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Focus Issue on the Grasses: Foxtail Millet: A Sequence-Driven Grass Model System
Journal ListPlant Physiolv.149(1); 2009 JanPMC2613750
Plant Physiol. Jan 2009; 149(1): 137–141. doi: 10.1104/pp.108.129627PMCID: PMC2613750Focus Issue on the GrassesFoxtail Millet: A Sequence-Driven Grass Model System[W]Andrew N. Doust,* Elizabeth A. Kellogg, Katrien M. Devos, and Jeffrey L. BennetzenDepartment of Botany, Oklahoma State University, Stillwater, Oklahoma 74078 (A.N.D.); Department of Biology, University of Missouri, St. Louis, Missouri 63121 (E.A.K.); and Department of Crop and Soil Sciences and Department of Plant Biology (K.M.D.), and Department of Genetics (J.L.B.), University of Georgia, Athens, Georgia 30602*Corresponding author; e-mail ude.etatsko@tsuod.werdna.Author information ► Article notes ► Copyright and License information ►Received September 8, 2008; Accepted November 6, 2008.Copyright © 2009, American Society of Plant BiologistsThis article has been corrected. See Plant Physiol. 2009 March; 149(3): 1619.This article has been cited by other articles in PMC.Foxtail millet (Setaria italica) is a small diploid C4 panicoid crop species, whose genome is being sequenced by the Joint Genome Institute (JGI) of the Department of Energy. The rationale for sequencing foxtail millet is that it is closely related to the bioenergy grasses switchgrass (Panicum virgatum), napiergrass (Pennisetum purpureum), and pearl millet (Pennisetum glaucum), yet is a more tractable experimental model because of its small diploid genome (1C genome size = 490 Mb) and inbreeding nature. This compares to the larger genomes of the outbreeding species pearl millet (diploid, 1C = 2,352 Mb), napiergrass (tetraploid, 1C = 2,254 Mb), and switchgrass (tetraploid, 1C =1,372–1,666 Mb, octaploid 1C = 2,352–3,136 Mb; Bennett et al., 2000). Switchgrass is largely self-incompatible, which significantly complicates forward or reverse genetics, trait mapping, genetic engineering, and genome sequencing. Foxtail millet is self-compatible and has a highly conserved genome structure relative to the ancestral grass lineage (Devos et al., 1998). Some genetic resources such as genetic maps (Devos et al., 1998; Wang et al., 1998) and a small collection of ESTs (Zhang et al., 2007) are already available for foxtail millet, but most of the tool development and genetic research in foxtail millet will be sequence driven.Foxtail millet will be useful as an experimental crop to investigate many aspects of plant architecture, genome evolution, and physiology in the bioenergy grasses. Foxtail millet, pearl millet, napiergrass, maize (Zea mays), and sorghum (Sorghum bicolor) differ from switchgrass in the decarboxylation enzyme used during C4 photosynthesis (NADP-malic versus NAD-malic; Watson and Dallwitz, 1992), but are otherwise similar in C4 photosynthesis, making foxtail millet of value in understanding this important trait. Last, but not least, the draft foxtail millet genome will provide an assembly guide for any future switchgrass sequencing project, as ongoing work in switchgrass indicates that there is good colinearity between switchgrass and foxtail millet (C. Tobias, personal communication).ORIGIN AND RELATIONSHIPS OF FOXTAIL MILLETThe earliest archeological remains of foxtail millet are found in northern China, in the Cishan and Peiligang ruins in the Yellow River Valley, approximately 7,400 years before present (BP) and 7,935 years BP, respectively (Li and Wu, 1996). A succession of sites in the Yiluo valley of northern China show that foxtail millet was the dominant grain for 4 millennia, before becoming part of the current North China farming tradition of millets, wheat (Triticum aestivum), and legume rotations by 3,600 to 3,300 years BP (Lee et al., 2007). Foxtail millet is also known from archeological remains in Europe and the Middle East, but these are younger, dating from around 4,000 years BP (Austin, 2006).Phylogenetic analyses using both chloroplast and nuclear genes show foxtail and green millet (Setaria viridis) as close relatives (Giussani et al., 2001; Doust et al., 2007), consistent with the hypothesis that foxtail millet is a domesticated version of green millet (Li et al., 1944, 1945; de Wet and Harlan, 1975; Wang et al., 1995; Le Thierry d'Ennequin et al., 1999). The two species are part of a larger monophyletic clade of approximately 300 species that all share the characteristic of inflorescences with both spikelets and sterile branches (bristles; Zuloaga et al., 2000). Pearl millet and napiergrass also belong to this group, as do the sand burrs (Cenchrus spp.).Isozyme and DNA analyses of green and foxtail millet have shown a high level of diversity (Wang et al., 1995; Le Thierry d'Ennequin et al., 2000), and revealed that the genetic distance between green and foxtail accessions from the same region are sometimes smaller than the genetic distance between two foxtail millet accessions from different geographical locations. This has been interpreted as supporting multiple domestications of foxtail millet (Kawase and Sakamoto, 1984, 1987; Jusuf and Pernes, 1985; Fukunaga et al., 1997, 2002, 2005, 2006; Li et al., 1998; Nakayama et al., 1998; Le Thierry d'Ennequin et al., 2000; Benabdelmouna et al., 2001; Kawase et al., 2005), followed by episodes of genetic introgression from green millet (Darmency et al., 1987; Wang et al., 1995; Jarvis and Hodgkin, 1999). Green millet is a serious weed, not only of foxtail millet, but of wheat and other temperate cropping systems worldwide (Defelice, 2002; Dekker, 2003).Foxtail millet and switchgrass last shared a common ancestor about 13 ± 3 million years ago (Vicentini et al., 2008), about the same degree of divergence as seen between maize and sorghum (Swigonova et al., 2004). This is much less than the approximately 60 million years since the divergence of switchgrass and Brachypodium, a grass model species being sequenced by the JGI for bioenergy traits (Garvin et al., 2008; Vicentini et al., 2008). Relative divergence time may be an important consideration when choosing model species for the genetic dissection of traits such as vegetative branching or C4 photosynthetic traits in bioenergy grasses, as closer phylogenetic relationships may indicate more genetic, genomic, and physiological similarity. However, the very small size and fast generation time of Brachypodium make it the grass system of choice for the genetic dissection of traits that are more highly conserved, such as genes involved in cell wall synthesis (Garvin, 2007).MORPHOLOGYFoxtail millet has a relatively