promotes the stabilization and assembly of microtubule primarily in the axon, it is unlikely that tau loss-of-function is exclusively responsible for dendritic degeneration in tauopathies. It is plausible that a gain-of-function for pathological tau may lead to dendritic retraction by indirectly contributing to microtubule destabilization in dendrites. Loss of microtubules occurs in dendrites into which tau is missorted, suggesting a mechanistic link between the presence of abnormal tau species and degenerative changes to dendrites [59,29]. Indeed, previous studies have shown that hyperphosphorylated tau sequesters endogenous tau and other microtubule-binding proteins, including those responsible for dendritic microtubule stabilization [21,1,53]. Microtubule destabilization alone would be expected to lead to perturbations in intracellular microtubule-dependent transport, but there is evidence that abnormal tau may cause transport deficits by other mechanisms as well. These proposed mechanisms have been generated in large part from studies focusing on transport deficits in axons during tau pathology. However, many of these mechanisms may also underlie impaired transport in dendrites, as has been shown to occur in tau-transfected hippocampal neurons [56], and has been proposed as a principal underlying cause for early synaptic dysfunction in rTg4510 mice [19]. For example, there is evidence that excess tau binding to microtubules displaces motor proteins [11] and may prevent access of motors to microtubules [52,56]. In addition, high levels of tau may cause microtubule bundling leading to transport blockage [56]. Given that tau itself is transported as a kinesin cargo, increased levels of tau may lead to increased competition between tau and other kinesinbinding cargoes [11]. Pathological tau may also play a role in preventing the successful docking of cargoes to molecular motors [22]. Thus, as a direct consequence of impaired transport, dendrites (as well as axons) are likely deprived of proteins, vesicles and organelles that support maintenance of their structural integrity. Lastly, given that ~50% of neurons are lost in 9-month-old rTg4510 mice [49], surviving neurons examined in the present study are likely significantly deafferented; this loss of presynaptic inputs may exacerbate dendritic regression [14,24].
One of the most intriguing findings of this study was that almost half of the advanced-stage TG neurons examined possessed intact dendritic arbors. It is theoretically possible that these TGintact neurons do not express the mutant (P301L) tau transgene or contain hyperphosphorylated tau, however the latter is unlikely given that they commonly contained NFTs. Future studies will attempt to identify the mechanisms underlying selective vulnerability or preservation of TG neurons in progressive tauopathy. In addition to preservation of the apical tuft, half of the TGintact neurons demonstrated proliferative sprouting of oblique apical dendritic branches. A similar phenomenon was previously reported for layer 3 frontal cortical neurons from htau mice that show increased proximal apical dendritic length and complexity [10]. The dendritic proliferation seen in some neurons in these mouse models of tauopathy are reminiscent of those that have been described in normal human aging [8,13] and in a variety of neuropathological conditions including AD (review [55]), Huntington's disease [12], and epilepsy [34]. Dendritic sprouting in tauopathies could plausibly be triggered by deafferentation [20,54] or by reactivation of developmentally regulated programs (review [3]).
There is an extensive literature demonstrating alterations in dendritic spines in neurodegenerative disease (review [38,31]). In the present study we found that in early tauopathy, spine density was not different in TG compared to NT neurons. This finding is consistent with a previous study showing that postsynaptic density (PSD) 95 levels are the same in PSD fractions of forebrain lysates prepared from young (4.5-month-old) rTg4510 relative to control mice [19]. Further, densities of spines are not different in PSD95-labeled primary hippocampal neurons from rTg4510 mice or in DsRed-labeled rat primary hippocampal neurons transfected with P301L human mutant tau compared to control neurons [19]. In addition, a recent study by Kremer and colleagues [26] showed that spine density is not significantly different in the cortex of young (1- to 2-month-old) Tau.P301L mice, but is increased at a later age (4- to 6-month-old) versus age-matched control mice. Taken together, these findings provide supporting evidence that spine density is not reduced in these tau P301L mutant mice at relatively early stages of disease pathogenesis. Our spine classification analyses revealed that the relative proportions of mushroom-, stubby-, thin-, and filopodia-type spines were the same in TG versus NT neurons in early-stage tauopathy, although subtle changes in spine morphology (such as head diameter or neck length) have been reported in early stage Tau.P301L mouse neurons [26]. In advanced tauopathy, a ~20–30% reduction in dendritic spine density occurred in TG relative to NT neurons [reported here and in: 43,9], and to a greater extent in the TGatrophic than in the TGintact neurons. Spine loss in surviving neurons in advanced tauopathy may be due to a combination of tau-mediated deficits in the transport of synaptic cargoes to pre- and postsynaptic targets and loss of afferent inputs due to substantial neuronal loss. Here and in previous work [9], we found that the relative proportion of mushroom spines was reduced, while the relative proportion of filopodia is increased in advanced TG neurons. In the present study, we demonstrated that these changes occur in both TGintact and TGatrophic populations. We propose that the increase in filopodia in TG neurons at the advanced stage may be a compensatory response to decreased excitatory input [9]. Indeed, previous studies in hippocampal [39] and neocortical [40] neurons have shown that proliferation of filopodia occurs following functional deafferentation.
We have previously demonstrated that cortical pyramidal neurons in ~9-month-old rTg4510 mice exhibit markedly increased excitability that is primarily a consequence of the significant depolarization of these neurons [9,43]. An unexpected finding of the present study was that this increase in excitability was also prominent in the early stage of tauopathy, prior to NFT deposition and neuronal death. Thus at both the early and the advanced stage, TG neurons exhibited a significantly depolarized resting membrane potential, increased depolarizing sag potential and increased evoked action potential firing rates. Threshold was unchanged at both stages of tauopathy, and therefore was not a major determinant of increased excitability in TG neurons. However, there was a reduction in rheobase in TG neurons, which reached the significance level for early TG and TGatrophic groups.
