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This property allows you to book online with a credit card. After submitting a reservation request, the owner or manager has 24 hours to accept, at which time your card will be charged and you will receive an email confirmation
Water Front Log Home#4269970ha4 BR 3 BA Sleeps 12$407 avg/night3-30 night min stay
2 BR 1 BA Sleeps 5$229 avg/night7 night min stay
3 BR 2 BA Sleeps 12$191 avg/night2-90 night min stay
4 BR 2 BA Sleeps 10$450 avg/night3 night min stay
4 BR 2.5 BA Sleeps 10$350 avg/night2-7 night min stay
4 BR 2 BA Sleeps 8$371 avg/night7 night min stay
23 BR 9 BA Sleeps 56$514 avg/night1 night min stay
3 BR 3.5 BA Sleeps 8$250 avg/night2 night min stay
4 BR 3 BA Sleeps 12$399 avg/night2-7 night min stay
Walk or Snowmobile to Everything!#4223758ha4 BR 3 BA Sleeps 12$280 avg/night2-7 night min stay
2 BR 1 BA Sleeps 6$150 avg/night2 night min stay
Waterfront Adirondack Cabin#4455542 BR 1 BA Sleeps 4$175 avg/night3 night min stay
3 BR 3 BA Sleeps 6$189 avg/night2-6 night min stay
6 BR 5 BA Sleeps 15$650 avg/night4-5 night min stay
2 BR 1 BA Sleeps 6$167 avg/night1 night min stay
3 BR 2 BA Sleeps 8$200 avg/night3-7 night min stay
4 BR 1 BA Sleeps 8$289 avg/night2-7 night min stay
Moosehaven Old Forge NY#5949103 BR 2 BA Sleeps 8$223 avg/night2-90 night min stay
Adirondack "Big House"#8105126 BR 3.5 BA Sleeps 18$400 avg/night6 night min stay
4 BR 2 BA Sleeps 9$217 avg/night2-7 night min stay
Cottage on 4th Lake Eagle Bay#7320504 BR 2 BA Sleeps 8$282 avg/night3-6 night min stay
Where Memories Are Made#8103754 BR 1.5 BA Sleeps 8$258 avg/night3 night min stay
5 BR 4 BA Sleeps 10$267 avg/night2-7 night min stay
2 BR 2 BA Sleeps 8$450 avg/night2 night min stay
Studio 2.5 BA Sleeps 8$414 avg/night2 night min stay
5 BR 2 BA Sleeps 8$400 avg/night7 night min stay
4 BR 4 BA Sleeps 13$250 avg/night1 night min stay
Fourth Lake Waterfront House and 2 Cottages#3848178ha7 BR 4 BA Sleeps 16$552 avg/night3-7 night min stay
3 BR 2 BA Sleeps 6$245 avg/night2-7 night min stay
12 BR 5 BA Sleeps 40$160 avg/night7 night min stay
Adirondack Vacation Home#7046742 BR 2 BA Sleeps 6$195 avg/night2 night min stay
5 BR 4 BA Sleeps 14$800 avg/night2-7 night min stay
Lake Tekini - Okara Lakes#4978345 BR 1.5 BA Sleeps 10$240 avg/night2 night min stay
6 BR 4.5 BA Sleeps 10$656 avg/night3-7 night min stay
Old Forge Vacation Cabin#5969862 BR 1 BA Sleeps 4View listing for price2 night min stay
Cozy Old Forge Condo 1st Lake#7098342 BR 2 BA Sleeps 8$214 avg/night2 night min stay
Adirondack Cabin Mountain Retreat#4053243ha1 BR 1 BA Sleeps 5$253 avg/night4-5 night min stay
Waterfront 3 Bedroom Cottage#6106853 BR 1 BA Sleeps 8View listing for price
3 BR 2 BA Sleeps 10$133 avg/night2-180 night min stay
4 BR 1.5 BA Sleeps 10$150 avg/night2-3 night min stay
4 BR 3 BA Sleeps 10$256 avg/night2-7 night min stay
3 BR 2.5 BA Sleeps 10$309 avg/night3-7 night min stay
2 BR 2 BA Sleeps 4$386 avg/night2 night min stay
4 BR 2 BA Sleeps 10$415 avg/night3-7 night min stay
3 BR 1 BA Sleeps 6$300 avg/night2-7 night min stay
3 BR 3.5 BA Sleeps 6$561 avg/night3-5 night min stay
1 BR 1 BA Sleeps 2$175 avg/night2 night min stay
4 BR 2 BA Sleeps 8$329 avg/night2-90 night min stay
7 BR 3 BA Sleeps 16$473 avg/night3-7 night min stay
4 BR 1.5 BA Sleeps 8$271 avg/night7 night min stay
Recent Old Forge reviews
"Once again our stay at this lovely home was truly enjoyed. The home is clean and fully equipped with just about everything you can ask for. Pretty much bring your food, clothes and an attitude to sit back and enjoy Old Forge. We are planning on making this trip again next year."Guest: Sue Date of Stay: July 2016 Review Submitted: August 3, 2016View listing
"From the moment we walked into the home, we knew we were in for a fantastic holiday week. The house was exactly as shown in the pictures. The rooms were spacious and the home immaculate. Through no fault of the owners whatsoever our dreams of a white Christmas were dashed by the unseasonably warm weather. That did not stop us from checking out neighboring towns (Utica, Lake Placid, Warrensburg), cooking some wonderful meals in the chef-style kitchen, grilling steaks (in December??), and relaxing with good books and wine. The surrounding area was beautiful, with great views of lakes and mountains. Although we did not need our snow shoes this time, we can't wait to go back and explore the trails! This was our first ever house rental, and First Lake Retreat sets a high bar for the future by having met every expectation we had!"Guest: Frances S. Date of Stay: December 2015 Review Submitted: December 28, 2015View listing
"Our extended family stayed in this house for a week over the 4th of July. Our experience right from the start with Doug and Barb was exceptional. They fully accommodated our initial arrangements when we had a few changes. They welcomed us with "open arms" and a bottle of wine upon our arrival! And they were available during the week if we needed them. The house is just lovely- very well appointed and equipped. There was nothing we needed. The yard was wonderful for the kids. And we all loved the visit from the deer ever morning and afternoon. Parking was adequate for the guests we had during the week. And the location is ideal- a couple blocks from the main street that goes through Old Forge. We could get to everything without getting in the car. I fully recommend this property. We stay there again in a heart beat. If felt like home in more ways than one."Guest: Sherry P. Date of Stay: July 2016 Review Submitted: July 23, 2016View listing
"Lovely, clean, well maintained home with everything you need to stay a week. Grocery store is two blocks away! The three full bathrooms and bedrooms were perfect for three couples. Our goal of just relaxing with family was met! We enjoyed Old Forge on foot , didn't need to drive. Working with John throughout the entire rental process was friendly and proficient. We all thoroughly enjoyed our stay at Four Seasons and recommended staying here."Guest: Joan D. Date of Stay: July 2016 Review Submitted: August 9, 2016View listing
Featured Destinations: Dewey Beach, Poconos Rentals, Ocean City Maryland Rentals, Ocean City NJ Rentals


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Home Clinic; HOW TO GET BRUSHES CLEAN WITH MINIMUM EFFORT
By BERNARD GLADSTONE
Published: August 16, 1981
PROBABLY the messiest and most annoying part of any paint job is cleaning out the brushes after the job is done - at least when working with an oil or alkyd base paint. With latex paint, cleaning tools is much less of a problem; you can simply rinse them out in running water. But cleaning brushes, rollers and other tools that have been used in a solvent-base paint seems to be more of a challenge for most people - and that is why so many buy cheap ''throwaway'' brushes that they can discard when the job is done.
Unfortunately, this is very seldom the answer to the problem. Aside from the fact that throwing a brush away each day gets kind of expensive, even if you buy cheap ones, there is also the fact that these so-called bargain brushes really cannot do a decent job of spreading the paint on smoothly, particularly if you are working with a good grade of varnish or enamel.
Yet cleaning a brush is really not a difficult job - if you tackle it promptly and if you are properly prepared for the chore. You will need a quantity of the appropriate thinner for the particular type of paint you have been using (turpentine or mineral spirits for oil paints and varnishes, lacquer thinner for lacquers, etc.), a small can or jar a little larger than the brush you have to clean and a stack of old newspapers.
As soon as you are done for the day, immediately wipe out as much excess paint from the brush as you can. Start by wiping across the rim of the can, then rub the brush out vigorously on some scrap surface such as old cardboard, or some scrap pieces of wood.
Next, pour about an inch of thinner or solvent into the jar, then press the brush down into this, working it hard against the bottom as shown in the accompanying illustration. This insures that you are working the thinner up into the heel of the brush where excess paint most often accumulates.
After doing this for a minute or two, pour the thinner out into another container, then wipe the brush hard on a stack of old newspapers as shown. As the top sheet becomes saturated, tear it off and start wiping on the next sheet. After three or four sheets, pour a small amount of fresh thinner into the jar (about half an inch will do this time) then rinse again by working the bristles against the bottom again. Turn the brush over every few seconds.
Repeat this rinsing and wiping process two or three more times - until no more color comes out when you wash it in fresh thinner, and until wiping leaves the bristles clean. All told, three or four rinses will usually do the trick, yet only a small amount of thinner will be used each time, and most of that can be saved. Each time you pour the dirty thinner into a second container (preferably a glass jar), allow this to settle for several hours. You can then save the top part by pouring it off carefully into the original container.
Cleaning in this manner is usually all that is needed to insure the brush staying soft until the next time it is needed. However, if a really good brush is to be stored for some time, then many prefer to go one step further and give it a final wash in a mild detergent solution. Rinse in plain water and shake dry, then comb smooth (use a metal brush comb or an old wide-tooth hair comb) and wrap in brown paper to help it retain its shape. Store by laying flat on a shelf or inside a drawer. Answering the Mail
Q. There is a sloping cement apron in front of my garage that connects the garage floor to the driveway surface. This apron is in excellent condition except in one corner, where it abuts the garage floor. There, a section of the concrete about two feet square has dropped over an inch and requires filling in or raising. I cannot get under this section to raise it because of a large shrub and a nearby cement walk. Can I add a topping and taper this around the edges so it will blend in smoothly with the rest of the area? - R.B.T., West Hempstead, L.I.