small stature, with plants of different accessions varying from 20 to 215 cm tall (Reddy et al., 2006). Foxtail millet has a short generation time (depending on the accession, approximately 5–8 weeks from planting to flowering, 8–15 weeks from planting to seed maturity) and can produce hundreds of seeds per inflorescence (Fig. 1). Seed of foxtail millet are generally not dormant and can easily be cultivated in the glasshouse at densities of up to 100 plants/m2 or in the field in temperate or tropical regions. A recent review details many of the growing requirements and other biology of foxtail and other millets (Dekker, 2003).Figure 1.A, Habit, foxtail millet variety Yugu1 (scale = 6 cm). B, Yugu1 mature inflorescence (scale = 4 cm). C, Green millet variety A10 (scale = 4 cm).Foxtail millet has a typical domesticated plant architectural form consisting of a single stalk or a few tillers, with large inflorescences that mature more or less at the same time (Fig. 1). Its wild progenitor, green millet, has tillers and axillary branches that appear over the life of the plant, and inflorescences that readily shatter to release the seed at maturity (Defelice, 2002). Segregating populations of crosses between the two species produce a wide range of plant architecture, suggesting that an understanding of the genetic control of plant architecture in foxtail millet may provide additional insights into branching patterns, an important component of biomass production (Doust et al., 2004; Doust and Kellogg, 2006).Foxtail millet varieties can also be remarkably drought and salt tolerant, and further breeding for these characteristics can increase the usefulness of foxtail millet for semiarid and marginal lands (Li and Wu, 1996; Dekker, 2003).GERMPLASM RESOURCESCollection of foxtail millet landraces in China started in the 1920s and more than 26,000 accessions, 90% of which are landraces, are currently preserved at the Chinese National Genebank in Beijing. Smaller germplasm collections are maintained at the International Crops Research Institute for the Semi-Arid Tropics, Patancheru, India (1,534 accessions from 26 countries), the National Institute of Agrobiological Sciences, Tsukuba, Japan (1,279 accessions), and the Plant Genetic Resources Conservation Unit of the U.S. Department of Agriculture-Agricultural Research Service (766 accessions; Supplemental Table S1). Phenotypic characterization of the collected materials has indicated that foxtail millet germplasm is highly diverse (Li and Wu, 1996; Reddy et al., 2006).GENETIC MAPSTwo complete genetic maps were created in the late 1990s using RFLP markers (Devos et al., 1998; Wang et al., 1998). The first was generated from an intervarietal cross within foxtail millet, whereas the second was generated in an interspecific cross between foxtail millet and green millet. Although wide crosses often lead to a reduction in recombination, map lengths were very similar in the intra- and interspecific crosses (Wang et al., 1998). The interspecific map used foxtail millet markers plus 111 rice (Oryza sativa) and five wheat RFLPs. The comparison of the position of common markers on the foxtail millet and rice genetic maps showed that the two genomes were highly colinear, with six foxtail millet chromosomes that were orthologous to single rice chromosomes and three foxtail millet chromosomes that were each orthologous to two rice chromosomes (Devos et al., 1998).CURRENT DEVELOPMENTSYugu1, a highly inbred commercial line from China, was selected for genomic sequencing. This was instead of the accession of foxtail millet used to generate the interspecies genetic map, because that accession showed partial male cytoplasmic infertility (Wang et al., 1998). Genomic sequence of foxtail millet inbred Yugu1 will be generated with a shotgun approach by the JGI, and the first (approximately 4×) draft is projected to be available soon after April, 2009. An approximately 11× bacterial artificial chromosome library has recently been constructed from Yugu1 by the Clemson University Genomics Institute (C. Saski and J.L. Bennetzen, unpublished data), and sequences of Yugu1 bacterial artificial chromosome ends will help assemble and orient scaffolds of the shotgun sequence. In addition, several hundred thousand ESTs are being sequenced with RNA from >15 different developmental stages, tissues, or treatment types, including whole seedling, inflorescence, root, and drought-stressed material. These sequences will help annotate the genomic sequence and provide the necessary genes for microarray chip construction.The genome of green millet inbred A10 will be sequenced using next generation sequencing technology and compared with the Yugu1 sequence to identify a set of >1,500 single nucleotide polymorphism markers for the construction of a high-density genetic map in a newly generated cross between Yugu1 and green millet accession A10. The single nucleotide polymorphism-based genetic map will be used to anchor the genomic sequence, facilitate comparative genomic analysis, and allow quantitative trait loci (QTL) analyses of biomass traits. An approximately 8.1× fosmid library from green millet inbred A10 has recently been constructed, and five randomly selected clones from this library have been sequenced (A.C. Pontaroli and J.L. Bennetzen, unpublished data). One of the clones contained tandem ribosomal DNA repeats, while the other four clones contained numerous nuclear genes that exhibited excellent colinearity with the rice genome (A.C. Pontaroli and J.L. Bennetzen, unpublished data).Foxtail millet has been adopted as a high-throughput platform for virus-induced gene silencing to model gene inhibition results that have been technically more difficult in switchgrass (R. Nelson, personal communication). Progress is also being made in developing an efficient transformation system for foxtail millet, necessary for testing mechanistic hypotheses of gene action. Embryogenic callus cultures (Rao et al., 1988; Vishnoi and Kothari, 1996) and direct somatic embryos (J. Zale, personal communication) have been established in vitro for foxtail millet. However, only one lab has reported successfully regenerating transgenic plants, using Agrobacterium, and with an efficiency of 6.6% (Liu et al., 2005). In a comprehensive publication, Liu et al. (2007) investigated the parameters affecting Agrobacterium-mediated transformation of foxtail millet and reported that plant genotype, explant source, time of inoculation, and length of cocultivation influenced gene transfer (Liu et al., 2007).CONCLUSIONFor the familiar model systems such as Arabidopsis (Arabidopsis