From a mechanistic standpoint the large increase in depolarizing sag potential in TG neurons is particularly intriguing. This sag potential is due to the activation of the mixed cationic H-current, which is carried through hyperpolarization-activated cyclic nucleotide-gated (HCN) channels. The H-current is active at rest and causes a tonic 4 to 8 mV depolarization of the membrane potential [42]. Thus the increased sag potential seen in TG neurons may underlie the depolarized resting potential and increased action potential firing rates in these neurons. By what mechanism might the sag potential be increased in TG neurons? HCN channels are rapidly trafficked along microtubule and actin cytoskeletal networks in dendrites [36] and are particularly targeted to the distal dendritic arbor [30,32]. These networks are disrupted in neurodegenerative tauopathies (review [51]), and trafficking abnormalities occur early in the time course of the disease in rTg4510 mice [25]. Early and persistent impairment of HCN channel trafficking may result in accumulation of HCN channels at the soma and resultant increased sag potential and depolarization. Given the significantly increased sag potential, an unchanged or even increased input resistance in TG neurons is unexpected. While an increase in HCN channels may lead to a decrease in input resistance, findings from previous studies suggest that this may not always be the case. For example, in olfactory bulb mitral cells an increase in the amplitude of the sag potential is correlated with an increase in input resistance [2], while HCN channel block in CA3b pyramidal neurons does not lead to alterations in input resistance [18].
The early and persistent increase in excitability that occurs in this mouse model may be an initial pathological event that leads to downstream alterations in neuronal structure on the one hand and/or may represent early functional changes that occur as a consequence of altered trafficking of ion channels in these neurons. Understanding the time course and mechanisms underlying increased excitability during progressive tauopathy is important given evidence for reduced seizure threshold in human neurodegenerative disease [37,35,58].
In early tauopathy, glutamatergic spontaneous excitatory postsynaptic currents (sEPSCs) were unchanged in mean frequency, amplitude and kinetics in TG neurons. However, while the mean amplitude of sEPSCs did not differ, there were a significantly higher proportion of small amplitude events in TG neurons at this stage. This difference in amplitude distribution suggests that changes in glutamatergic transmission occur prior to spine loss. Indeed, a recent study by Hoover et al. [19] demonstrated that reduced surface expression of postsynaptic GluR1 AMPA receptors, related to mislocalization of hyperphosphorylated tau to spines, is associated with impaired basal synaptic transmission and LTP in the hippocampus of early stage rTg4510 mice.
In advanced tauopathy, neither the mean frequency nor the kinetics of sESPCs of TGintact or TGatrophic neurons differed from those of NT neurons. Thus, it is possible that surviving neurons at this stage may compensate for decreased input from dying neurons within the cortical network by increasing insertion or turnover of AMPA receptors at postsynaptic sites and/or by increasing presynaptic neurotransmitter release or the number of presynaptic terminals [9]. Indeed, we previously reported that in addition to increased firing rates, TG neurons from 9-month-old mice exhibit sprouting of new axonal boutons and filopodia; evidence for the formation of new, albeit shorter, excitatory synapses was also observed in TG frontal cortical neuropil [9]. These small synapses could account for the significantly higher proportion of small amplitude sEPSCs observed in TGatrophic versus NT neurons. Further, the mean amplitude of sEPSCs is significantly higher in TGintact neurons versus NT neurons and cumulative distribution histograms reveal a significantly higher proportion of large amplitude events in TGintact neurons compared to all other groups. This phenomenon could be due to the increased number of oblique apical branches observed in these neurons. Previous studies have shown that in healthy neurons, oblique dendrites are responsible for ~20% of the total area of the apical dendritic shaft of layer 2/3 neurons [27] and that these dendrites, together with basal branches, contribute ~2/3 of the total number of spines for an individual neuron [28]. The increased number of apical oblique dendritic branches and their relatively close proximity to the soma could result in less attenuation and greater summation of synaptic currents along the main apical dendritic trunk. Thus, dendritic sprouting may be a mechanism by which neurons in advanced tauopathy maintain synaptic gain for a time during progressive tauopathy. Additionally, a larger proportion of axodendritic versus axospinous synapses would be expected to lead to larger amplitude events in TGintact neurons. While we previously found no evidence for an increase in the proportion of axodendritic synapses in TG neurons from 9-month-old mice, it is possible that axodendritic synapses could increase with disease progression, and/or that TGintact neurons in particular exhibit increased axodendritic synapses. Studies are currently underway to determine if this is the case.
The principal findings of this study were that TG neurons exhibit increased excitability both in early and advanced stages of tauopathy, while dendrite and spine retraction occurs only late in disease progression. Initial changes in electrophysiological properties of TG neurons are likely due to impaired trafficking of ion channels, such as the HCN channel, whereas slower subsequent changes to morphology may be a consequence of cytoskeletal destabilization, deafferentation, and neuronal death. Whether early electrophysiological alterations contribute to subsequent morphological changes and/or to neuron death, or whether these events are independent of one another remains to be determined.
The authors declare no competing financial interests. This work was supported by NIH/NIA grants R01-AG025062. The authors are grateful to Dr. Jada Lewis for generously providing us with rTg4510 mice, Joseph Amatrudo for assistance with data collection, Dr. Christina Weaver for assistance with statistical analyses, Dr. Maria Medalla for careful reading of the manuscript, and Dr. Tara Spires-Jones for helpful discussions throughout the course of this project.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


text:
Welcome to Peak Education and Training's dental continuing education program. We offer quality continuing education, compliance training, products and services to build successful practices and encourage career development. Our staff believes that each dental team member is a value to the whole. Peak is dedicated to excellence in dentistry, your professional development and personal fulfillment.
We know that you are committed to the profession of dentistry and are seeking courses that fulfill your continuing education licensure requirements. At Peak Education and Training, we strive to be the most reliable resource that offices have for information concerning continuing education, dental licensure renewal and OSHA/HIPAA compliance. We offer a wide variety of classes for the entire dental team. All classes are selected through current research, collaboration with dental professionals and attendee requests. Every speaker is hand-selected for their expertise. We are proud of the reputation we have earned and strive to exceed your expectations!
Click Here to View Our Current Classes
Our Mission…
To be your trusted provider of dental continuing education,
encouraging excellence in dentistry through
the career development of dental professionals.