A. You can, but it won't help if there is a hollow space under the concrete that will allow it to continue sinking. I would advise breaking up that section of old concrete and allowing the pieces to fall into the void under them, then wait a few months to make sure the concrete will not sink further. Then you can fill in with new concrete. To help it adhere where you feather it out around the edges it is best to chop away some of the old concrete, and then use a masonry adhesive liquid mixed in with the concrete.
Q. I have a light-colored leather couch that has got dirty from newsprint rubbing off when newspapers were placed on the surface. Can you tell me what I can use to clean this leather? - J.J.B., Verona, N.J.
A. A nonflammable dry-cleaning fluid should do the trick. Put clean rags or paper towels into the fluid and wipe a small section, then turn the cloth or use a fresh piece to keep from spreading the soil around. Stores that sell leather goods also sell cleaners and conditioners for this, so it might pay to use one of these afterward. --------------------------------------------------------------------- Questions about home-repair problems should be addressed to Bernard Gladstone, The New York Times, 229 West 43d Street, New York, N.Y. 10036. Questions of general interest will be answered in this column; unpublished letters cannot be answered individually.
Illustrations: diagram of procedure involved in cleaning brushes

subset: commoncrawl


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Bivouac Canteen & Bar
Perth Institute of Contemporary Arts
PICA (pee-kah) may look traditional – it's housed in an elegant 1896 red-brick former school – but inside it's one of Australia's...
Ezra Pound
Down a much-graffitied lane leading off William St, Ezra Pound is favoured by Northbridge's bohemian set. It's the kind of place where...
Kakulas Bros
Deli and provisions store overflowing with dirt-cheap legumes, nuts and olives.
198 William St · interesting places nearby
Bivouac Canteen & Bar information
Mediterranean-style cuisine partners with a good wine list, and gourmet pizzas go well with boutique beers and artisan ciders. The coffee is excellent, and Bivouac's utilitarian decor is softened with a rotating roster of work from local artists.


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Home > Supernatural > Season 9 > Episode 2
Devil May Care
Episode Info
The ninth season of the US fantasy drama continues as demon hunters Sam and Dean Winchester (Jared Padalecki and Jensen Ackles) combat more supernatural forces. Dean shocks Sam by opening the boot of the Impala and showing him that he has kidnapped Crowley. Kevin is horrified when he sees the King of Hell, but Sam and Dean convince him that they need Crowley to find the demons on Earth. Meanwhile, Abaddon re-emerges and plans to take over Hell.
|Air Date:
|Oct 15, 2013
|Directed By:
|Guy Norman Bee
|Written By:
|Andrew Dabb
as Sam Winchester
as Dean Winchester
as Crowley
as Crowley
as Kevin
as Abaddon
News & Interviews for Devil May Care
Devil May Care Trailers & Photos
Discussion Forum
Discuss Supernatural on our TV talk forum!Go to Forum


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CV & Interview Skills£29 instead of £156 (from Vizual Coaching) for a CV and interview skills online package - save 81%
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Recently I wrote about an inspiring group of women recent who are fighting for a better future for their children. You can read more about their story here. World Vision's work in this area helped this group of women break out of the cycle of debt, leading to financial independence. Another way World Vision assists in this […]
The EKKA 2016
One of the things I was looking forward to this year (after moving from a year in New Zealand back to Brisbane) was The Ekka. The Ekka is short for the word exhibition in Brisbane Exhibition, and we don't think of it as anything else! I have extremely fond memories of attending The Ekka as a child with my […]
A Fight
A Cow
It was still light; the sky was deepening into dusk. Cow bellows interrupted the quiet vibe of village life; a few pushbikes lined the streets; spots of colourful saris painted the countryside. I like to watch their faces. It's a good way for me to get a feel for what they are saying, even though I can't understand the […]
A School Garden
Left Part of My Heart in India
Finding Dory – The Reef in Real Life
Travel First Aid Kit Essentials
Doing a Family Holiday This Year?
Are you doing travelling or holidaying as a family this year? I found a gorgeous, gorgeous product for you! This Travel Journal from Caravanning With Kids is a wonderful way to document family travels. You can do things like a daily journal, add photos or draw pictures; it's something both adults and kids can enjoy doing […]
Pan for Gold
New Playground – South Bank Parklands Brisbane
Sponsored by Nuffnang and South Bank When people ask me about visiting my hometown Brisbane and where to go, one of my top picks is South Bank because it's such a beautiful spot for families. I'm excited to announce there's a new playground being built that will make this already fab spot even more awesome. […]
Stationery Travel Packs for Kids
Whale Watching on the Gold Coast
Being a Blogger: Behind the Scenes
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Sweet-and-Sour Chicken Wings
Total Time
Prep 20 mins
Cook 30 mins
These chicken wings, sweetly glazed with pineapple juice, catsup and soy sauce, are a can't-miss choice to make for Super Bowl parties or to take to potlucks. From BH&G
Ingredients Nutrition
- 1133.98 g chicken wings (12 to 15 wings)
- 118.29 ml all-purpose flour
- 1.23 ml garlic salt
- 1.23 ml lemon pepper seasoning
- 44.37 ml cooking oil
- 177.44 ml sugar
- 118.29 ml vinegar
- 59.14 ml unsweetened pineapple juice
- 59.14 ml catsup
- 4.92 ml soy sauce
- Cut off and discard wing tips from chicken wings.
- Cut each wing at joint to make 2 sections.
- In a shallow dish, combine flour, garlic salt and lemon pepper seasoning; coat chicken pieces with flour mixture.
- Heat oil in a 12-inch skillet; add coated chicken pieces and cook until browned, turning occasionally.
- Remove chicken from skillet and arrange pieces in a shallow baking dish.
- In a medium saucepan, combine sugar, vinegar, pineapple juice, catsup and soy sauce; heat to boiling, stirring to dissolve sugar.
- Pour over chicken.
- Bake, uncovered, in a 350 degree F oven about 30 minutes or until chicken is no longer pink, turning pieces over after 15 minutes.
- Makes 24 to 30 pieces.
Most Helpful
Bev these are amazing wings! I thought I had pineapple juice but didnt so used orange juice instead. The sweet sour sauce is so good and the meat fell off the bones. I did not get the wings seperated in two but they were still so very good! Thank you for posting this recipe!


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Play naughty pranks on your classmates and teachers! As a silly prankster, your job is to wreak havoc on others. You can spill soda, let air out of bus tires, and embarrass the cute girls. Turn the field trip into a hilarious nightmare!
Naughty College Days is one of our selected Point and Click Games . Also very popular on this website right now are Monkey GO Happy Ninjas 2, Christmas Slacking, Fleeing the Complex and Monkey Go Happy Valentines.
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- CaMKII
Ca2+/calmodulin-dependent kinase II
complete Freund's adjuvant
cAMP response element-binding
- fEPSP
field EPSP
- GluN3A
glutamate NMDA receptor subunit 3A or NR3A
high-frequency stimulation
low-frequency stimulation
long-term depression
long-term potentiation
NMDA receptor
protein kinase A
protein kinase C
paired-pulse facilitation
postsynaptic density
Regulatory roles of the NMDA receptor GluN3A subunit in locomotion, pain perception and cognitive functions in adult mice
O. Mohamad and M. Song made equivalent contributions to this work.
S. P. Yu: 101 Woodruff Circle, Suite 620, Emory University School of Medicine, Atlanta, GA 30322, USA. Email: firstname.lastname@example.com
Key points
- • Adult glutamate NMDA receptor subunit 3A (GluN3A) knockout (KO) mice showed slow locomotor activity, motor deficits and increased pain sensation.
- • Hippocampal slices from juvenile and adult GluN3A KO mice showed greater long-term potentiation compared with wild-type slices.
- • Adult GluN3A KO mice showed enhanced abilities in learning and memory tasks.
- • GluN3A deletion resulted in increased expression and/or phosphorylation of Ca2+/calmodulin- dependent kinase II in the brain.
- • This is the first investigation showing that deletion of GluN3A in the embryonic stage has imperative impacts on multiple behaviours in adults.
Abstract Since the discovery of the glutamate NMDA receptor subunit 3A (GluN3A), the functional role of this unique inhibitory subunit has been largely obscure. GluN3A expression is high in the neonatal brain but declines to a low level in the adult brain; it is thus commonly believed that GluN3A does not have a major functional impact in adulthood. Using wild-type (WT) and GluN3A knockout (KO) mice, we show here that deletion of GluN3A affected multiple behavioural functions in adult animals. GluN3A KO mice showed impaired locomotor activity on a variety of motor function tests, and increased sensitivity to acute and sub-acute inflammatory pain. GluN3A KO mice also showed enhanced recognition and spatial learning and memory functions. Hippocampal slices from juvenile and adult GluN3A KO mice showed greater long-term potentiation (LTP) compared with WT slices. GluN3A deletion resulted in increased expression of Ca2+/calmodulin-dependent kinase II (CaMKII) in the forebrain, and the phosphorylated CaMKII level upon LTP induction was significantly higher in the GluN3A KO hippocampus compared with WT controls. CaMKII inhibition abrogated the enhanced LTP in GluN3A KO slices. These data reveal for the first time that the presence of GluN3A may have profound impacts on several functional/behavioural activities in adult animals, and could be a therapeutic target for neurological disorders associated with NMDA receptor functions.
NMDA receptors (NMDARs) are key components in neuronal development and functional activities in the nervous system, including membrane excitability, synaptic plasticity, pain sensation and neuronal excitotoxicity (Barkus et al. 2010). NMDARs are hetero-multimers that are composed of at least one GluN1 (NR1) plus one or more GluN2A-D (NR2A-D) subunits (Sheng et al. 1994). GluN2 subunits, especially GluN2B, play important roles in the induction and maintenance of long-term potentiation (LTP) and long-term depression (LTD) that are likely the underlying mechanisms of learning and memory (Bliss & Collingridge, 1993). The lately discovered and characterized glutamate NMDAR subunit 3A (GluN3A) (NMDAR-L, χ−1 or NR3A) and GluN3B (NR3B) subunits are unique subunits of NMDARs. GluN3A is widely expressed in different brain regions, such as the cortex, hippocampus, thalamus, hypothalamus, brainstem and spinal cord (Sucher et al. 1995). Unlike GluN2B whose expression gradually decreases after birth but remains highly expressed in adulthood (Wenzel et al. 1997), GluN3A mRNA and protein levels peak at the end of the first postnatal week, but decrease from P12 to a low level (∼15% of GluN1) by adulthood in rodents (Ciabarra et al. 1995). This sharp reduction in GluN3A expression has led most investigations on GluN3A to focus on neonatal stages (Roberts et al. 2009).