thaliana), rice, or maize, a fully sequenced genome was the culmination of decades of genetic study, and was the tool that laced together many disparate strands of research. In contrast, for foxtail millet, as for Brachypodium and other novel model systems, the fully sequenced genome is the foundation on which genetic resources will be built. Foxtail millet and Brachypodium illustrate a novel way to create a model, in which the basic genome structure is given and can be interpreted by comparison to the other grasses. QTL studies, forward genetics, and reverse genetics can then proceed rapidly, and in a comparative context from the outset.Foxtail millet provides a valuable tool for investigating the C4 grasses, particularly those that are being developed as biomass sources for biofuel production. It was chosen as a target model system as much for its phylogenetic position and its photosynthetic pathway as for its ease of growth and small genome size. Such choices will become increasingly common in the future, as sequencing costs continue to drop. Model systems can be chosen for their ability to address particular questions or to represent particular phylogenetic groups. This approach is especially promising in grasses, where the presence of several completed genomes (and the promise of more) provides an evolutionary genomic context for each new species sequenced. Multiple sequenced genomes, coupled with the ease of comparative analysis among the highly colinear chromosomes of grasses, allow new genomic sequences to be successfully annotated and related to genetic information from other grass species.The foxtail millet genome will be useful not only for understanding the genetics of biomass production with application to biofuel grasses, but also for the favorable agronomic characteristics of foxtail millet itself. Although foxtail millet is a minor North American grain, it is grown extensively in northern China, Mongolia, India, and Russia, and is also grown for hay in the northern and western Great Plains in the United States (Baltensperger, 1996, 2002). The genome sequence will advance foxtail millet breeding as well as gene discovery (Diao, 2005, 2007).However, even with the planned genomic sequence analyses, EST discovery process, genetic map improvements, and QTL analyses, foxtail millet lacks some essential tools needed to become an ideal model system. Mutagenized populations are few, as are reverse genetics tools for characterization of gene function. An expression array has been constructed to examine gene expression under drought stress, but, to our knowledge, neither additional expression arrays nor metabolomic and proteomic tools are yet under development. These will be needed to provide comprehensive analyses of gene function. However, the presence of a completed genome sequence and the current perception of an urgent need to develop herbaceous sources of biomass for biofuel production will provide a powerful impetus to create a full genetic and genomic toolkit in foxtail millet. In response to this challenge, researchers from the United States, China, France, and Japan are beginning to collaborate to fast track tool development in this promising model species.Supplemental DataThe following materials are available in the online version of this article.Supplemental Table S1. Foxtail millet germplasm resources.Supplementary Material
NotesThe author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Andrew N. Doust (ude.etatsko@tsuod.werdna).[W]The online version of this article contains Web-only data.www.plantphysiol.org/cgi/doi/10.1104/pp.108.129627ReferencesAustin DF (2006) Fox-tail millets (Setaria:Poaceae)—abandoned food in two hemispheres. Econ Bot 60 143–158Baltensperger DD (1996) Foxtail and proso millet. In J Janick, ed, Progress in New Crops. ASHS Press, Alexandria, VA, pp 182–190Baltensperger DD (2002) Progress with proso, pearl, and other millets. In J Janick, A Whipley, eds, Trends in New Crops and New Uses. ASHS Press, Alexandria, VA, pp 100–103Benabdelmouna A, Abirached-Darmency M, Darmency H (2001) Phylogenetic and genomic relationships in Setaria italica and its close relatives based on the molecular diversity and chromosomal organization of 5S and 18S-5.8S-25S rDNA genes. Theor Appl Genet 103 668–677Bennett MD, Bhandol P, Leitch IJ (2000) Nuclear DNA amounts in angiosperms and their modern uses—807 new estimates. Ann Bot (Lond) 86 859–909Darmency H, Zangre GR, Pernes J (1987) The wild-weed-crop complex in Setaria—a hybridization study. Genetica 75 103–107de Wet JMJ, Harlan JR (1975) Weeds and domesticates: evolution in the man-made habitat. Econ Bot 29 99–107Defelice MS (2002) Green foxtail, Setaria viridis (L.) P. Beauv. Weed Technol 16 253–257Dekker J (2003) The foxtail (Setaria) species-group. Weed Sci 51 641–656Devos KM, Wang ZM, Beales J, Sasaki T, Gale MD (1998) Comparative genetic maps of foxtail millet (Setaria italica) and rice (Oryza sativa). Theor Appl Genet 96 63–68Diao X (2005) Advances in foxtail millet biotechnology and its future directions. Journal of Hebei Agricultural Sciences 9 61–68Diao X (2007) Foxtail millet production in China and its future development tendency. In Y Chai, FS Wan, eds, The Industrial Development of China Special Crops. Chinese Agricultural Science and Technology Press, Beijing, pp 32–43Doust AN, Devos KM, Gadberry MD, Gale MD, Kellogg EA (2004) Genetic control of branching in foxtail millet. Proc Natl Acad Sci USA 101 9045–9050 [PMC free article] [PubMed]Doust AN, Kellogg EA (2006) Effect of genotype and environment on branching in weedy green millet (Setaria viridis) and domesticated foxtail millet (Setaria italica) (Poaceae). Mol Ecol 15 1335–1349 [PubMed]Doust AN, Penly AM, Jacobs SWL, Kellogg EA (2007) Congruence, conflict, and polyploidization shown by nuclear and chloroplast markers in the monophyletic “Bristle clade” (Paniceae, Panicoideae, Poaceae). Syst Bot 32 531–544Fukunaga K, Domon E, Kawase M (1997) Ribosomal DNA variation in foxtail millet, Setaria italica (L.) P. Beauv., and a survey of variation from Europe and Asia. Theor Appl Genet 95 751–756Fukunaga K, Ichitani K, Kawase M (2006) Phylogenetic analysis of the rDNA intergenic spacer subrepeats and its implication for the domestication history of foxtail millet, Setaria italica. Theor Appl Genet 113 261–269 [PubMed]Fukunaga K, Ichitani K, Taura S, Sato M, Kawase M (2005) Ribosomal DNA intergenic spacer sequence in foxtail millet, Setaria italica (L.) P. Beauv. and its characterization and application to typing of foxtail millet landraces. Hereditas 142 38–44 [PubMed]Fukunaga K, Wang ZM, Kato K, Kawase M (2002) Geographical variation of nuclear genome RFLPs and genetic differentiation in foxtail millet, Setaria italica (L.) P. Beauv. Genet Resour Crop Evol 49 95–101Garvin DF (2007) Brachypodium: a new monocot model plant system emerges. J Sci Food Agric 87 1177–1179Garvin DF, Gu YQ, Hasterok R, Hazen SP, Jenkins G, Mockler TC, Mur LAJ, Vogel JP (2008) Development of genetic and genomic research resources for Brachypodium distachyon, a new model system for grass crop research. Crop Sci 48 S69–S84Giussani LM, Cota-Sanchez JH, Zuloaga FO, Kellogg EA (2001) A molecular phylogeny of the grass subfamily Panicoideae (Poaceae) shows multiple origins of C4 photosynthesis. Am J Bot 88 1993–2012 [PubMed]Jarvis DI, Hodgkin T (1999) Wild relatives and crop cultivars: detecting natural introgression and farmer selection of new genetic combinations in agroecosystems. Mol Ecol 8 S159–S173Jusuf M, Pernes J (1985) Genetic-variability of foxtail millet (Setaria-Italica P Beauv)—electrophoretic study of 5 isoenzyme systems. Theor Appl Genet 71 385–391 [PubMed]Kawase M, Fukunaga K, Kato K (2005) Diverse origins of waxy foxtail millet crops in East and Southeast Asia mediated by multiple transposable element insertions. Mol Genet Genomics 274 131–140 [PubMed]Kawase M, Sakamoto S (1984) Variation, geographical-distribution and genetic-analysis of esterase isozymes in foxtail millet, Setaria-Italica (L) P-Beauv. Theor Appl Genet 67 529–533 [PubMed]Kawase M, Sakamoto S (1987) Geographical-distribution of landrace groups classified by hybrid pollen sterility in foxtail millet, Setaria-Italica (L) Beauv,P. Jpn J Breed 37 1–9Le Thierry d'Ennequin M, Panaud O, Toupance B, Sarr A (2000) Assessment of genetic relationships between Setaria italica and its wild relative S. viridis using AFLP markers. Theor Appl Genet 100 1061–1066Le Thierry d'Ennequin M, Toupance B, Robert T, Godelle B, Gouyon PH (1999) Plant domestication: a model for studying the selection of linkage. J Evol Biol 12 1138–1147Lee GA, Crawford GW, Liu L, Chen XC (2007) Plants and people from the early neolithic to Shang periods in North China. Proc Natl Acad Sci USA 104 1087–1092 [PMC free article] [PubMed]Li HW, Li CH, Pao WK (1944) Cytological and genetical studies of the interspecific cross of the cultivated foxtail millet, Setaria italica (L.) Beauv., and the green foxtail millet, S. viridis L. J Am Soc Agron 9 32–54Li HW, Li CH, Pao WK (1945) Cytological and genetical studies of the interspecific cross of the cultivated foxtail millet, Setaria-Italica (L) Beauv, and the green foxtail millet, S-Viridis L. J Am Soc Agron 37 32–54Li Y, Jia JZ, Wang Y, Wu SZ (1998) Intraspecific and interspecific variation in Setaria revealed by RAPD analysis. Genet Resour Crop Evol 45 279–285Li Y, Wu SZ (1996) Traditional maintenance and multiplication of foxtail millet (Setaria italica (L) P. Beauv.) landraces in China. Euphytica 87 33–38Liu Y, Yu J, Zhao Q, Zhu D, Ao G (2005) Genetic transformation of millet (Setaria italica) by Agrobacterium-mediated. Chin J Agr Biotechnol 13 32–37Liu YH, Yu JJ, Ao GM, Zhao Q (2007) Factors influencing Agrobacterium-mediated transformation of foxtail millet (Setaria italica). Chin J Biochem Mol Biol 23 531–536Nakayama H, Afzal M, Okuno K (1998) Intraspecific differentiation and geographical distribution of Wx alleles for low amylose content in endosperm of foxtail millet, Setaria italica (L.) Beauv. Euphytica 102 289–293Rao AM, Kishor PBK, Reddy LA, Vaidyanath K (1988) Callus induction and high-frequency plant-regeneration in Italian millet (Setaria-Italica). Plant Cell Rep 7 557–559 [PubMed]Reddy V, Upadhyaya, Gowda C (2006) Characterization of world's foxtail millet germplasm collections for morphological traits. Journal of SAT Agricultural Research 2: 1–3Swigonova Z, Lai JS, Ma JX, Ramakrishna W, Llaca V, Bennetzen JL, Messing J (2004) Close split of sorghum and maize genome progenitors. Genome Res 14 1916–1923 [PMC free article] [PubMed]Vicentini A, Barber JC, Aliscioni AA, Giussani LM, Kellogg EA (2008) The age of the grasses and clusters of origins of C4 photosynthesis. Glob Change Biol 14 2693–2977Vishnoi RK, Kothari SL (1996) Somatic embryogenesis and efficient plant regeneration in immature inflorescence culture of Setaria italica (L) Beauv. Cereal Res Commun 24 291–297Wang R-L, Wendel JF, Dekker JH (1995) Weedy adaptation in Setaria spp. I. Isozyme analysis of genetic diversity and population genetic structure in Setaria viridis. Am J Bot 82 308–317Wang ZM, Devos KM, Liu CJ, Wang RQ, Gale MD (1998) Construction of RFLP-based maps of foxtail millet, Setaria italica (L.) P. Beauv. Theor Appl Genet 96 31–36Watson L, Dallwitz M (1992) The grass genera of the world: descriptions, illustrations, identification, and information retrieval; including synonyms, morphology, anatomy, physiology, phytochemistry, cytology, classification, pathogens, world and local distribution, and references. http://delta-intkey.com (June 6, 2008)Zhang JP, Liu TS, Fu JJ, Zhu Y, Jia JP, Zheng J, Zhao YH, Zhang Y, Wang GY (2007) Construction and application of EST library from Setaria italica in response to dehydration stress. Genomics 90 121–131 [PubMed]Zuloaga FO, Morrone O, Giussani LM (2000) A cladistic analysis of the Paniceae: a preliminary approach. In SWL Jacobs, J Everett, eds, Grasses: Systematics and Evolution. CSIRO, Collingwood, Victoria, Australia, pp 123–135Articles from Plant Physiology are provided here courtesy of American Society of Plant Biologists
Formats:Article | PubReader | ePub (beta) | PDF (197K)Related citations in PubMedGenome sequence of foxtail millet (Setaria italica) provides insights into grass evolution and biofuel potential.[Nat Biotechnol. 2012]Zhang G, Liu X, Quan Z, Cheng S, Xu X, Pan S, Xie M, Zeng P, Yue Z, Wang W, et al. Nat Biotechnol. 2012 May 13; 30(6):549-54. Epub 2012 May 13.Foxtail millet: a model crop for genetic and genomic studies in bioenergy grasses.[Crit Rev Biotechnol. 2013]Lata C, Gupta S, Prasad M. Crit Rev Biotechnol. 2013 Sep; 33(3):328-43. Epub 2012 Sep 18.Genetic structure of landraces in foxtail millet (Setaria italica (L.) P. Beauv.) revealed with transposon display and interpretation to crop evolution of foxtail millet.[Genome. 2011]Hirano R, Naito K, Fukunaga K, Watanabe KN, Ohsawa R, Kawase M. Genome. 2011 Jun; 54(6):498-506. Epub 2011 May 30.Effect of genotype and environment on branching in weedy green millet (Setaria viridis) and domesticated foxtail millet (Setaria italica) (Poaceae).[Mol Ecol. 2006]Doust AN, Kellogg EA. Mol Ecol. 2006 Apr; 15(5):1335-49. Methods for linkage disequilibrium mapping in crops.