Thanks for the webinar! It was timely and very informative. I am happy to have tuned in!Let's Talk about Social
Steven Keltsch Webinar participant

subset: commoncrawl


text: >Alessio, > >It is ok for the delay, I am sure you have plenty of things to do, and >not only kernel work, of course. I will try to test that this evening if >possible, but be sure that I provide you some feedback. Thanks for your >work. > >Toine >Alessio Igor Bogani a écrit : >>* Hi, *>>* >*>* 2010/2/7 ttoine at ttoine.net <https://lists.ubuntu.com/mailman/listinfo/ubuntu-studio-devel> <ttoine at ttoine.net <https://lists.ubuntu.com/mailman/listinfo/ubuntu-studio-devel>>: *>>* [...] *>>* * >>>* As soon as you can provide a way to test closed source, I could report* >>>* using a RME hdsp Multiface II sound card of my workstation. *>>>* *>>* >*>* Antoine excuse me for long delay in reply. *>>* >*>* In this PPA you can find -headers packages necessary for enable closed drivers: *>>* https://launchpad.net/~abogani/+archive/ppa/+packages <https://launchpad.net/%7Eabogani/+archive/ppa/+packages> >*>* >*>* Thanks a lot for you feedback! *>>* >*>* Ciao, *>>* Alessio *>>* >*>* * A few items that I have been asked to mention: 1. We now have Abogani's -lowlatency kernel in the Ubuntu Studio Developers PPA. The PPA is available at https://launchpad.net/~ubuntustudio-dev/+archive/ppa 2. We are asking for volunteers to compare the -lowlatency kernel with the -preempt kernel on amd64 machines If you have questions or need additional information please respond to this email and we will be happy to assist you. Thank you for your time and efforts, they are tremendously helpful to both Ubuntu Studio and Abogani. Regards, ScottL -------------- next part -------------- An HTML attachment was scrubbed... URL: https://lists.ubuntu.com/archives/ubuntu-studio-devel/attachments/20100212/22a2c283/attachment-0001.htm


text:
If this is your first visit, be sure to check out the FAQ. You need to Register and Activate your account before you can Search add-ons, Download add-ons. Once activated you must request permission to Post in the forums or use the PM system. If you have problems accessing SOH, find your IP blocked or can't Register due to your Email Address please contact an administrator. Use the Contact Administrator link at the bottom of this page and We can help you!
you must add the 8 AAA on top of the new gsl i just sent you to get them firing as part of the mission.
Avsim - Flightsim - Simviation - Quarter Moon Saloon - Com-Central


text:
Free personalized radio that
plays the music you love
Songs (6) For Voice & Piano, Op. 90: II. Her Message (Hennes Budskap)
Features of This Tracka subdued, expressive aesthetic
chromatic harmony
a well-known composer
a Romantic-Era style
a mezzo-soprano
tonal harmony
minor key tonality
a broad tempo
These are just a few of the hundreds of attributes cataloged for this track by the Music Genome Project.show more
Similar Tracks
Don't have a Pandora account? Sign up
Already have a Pandora account? Log In


text:
In conjunction with the momentous one-time-only appearance of actress Lalla Ward — who is making her very first visit to a North American convention in over 20 years, and being the final living principal cast member of the classic series available for conventions in North America that we haven't yet hosted — we are thrilled to announce that Gallifrey One 2017's charity beneficiary (for our annual Bob May Memorial Charity Auction) will be for Denville Hall. Denville Hall is an actors care home in Britain, for which Ms. Ward is the chairman of its board of trustees, and we were very pleased to offer to proceeds of our 2017 charity to such a worthwhile cause, as she has been tireless in her championing of this and other care homes for many years. We will have more details about our charity auction in the months to come.
Our list of guests announced in this first block is as follows. (IMPORTANT NOTE: Our website has been reverted to the previous incarnation, before our April site makeover, due to some unforeseen issues we had with the template we were using, and will be updated again when we are able to fix them.)
Paul McGann portrayed the Eighth Doctor in the 1996 Doctor Who TV Movie, a role he later continued through audio with Big Finish Productions. To the astonishment of fans worldwide, McGann was the headliner in "The Night of the Doctor," a special mini-episode filmed in 2013, and saw him regenerate into his next incarnation – an event most everyone thought would never be seen on television. McGann is an actor, director and performer well known for his appearances in "Withnail & I," "Luther," "Collision," "Alien 3," "The Monocled Mutineer" and a series of TV films about Captain Horatio Hornblower. McGann made his first-ever convention appearance at Gallifrey One in 2004 and this will be his third appearance at our event, courtesy Showmasters Events.
Lalla Ward first appeared on Doctor Who in 1979 as Princess Astra in the serial "The Armageddon Factor"; it was this appearance that led to her taking on a regular role, replacing the outgoing Mary Tamm as the second incarnation of Romana, traveling companion to Tom Baker's Fourth Doctor, the following season. After leaving the program in 1981, she later retired from acting (and married author Richard Dawkins), though she has returned to reprise the role of Romana in Big Finish's audio series, primarily the "Gallifrey" audio series in which Romana becomes Lord President of Gallifrey. Her last appearance in North America was 21 years ago; Ward is the final principal living cast member of the classic series able to travel to North America whom Gallifrey One hasn't yet hosted, and we're absolutely thrilled that she will be joining us for our 2017 convention for this one-time-only event.
In addition, Ward is chairman of the trustees of Denville Hall, a UK actors' care home and the beneficiary of the proceeds of our 2017 charity auction at Gallifrey One. Our special thanks to Showmasters Events for arranging such a memorable appearance at our convention.
Louise Jameson played Leela, the leather-clad barbarian warrior companion of the Fourth Doctor, from "The Face of Evil" to "The Invasion of Time." She later reprised the role for the anniversary special, "Dimensions in Time" and for several Big Finish Productions audio adventures. She also appeared in three "direct-to-video" adventures of the investigating organization P.R.O.B.E., with Caroline John as Liz Shaw. She also became well known in "Tenko" and "EastEnders," and also appeared in "Bergerac," "The Bill," "Casualty," "Doctors," "Space: 1999" and "The Omega Factor." She is also very active in British theatre, including touring with her one-woman show "Face Value" and "Pulling Faces" (the latter she performed live at our convention on her last visit), and is currently touring in "The Mousetrap". We're delighted to welcome Ms. Jameson back for our 2017 event, courtesy Showmasters Events.
William Russell played Ian Chesterton in Doctor Who from the series pilot "An Unearthly Child" to "The Chase." As one of Susan Foreman's teachers, he was one of the first of the Doctor's companions to appear in the series. In 1999 he returned for the video release of the Doctor Who serial "The Crusade." He has also provided audio commentaries for various Doctor Who DVD releases, read audio book versions of three early Doctor Who novelisations and provided linking narration for CD releases. Russell had originally been intended to reprise his character of Ian for the 1983 story, "Mawdryn Undead," but scheduling conflicts required the story to be rewritten to instead bring back Nicholas Courtney as another, later regular character, the Brigadier. Russell made his first appearance at Gallifrey One in 2012 and we are thrilled to welcome him back for his second visit.