GluN3A co-localizes with other subunits to form functional channel complexes (Perez-Otano et al. 2001). Co-expression of GluN3A with GluN1 and GluN2 reduces NMDAR-mediated Ca2+ influx and whole-cell currents, resulting in smaller unitary conductance of NMDAR channels. Because of this property, GluN3 subunits (GluN3A and GluN3B) have been regarded as inhibitory subunits in the NMDAR complex. The smaller conductance is also seen in cortical neurons isolated from wild-type (WT) P8 rodents when compared with age-matched neurons from GluN3A knockout (KO) mice (Sasaki et al. 2002). These small NMDAR inward currents are most likely due to the relatively reduced Ca2+ permeability and reduced Mg2+ sensitivity in the GluN3A-containing NMDARs (Matsuda et al. 2002). Mice lacking GluN3A have increased dendritic spine densities in cortical neurons. This difference is more pronounced in cortical neurons of P19 mice compared with the adults (Das et al. 1998). Supporting a role in early brain development, the elimination of GluN3A from active synapses in the first 2 weeks in rodents is required to increase NMDAR conductance, and to form stronger and larger synapses. Recent evidence also suggests that presynaptic GluN3A receptors may participate in regulation of glutamate release (Larsen et al. 2011). At the behavioural level, GluN3A downregulation is required to form long-term memories (Roberts et al. 2009). Thus, GluN3A downregulation appears critical for synapse maturation and memory formation during development. On the other hand, whether GluN3A plays a potential role in the adult brain has been largely ignored and remains poorly understood.
A previous investigation showed that enhanced LTP was only observed in hippocampal slices from P6–P8 GluN3A KO mice, but not in juvenile and adult slices (Roberts et al. 2009). These observations have enforced the general assumption that GluN3A is not a major player in NMDAR-dependent activities in the adult brain. Different from previous observations and assumptions, here we demonstrate for the first time that after embryonic deletion of GluN3A, markedly significant differences in locomotor functions, pain sensation, learning and memory and synaptic plasticity (LTP) can be detected between adult WT and GluN3A KO mice. These novel findings indicate that the GluN3A subunit, despite its relatively low expression, has a surprisingly imperative functional role in adult animals and could be a target for therapeutic interventions.
WT and GluN3A KO mice
Animal handling and all experiments were approved by the Institutional Animal Care and Use Committee (IACUC) at Emory University. The GluN3A KO mice and WT counterparts were originally provided by Nobuki Nakanishi and Stuart A. Lipton at Sanford-Burnham Medical Research Institute (La Jolla, California, USA). Detailed information about the background of these mice has been described before (Das et al. 1998). In brief, embryonic stem cells derived from 129/SvJ were electroporated with DNA carrying disrupted GluN3A gene and then injected into blastocysts from C57BL/6 mice. The resulting chimeric males were crossed with BlackSwiss or 129SvEv females to produce F1 heterozygotes. GluN3A KO homozygote mice were then produced by cross-breeding F1 mice. In our lab, homozygote colonies of either WT or GluN3A KO mice were maintained under the same conditions with identical room temperature (22°C), same food and water supply, and animal care environment.
DNA for genotyping was extracted from tail snips. Two separate sets of primers were used for the GluN3A KO and WT mice, respectively. For the WT reaction, forward primer: 5′-CCACGGTGAGCTTGGGGAAG-3′ and reverse primer 5′-TTGGGGAGCGCCCTGCATGG-3′. For the KO reaction, forward primer: 5′-CCACGGTGAGCTTGGGGAAG-3′ and reverse primer 5′-GCCTGAAGAACGAGATCAGG-3′. DNA (2 μl) was amplified on a thermocycler (MJ Mini, Personal Thermal Cycler, Bio-Rad, CA, USA) for 40 cycles (2 min, 94°C; 30 s, 60°C; 10 min, 72°C for the WT reaction; and 2 min, 94°C; 30 s, 58°C; 10 min, 72°C for the KO reaction).
Behavioural tests
For all behavioural tests, mice were handled for 3 days and acclimated to the testing location at least 1 h before starting any test.
SHIRPA primary screen The screen test was performed to identify gross phenotypic and behavioural characteristics. We followed similar protocol procedures as described online by the MRC Harwell centre for mouse genetics (http://empress.har.mrc.ac.uk/; Lalonde et al. 2005). Mice were placed inside a glass beaker and inspected for the following phenotypes: body position (0 = inactive, 1 = active, 2 = excessively active); tremor (0 = absent, 1 = present); palpebral closure (0 = eyes open, 1 = eyes closed); coat appearance (0 = tidy, well groomed, 1 = irregular); whiskers (0 = present, 1 = absent); and defecation (0 = present, 1 = absent). The mice were then transferred to a new non-familiar arena and observed for the following responses: transfer arousal (0 = extended freeze >5 s, 1 = brief freeze then move, 2 = immediate movement); gait (0 = fluid movement and about 3 mm pelvic elevation, 1 = lack of fluidity); and tail elevation (0 = dragging, 1 = horizontal extension, 2 = elevated). Finally, the mice were held by their tails and assessed for the following: positional passivity (0 = struggles when held by tail, 1 = struggles when held by neck, 2 = struggles when laid supine, 3 = does not struggle); limb grasping (0 = absent, 1 = present); corneal reflex (0 = present, 1 = absent); biting (0 = none, 1 = biting with handling); and vocalization (0 = none, 1 = vocal).
Locomotor and anxiety tests The open field test was conducted to measure baseline locomotor activity. In the open field locomotion test, mice were allowed to freely explore an open field container (45 × 35 × 25 cm; divided into 12 equal squares) for 5 min. Activity was recorded on a video camera. The average distance and speed, and the total time spent in the central zone were recorded using TopScan CleverSys (CleverSys, Inc., Reston, VA, USA). The Rota-rod test was performed to assess the ability of mice to maintain balance and coordination on an accelerating rotating rod (UGO Bacile 7650). The Rota-rod was set to accelerate from 4 to 40 rpm in 300 s. In each session, mice were placed on the rotating drum and the time to fall off was recorded. Each session was stopped after 6 min even if the mouse was still on the drum. The timer was also stopped if the mouse clings to the rotating drum and completes a full rotation. Drums were cleaned with ethanol after each trial. On each testing day, mice were tested three times separated by at least 15 min inter-trial interval. The elevated plus maze test was performed to measure anxiety levels. The elevated plus maze was built in the lab using black Plexiglas material with the following dimensions: arm's length 30 cm; width 5 cm; and wall's height 15 cm. The central compartment is 5 × 5 cm, and the stage is elevated 45 cm above the floor. Each mouse was placed in the central platform facing an open arm, and its behaviour was videotaped for 5 min. The maze was cleaned with ethanol between trials. We recorded the number of entries and total time spent in the open and closed arms. The defensive withdrawal test was performed to measure exploratory behaviour. In the defensive withdrawal test, animals were placed in a dark chamber (10 × 15 × 10 cm high) placed along the wall of a large box (50 × 50 × 50 cm high). Animals were habituated to the open field 24 h prior to behavioural testing by allowing them to explore it for 15 min without access to the sheltered chamber. The 'latency to exit' was measured as the time it takes the animal to place his four paws outside the small chamber. The 'latency to cross' was measured as the time to cross a square drawn to represent the central zone of the box. The experiment was video-taped and analysis was performed by a researcher blinded to the identity of the groups.
Pain tests To test acute pain, we measured the latency of pain responses on the hot-plate and tail-flick tests. For the hot-plate test, mice were placed on a hot plate (55°C), and the latency to forepaw licking or jumping was recorded for each mouse in three trials separated by at least 15 min. For the tail-flick test, the latency to withdraw the tail from water (50°C) was recorded for three trials separated by 15 min. To test inflammatory pain, the response to subcutaneous formalin injection was recorded. In brief, 10 μl of 5% formalin was subcutaneously injected in the plantar surface of a forepaw. Pain scores were collected as detailed before (Roussy et al. 2009). In brief, the total time spent licking or biting the injected forepaw was recorded over a 3 min interval for each mouse for 1 h.
Memory tests The Novel Object Recognition and Morris water maze tests were performed and analysed as previously described (Lalonde et al. 2002; Bevins & Besheer, 2006). All trials were videotaped and performance was analysed using TopScan or MazeScan (CleverSys, Inc.).
The Novel Object Recognition test was performed during the dark cycle. On the 2 days preceding the training session, mice were individually habituated to the testing environment (30 cm × 30 cm × 20 cm; polycarbonate box) for 10 min. During the training session, mice were allowed to explore a box in which two identical objects were placed in adjacent corners for 5 min. During the retention sessions (1 h, 1 day and 7 days after training), one of the objects was replaced by a novel object and mice were allowed to explore for 5 min. The ratio of the time spent exploring any of the identical objects during the training session or the time spent exploring the novel object during the retention session to the time spent exploring the familiar object was calculated.
The Morris water maze test was performed by the animal behaviour core lab at Emory University. The water maze apparatus is a round, water-filled tub (52 inch or 132.1 cm diameter filled with white tempura paint) placed in a room rich with extra maze cues. Mice were placed in the maze starting from four different positions (N, S, E and W). An invisible escape platform was located in the same spatial location 1 cm below the water surface independent of the starting position on a particular trial. In this manner, subjects were able to utilize extra maze cues to determine the platform's location. Each subject was given four trials per day (N, S, E and W) for 5 days with a 15 min inter-trial interval. The maximum trial length was 60 s, and subjects were manually guided to the platform if they do not reach it in the allocated time. Upon reaching the invisible escape platform, subjects were kept on it for an additional 5 s to allow them to survey the spatial cues in the environment to guide future navigation to the platform. After 5 days of task acquisition, a probe trial was carried out during which the platform was removed. The time spent and distance travelled in the quadrant that previously contained the escape platform during task acquisition was measured over 60 s.