[Trends Plant Sci. 2007]Mackay I, Powell W. Trends Plant Sci. 2007 Feb; 12(2):57-63. Epub 2007 Jan 16.See reviews...See all...Cited by other articles in PMCIdentification and Molecular Characterization of MYB Transcription Factor Superfamily in C4 Model Plant Foxtail Millet (Setaria italica L.)[PLoS ONE. ]Muthamilarasan M, Khandelwal R, Yadav CB, Bonthala VS, Khan Y, Prasad M. PLoS ONE. 9(10)e109920An ABA-responsive DRE-binding protein gene from Setaria italica, SiARDP, the target gene of SiAREB, plays a critical role under drought stress[Journal of Experimental Botany. 2014]Li C, Yue J, Wu X, Xu C, Yu J. Journal of Experimental Botany. 2014 Oct; 65(18)5415-5427Genome and transcriptome sequencing identifies breeding targets in the orphan crop tef (Eragrostis tef)[BMC Genomics. ]Cannarozzi G, Plaza-Wüthrich S, Esfeld K, Larti S, Wilson YS, Girma D, de Castro E, Chanyalew S, Blösch R, Farinelli L, Lyons E, Schneider M, Falquet L, Kuhlemeier C, Assefa K, Tadele Z. BMC Genomics. 15(1)581Mapping of Quantitative Trait Locus (QTLs) that Contribute to Germination and Early Seedling Drought Tolerance in the Interspecific Cross Setaria italicaxSetaria viridis[PLoS ONE. ]Qie L, Jia G, Zhang W, Schnable J, Shang Z, Li W, Liu B, Li M, Chai Y, Zhi H, Diao X. PLoS ONE. 9(7)e101868Methods to Promote Germination of Dormant Setaria viridis Seeds[PLoS ONE. ]Sebastian J, Wong MK, Tang E, Dinneny JR. PLoS ONE. 9(4)e95109See all...LinksBioProjectBioProjectBioProject linksPubMedPubMedPubMed citations for these articlesTaxonomyTaxonomyRelated taxonomy entryTaxonomy TreeTaxonomy TreeRecent ActivityClearTurn OffTurn OnFocus Issue on the Grasses: Foxtail Millet: A Sequence-Driven Grass Model SystemFocus Issue on the Grasses: Foxtail Millet: A Sequence-Driven Grass Model SystemPlant Physiology. Jan 2009; 149(1)137PMCYour browsing activity is empty.Activity recording is turned off.Turn recording back onSee more...


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Expensive, but well worth a visit - Review of Restaurant Cornelius, Bergen, Norway - TripAdvisor
Expensive, but well worth a visit - Restaurant Cornelius
“Expensive, but well worth a visit”
Ranked #8 of 398 Restaurants in Bergen
Price range: $69 - $94
Expensive, but well worth a visit
This day I heard a lot of good stories, ate a lot of new and good food, and was served some excellent wines. What else can I ask for..
Located far out in the "fjords", this restaurant is as much of an experience in life as a good meal. We took the "company tour" which included a boat trip out (no other way that I know of). On arrival we were told the story behind the place, and were served fresh seafood picked from the bottom of the ocean right outside the restaurant.
I very rarely order anything but beef when I eat at a restaurant, but this time I went for some local sea delicacies. I'll do that again the next time I'm there. The food was great (though I prefer meat), the surroundings great and the people working there were great. All in all well spent time and money.
If you've got some money to burn and you're in Bergen I'd strongly recommend visiting.
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Donny150
Fresh Catch of the Day By the Ocean
We were ferried to this tiny island off of a small island near Bergen. The ferry ride is beautiful and relatively fast. We all took in the scenery then had some welcome bubbly drinks before taking our tables. The restaurant grows its own herbs and keeps some live sea animals caught off the island in their holding tanks for the ultimate fresh taste. We were told about the history of the island by the charming and witty owner while enjoying some fantastic house-smoked salmon and trying fresh raw escalopes including the roe sac which is very mild and sweet and some Scotch oysters. The meals included a freshly caught escalope seared with butter along with some vegetables. The main dish was a typical and very delicious Norwegian Summer white fish (I forgot the name) freshly caught in a zesty cream sauce. The desert was a rhubarb and strawberry cold soup with a berry sorbet. The wine cellar holds a good variety of wines too. Finally, the servers were all excellent and very international (spoke very fluent English). I would highly recommend trying this restaurant if your pockets would allow. Of course, booking before hand is a must to catch the ferry in time.
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Thank Donny150
Fantastic fresh seafood and the best fish soup.
Arriving by boat, all the restaurant staff outside to welcome clients with a smile. Super fresh Oysters and scallops where served immediately. Then inside after a quick explanation of what will be served, we could taste a sensational fish soup followed by other tasteful specialities. Besides the view is amazing, the staff super nice. I loved it!
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1 Thank Teresa110
Loved the place, the interior and the exterior,
Loved what we had for drinks , the wines and the wine cellar.
The Swedish waiter was wonderful, polite, efficient.
The view is spectacular as well.
Ask Jenta about Restaurant Cornelius
If you love seafood this is THE best in Norway.
This was my first visit to this incredible place, carved into the rocky sides of an island around 15 minutes by boat from Bergen. It is almost like the setting from a Bond Movie, exclusive and alluring, a mysterious location that seems to promise either a wonderful meal, or possible kidnap!
Thankfully the focus is on food, something the owner, chef and sommelier will talk to you at length about, not in an oppressive and dull way but with a gift for humour that would grant them a TV series in the UK. Alf, the Crocodile Dundee owner will regale you with tales of diving for Scallops in between explaining the benefits to the libido.
Later on the Swedish sommelier joined in with a full run down of how the place really came together in spite of some bad luck, with loads of humour.
The food, well, the focus is on seafood, locally caught and sourced, much of it dived for by the owning team. There are tanks all over the place filled with Lobster, Scallops and oysters all kept in perfect conditions to give you, the customer, maximum taste and enjoyment. And my goodness, what quality, it is utterly superb and must rank amongst the best I have ever enjoyed.
Of course, good food can become great food with the right wines and this is where the sommelier does his thing, the cellar is extensive but filled with personal choices, exclusive wines found in out of the norm places and the effect is astounding. Rielsing is not normally a favourite of mine, but their choice was just perfect.