After portraying bumbling UNIT operative Josephine "Jo" Grant during the Jon Pertwee era in the early 1970s to rave reviews, Katy Manning later returned to the role (as the married Jo Jones) in the Sarah Jane Adventures episode "Death of the Doctor". She also plays both Jo as well as the popular spinoff audio character Iris Wildthyme on audio adventures from Big Finish Productions. She's also appeared in "Casualty," "All Saints," "Target," "Whodunnit" and "Man at the Top". A worldwide favorite at conventions, Katy has appeared at three Gallifrey One events and we are delighted to welcome her back to LA for her fourth visit, in an appearance sponsored by Showmasters.
Anneke Wills' last visit to Gallifrey One was all the way back in 2010, after appearances in 1997 and 2002, and we are delighted to welcome her back to Los Angeles once more! Best known as Polly, the Swinging Sixties' secretary who alongside Michael Craze's Ben Jackson became the final William Hartnell companions and continued through Patrick Troughton's first season, Anneke Wills also appeared in "The Avengers" and "Strange Report," and most recently has done considerable work on audio for Big Finish and BBC Audio, as well as penned two autobiographies, "Self Portrait" and "Naked". She also recently published the photo collection "In Focus: A Life in Images" with Paul W.T. Ballard. We are very pleased to bring Anneke back for a rare appearance in LA.
Peter Purves starred as companion Steven Taylor, the first new male companion character after the series' initial debut leads, in 1964, and remained a popular co-star of Willam Hartnell throughout his later tenure. Purves later went on to fame as a host on the series Blue Peter (so closely associated with the programme that he later found his association difficult to shake off), and he also appeared in the UK original "The Office," "EastEnders," "Z Cars" and "Dixon of Dock Green". More recently, Purves returned via Big Finish narrating some of their 'lost era' and Companions audio series. This will be Purves' second visit to Los Angeles (after his appearance in 2013) and we're glad to have him back!
Frazer Hines – aka Jamie McCrimmon opposite Patrick Troughton's Doctor – is not only as synonymous with Doctor Who as the Doctor himself, but returns for his unprecedented ninth appearance in a row at Gallifrey One, where he's become an annual tradition! Well known for his work on Doctor Who as well as his long-running role on the British soap Emmerdale and his recent appearances on Outlander (as Sir Fletcher Gordon), Frazer has also been reprising his Jamie role for Big Finish on their many audio releases – and more recently has also been featured as the Second Doctor, his impression of his long-time friend and colleague Patrick Troughton is spot on – and he's also narrated the BBC Audio soundtracks as well. Beloved by fans and beloved by us, it just wouldn't be Gallifrey One without him!
Considered to be one of the classic series' finest and most important producers, Philip Hinchcliffe oversaw the first three seasons of the Tom Baker era – an era noted for many of its important classics (such as "Genesis of the Daleks," "The Seeds of Doom," "Pyramids of Mars" and "The Talons of Weng-Chiang")… and for its now-legendary battle with Mary Whitehouse and the National Viewers & Listeners' Association over the end of part 3 of "The Deadly Assassin". Hinchcliffe guided the series through one of its most successful eras. He joined us in 2013 and we are delighted to welcome him back for a second visit.
Popular award-winning stand-up comic, actress and author Hattie Hayridge makes her Gallifrey One debut in 2017! She is of course best known as the female version of Holly on the long-running comedy series "Red Dwarf," on behalf of which she has become a popular guest at conventions around the world, and she has also appeared in "Jonathan Creek," "Have I Got News for You" and "Celebrity Squares". Her autobiography "Random Abstract Memory" was published by Penguin. The opportunity to bring Ms. Hayridge to Los Angeles for Gallifrey One 2017 was too good to pass up (and we know a lot of you are Red Dwarf fans) so we're thrilled to bring her to our convention this coming February!
Long-time friend of Gallifrey One, Daphne Ashbrook has appeared in many television and film roles including Star Trek: Deep Space Nine (as the titular character in "Melora") and the Doctor Who 1996 movie, in which she played the role of Doctor Grace Holloway. In 2004, Ashbrook traveled to the United Kindgom, which was filmed and later released on DVD in 2005, "Daphne Ashbrook in the UK". She performed in the Big Finish Productions audio play The Next Life opposite Paul McGann as the Eighth Doctor, but playing a different character named Perfection (this was the first time the two stars had worked together again since the 1996 telemovie). Ashbrook made her debut on the convention scene right here at Gallifrey One back in 1999, and we are always thrilled to welcome her back to our event.
June Hudson is best remember by our audience as the costume designer during one of the series' most iconic periods: the latter half of the Tom Baker era, where she was responsible for, among other things, the designs of Baker's "bohemian" look and of course the widely varying costume work for Lalla Ward's Romana. Her work also included a stint as costume designer on Blake's 7 and Survivors. She left the BBC in 1990 to diversify her portfolio still further. Her design drawings have been exhibited at the National Theatre and Chichester Theatre and her work is included in the RSC's permanent exhibition at Stratford, and she has lectured at many venues including the University of Redlands here in Southern California teaching intensive courses on design for science fiction television. She was with us in 2013 and we are happy to welcome her back for her second visit.
Prentis Hancock is best known to television viewers as Paul Morrow, second-in-command of Moonbase Alpha on Space: 1999, and to our attendees from his appearances in Doctor Who, most notably in "The Ribos Operation" (as the Captain), "Planet of Evil" (as Salamar) and "Planet of the Daleks" (as Vaber). His many other television appearances include "Z-Cars," "Colditz," "Survivors," "The New Avengers," "Secret Army," "Return of the Saint," "Minder," "Chocky's Children," "The Professionals," and "The Bill". He recently appeared in "Outlander" as Uncle Lamb in the episode "Sassenach". We are excited to welcome Mr. Hancock to LA for a rare appearance.
Actor and writer Michael Troughton, the son of the late Doctor Who star Patrick Troughton, was with us once before in 2012, and we're happy to welcome him back! He is best known for his roles in TV including Testament of Youth (1979), Minder (1984-89), The Heart Surgeon (1997), The New Statesman (1987-92) and the film Enigma (2001), and recent appearances include played Professor Albert in the recent Doctor Who episode "Last Christmas," as well as "Birds of a Feather," "Jonathan Creek" and "Restless". His new book "Smile on Your Face" was recently published, and Mr. Troughton will be signing copies at our 2017 convention as well as participating on our program.