Hippocampal slice preparation and electrophysiological recording
WT and GluN3A KO mice were anaesthetized with isoflurane, decapitated, and their brains were dropped in ice-cold artificial cerebrospinal fluid (aCSF) containing (in mm): NaCl, 124; KCl, 3; NaH2PO4, 1.25; MgCl2, 6.0; NaHCO3, 26; CaCl2, 2.0; glucose, 10. The high Mg2+ concentration here was used to suppress NMDAR activation and associated excitotoxicity during the cutting procedure. The aCSF solution was saturated with 95% O2 and 5% CO2, at pH 7.4. The hippocampi were cut into 400 μm-thick transverse slices with a vibrating blade microtome (Vibratome 1000 Plus Sectioning System, St Louis, MO, USA). After incubation at room temperature (23–24°C) in another aCSF containing 1 mm MgCl2 for 60–90 min, slices were placed in a recording chamber (RC-22C, Warner Instruments, Hamden, USA) on a stage of an upright microscope (Olympus CX-31, Pittsburgh, USA) and perfused at a rate of 3 ml per min with aCSF (1 mm MgCl2) at 23–24°C. A 0.1 MΩ tungsten monopolar electrode was used to stimulate the Schaffer collaterals. The field (f)EPSPs were recorded in CA1 stratum radiatum by a glass microelectrode filled with aCSF with resistance of 3–4 MΩ. The stimulation output (Master-8; AMPI, Jerusalem) was controlled by the trigger function of an EPC9 amplifier (HEKA Elektronik, Lambrecht, Germany). fEPSPs were recorded in the current-clamp mode. Data were filtered at 3 kHz and digitized at sampling rates of 20 kHz using Pulse software (HEKA Elektronik). The stimulus intensity (0.2 ms duration, 16–30 μA) was set to evoke 40% of the maximum fEPSP initial slope, and the test pulse was applied at a rate of 0.033 Hz.
To induce LTP, high-frequency stimulation (HFS) was applied at Schaffer collaterals at 100 Hz for 1 s once or repeated three times with 10 s intervals. Schaffer collaterals are axonal collaterals extending from the CA3 pyramidal neurons in the hippocampus, and play a critical role in learning and memory. Paired-pulse facilitation (PPF) was examined by applying pairs of pulses that were separated by 10–500 ms intervals. Low-frequency stimulation (LFS; 1 Hz, 15 min) or NMDA (30 μm) application were used for the induction of LTD in both WT and KO slices. To perform the low Mg2+ experiments, no HFS was applied. Normal aCSF was changed to low Mg2+ aCSF (containing 0.2 mm Mg2+), slices were perfused and fEPSPs were recorded for 60 min (Townsend et al. 2006). The magnitudes of LTP and LTD were expressed as the mean percentage of baseline fEPSP initial slope.
To monitor membrane potential changes (depolarization) in response to stimulation at the Schaffer collaterals, we conducted blind whole-cell patch-clamp recording of pyramidal neurons in the CA1 region of the hippocampus. The recording glass pipettes (Sutter Instrument, USA) had a tip resistance between 6 and 8 MΩ when filled with an internal solution containing (in mm): KCl, 140; MgCl2, 2; CaCl2, 1; Na2ATP, 2; EGTA, 10; Hepes, 10; at pH 7.2. The stimulus intensity applied to the Schaffer collaterals was 16 μA for 0.2 ms. The distance between stimulation electrode and recording pipette was about 300 μm.
Western blot
Western blot analysis was performed to analyse protein expression in brain tissues from WT and GluN3A KO mice following previous procedures (Manzerra et al. 2001). In brief, brain tissue was lysed in lysis buffer containing (in mm): Tris–HCl pH 7.4, 25; NaCl, 150; EDTA, 5; 0.1% SDS; sodium orthovanadate, 2; NaF, 100; 1% Triton X-100, leupeptin, aprotinin and pepstatin with continuous manual homogenization. Forebrain or hippocampal tissue lysate was then spun at 14,000 × g for 15 min and supernatant was collected. Protein concentration was determined using BCA protein assay (Pierce, Rockford, IL, USA). Equal amounts of protein (30 μg) were resolved on SDS–PAGE using gradient gels (6–18%), and gels were blotted onto polyvinylidene fluoride (PVDF) membranes (Amersham, Buckinghamshire, UK) blocked with 5% BSA in TBST buffer (20 mm Tris, 137 mm NaCl and 0.1% tween) and incubated overnight with primary antibodies against rabbit polyclonal anti-GluN3A (1:1000; Millipore, Billerica, USA), rabbit anti-GluN3B (1:1000; Millipore), mouse monoclonal anti-GluR1 (1:1000; Millipore), rabbit anti-cAMP response-element binding (CREB; 1:1000; Cell Signaling, Danvers, USA), rabbit anti-phospho-CREB (1:1000; Cell Signaling), rabbit anti-Akt (1:1000; Cell Signaling), rabbit anti-phospho-Akt (1:1000; Cell Signaling), rabbit anti-protein kinase C (PKC)α (1:1000; Cell Signaling), rabbit anti-protein kinase A (PKA) C-α (1:1000; Cell Signaling), rabbit anti-Ca2+/calmodulin-dependent kinase II (CaMKII)α (1:1000; Cell Signaling), rabbit anti-phospho-CaMKII (Thr286; 1:1000; Cell Signaling) and mouse anti-actin (1:5000; Sigma, St Louis, MO, USA). After three washes with TBST, blots were incubated with HRP-conjugated secondary antibodies (anti-rabbit or anti-mouse, 1:2000; Bio-Rad) in 5% BSA for 1 h. Blots were developed using Pierce ECL Western Blotting Substrate (Thermo Scientific, IL, USA). The level of protein expression was normalized to β-actin controls.
For synaptosomes and postsynaptic density (PSD) preparations, we used a modification of previous protocols (Dunkley et al. 2008). In brief, total forebrains were homogenized in 0.32 m sucrose solution using Teflon-coated homogenizers. The homogenate was centrifuged to collect the supernatant S1 and the pellet P1. S1 was further centrifuged to collect S2 and P2. P2 was resuspended in Percoll, and the P2/Percoll solution was differentially centrifuged in a Percoll gradient. The band corresponding to the heavy synaptosomes was resuspended in Hepes/EDTA/Triton-X buffer and centrifuged (32,000 × g; 70.1 Ti rotor) to collect the first PSD fraction (PSD-1T). PSD-1T was resuspended in Hepes/EDTA/Triton-X buffer and centrifuged again (200,000 × g; 70.1 Ti rotor) to collect the second PSD fraction (PSD-2T). The following antibodies were used: mouse monoclonal anti-GluN1 (1:1000; Millipore), rabbit polyclonal anti-GluN2B (1:1000; Millipore), rabbit polyclonal anti-GluN2A (1:1000; Millipore) and rabbit anti-PSD-95 (1:1000; Cell Signalling). The level of protein expression was normalized to PSD-95 controls.
Statistical analysis
Results were expressed as the mean ± SEM. Statistical comparisons were made with unpaired two-tailed Student's t test and one- or two-way analysis of variance (ANOVA), with the Bonferroni's post hoc analysis to identify significant differences. A value of P < 0.05 was considered significant difference for all comparisons.
General phenotypic and behavioural features of adult GluN3A KO mice
Adult GluN3A KO mice (2–4 months old) appear indistinguishable from WT mice either visually (Fig. 1K) or as shown by the SHIRPA test (Table 1), which is a primary screen for gross phenotypic and behavioural characteristics and neurological dysfunctions. Adult WT and GluN3A KO mice showed no increased seizures or convulsions.
|WT mice
|GluN3A KO mice
|WT mice
|GluN3A KO mice
|Body position
|Tail elevation
|Palpebral closure
|Positional passivity
|Coat appearance
|Limb grasping
|Corneal reflex
|Transfer arousal
We noticed, however, that there were some body weight differences between WT and GluN3A KO pups (Table 2). While WT and GluN3A KO pups weighed similar at birth, GluN3A KO mice became heavier than WT animals starting day 10 of the first month after birth. The difference in weight, however, disappeared at early adulthood (1 month old). This observation suggested that GluN3A KO mice initially grew faster than the WTs. Given the published reports on the influence of maternal care on animal phenotypes (Kappeler & Meaney, 2010; Cameron et al. 2011), we measured the body weight of WT and GluN3A KO mice that were raised in the same cage. GluN3A KO mice still weighed more during development, and the difference in weight similarly disappeared in early adulthood.
|Weight (g) of WT and KO litters separate
|Weight (g) of WT and KO litters combined
|Day 1
|1.41 ± 0.08 n = 17
|1.55 ± 0.17 n = 9
|1.23 ± 0.07 n = 13
|1.50 ± 0.07 n = 10
|Day 5
|3.31 ± 0.11 n = 16
|3.64 ± 0.13 n = 14
|2.58 ± 0.14 n = 12
|3.30 ± 0.15 n = 10
|Day 10
|4.72 ± 0.19 n = 18
|6.52 ± 0.14 n = 21
|4.58 ± 0.19 n = 12
|5.10 ± 0.17 n = 10
|Day 15
|6.66 ± 0.16 n = 9
|8.80 ± 0.20 n = 10
|6.25 ± 0.17 n = 12
|7.30 ± 0.26 n = 10
|Day 20
|7.91 ± 0.25 n = 24
|11.90 ± 0.87 n = 10
|7.50 ± 0.28 n = 12
|8.70 ± 0.33 n = 10
|Day 30
|17.27 ± 0.62 n = 15
|23.33 ± 0.61 n = 6
|13.64 ± 0.50 n = 11
|15.10 ± 0.60 n = 10
|Young adult
|24.38 ± 0.52 n = 13
|25.14 ± 0.68 n = 14
|21.18 ± 0.62 n = 11
|22.00 ± 1.75 n = 6
Impaired locomotor activity and normal stress/anxiety levels in adult GluN3A KO mice
The role of GluN3A in locomotion is completely unknown. We measured locomotion and locomotor learning using a variety of motor tests. We started with the open field test, which assesses the baseline locomotor activity. Adult GluN3A KO mice walked slower and travelled less distance than WT mice (Fig. 1A and B). However, the total time GluN3A KO mice spent in the central zone of the test box was similar to WT mice (Fig. 1C). Thus, GluN3A KO mice were less active in the open field test but did not show any sign of increased anxiety. In order to test for gross motor skills including balance, coordination and motor learning abilities, animals were trained everyday for 5 days on the Rota-rod test. On day 1, despite their reduced activity in the open field, both WT and GluN3A KO mice performed similarly on the Rota-rod, indicating that in forced movement, KO mice were able to perform as well as WT mice. With continuous training, adult WT mice improved their performance better (stayed longer on the rotating beam) than GluN3A KO mice. By day 5, however, GluN3A KO mice caught up with their WT counterparts. The animals were then rested for a week before being tested again at days 12 and 16. The performance of the KO mice declined at day 12, but improved again at day 16 to levels similar to the WTs (Fig. 1D). This test suggested that GluN3A KO mice had some motor deficits, as shown by their slowed ability to reach their plateau performance during the five training days and by the drop in performance at day 12. However, it was evident that this decreased motor ability was compensated for by prolonged training.