I can honestly say that in terms of dining experience, this ranks amongst the best I have ever enjoyed. If it was in Central London or Paris, it would be booked up for years in advance, it really is that good.
The price is perfectly acceptable for this kind of experience. I'd say it suits conference, incentive and just about any group and would work a treat for a romantic evening with a twist. Certainly, as Alf will tell you, the effects of that seafood will kick in just about the time you leave the return boat. A good night therefore, need not end when you leave...
Ask Mitchurchin about Restaurant Cornelius
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Curs Eficienta Sistemelor de Energie
P. 1Curs Eficienta Sistemelor de EnergieCurs Eficienta Sistemelor de EnergieRatings: (0)|Views: 15
|Likes: 0Published by Lavinia Cristina DreghiciuCurs energeticaCurs energeticaMore info:Categories:Types, School WorkPublished by: Lavinia Cristina Dreghiciu on Jul 28, 2013Copyright:Attribution Non-commercialAvailability:Read on Scribd mobile: iPhone, iPad and Android.download as PDF, TXT or read online from ScribdFlag for inappropriate content|Add to collectionSee moreSee lesshttps://www.scribd.com/doc/156503095/Curs-Eficienta-Sistemelor-de-Energie03/20/2014pdftextoriginal You're Reading a Free Preview
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SSP Subiecte Pe Larg
Energia Hidrogenului - Hydrogen Properties Course 2
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Subiectele Dd Rezolvate
Tema 3 Dreghiciu Lavinia-Solicitari Termice in Celule FV
Curs 5 CircuiteEchivalent-recov

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Clinical Management of Chronic Obstructive Pulmonary Disease by Stephen I. Rennard | 9780849375873 | Hardcover | Barnes & Noble
Since the publication of the first edition chronic obstructive pulmonary disease (COPD), as a public health issue, has continually increased in line with the predictions of the World Health Organization - unfortunately, this trend is increasing, and by the year 2020 COPD will become one of the main killers of human life. With several important large scaled trials…
Reviewer: Joel C Seidman, MD (William Beaumont Hospitals)Description: This is the second edition of a book, significantly revised from its first edition published in 2002, which broadly addresses all aspects of chronic obstructive pulmonary disease (COPD). Both editions were inspired by the Global Initiative for Obstructive Lung Disease (GOLD) guidelines published in 2001.Purpose: The book is an attempt to adopt the observations of authorities from diverse experience, providing a comprehensive review of the current state of the art respecting COPD.Audience: The audience is large and varied, but would include primary care internists, pulmonologists, and intensivists. Nurses, respiratory therapists, and rehabilitators would undoubtedly find the reference informative. It would also provide a real-life reference for basic science researchers in putting a face on applications carried out in the laboratory. There are nearly 50 contributors of international renown, although primarily from the United States, Canada, and Western Europe.Features: This is one of the most comprehensive and up-to-date treatises that I have surveyed covering basic molecular biology and genetics underlying the disease for the clinician, epidemiology, early diagnosis, clinical presentation, and management from mildly afflicted to those requiring assisted ventilation and end-of-life palliation. The book focuses exceptionally upon the speculated growing role for pharmacotherapeutics, originally relegated to symptom palliation; that is, it makes inferences upon disease modification and impact upon mortality. In the latter respect, it offers evidence-based medicinal reviews and practice guidelines, constituents often lacking in similar undertakings. Particular emphasis is placed on objective measures of monitoring disease progress, an exercise becoming more important as survival potentially increases and upon hearing about management that might be rendered from multidisciplinary perspectives; experiences, in reality, not always afforded patients and/or their caregivers firsthand. This edition augments substantially the presentation of the last edition. Illustrations and graphics are limited, but sufficient to carry out the ends of each chapter. The bibliographies at each chapter end are extensive and relevant.Assessment: This book achieves its likely intended goals well. For an average practitioner, it comes across as very accessible. I would personally consider it a very practical resource and would be intent upon adding it to my personal library.
Lung Biology in Health and Disease Series, #222

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Great stay, highly recommendable - Review of Hotel Tres Reyes, Pamplona, Spain - TripAdvisor
Hotels near Palacio de Ezpeleta
Great stay, highly recommendable - Hotel Tres Reyes
The price you found is 35% lower than this hotel's average rate of $133/night.
Official website$96
Great stay, highly recommendable
We stayed at this hotel for the weekend with my wife and son, and were give a junior suite. Really beautiful & with space, nice design, comfortable bed. Perfect location, at a walking distance of all the hot spots that Pamplona offers. You can park your car for free at a walking distance, just off the wall, accessible with an urban elevator. Great value for money, will repeat on my next visit.More
Room tip: sunny junior suites situated at the corner of the hotel
Ask marknuyten about Hotel Tres Reyes
VSANCHEZ3REYES, Director General at Hotel Tres Reyes, responded to this reviewResponded April 8, 2019
See you soon and thank you so much for your visit, and your words.
0.8 miles from Hotel Tres Reyes
0.2 miles from Hotel Tres Reyes
The price you found is 43% lower than this hotel's average rate of $138/night.
0.4 miles from Hotel Tres Reyes
The price you found is 47% lower than this hotel's average rate of $171/night.
The price you found is 51% lower than this hotel's average rate of $309/night.
1.8 miles from Hotel Tres Reyes
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2.5 miles from Hotel Tres Reyes
The price you found is 38% lower than this hotel's average rate of $87/night.
2.2 miles from Hotel Tres Reyes
The price you found is 36% lower than this hotel's average rate of $84/night.
0.9 miles from Hotel Tres Reyes
All reviews old town junior suite breakfast was delicious fresh orange juice easy access lovely hotel spent two nights hotel bar loved this hotel excellent location reception staff city parking run edge citadel bull
2 - 7 of 882 reviews
Clean and brightt
We were accommodated one night only. Front desk offer the possibility to upgrate room. €15 extra only. Very raisonable rack price. May be because is January. So we got a clean and bright room at 6th. floor. Nice views over old town of Pamplona, just in front of San Lorenzo church. Confortable mattresses and linen. Nespresso machine. Parking just in front of main entrance.
Ask joanbabot about Hotel Tres Reyes
VSANCHEZ3REYES, Director General at Hotel Tres Reyes, responded to this reviewResponded January 8, 2019
We are very happy to know that your stay has been satisfactory and that you liked our Executive Room.
536orlak
Lovely central clean hotel
Stayed in this hotel last night. It’s a perfect location for exploring Pamplona. We didn’t get a taxi for any reason as everything is within walking distance. Hotel was super clean and breakfast was nice.