Actor, writer, producer and director Nicholas Briggs is known equally for his voice talents – among others, he's the principal voice of the Daleks and Cybermen in modern-day "Doctor Who" on television – as well as his long work for Big Finish Productions, where he now the co-executive producer and has written, produced, directed and starred in countless audio dramas in the "Doctor Who" and other ranges, including as the title character in their "Sherlock Holmes" range. Nick is a wonderful guest at conventions around the world who has made many appearances at Gallifrey One in the past, and we're thrilled to welcome him back in 2017!
Writer and actor Barnaby Edwards has been one of Doctor Who's principal Dalek operators since the series returned to the air in 2005, as well as both in front of and behind the microphone for Big Finish's audio adventures, including as Paul Hunt in the "Cyberman" series and penning audios including "The Bride of Peladon," "The Wreck of the Titan" and "The Emerald Tiger". He has also provided voices for commercials, video games, animated series and awards ceremonies, and has painted, lectured and exhibited widely both within the UK and abroad. His works hang in private collections around the world and he regularly undertakes portrait and design commissions. He also has a range of postcards, greetings cards and T-shirts! He was last with us in 2012 and we're very happy to have him back.
Nicholas Pegg has appeared as one of Doctor Who's primary Dalek operators since 2005, like Edwards, as well as a lengthy stage career including in productions for Nottingham Playhouse, Scottish Opera, Birmingham Repertory Theatre and the Theatre Royal, Plymouth, and on television in "EastEnders" and "Doc Martin". Pegg is a noted authority on the life and work of David Bowie, and he is the author of "The Complete David Bowie" and was a consultant on the 2013 BBC TV documentary "David Bowie: Five Years". He has appeared in a variety of roles for Big Finish Productions as well as wrote the audio play "The Spectre of Lanyon Moor" (Colin Baker & Nick Courtney's first appearance together on audio) and directed several releases, too. Pegg joined us in 2013 and we welcome him back for his second appearance.
No one who was with us in 2012 will ever forget Simon Fisher-Becker's memorable pronouncement about "The question that must never be answered" during our Closing Ceremonies, and we're thrilled to welcome him back in 2017 for a return visit! Simon of course guest starred as Dorium Maldovar in series 5 & 6 of Doctor Who; his more notable roles include Tony Fazackerley in "Puppy Love" for the BBC, and the Fat Friar in "Harry Potter and the Philosopher's Stone," and he's recently been featured in several audios for Big Finish Productions. We look forward to seeing him again next February!
Veteran film and television composer Dominic Glynn has written music for such varied productions as Dead Like Me, Coronation Street, Emmerdale, Ramsey's Kitchen Nightmares, Red Dwarf, Eerie Indiana, The Simpsons, the Blake's 7 audio revival and several video games, but is best known to fans as the composer of the surreal Doctor Who Season 23 (Trial of a Time Lord) theme music, along with the incidental music for the season, still used today on audios from Big Finish Productions. Dominic returns for his second visit, having been last with us in 2014.
Writer Paul Cornell's credits including "Human Nature/The Family of Blood" and "Father's Day" from the Doctor Who series revival, a lengthy list of Doctor Who novels and audio plays which include the creation of the popular character Bernice Summerfield, as well as TV series such as his own CITV show Wavelength as well as Robin Hood, Primeval, Coronation Street and Casualty, and a list of comics for Marvel, DC, Vertigo and 2000AD on titles such as Captain Britain and MI-13, Young Avengers, Wolverine, Knight and Squire, Batman and Robin, Action Comics, Demon Knights, XTNCT, Pan-African Judges and Saucer Country. He's also written the novels Something More, British Summertime, London Falling and The Severed Streets and he is penning Titan Publishing's forthcoming Third Doctor comic series. We welcome Paul back for another visit.
Gary Russell is the former script editor of Doctor Who, The Sarah Jane Adventures, Torchwood and Wizards vs. Aliens, as well as the former editor of Doctor Who Magazine, and one of the most prolific writers, directors and producers of the spinoff Doctor Who media from Virgin Publishing, BBC Books and Big Finish Productions, including continuing to function as the senior producer of the Gallifrey audio series. He is currently part of the team at Planet 55 Studios in Australia producing the new science fiction animated series Prisoner Zero for ABC. Gary joins us for his nineteenth Gallifrey One appearance.
Comics writer, screenwriter and playwright Tony Lee has written for IDW's Doctor Who comics series, Big Finish's audio productions (for the Doctor Who, Bernice Summerfield and Dorian Grey series) and has written for Marvel Comics, Panini, DC Comics, Markosia Comics and Titan Comics, among others. Among his most recent works are a MacGuyver series for Image Comics and a Battlestar Galactica miniseries for Dynamite Entertainment, as well as several recent teen novels, Stalker, Mister Scratch, Jigsaw Lady, and Noticed and he has been announced as penning a new Robin Hood film. Tony returns to Gallifrey One in 2017 as both a program guest as well as the Master of Ceremonies of our 2016 Masquerade.
Long-time Gallifrey One guest and friend Jason Haigh-Ellery is the Managing Director of Big Finish Productions. He founded and owns the company. As well as attending regular production and company meetings and making crucial decisions about how the company runs, Jason also occasionally directs and is involved in the commissioning of new projects (such as Graceless and The Mervyn Stone Mysteries). He has also been responsible for many key, creative decisions in Big Finish (such as the casting of India Fisher and Sheridan Smith). Nick Briggs has described him as 'the best boss you could ever wish for'. Jason also produces theatre worldwide including Footloose and Never Forget, work with Idina Menzel, New Boy, and Ladykillers.
Screenwriter, novelist, and audio playwright Richard Dinnick is a long-time guest of Gallifrey One who co-hosted our charity auction in 2016. He is the winner of the 2012 BBC Writersroom opportunity to create a new show and to write for TV on the BBC and is now writing on the CBeebies TV shows Tree Fu Tom. Dinnick has written prose, scripts and comics for many media properties including MGM's Stargate and the BBC's Doctor Who, as well as adapting the Sherlock Holmes stories of Sir Arthur Conan Doyle – including The Hound of the Baskervilles – for CD release. His first novel Alien Adventures was published by BBC Children's Books in 2010 and he has since gone on to write books and short stories for Penguin UK, Titan Publishing, Black Library, Running Press and Snow Books. He has also written comic strips for IDW and BBC Magazines and his first original graphic novel is in the works. We welcome back Richard in 2017.