Consistent with the results from the Rota-rod test, in a 5 day Morris water maze testing, the swimming speed of adult GluN3A KO mice was similar to WT mice at day 1. In the following days, GluN3A KO mice swam slower than WT mice, and statistically significant difference was detected at days 2 and 5 (Fig. 1E). While the average swimming speed of WT mice remained about the same, the average speed of the KO mice during day 2–5 was significantly slower than that of WT mice (219.0 ± 6.7 mm s−1 and 193.0 ± 5.4 mm s−1 for WT and KO mice, respectively, n = 10, P < 0.05).
Because some of these locomotor phenotypes could alternatively be explained by increased anxiety levels, we specifically evaluated anxiety using the elevated plus maze (Takahashi et al. 1989). In this test, there was no difference in the number of entries to the open (Fig. 1F) or closed arms (Fig. 1G), or the total time spent in the closed arms (Fig. 1H). In testing exploratory behaviour using the defensive withdrawal test (Takahashi et al. 1989), GluN3A KO mice showed no difference in the time to leave the sheltered chamber and the time to cross into the central zone compared with WT mice (Fig. 1I and J). These data collectively indicated that GluN3A KO mice were not more stressed/anxious than their WT counterparts.
Enhanced pain sensation in adult GluN3A KO mice
NMDARs play a key regulatory role in pain sensation. Hot-plate and tail-flick tests were performed to elucidate whether adult GluN3A KO mice had altered acute nociception. The hot-plate test reflects supraspinal sensory integration (Rubinstein et al. 1996), while the tail-flick test is a measurement of spinal pathways of acute nociception (Tham et al. 2005). Adult GluN3A KO mice showed enhanced responses (shorter latency) on the hot-plate test (Fig. 2A). On the other hand, the tail-flick test showed no difference in the responses elicited in WT and GluN3A KO mice (Fig. 2B). Taken together, these results are consistent with a role of GluN3A in mediating an analgesic response in supraspinal but not spinal pain pathways.
The established formalin inflammatory pain model was used to investigate the response to inflammatory nociception and tissue injuries. Formalin was subcutaneously injected into the plantar side of the right forepaw and the pain score was determined (Roussy et al. 2009). There was no obvious difference in the forepaw oedema between the WT and GluN3A KO genotypes 1 day after the formalin insult (data not shown). We analysed the duration of pain responses (licking and biting the injected forepaw) during the first hour, and pooled the data into two distinct phases: phase I (0–5 min); and phase II (6–60 min). In phase I, there was no difference in the responses between WT and KO mice (Fig. 2C). In phase II, GluN3A KO mice exhibited a more pronounced pain response compared with WT controls (Fig. 2D), indicating an enhanced response to inflammatory pain stimuli with GluN3A deletion.
Enhanced object recognition and spatial memory in adult GluN3A KO mice
Because learning and memory are intimately NMDAR dependent, adult WT and GluN3A KO mice were tested in the Novel Object Recognition and Morris water maze tests for assessing visual and spatial memory, respectively (Bevins & Besheer, 2006; Vorhees & Williams, 2006). In the Novel Object Recognition test, which requires acquisition, storage and recall of features of familiar objects, WT and GluN3A KO mice showed similar preference to the 'familiar' objects during the training session (Fig. 3A), indicating that these two genotypes had similar exploratory behaviours, curiosity and motivation. During the retention tests where one of the objects was replaced with a novel object, both WT and GluN3A KO mice showed more preference for the novel object 1 h after training. GluN3A KO mice, however, showed even more preference for novelty compared with WT controls. Different animal groups were tested with the novel object 1 or 7 days after training, respectively. WT mice lost preference for novelty in these tests, while the GluN3A KO mice still showed preference for the novel object at day 1 after training (Fig. 3A), indicating better recognition memory compared with WT mice.
In the Morris water maze test in which animals use spatial cues to swim towards an escape platform (Vorhees & Williams, 2006), adult WT and GluN3A KO mice were trained to find the submerged platform four times a day for 5 days. On day 1, GluN3A KO mice needed more time to get to the platform (Fig. 3B) likely due to their locomotor deficit. The latency and distance travelled to escape to the platform decreased in both WT and GluN3A KO mice during the 5 day training (Fig. 3B and C). Analysis on the latency and distance covered by WT or KO mice during the 5 day training showed that there is a significant learning curve in both WT (day 3) and KO mice. By averaging the latency in the last three training days, GluN3A KO mice performed significantly better than WT controls (27.7 ± 3.0 s and 19.0 ± 2.2 s for WT and KO mice, respectively; n = 10, P < 0.05; Fig. 3D). When animals were tested after removal of the platform on day 6, both WT and GluN3A KO mice spent significantly more time in the target quadrant than in other quadrants (Fig. 3E; P < 0.0001 comparing the time spent in the target quadrant with all other quadrants in WT and GluN3A KO mice separately using one-way ANOVA). GluN3A KO mice, however, significantly spent even more time in the target quadrant compared with WT controls (41.9 ± 5.0% and 57.3 ± 4.6% for WT and KO mice, respectively; n = 10, P < 0.05), indicating better spatial memory of the KO mice.
Enhanced LTP in the juvenile GluN3A KO brain
LTP is a cellular and molecular event that is believed to be the electrophysiological basis of synaptic plasticity associated with learning and memory (Bliss & Collingridge, 1993). NMDARs are major regulators in the induction and maintenance of LTP (Coan et al. 1987). LTP was examined in the CA1 region of hippocampal slices from juvenile (postnatal day 10 and 20) WT and GluN3A KO mice. The fEPSPs were recorded before and after a single episode of HFS (100 Hz, 1 s) at the Schaffer collaterals. In this test, the average magnitude of HFS-induced LTP was similar between P10 WT and GluN3A KO mice (Fig. 4A), and between P20 WT and GluN3A KO animals (Fig. 4B). However, it is commonly known that the activation of AMPARs and NMDARs by glutamate released from presynapses contributes to postsynaptic membrane depolarization, and that LTP induction relies on sufficient activation of AMPARs that removes the voltage-dependent Mg2+ block on NMDARs (Premkumar & Auerbach, 1996). Because AMPAR expression in the developing brain is much lower than in adults (Pickard et al. 2000; see Fig. 6D) and NMDARs in the absence of GluN3A are more sensitive to the Mg2+ block than GluN3A-containing NMDARs (Chatterton et al. 2002), we hypothesized that LTP induction in P10/P20 mice might be greatly affected by the nature of HFS. Specifically, we proposed that a single HFS in GluN3A KO brain slices could not induce sufficient AMPAR-mediated membrane depolarization to remove the Mg2+ block as it can do in adult brains. Thus, sufficient postsynaptic depolarization might be needed to reveal the LTP capability in the P10 and P20 GluN3A KO brain.
To examine our hypothesis, we then tested the effect of low Mg2+ (0.2 mm instead of 1 mm in aCSF) on membrane depolarization in hippocampal slices from P10 WT and GluN3A KO mice. Whole-cell current-clamp recording of pyramidal neurons in the CA1 region showed that stimulation at the Schaffer collaterals induced membrane depolarization that triggered action potential spikes (Fig. 4C and D). To quantify the depolarization response, we measured the width (duration) of the response at the half depolarization level. When brain slices from WT and GluN3A KO mice were perfused with the low Mg2+ aCSF, the depolarization response was much longer compared with the response in the normal aCSF solution (Fig. 4C and D). Moreover, the increased duration of membrane depolarization in low Mg2+ was greater in GluN3A KO than that in WT mice (Fig. 4D). This is consistent with the previous observation that NMDARs lacking GluN3A are more sensitive to Mg2+ block (Chatterton et al. 2002; Matsuda et al. 2002). The resting membrane potential did not change significantly in the low Mg2+ solution.
To ensure that sufficient membrane depolarization can overcome the voltage-dependent Mg2+ block of NMDARs, LTP was tested using either a 3× HFS protocol or the 0.2 mm Mg2+ external solution without HFS in young brain slices of P10 and P20 mice (Townsend et al. 2006). In the 3× HFS protocol, fEPSP recordings showed significantly greater LTP in P10 but not in P20 GluN3A KO slices (Fig. 4E and F). In the low Mg2+ solution, fEPSPs were gradually potentiated in P10 and P20 slices from both WT and KO animals. However, the increases were greater in GluN3A KO slices (Fig. 4G and H). Therefore, using protocols that provoke additional glutamate release and/or sufficient postsynaptic depolarization that can compensate for insufficient AMPAR expression/activation or the difference in Mg2+ sensitivity, we demonstrate that GluN3A can be a key regulator of synaptic plasticity in the developing brain.