Ask 536orlak about Hotel Tres Reyes
1 Thank 536orlak
VSANCHEZ3REYES, Director de Recepción at Hotel Tres Reyes, responded to this reviewResponded December 31, 2018
Thanks a lor four your visit and four your nice words.
See you in 2019 and Happy New Year.
Respite from the Camino albergues
1st class hotel with breakfast included. Located right in the heart of the city. What a great R & R break from the Camino. Staff was friendly and helpful to bedraggled Pilgrims. Spotless room and CLEAN with lots of hot water!!
Ask Donald M about Hotel Tres Reyes
VSANCHEZ3REYES, Director General at Hotel Tres Reyes, responded to this reviewResponded December 18, 2018
Thank you for your visit and your wonderful words.
See you soon in "El Camino".
jtammi
Love this place! Great and attentive service in both English and Spanish (and likely other languages...very professional). The huge bathtub was heavenly. The rooms are comfortable and immaculate. Room service is good. 420'd on the balcony overlooking Old Town Pamplona with no hassles. Gym is satisfactory for getting a workout. Pool water is somewhat cold. Bar area is fun and relaxing to lounge, but the service can be somewhat slow. All in all, my favorite hotel in Spain. On the edge of old town Pamplona and a quick walk to plenty of bars and restaurants. 10 minute walk to Plaza del Castillo.
Ask jtammi about Hotel Tres Reyes
Thank jtammi
VSANCHEZ3REYES, Director General at Hotel Tres Reyes, responded to this reviewResponded December 10, 2018
We love clients, we love our clients.
Merry Christmas and see you soon in Spain.
RguezFdezOscar
Rest like a king
We spent two nights and we really enjoyed our stay. The breakfast is delicious: fresh fruit, tortilla, fresh orange juice, cakes, cheese… They also offer gluten free products such as muffins. The hotel is really quiet, perfect to work and sleep well. It is located in the city centre, next to one of the main parks. They offer parking service for a reasonable price. Our room was big, clean and luminous.
Ask RguezFdezOscar about Hotel Tres Reyes
Thank RguezFdezOscar
VSANCHEZ3REYES, Director General at Hotel Tres Reyes, responded to this reviewResponded November 23, 2018
You are our King.


text:
Opening a door towards Spirit: a parser framework - CodeProject 12,693,254 members (32,334 online)
TonyRMarshall28-May-04 7:28 I tried your sample code on the following string:cmd -key1=value1 -key2="some value for key2"The value of the first key/value pair is "value1 " - with a trailing space.Is this deliberate? How do you adjust the parser to ignore trailing spaces?RegardsTony Last Visit: 31-Dec-99 19:00 Last Update: 18-Jan-17 5:37Refresh1

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Dynamic, yet structured: The cell membrane three decades after the
Singer–Nicolson model
Journal ListProc Natl Acad Sci U S Av.100(14); Jul 8, 2003PMC166180
Proc Natl Acad Sci U S A. 2003 July 8; 100(14): 8053–8058. Published online 2003 June 27. doi: 10.1073/pnas.1332550100PMCID: PMC166180Dynamic, yet structured: The cell membrane three decades after the
Singer–Nicolson modelG. Vereb,*† J. Szöllősi,*†‡ J. Matkó,§ P. Nagy,* T. Farkas,¶ L. Vígh,¶ L. Mátyus,* T. A. Waldmann, and S. Damjanovich*‡***Department of Biophysics and Cell Biology and ‡Cell Biophysical Research Group of the Hungarian Academy of Sciences, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, H-4012, Debrecen, Hungary; §Department of Immunology, Loránd Eötvös University, H-1117, Budapest, Hungary; ¶Institute of Biochemistry, Biological Research Center, Hungarian Academy of Sciences, H-6701, Szeged, Hungary; and Metabolism Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-1374**
To whom correspondence should be sent at the * address. E-mail:
dami/at/jaguar.dote.hu.
†G.V. and J.S. contributed equally to this work.Contributed by T. Farkas, April 29, 2003Author information ► Copyright and License information ►Copyright © 2003, The National Academy of
SciencesThis article has been cited by other articles in PMC.AbstractThe fluid mosaic membrane model proved to be a very useful hypothesis in
explaining many, but certainly not all, phenomena taking place in biological
membranes. New experimental data show that the compartmentalization of
membrane components can be as important for effective signal transduction as
is the fluidity of the membrane. In this work, we pay tribute to the
Singer–Nicolson model, which is near its 30th anniversary, honoring its
basic features, “mosaicism” and “diffusion,” which
predict the interspersion of proteins and lipids and their ability to undergo
dynamic rearrangement via Brownian motion. At the same time, modifications
based on quantitative data are proposed, highlighting the often genetically
predestined, yet flexible, multilevel structure implementing a vast complexity
of cellular functions. This new “dynamically structured mosaic
model” bears the following characteristics: emphasis is shifted from
fluidity to mosaicism, which, in our interpretation, means nonrandom
codistribution patterns of specific kinds of membrane proteins forming
small-scale clusters at the molecular level and large-scale clusters (groups
of clusters, islands) at the submicrometer level. The cohesive forces, which
maintain these assemblies as principal elements of the membranes, originate
from within a microdomain structure, where lipid–lipid,
protein–protein, and protein–lipid interactions, as well as sub-
and supramembrane (cytoskeletal, extracellular matrix, other cell) effectors,
many of them genetically predestined, play equally important roles. The
concept of fluidity in the original model now is interpreted as permissiveness
of the architecture to continuous, dynamic restructuring of the molecular- and
higher-level clusters according to the needs of the cell and as evoked by the
environment.Scientific dogmas, let alone models, rarely survive more than a quarter of
a century without significant modifications. Around its 30th anniversary, the
time seems to be ripe for at least a modest modification of an old paradigm.
The Singer–Nicolson fluid mosaic membrane model (S-N model)
(1) predicts lateral and
rotational freedom and random distribution of molecular components in the
membrane. Membranes had been considered by the S-N model as “a
two-dimensional oriented solution of integral proteins... in the viscous
phospholipid bilayer”
Now it is known, however, that this freedom of protein (and lipid) mobility is
far from being unrestricted. One of the earliest indications of a nonrandom
distribution of proteins was provided by the discovery of cocapping
(4). The emerging evidence on
hierarchically built supramolecular protein complexes
hindered diffusion of proteins in the membrane
(8–10),
and the existence of distinct membrane domains termed “rafts”
(11) also contradicts the S-N
model. Therefore, Jacobson et al.