Writer, director and producer Scott Handcock worked for BBC Wales as a production runner on the third/fourth series of Doctor Who Confidential, the animated Doctor Who adventure, The Infinite Quest, the second series of the BBC's The Sarah Jane Adventures, and as production secretary on the sixth series of Doctor Who, beginning with Matt Smith's first Christmas special, A Christmas Carol. He has also directed The Confessions of Dorian Gray series for Big Finish as well as co-write part of the "Gallifrey" audio series. Scott will be joining us again to participate in programming as well as be one of our featured interviewers.
Longtime friends of Gallifrey One David J. Howe and Sam Stone once again return to LA to participate in convention programming. David has been a cornerstone of Doctor Who publishing for decades, as a popular nonfiction writer and proprietor of Telos Publishing, while Sam Stone is one of today's most prolific gothic, horror and fantasy fiction novelists and short fiction writers, and co-writer of the Doctor Who anthology film "The White Witch of Devil's End". We are always thrilled to have them join us for Gallifrey One and welcome them back to LA.
And there are MANY more guests to be announced in the weeks and months to come…]]>
Nobody is happier than we are that we were able to keep ticket sales open for a full two hours this year as EventBrite was able to release groups of tickets back into the queue… as many people as possible who paid attention to the ticket sale date were able to get tickets to next year's event. For those who purchased guest-of tickets, we'll have details about how you can add individuals' names on those tickets a bit further down the road.
We have some amazing guests and programming planned for next year, and we'll be previewing some of that in the months to come.
Also… HOTEL ROOMS at the Marriott LAX Hotel are also SOLD OUT.
This year there is no 'official' overflow hotel but there are rooms available at plenty of hotels beyond the Marriott; among the closest are the LAX Hilton, Four Points Sheraton, Renaissance LAX, Travelodge LAX, Residence Inn LAX, Crowne Plaza LAX and others. If you do not get a hotel room at the Marriott, try reserving via one of the online tools for a hotel in the area like Expedia. Note that there are other events in the area on Presidents' Day Weekend so reserve your hotel space early!
Thanks for your continued support of Gallifrey One!]]>
Facts & Figures: Gallifrey One 2016 welcomed 3974 attendees, staff and guests. All of our major guests appeared at the event (only Rachel Talalay was a last minute cancellation due to extended filming and family illness; Samuel Anderson arrived late on Saturday but participated in Saturday night and Sunday programming.) Last minute guests included Will Thorp, Andy Lane, and Vic Mignogna, who all joined our programming. Panels were almost always full, but only one event (the Gomez/Oliver panel on Saturday) filled the room beyond capacity requiring seating assistance; meanwhile many people did acknowledge our request to attend one of the two John Hurt events. Photo ops were done with all of our major guests (with some added on site including Mr. Davison.)
Auction Update: The 2016 Bob May Memorial Charity Auction raised a record $11,236 for the LUMOS Foundation. Our Sunday charity auction, originally hoped to go three hours, went almost six… but beat the previous charity record by nearly $3000! Well done! Joe Morgan, the representative from LUMOS who we were put in touch with through the kind assistance of Nerder Mysteries, says that they were "speechless" and are very thankful for the donation. We'd like to thank everyone who participated in our charity auction, both bidders as well as those who donated merchandise to auction at the event. Our check to LUMOS has now been mailed to them. (Also, our very special thanks to Tony Lee and Richard Dinnick for doing such a terrific job hosting our auction this year!)
What Worked & What Didn't: This year we have seen enormously positive responses from our attendees on the convention. The biggest hit of the weekend: our new electronic schedule, widely lauded throughout the event (we even kept it fully updated on site!) We had one issue, primarily on Saturday, having to do with the photo op lines which became very unwieldy; we're looking at potential solutions for 2017. The only major complaint for the year was of course the ticket sales last May; at the convention this year, we announced that we are going with a new provider, EventBrite, which handles very large sized events with relative ease. Programming ran on time, though we did have a few delays in one room on Sunday requiring a shift of about 15 minutes; the only major program shifts involved Samuel Anderson (moving his interview to Sunday due to late arrival) and Rachel Talalay (we had to cancel her Sunday commentary; in its place was an encore presentation about "Prisoner Zero" on Sunday morning.) Parking continues to be an issue but fewer people reported issues, perhaps because of the information we armed our attendees with about parking in the area. One major change we will make for next year is more reliance on the electronic schedule and signage and less on the printed grids.
Other things to report: We would like to thank Katie Aiani, Isaac Pierce and Brian Cook from Nerder Mysteries for running such a terrific convention-wide game for us this year, Let Zygons Be Zygons, and we hope to have them back in 2017. Speaking of Nerder Mysteries, we'd like to congratulate attendee Jon Sipes, who won a pair of free tickets to our 2017 convention as part of the game's final raffle at Closing Ceremonies. We've also had word from Troop 15595 of the Girl Scouts of Greater Los Angeles, who were a part of this year's convention selling cookies in the Dealers Room. "Because of your support all the Brownies that participated will be given the opportunity to attend a leadership camp over the summer," writes leader Alexandra Lewinson. "A wonderful achievement. We will also be making a very generous cash donation to our Soldiers Overseas and the Amerman Foundation Dog Therapy Program of Children's Hospital Los Angeles." Gallifrey One is thrilled we were able to help the Brownies of Troop 15595 and we'd like to thank them for joining us this year, and especially Ms. Lewinson for arranging the visit.
Photo JPGs: Convention Photo Shoots will shortly be posting photo ops taken on their website free of charge; we expect these photos to be made available sometime in late February.
Post-Convention Feedback: If you'd like to let us know what you think, we welcome your feedback via email.
Pictures: We'd love to see your pictures of the convention! Send us email if you'd like to share them (either a link to your Flickr/online album, or let us know you have some you'd like to send over and we'll give you an email address to mail them to!)
Our Thanks: As always, our convention never happens without tremendous hard work from our staff, all of whom we thank for their support and assistance both before and at the convention. Gallifrey One wishes to offer our very special thanks to Jason Joiner, Paul Jones, Laurence Wreford, Gene Smith, Paul Cornell, Chase Masterson, Mary Lee Holzheimer, Jason Haigh-Ellery, Steven Schapansky, Gary Russell, Derek Maki, Emily Danyel, James Kerwin, Christopher Jones, Craig Miller, Mike Horner, Bob Mitsch, Zach McGinnis, Scott Ray, Steve Himber, David Howe & Sam Stone, Barnaby Edwards, Tony Lee, Charlie Kirchoff, James @ the Talented Agency, Howard Hayes, Tillman Smoot, Michelle Thaller, Phil Plait, Lee Whiteside, Eric Hoffman, Erik Engman, Katie Alani, Isaac Pierce, Brian Cook and Lindsay Henderson for their assistance with this year's guest lineup and program.