Enhanced LTP in the adult GluN3A KO brain
In adult brain slices from 2- to 4-month-old mice, the role of GluN3A in LTP regulation was studied using normal aCSF (1 mm Mg2+) because a higher level of AMPARs exists in the adult brain (see Fig. 6D) and HFS should induce sufficient postsynaptic depolarization. Using the previously reported HFS protocol (100 Hz, 1 s, 3×; Roberts et al. 2009), we similarly observed that there was no significant difference in LTP between adult WT and GluN3A KO mice (Fig. 5A; P > 0.05, n = 6 in each group). However, a single HFS episode (100 Hz, 1 s, 1×) of a smaller depolarizing action revealed significantly greater LTP in adult GluN3A KO slices compared with WT slices (Fig. 5B). We then measured PPF, which is a specific indicator of presynaptic plasticity. PPF induced by various inter-stimulus intervals (ranging between 10 and 500 ms) was similar between WT and GluN3A KO hippocampal slices (Fig. 5C), indicating that the presynaptic function of GluN3A KO neurons was not different from that of WT neurons. As a control, we checked whether the enhanced LTP in the adult KO mice was dependent on NMDAR activation. The NMDAR antagonist d(−)2-amino-5-phosphonovalerate (AP-5; 100 μm) was bath applied 20 min prior to HFS. LTP induction was almost fully blocked in both WT and GluN3A KO slices (Fig. 5D). On the other hand, LTP induction was not affected when AP-5 was applied after HFS (Fig. 5E).
LTD is another form of synaptic plasticity. We observed that LFS (1 Hz, 15 min) induced similar LTD in hippocampal slices of adult WT or GluN3A KO mice (Fig. 5F). We also tested a chemical induction protocol to induce LTD in the CA1 region of the hippocampus (Lee et al. 1998). A brief (4 min) bath application of NMDA (30 μm) transiently abolished the evoked field responses followed by stable responses that were depressed below the baseline. The magnitude of this chemically induced LTD in KO slices was also not different from that of the WT controls (Fig. 5G). Thus, LTD induced by two distinct inducers was not affected by deletion of GluN3A, suggesting that some GluN3A-independent mechanisms might regulate the induction of LTD.
Expression profile of glutamate receptors in the GluN3A KO brain
In Western blot examination, the expression of GluN3A in the forebrain of WT mice increased from postnatal day 1 to day 10, reached a maximal level between day 10 and 20, and then significantly dropped in adulthood (Fig. 6A; Wong et al. 2002). We evaluated the expression levels of major ionotropic glutamate receptor subunits in adult GluN3A KO mice. The expression levels of the AMPAR subunit GluR1 and NMDAR subunit GluN3B were similar in the adult brains of WT and GluN3A KO mice (Fig. 6B and C). As shown previously (Petralia et al. 1999), expression of GluR1 was low in postnatal day 10, and increased significantly in the adult brain for both WT and GluN3A KO mice (Fig. 6D). There was no difference in the expression of other NMDAR subunits (GluN1, GluN2A and GluN2B) measured in PSD-2Ts of adult WT and KO forebrains (Fig. 6E).
Enhanced CaMKII activity in the adult GluN3A KO mice
Western blot analysis of forebrain tissue lysates showed similar expression levels of different LTP regulators, including Akt, CREB, PKA and PKC (Fig. 7A and B). Moreover, there was no difference in the expression of phosphorylated Akt (p-Akt) and phosphorylated CREB (p-CREB; Fig. 7C).
The NMDA-dependent LTP induction requires NMDAR-mediated Ca2+ influx and rapid rise of intracellular Ca2+ concentration, which serves to activate CaMKII that acts as a major mediator in the induction of LTP (Barria et al. 1997; Giese et al. 1998). The expression of CaMKII was significantly higher in the GluN3A KO brain than in the WT brain, while no significant difference was seen in the level of phosphorylated CaMKII (p-CaMKII; Fig. 8A). To more closely assess this LTP key regulator in the brain region associated with LTP induction, CaMKII as well as the activated form of p-CaMKII were measured in the hippocampal region of adult animals before and 10 min after the LTP-inducing stimuli (1× HFS). We first calculated the ratio of p-CaMKII against CaMKII (p-CaMKII/CaMKII), and observed an expected trend of increased conversion to p-CaMKII after HFS in both WT and KO mice (Fig. 8B). Comparisons of the basal level of CaMKII and p-CaMKII, respectively, showed no difference before HFS between WT and GluN3A KO mice (Fig. 8C). However, and importantly, 10 min after the HFS, the level of p-CaMKII in the GluN3A KO hippocampus was significantly higher compared with that in the GluN3A KO slices (Fig. 8D). To understand whether the CaMKII signalling in the adult GluN3A KO mice could explain the differences in LTP and the increased memory performance, the selective CaMKII inhibitor, 1-[NO-bis-1,5-isoquinolinesulphonyl]-N-methyl-l-tyro-syl-4-phenylpiperazine (KN-62; 5 μm; Sigma), was applied into the bathing solution 15 min prior to and during the first 5 min after HFS. KN-62 suppressed LTP induction in hippocampal slices from both WT and GluN3A KO mice, and abrogated the LTP difference between them (Fig. 8E).
The present investigation reveals a potential novel role for the NMDAR subunit GluN3A in locomotion, pain sensation and learning/memory in adult animals. Previous investigations on the functional role of the GluN3A subunit have focused on neonates due to the high expression level of this subunit during the brain development. The marked impacts of GluN3A deletion on functional/behavioural activities of adult animals were previously unknown and unexpected. This is a surprising discovery in adult animals considering previous reports on adult GluN3A KO mice and that only low levels of GluN3A expression exist in the adult brain (Wong et al. 2002). The results from our investigation suggest that, even at relatively low expression levels, the functional impact of GluN3A in the adult brain should not be underestimated.
Adult GluN3A KO mice were morphologically and phenotypically indistinguishable from adult WTs. The KO animals grew faster than WT mice during the first month. The difference in body weight, however, disappeared in early adulthood as GluN3A expression declined. This is an interesting and previously unknown phenomenon. We suggest that the role of GluN3A in affecting body weight deserves a further detailed investigation. GluN3A KO mice were generally hypoactive in the open field test compared with the WT counterparts. Under forced circumstances, as in the Rota-rod or the swim test, the locomotor deficits were abrogated in the first trial day. Additionally, GluN3A KO mice showed reduced motor ability and motor skills/coordination on the Rota-rod test, but this deficit was compensated by training. At the brain structural and molecular level, GluN3A is moderately expressed in the striatum and the cerebellum (Wong et al. 2002); both regions heavily implicated in motor learning and coordination (Paulin, 1993; Dang et al. 2006).
Western blot analysis of different LTP-related regulators revealed that GluN3A KO mice have increased basal expression of CaMKII in the forebrain, and increased phosphorylation of CaMKII after HFS in the hippocampus. It is possible that GluN3A-deficient neurons with higher NMDAR Ca2+ permeability permit a facilitated intracellular Ca2+ accumulation that leads to a constitutively higher expression as well as activation of CaMKII. Transgenic mice overexpressing CaMKII in the forebrain show reduced locomotor activity in novel environments (Hasegawa et al. 2009). Thus, higher CaMKII expression in adult GluN3A KO mice might explain the reduction in locomotor activity. In contrast to the CaMKII overexpressing mice that display increased aggression and anxiety-like behaviours (Hasegawa et al. 2009), GluN3A KO mice exhibit unaltered anxiety levels as shown by the elevated plus maze. CaMKII is a major constituent of the PSDs, and plays an important role in initiating a cascade of events that regulate LTP and potentiate synaptic transmission (Lisman et al. 2002). Pharmacological inhibition of CaMKII blocks hippocampal LTP (Stanton & Gage, 1996), and impairs hippocampus-dependent memory and learning (Tan & Liang, 1996). Similar results were also found in transgenic mice with deletion of CaMKII (Silva et al. 1992). CaMKII also regulates nociception. Acute pharmacological inhibition of CaMKII with KN-62 reversed complete Freund's adjuvant (CFA)-induced inflammatory pain responses (Luo et al. 2008). On the other hand, forebrain overexpression of CaMKII reduces CFA-induced allodynia and hyperalgesia without affecting pain responses to heat, mechanical pressure and formalin injection (Wei et al. 2006). While the exact role of CaMKII in mediating LTP and pain sensation may be affected by multiple co-factors, the enhanced expression of CaMKII in the forebrain of the GluN3A KO mice is probably a molecular mediator in enhanced LTP and nociceptive responses.
The enhancement of learning and memory in GluN3A KO mice is similar to that seen in the 'smart mice' overexpressing GluN2B (NR2B; Tang et al. 1999). GluN2B, however, has been identified as a pro-death NMDAR subunit (Liu et al. 2007), whereas deletion or downregulation of GluN3A in the adult brain does not increase the risk of excitotoxicity or ischaemic injury (Nakanishi et al. 2009). In this respect, GluN3A appears to be a more realistic target for learning and memory modulation than GluN2B. To this end, the role of GluN3A in adult behaviour should be confirmed with conditional GluN3A manipulation studies. Supporting our observations, prolonged GluN3A expression in the forebrains of adult mice attenuates LTP and disrupts long-term memory (Roberts et al. 2009). Both memory tests used (Novel Object Recognition and Morris water maze) in this paper require locomotion and, given the difference in locomotor abilities between the two genotypes, it is possible that the difference seen in memory tests may not be due to memory and learning abilities alone. However, we were not able to objectively study emotional memory using contextual and cued fear conditioning, which does not require the measurement of movement because of the difference in acute pain responses. On the other hand, it is possible that the actual memory enhancement is much greater than what we observed in these tests because of the reduced locomotion in GluN3A KO mice. For example, despite their significantly reduced swim speed, GluN3A KO mice still managed to reach the platform in the water maze test faster than WT controls. Moreover, the expression of GluN3A in adult non-human primates is higher in contrast to the rodent brain (Mueller & Meador-Woodruff, 2005), suggesting that the GluN3A role observed in mice could be more prominent in primates.
Our data demonstrate that the GluN3A regulation of LTP is present in immature as well as adult brains. The observed facilitation in LTP is similar to what is seen in PSD-95 KO mice, except that GluN3A KO mice do not show any disruption in LTD (Carlisle et al. 2008). A previous study using the 3× HFS protocol could not detect any LTP difference between adult WT and GluN3A KO mice (Roberts et al. 2009). As we hypothesized in the current investigation, the strong stimulation protocol (3× HFS) used in the earlier study likely overcame the inhibitory effect of GluN3A in the WT mice, and thus prevented the detection of LTP differences between the two genotypes. That same study detected a difference in LTP between P6–P8 WT and GluN3A KO mice with a 3× HFS protocol. This is expected because 3× HFSs are strong enough to stimulate/depolarize the postnatal slices despite the low GluR1 expression at this developing stage. It is also important to mention that, as shown in Fig. 5A, upon more intensive stimuli (3× HFS), LTP in WT slices increased to the same level of KO slices that abrogated the LTP difference between WT and GluN3A KO mice seen with 1× HFS. Similar to our work and as shown in a study published while this work was in revision, there was no difference in the expression of AMPARs and other NMDARs between the WT and GluN3A KO adult brain (Henson et al. 2012). While it is possible that the behavioural differences in adults are due to the difference in GluN3A expression level between the WT and KO mice, a possibility that requires further investigation is that the phenotypic differences found in this study are inherited from the early knocking out of GluN3A during the embryonic and neonatal stages. There is a possibility that GluN3A deletion during brain development may cause subsequent gene regulation and/or synaptic modification that could have impacts on brain functions and behaviour in adulthood. To address the question whether early deletion of GluN3A is necessary for the observed changes in adults, the best approach is a conditional KO or shRNA knockdown in adult animals. As far as we know, successful knockdown or conditional KO of GluN3A in adult animals has not been reported by any group. This issue, therefore, remains to be specifically tested in a future investigation.
Patients suffering from schizophrenia exhibit impaired memory (Aleman et al. 1999), pain insensitivity (Dworkin, 1994) and disrupted motor functions (Marsden & Jenner, 1980). Animal models of schizophrenia also show decreased expression of CaMKII (Matsuo et al. 2009; Novak & Seeman, 2010). Noticeably, these symptoms and CaMKII expression are more or less the opposite phenotypes to what we observed in GluN3A KO mice. Indeed, GluN3A transcription level is elevated by 32% in schizophrenic patients and has been implicated in the pathogenesis of schizophrenia (Mueller & Meador-Woodruff, 2004). However, the association between the GluN3A expression level or polymorphisms and schizophrenia is still controversial (Gulli et al. 2007; Henson et al. 2008; Shen et al. 2009). The present work supports the merit for further investigations on the relationship between GluN3A and schizophrenia. It may additionally indicate a novel target whose modulation in adults may reverse phenotypes in disorders that involve impaired synaptic structures and dysfunctions such as those found in patients with schizophrenia.
Author contributions
O.M.: design and perform experiments, data analysis and manuscript preparation/revision. M.S.: design and perform experiments, data analysis and manuscript preparation/revision. L.W.: concept development, data analysis and manuscript preparation, research grant support. S.P.Y.: concept development, design experiments, data analysis and manuscript preparation/revision, research grant support.
This work was supported by NIH research grants NS057255, NS058710, NS062097 and the AHA Pre-doctoral Fellowship 10 PRE4430032. We thank G. Neigh for providing the behavioural analysis program CleverSys; J. Larimore, V. Faundez, J. McCoy III, Y. Zhu and D. Lambeth for helping with subcellular fractionation; the Emory Rodent Behavioural Core for performing the Morris water maze test; and K. Ressler for advice and comments.


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Wifi Unlocker
Wifi Unlocker its based on the great Work of Rui Araújo, this app its intended to check sucurity bugs only, this should not be used in other way. This app will try get DEFAULT passwords from vulnerable modems, sometimes we can get false positives or even lack of vulnerable wifi signals. This is not problem of the app maybe in your area its not work…
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Create a free account to check out faster, store multiple shipping addresses, view and track your orders in your account, and much more.
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Unlike the charcoal you toss in a grill, activated charcoal—also known as activated carbon—has been treated with oxygen to remove dirt, debris, and pollution deep beneath the skin's surface. And that's just the magical ingredient in this hard-working mask. Harnessing the purifying power of natural charcoal, it gets rids of pore-clogging impurities revealing a more radiant complexion. Still not totally convinced? Read more about its good-for-you benefits people have been using for ages.
I have acne-prone skin, even still at 28 years-old. (Genetics are swell.) Anyway, I'm always on the lookout for any skincare product that will improve my acne without drying my skin out. (It's just a cruel joke to have acne and forehead wrinkles at the same time.) Anyway, I put this bad boy on last night and the first thing I noticed was the intense tingle. It feel like someone was blowing their Listerine breath on my face. (That was meant to sound more pleasant than it probably did.) It got a little intense, but once the product started to dry the intensity came down a notch. And after the rinse, I could SEE the tiny blackheads between my eyebrows had vanished. Long story short, this works. Be sure to throw a good moisturizer on overnight. You will wake up with brand new(ish) skin. You're welcome.5
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I started using this mask a few times in one week and in a couple days I started seeing way more new breakouts than before (and my skin had been pretty clear for a while). I took a break and waited for it to clear up and then several weeks later when I saw the beginnings of a new breakout, I gave this product another shot BUT as a spot-treatment tool (so not all over my face) and it did a really good job for treating that one area.
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Woodbridge Penn Foster
1. Penn Foster
I have two maltase dogs males they use to get along but now all they want to do is fight I believe its jealousey @ need help don't no what to do nex…
2. Univ of Penn
88 Afterglow AveVerona, NJ 07044
3. Lincoln Tech
Serving the Woodbridge Area.
From Business: Lincoln's practical nursing program focuses on providing hands-on training so you can develop the tasks you'll perform as a practical nurse. From taking and reading…
4. Sollers Institute
100 Menlo Park BlvdEdison, NJ 08837
From Business: Sollers education programs are founded on providing advanced level training in the field of clinical research. They are based on a tried and tested curriculum which…
5. San Joaquin Valley College
can you pull teeth I need help
6. Impetus Systems & Careers
850 US Highway 1Edison, NJ 08817
7. Wholeness Wellness Meditation
54 Colleen CtKendall Park, NJ 08824
8. May Drving School
Serving the Woodbridge Area.
From Business: Want to get that permit or license? Do you just need more practice? Either way U.S Driving School can help you achieve your goal in an enjoyable and affordable way.…
9. Essence of You Wellness Center
12 Soltes AveCarteret, NJ 07008
10. Angelic Healing
Middlesex, NJ 08846
11. Lamb of God Full Gospel
Serving the Woodbridge Area.
From Business: Reaching out to the world through a ministry of sound doctrine. Building communities while promoting Jesus Christ. Spiritual Leadership driven by God.
12. Make The Grade Academics
Serving the Woodbridge Area.
From Business: Small group SAT prep, strategies, individual planning, and unique methods for scoring high. Summer program for reading and math for 5, 6, and 7 year olds using Orte…
13. Academic Testing Solutions
120 Perry DrNorth Brunswick, NJ 08902
From Business: ATS offers tutoring support services and Nursing workshops for PN and RN students. ATS offers NCLEX Review workshops in addition to Adult Health I & II Refresher wo…
14. Ultrasound Diagnostic School
675 US Highway 1 SIselin, NJ 08830
15. The School of Medical Technology
223 State Route 18 Ste 204East Brunswick, NJ 08816
16. Central Career Institute
126 Corporate BlvdSouth Plainfield, NJ 07080
From Business: Are you ready to find a career that is meaningful and rewarding to you? Are you ready to make a change to a more stable and more gratifying career path? Central Car…
17. Anthem Institute
Best CAD program in the state, recommend anyone thinking of upgrading their education to consider. Program Manager Frank Minnella is knowledgeable a…
18. Sanford-Brown Institute-Iselin
675 US Highway 1 S 2nd FloorIselin, NJ 08830
From Business: Sanford-Brown Institute in Iselin - Medical Billing, Massage Therapy & Medical Assistant The Sanford-Brown Institute opened in 1933 and is a private, career-oriente…
19. Com-Tec Institute
44 Glenwood AveEast Orange, NJ 07017
From Business: Com-Tec Institute is a full service Healthcare & Technical Training Center Our goal at Com-Tec Institute is to give you the absolute best Healthcare & Technical edu…
20. Medical Technical Institute
60 Park PlNewark, NJ 07102
21. Christ Hospi School-Radiologic
179 Palisade AveJersey City, NJ 07306
From Business: Christ Hospital is a 381-bed acute care facility located in Jersey City, New Jersey. A modern, state-of-the-art facility, Christ Hospital offers a broad range of se…
22. Leeza's Healthcare Consultant,Inc.
24117 44th Ave 1Little Neck, NY 11363
23. All Care Health Institute Center - CLOSED
17 Muhammad Ali AveNewark, NJ 07108
24. Iron Will Kids
910 Sylvan AveEnglewood Cliffs, NJ 07632
25. Far Hills Bedminster First Aid Squad
500 Main StChester, NJ 07930
26. Aureus University
21-00 State Rt 208 # Ste 220Fair Lawn, NJ 07410
27. Empower Yourself of NJ
1 Gateway CtrNewark, NJ 07102
From Business: We offer tutoring and mentoring services online and in person. Participants will also have the opportunity to participate in educational trips such as attending Bro…
28. Manhattan Institute
I would recommend the Manhattan Institute which is located in New York, NY, to anyone interested in pursuing a career in allied health and nursing. …
29. Anderson Health Institute
305 BroadwayPaterson, NJ 07501
30. AACT Solutionc
1 Gateway CtrNewark, NJ 07102
From Business: Self awareness training, Professional development training, mentoring, public speaker, behaviorable training, rehabilitation training, career conseling, personal ad…


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Although the National Basketball Association and its players' union continued to quibble about the financial state of the league over the weekend, there was a glimmer of good news after the sides met to discuss their expiring collective bargaining agreement.
"They indicated they're prepared to talk about everything, and we're prepared to talk about everything," union chief Billy Hunter told ESPN.com on Friday.
What the NBA and its union need to talk seriously about is revamping the age requirement to enter the league. The so-called freshman rule was an 11th-hour concession by the union during the last negotiations in 2005, and it requires a non-international player to be at least one year out of high school and 19 years old to enter the NBA draft. That rule is damaging basketball - the NBA, and the college game.


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'The Profit' on GM: Calm consumers, placate dealers
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Yahoo Tech founder David Pogue never ever wants to get another kidney stone again. Here are some tips to prevent kidney stones.