And of course, we'd like to thank our amazing guests in 2016: Sir John Hurt, Peter Davison, Colin Baker, Michelle Gomez, Samuel Anderson, Nicola Bryant, Frazer Hines, Ingrid Oliver, Neve McIntosh, Sarah Dollard, Jamie Mathieson, Andy Pryor, Patricia Quinn, Julian Glover, Ian McNeice, Naoko Mori, Jessica Martin, Andrew Hayden-Smith, Clare Higgins, Sarah Douglas, India Fisher, Seán Carlsen, Will Thorp, Jami Reid-Quarrell, Matthew Doman, Chase Masterson, Nichelle Nichols, Jay Acovone, Andrew Cartmel, Cliff Simon, Vic Mignogna, Andy Lane, Paul Cornell, Gary Russell, Simon Guerrier, Nev Fountain, Phil Plait, Michelin Sisti, Nick Robatto, Marv Wolfman, Tony Lee, Ty Franck, Naren Shankar, Barbara Hambly, Matthew Jacobs, Richard Dinnick, Jason Haigh-Ellery, Nancy Holder, Charlie Kirchoff, Greg Weisman, Scott Tipton, David Tipton, J K Woodward, Michelle Thaller, Richard Starkings, David J. Howe, Sam Stone, Keri Bean, Hayden Black, Kipleigh Brown, Graeme Burk, Robert Smith?, Chris Doohan, Scott Edgington, Kevin Grazier, Lisa Hansell, Darin Henry, Steve Holland, Gillian Horvath, Christopher Jones, James Kerwin, Bridget Landry, Court Lewis, Sarah Milkovich, Craig Miller, Todd Nauck, Robert Napton, Larry Nemecek, Trina Ray, Cat Roberts, Steve Roberts, Anita Sengupta, Blair Shedd, Michele Specht, Arne Starr, Kim Steadman, Lary Stucker, Audry Taylor, David Wise, and Joseph Scrimshaw.
But most of all, thank you – all of our attendees, our cosplayers, our panelists, our tireless enthusiasts – for making this a year to remember!
Onward to Next Year: The countdown resets again! Gallifrey One Conventions cordially invites you to our next event, GALLIFREY ONE: 28 YEARS LATER, February 17-19, 2017, at the Marriott Los Angeles Airport Hotel, with pre-convention events beginning at 8:00pm on Thursday, February 16. Guests will be announced starting this summer. Tickets for our 2017 convention will go on sale will go on sale this April; an exact time and date will be announced very soon. We are now using EventBrite, which can handle much larger groups without website crashes and will also support Saturday ticket sales; please keep your eye on the very middle of April if you're making plans now. Ticket prices will be announced at the same time as the date of sale is announced; we will once again offer a limited number of one-day tickets. We encourage you to be vigilant and keep an eye out, because we now technically sell out within minutes. Hotel rooms at the Marriott will be available in early May; we have decided not to provide an 'official' overflow hotel this year but there are many hotels in the local area.
What can you expect next year? Amazing new guests — we know who you want to see, trust us! — with more programming and panels, tons of great evening events, and so much more. We are just getting started, so be sure to be with us for the next Gallifrey One event… the next iteration of the world's largest, longest-running and most beloved Doctor Who celebration of them all. Allons-y!]]>


text:
Hotels near Ruesselsheim Station
Search hotels near Ruesselsheim Station in Ruesselsheim
Deal of the day
Trending now in Ruesselsheim
Hotels within 0.5 miles of Ruesselsheim Station
- Most bookedArona Hotel Frankfurt Rüsselsheim
13 people looked at this hotel in the last hour
Most recent review
"The staffs are very nice, breakfast is excellent. "
A Traveller, Aug 2016, US
7 people looked at this hotel in the last hour
Most recent review
"This is an apartment more than a hotel. The owner/operator was incredibly kind and communicated well prior to our stay. There was plenty of room for our family (2 adults - not a couple - and two children). It was really nice to have the extra room. The set up of the room was not what we are used ..."
A Traveller, Aug 2016, US
Hotels within 2 miles of Ruesselsheim Station
212 people looked at this hotel in the last hour
Most recent review
"We use this hotel twice during our vacation. It was inexpensive. The breakfast buffet was great. For a break, we walked to Aldis Market for food, about a mile way from the hotel. The hotel is in a small village, very safe walking. This hotel is 5 star "in my book", but is not glamorous. We was on..."
A Traveller, Aug 2016, US
26 people looked at this hotel in the last hour
Most recent review
"Big room, spacious and with kitchenette, comfortable bed and so nice to have a balcony with a lovely view over the river. Excellent value."
A Traveller, Jul 2016, GB
- Best Western Hotel Rüsselsheim Frankfurt AirportNo Guest Reviews yet
23 people looked at this hotel in the last hour
21 people looked at this hotel in the last hour
Most recent review
"We were just staying 1 night then back to airport. Wish it were closer to airport."
A Traveller, Sep 2016, US
13 people looked at this hotel in the last hour
Most recent review
"I have been staying at the Columbia for over 20 years It is closing on the 15th July a great pity"
A Traveller, Jul 2016, GB
12 people looked at this hotel in the last hour
Most recent review
"Just stayed overnight to visit a friend and the Opel museum on the way to southern Germany from Frankfurt airport. In an semi-industrial part of town. I think its fair to say that goes without saying as this is right near the Opel complex and it covers most the area so that's just the way it is. ..."
A Traveller, Oct 2015, US
10 people looked at this hotel in the last hour
9 people looked at this hotel in the last hour
Hotels in Ruesselsheim near Ruesselsheim Station
Are you looking for a cheap Ruesselsheim Station hotel, a 5 star Ruesselsheim Station hotel or a family friendly Ruesselsheim Station hotel? You just landed in the best site to find the best deals and offers on the most amazing accommodations for your stay.
When you search for hotels near Ruesselsheim Station with Hotels.com, you need to first check our online map and see the distance you will be from Ruesselsheim Station, Germany.
Our maps are based on hotel search and display areas and neighborhoods of each hotel so you can see how close you are from Ruesselsheim Station and refine your search within Ruesselsheim or Germany based on closest public transportation, restaurants and entertainment so you can easily get around the city. All the hotels details page show an option for free or paid onsite parking.