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Raw: Deer Tries to Jump into SUV That Hit It
Police car video captured what happened after a motorist struck a deer in Howell Township, NJ. The driver stopped her SUV and was attempting to look at the damage when the buck ran back across the road and tried to jump into the driver's seat. (Sept. 28)
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Members of the House Judiciary Committee grilled FBI Director James Comey Wednesday over the Bureau's handling of the investigation into Hillary Clinton's use of her private email server as secretary of state. (Sept. 28)
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The Vermont senator joins the Democratic nominee for president at a Durham, New Hampshire rally to promote her campaign's education plan.
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Yahoo Tech founder David Pogue never thought he would get a kidney stone. But when he did, he was shocked to find how little we know about them, and how crude the treatments seem. So, based on his experience with kidney stones, Pogue has created a user's guide to kidney stones for stone sufferers everywhere.
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Young girl brings many at a Charlotte City Council meeting to tears as she address the council in the wake of the the September 20 shooting of Keith Lamont Scott by Charlotte-Mecklenburg police. (Sept. 28)
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ABC News' Charli James and Michael Rothman take us preview of the 2017 Kitten Bowl.
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Scotts Valley Campgrounds Recreational Vehicle Parks
1. Santa Cruz Ranch RV Resort
917 Disc DrScotts Valley, CA 95066
From Business: Nestled in the trees just minutes from Scotts Valley and Santa Cruz, our resort serves your RV vacation needs. Since the days of California's first inhabitants, Sco…
2. Santa Cruz Redwoods RV Resort
4980 Highway 9Felton, CA 95018
3. Cotillion Gardens RV Park
Bring the family for private sites a lodge with pool table/ping pong, plenty of trails, historic train, stream for fish or swim, close to golf and m…
4. Smithwoods RV Park
Went here July '07, really enjoyed our stay. It's close to Santa Cruz Boardwalk & many state parks. The owners of the park were very nice & the park…
5. Santa Vida RV Park
1611 Branciforte DrSanta Cruz, CA 95065
6. Bobs Pine Grove Campground
210 Golf Club DrSanta Cruz, CA 95060
7. Henry Cowell Redwoods State Park
101 Big Trees Park RdFelton, CA 95018
From Business: Minutes away from the city, this park has old redwoods.
8. California State Park
303 Big Trees Park RdFelton, CA 95018
9. Santa Cruz Harbor RV Park
Beautiful views of Harbor...nice campers, clean and safe. Beach close by near cool restaurants and nice bar. Nice for walking and bike rides in th…
10. Redwood Resort
I came for a short stay and don't want to leave. They go out of the way to make your stay fantastic in every way.
11. Beach Park RV
2505 Portola DrSanta Cruz, CA 95062
12. State Of Ca Parks & Rec
1500 Park AveCapitola, CA 95010
13. Friends Of Sc State Parks
201 State Park DrAptos, CA 95003
14. Seacliff Center Market
218 State Park DrAptos, CA 95003
15. Santa Cruz / Monterey Bay KOA Holiday
1186 San Andreas RdWatsonville, CA 95076
Such a fun place to "camp"! We stayed in a cabin which surprised us with the amenities. Loved the patio with fire pit and BBQ. The play area and bou…
16. Saratoga Springs Picnic & Campgrounds
This is a nice family type campground. My family and friends have been coming to Saratoga Springs Campgrounds for some years now and enjoy every …
17. Big Basin Redwoods State Park
21600 Big Basin WayBoulder Creek, CA 95006
From Business: Towering redwoods meet the sandy beach, offering something outdoorsy for everyone.
18. Pinto Lake Park
I LOVE THIS PLACE! It's simply beautiful at Pinto Lake RV Park. We recommend it to everyone traveling through! Dogs are welcome and the bass fishing…
19. Sunset State Beach
201 Sunset Beach RdWatsonville, CA 95076
20. J & S Surplus & Outdoor Store
HWY 1 & N Struve RdMoss Landing, CA 95039
From Business: * Camping - Hunting & Survival Products * Camouflage & Professional Clothing * Footwear - Backpacks - Sleeping Bags * 6-Pocket Military Pants In Many Colors & Flavo…
21. Coyote Valley Resrt & Recreational Vehicle
9750 Monterey RdCoyote, CA 95013
22. Visalia-Fresno South Koa
Serving the Scotts Valley Area.
From Business: Located near Sequoia and Kings Canyon National Parks. Our location provides opportunities for wonderful day trips.
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Dr. Shannon M. Lensing
Ratings for Dr. Jon R. Goldsmith
We are so happy to find him. He took excellent care of my daughter, even scheduling having her stitches removed around her prom. Now by weird coincidence I need surgery for a cyst on my foot and I wouldn't consider anyone else. Not only is he a great doctor he happens to have a pretty good sense of humor. Good bedside manner counts!
He and his staff are definitely the "A-Team"! I have a rather complicated ankle issue and am so glad I was referred to him. I have to travel over 3 hours for my appointments but it is so worth it. He and his staff are genuinely caring and I do believe Dr. Goldsmith is on top of his game.
My aching bunion was removed by Dr Goldsmith after a thorough discussion with him and his staff informing me of everything to expect before and after. They also supplied me with x-ray photos to compare so I, too, could see the difference in my bone structure after the surgery and healing. There was never a waiting period when I went for my appointments and when I had a question, I picked up the phone and had an answer from the doctor or staff within minutes. I cannot recommend this doctor too highly. He knows his stuff and so does his staff! They are genuinely "good people"!
Great staff, great doctor! My issue was severe but the staff and doctor made it as easy as possible. I would definitely recommend!
Dr. Goldsmith performed surgery on my foot to remove a bunion. The pre-surgery appointment was very helpfull, and allowed me the opportunity to ask any questions I had. The surgery was fully explained ahead of time, with pictures/x-rays from previous similar surgeries. The post-operative care was also excellent. I had to go back for some (what I felt) excessive bleeding 2 days post-op. I was able to get in right away and the issue was taken care of. I'll be going back to Dr. Goldsmith when/if I decide that the other bunion is getting too painful.
Dr. Jon R. Goldsmith's Credentials
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- Dr. William M. Scholl College Of Podiatric Medicine (Grad. 2003)
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O4BO Founder presents at IBM Conference in Hong Kong
O4BO Marcações: saas smartcloud iaas video paas hong presentation o4bo kong ibm 2.667 Visualizações
This video http://tinyurl.com/cscrhey shows my presentation to an IBM Conference in Hong Kong. The conference was about the new release of the IBM SmartCloud which O4BO uses for its Infrastructure as a Service. This video and the PowerPoint Presentation...
do Blog: Michael Oliver
IBMs 10th cloud lab with LotusLive in Hong Kong with Outblaze
Check out this IBM announcement: http://www-03.ibm.com/press/us/en/pressrelease/27214.wss "First IBM cloud computing laboratory in Hong Kong. The new facilitywill provide a global hub for Web-based messaging services to supportIBM's emerging LotusLive...
do Blog: Social Business transformation
ODF Alliance: 350+ members, 50 countries and ODF adopters
AntonySatyadas Marcações: france kong germany malaysia croatia odf minnesota massachusetts india poland belgium extemadura brazil italy alliance denmark norway hong 3.001 Visualizações
The ODF alliance ( http://www.odfalliance.org ) recently applauded Brazil's decision to recommend ODF as the government's preferred format (e-Ping Interoperability Framework version 2.0); India's decision to use ODF at a major state government agency; Italy's...
do Blog: Social Business transformation


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Mesa pediatric dentist providing comprehensive, quality dental services in a compassionate and ch... Read More
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At McClintock Family Dental, we provide high quality, affordable dental care and individualized r... Read More
19. Paradise Ridge Dentistry - David Kloss DDSphoenixarizonadentistry.com
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Cosmetic dentist Scottsdale, Phoenix. Dr. Paul Douglas specializes in cosmetic dentistry includin... Read More
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23. Valley Dental Care - Stephen A Denbrock DDSwww.valleydentalcare.com
1300 N Mcclintock Dr,Chandler, AZ 85226 | Map
Dr. Varda has lived and practiced in the East Valley since 1993 and has completed more than 700 c... Read More
24. Jones Family Dentistry Inc - Paul L Jones DDSwww.jonesfamilydentistry.com
7350 S Mcclintock Dr,Tempe, AZ 85283 | Map
In Tempe Arizona Dentist Paul L. Jones DDS of Jones Family Dentistry,offers dental services to ch... Read More
14545 W Grand Ave,Surprise, AZ 85374 | Map
We know that professional dental care comes with anxiety, but there's no need to worry. John Kim ... Read More
7737 E Indian School Rd,Scottsdale, AZ 85251 | Map
27. Southwest Oral Surgery - William L Hull Iii DMDwww.southwestimplants.com
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The Center for Oral, Facial and Dental Implant Surgery opened in 1991 and has been in its present... Read More
28. Pima Dental Center for Family, Cosmetic & Implant Dentistry- Dr Bruce D Schwartz DDSwww.pimadental.com
10850 N 90th ST,Scottsdale, AZ 85260 | Map
10209 N 35th Ave,Phoenix, AZ 85051 | Map
At Smile for Life Dentistry, we understand that you're looking for high-quality, caring dental se... Read More
1661 S Val Vista Dr,Gilbert, AZ 85295 | Map
Our goal is to build a long-term relationship between our staff & our patients so that we can pro... Read More


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He was a stranger in the rain, needing help. . . only why did he seem so familiar?
Seven years ago this very night, Ciara MacIain lost her fiancé, Derek, in a tragic car accident. As she takes the curve in the storm—at the exact spot he died— Ciara sees a car off the road. A man flags her down asking for help, and he takes her on a journey of the souls, ending in a truth Ciara never expects.
Pearl Nominee 2006 for Best Novella
Rider in the Storm
Deborah MacGillivray
Published by Highland Press Publishing at Smashwords


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Dream trip for brave Maddy
Dream trip for brave Maddy
Team among the finest in the NHS
South Tyneside and Sunderland Healthcare Group
South Tyneside NHS Foundation Trust CQC Report 2015
Thank you for choosing to visit our website. South Tyneside NHS Foundation Trust provide a range of NHS services across Gateshead, South Tyneside and Sunderland. We provide our services in hospital, in the community and in patients' homes ..