If you wish to see the hotels with the highest featuring discounts and deals near Ruesselsheim Station, simply filter by price/ average nightly rate. We recommend you filter by star rating and read our genuine guest reviews so you can get the best quality hotel with the best discount.
One of the new features on Hotels.com guest reviews is that also show reviews from Expedia for Ruesselsheim Station hotels and the TripAdvisor Ruesselsheim Station hotels reviews so you can make sure that you checking with a reliable source. See the review scores on our Ruesselsheim hotel information pages.
Make the most out of your family vacation when you book your accommodation with Hotels.com – book your hotel near Ruesselsheim Station, Ruesselsheim after reviewing the facilities and amenities listed for each hotel.
After booking your hotel near Ruesselsheim Station, expect to receive your reservation confirmation in the mail in less than 10 minutes. The confirmation email contains more information on all nearby attractions, local directions and weather forecast, so you can better plan the days during your trip.
After getting the best hotel rates you can still save more by winning 1 free night! That's right, book 10 nights in any hotel near Ruesselsheim Station, Ruesselsheim and after you sign up for the Welcome Rewards program, you are eligible hotel you receive 1 night free*
The best hotel deals are here: We have Ruesselsheim Station hotel deals, Ruesselsheim Station last minute deals and offers to get you the cheapest Ruesselsheim Station hotel with our lowest price guarantee.


text:
Russia's ghastly Children's Rights Commissioner finally quits
Pavel Astakhov, Russia's children's rights commissioner and a man of many talents, is finally stepping down. Astakhov's resume is quite something: he's written crime novels, hosted TV shows
By: rockstarz | Comments: 26 | Views: 3056 | Votes: 4 | Shared: 1
Leaked: Jul-4-2016 in Ukraine
Lightning Strikes St Peter's Basilica as Pope Resigns 2013
Pope Benedict has shocked a billion Roman Catholics around the world, and his closest advisers, by announcing that he will resign at the end of this month. Within hours of Pope Benedict announcing his
By: th1sf8te | Comments: 660 | Views: 71938 | Votes: 12 | Shared: 2532
Leaked: Feb-12-2013 in Other News
Pope Resigns: Vatican Backlash Over Dossier Rumours Linking Church To 'Gay Network' And 'Sex Parties'
http://www.dailymail.co.uk/news/article-2283190/Pope-resigns-Vatican-backlash-dossier-rumours-linking-Church-gay-network-sex-parties.html Italian media says dossier alleges clergy were 'united by sex
By: OnlyLiarsBlock | Comments: 57 | Views: 3017 | Votes: 1 | Shared: 45
Location: Vatican City State (Holy See) | Leaked: Feb-24-2013 in World News, Other News, Religion
BREAKING - Pope Benedict To Resign Feb 28 2013
Rome (CNN) -- Pope Benedict XVI will resign on February 28, his spokesman Father Federico Lombardi told CNN Monday. The 85-year-old pope is resigning "because of advanced age," Benedict told t
By: MaKaElectric | Comments: 14 | Views: 1473 | Votes: 0 | Shared: 16
Leaked: Feb-11-2013 in World News
Gay Sex Rings, 'The Filth' Corrupting The Vatican...And Why The Pope REALLY Quit
http://www.dailymail.co.uk/news/article-2287074/Pope-resigns-2013-Gay-sex-rings-The-Filth-corrupting-Vatican--Pope-REALLY-quit.html Sickened by moral corrosion in his own shadowy cabal, Benedict can
By: OnlyLiarsBlock | Comments: 32 | Views: 7227 | Votes: 0 | Shared: 66
Location: Vatican City State (Holy See) | Leaked: Mar-5-2013 in World News, Other News, Religion
STATEMENT BY THE POPE Posted on 02-11 at 11:09:43 CST as released by the vatican Pope Benedict XVI on Monday said he plans on resigning the papal office on February 28th. Below ple
By: gregsto | Comments: 89 | Views: 1804 | Votes: 1 | Shared: 0
Location: Vatican City State (Holy See) | Leaked: Feb-11-2013 in World News, LiveLeaks, Religion
Pope will remain in Vatican to avoid prosecution for child sex abuse
After resigning Pope Benedict XVI will remain in the Vatican to avoid criminal prosecution for his role in the sexual abuse of children by clergy, according to Vatican sources. Church sources and
By: LoganTNY | Comments: 30 | Views: 1027 | Votes: 2 | Shared: 20
Leaked: Feb-17-2013 in Conspiracy
Pope resigned in wake of gay priest scandal
Pope transfers top official amid speculation about report on Vatican leaks scandal. The Italian media is reporting that Pope Benedict XVI resigned after receiving the results of an internal investigat
By: Nazel_Hut | Comments: 51 | Views: 2298 | Votes: 1 | Shared: 0
Leaked: Feb-25-2013 in World News, Religion
Pope Francis States Ideological Christians Have An "Illness"
Pope Francis recently made quite the stir after commenting on the ideological purity in religion, particularly with regards to Christianity, claiming that such ideological extremism is dangerous
By: dcmfox | Comments: 128 | Views: 1669 | Votes: 5 | Shared: 0
Leaked: Dec-9-2013 in Other News, Religion
Argentina's Cardinal Bergoglio Elected Pope Francis
Fresh start for New POPE Francis - 03-13-2013 Habemas Papam, "We have a pope." And the name of the man to emerge on the balcony in Vatican City is Cardinal Jorge Mario Bergoglio of Argentina. Th
By: NewspaperOnline | Comments: 44 | Views: 1391 | Votes: 3 | Shared: 0
Leaked: Mar-13-2013 in World News
A New Pope is Announced (
Obama: An American pope would 'preside just as effectively' As cardinals gather at the Vatican to elect a new pope, President Barack Obama said he believes U.S. politics wouldn't stand in the way
By: Tama7866 | Comments: 52 | Views: 1332 | Votes: 0 | Shared: 3
Location: Rome, Lazio, Italy | Leaked: Mar-13-2013 in Other News
Pope Benedict Quits! Bible Prophecy being fulfilled?
Pope Benedict XVI Resigns. Is the Vatican Preparing the World for First Black Pope? "The head of the Jesuits is called the "Black Pope"(NOT BLACK COLOR) and holds the title of General, just as in the
By: amy2x | Comments: 4 | Views: 1452 | Votes: 0 | Shared: 0
Leaked: Feb-13-2013 in Religion, Conspiracy
Search results for Pope Resigns 2013Sort by:
Item Menu