(2) have correctly stated,
“Most membrane proteins do not enjoy the continuous unrestricted lateral
diffusion.... Instead, proteins diffuse in a more complicated way that
indicates considerable lateral heterogeneity in membrane structure, at least
on a nanometer scale.” The great variety of phospholipid molecular
species, the differences in their molecular shapes and physical properties,
and their asymmetric distribution in the membrane bilayer all indicate a
molecular heterogeneity and the possible formation of membrane microdomains
(12).Advanced Cell Biophysical and Molecular Biological Methodology
Provides Quantitative Data on the Static and Dynamic Organization of
MembranesMembrane dynamics, i.e., the ever changing mobility and proximity
relationships of lipid and protein molecules in the plasma membrane, have a
significant impact on essential cellular processes, such as activation,
ligand-receptor recognition, antigen presentation, intercellular interactions
(e.g., between target and killer cells), etc. Quantitative measurements of
membrane dynamics are possible with fluorescence recovery after
photo-bleaching (13,
14), single-particle tracking
techniques (8,
16), and optical trapping by
laser tweezers (13,
18). Fluorescence correlation
spectroscopy, a method with tradition in the study of reaction kinetics and
molecular interactions in solution
20), also has been applied to
the study of cellular systems recently
22). The method allows the
determination of absolute molecular concentration, mobility, and comobility in
small, confocal volume elements of living cells
(23). Confocal laser-scanning
microscopy (24) at the verge
of its resolution limits proved to be successful in determining the uneven
cell-surface distribution of various antigens. Scanning near-field optical
microscopy (NSOM), a method ideal for assessing localization of membrane
proteins at the resolution of several tens of nanometers, also has been
gaining space in investigating the cytoplasm membrane
(25–28),
although, as Edidin (29)
points out, “while NSOM promises much, its application to biology is
about where electron microscopy was 40 or 50 years ago.” These
methodologies supported by digital image processing add valuable information
to the dynamic data about the spatial distribution and compartmentation of
membrane constituents. In general, these approaches provide evidence for the
domain-like distribution of lipids and proteins in biological membranes
31).Restrictions in the lateral mobility of both lipid and protein components
were studied extensively by using the
fluorescence-recovery-after-photobleaching technique, measuring the diffusion
of fluorescently labeled membrane components from non-bleached areas into a
small, bleached spot. Lateral diffusion parameters of MHC molecules
(9) were highly dependent on
the bleached-spot size. Because diffusion rate in a lipid bilayer is expected
to be independent of this size, one plausible explanation is the mosaic-like
domain structure of the biological membranes that restrict the barrier-free
path of proteins and can be partly responsible for the clustered arrangements
of membrane proteins.Single-particle tracking follows the “random walk” of a gold
particle fixed to a cell-surface protein. For the transferrin receptor and
E-cadherin, a barrier-free path of ≈400 nm was determined
(8), whereas for wild-type and
cytoplasmatically truncated mutant class I MHC, ≈600 and ≈1,700 nm were
measured, respectively (13),
indicating that, for membrane-spanning proteins, the barrier-free path is in
the same range and that barriers to the “free” diffusion are
present 2–3 nm below the membrane bilayer. As shown in
Fig. 1, cytoskeletal elements
and intracytosolic molecules with functional connections to cell-surface
receptors have the capacity either to slow down or to completely stop the
lateral motion of transmembrane proteins.Fig. 1.Proteins experience different types of restrictions to translational
diffusion in the plasma membrane. The view of the membrane is shown from
beneath. A, Proteins showing preferential accumulation in a lipid microdomain
may be confined to the area ...Fluorescence Resonance Energy Transfer (FRET) Highlights
Nanometer-Scale Associations in the Cell MembraneIt has been suggested that membrane proteins can be codistributed
nonrandomly, and this nonrandom yet dynamic codistribution pattern even may be
controlled genetically
This assumption has been supported by a significant amount of experimental
data (36,
37). A major asset in studying
these codistributions was the application of FRET to cellular systems. In
FRET, an excited fluorescent dye (donor) donates energy to an acceptor dye if
the separation distance between them is 1–10 nm. In the 1970s,
cell-surface lectins were the first to be investigated by FRET
39). Since the early 1980s,
more systematic and better established data acquisition and evaluation methods
of FRET were introduced in flow cytometry
40). A quantitative method was
introduced in digital fluorescence microscopy that exploited the differences
in photobleaching kinetics of donor dyes in the presence and absence of
acceptors (36,
41). Recently, a new method,
based on systematic and normalized FRET measurements between individual
receptor pairs yielding a receptor map by triangulation, has been introduced
to describe the exact two-dimensional topology of receptor clusters
37). The homo- and
heteroassociations of MHC class I and II
(42), the IL-2 receptor
α-subunit and intercellular adhesion molecule 1
(43–45),
MHC molecules and transferrin receptors
(46), CD4 and CD8 antigens
48), the T cell antigen
receptor (TCR)/CD3 complex
(49), tetraspan molecules
(CD53, CD81, CD82) and CD20 with MHC class I and class II
(50), the three subunits of
the multisubunit IL-2 receptor
(37), the tumor necrosis
factor receptor (51), Fas
(CD95; ref. 52), and many
others have been analyzed in detail. Many receptors were found to be oligomers
preassembled without ligand binding, including tumor necrosis factor receptor,
Fas, and TCR. Results for the IL-2 receptor indicated that its subunits are
preassembled in T cells and do not require cytokine-induced aggregation. This
colocalization was modulated significantly by binding of relevant ILs. In
addition, there is evidence that the IL-15 receptor α-subunit, which
shares the β- and γ-subunits with IL-2Rα, also can form
preassembled supramolecular structures with IL-2Rβ and the
“common” γ-chain
These data (37) have
challenged the frequently applied paradigm that multisubunit receptors are
assembled under the influence of their specific ligands and support an
alternative model in which preassembled receptor clusters facilitate faster
and stronger biological responses, because there is no need for lateral
diffusion of receptors to associate.Electron Microscopy and Scanning-Force Microscopy Reveal
Submicrometer Clusters of Membrane ReceptorsFRET measurements detect molecular associations in the 1- to 10-nm range.
Combined application of electron and scanning-force microscopy made possible
the discovery of a new, higher hierarchical level of receptor clustering in
lymphoid cells (36). The
distribution of ImmunoGold labels attached to receptors showed a nonrandom
pattern, differing from the Poisson distribution assumed for randomly
scattered molecules. The method also was used under near-physiological
conditions, where the gold particles were detected with scanning-force
microscopy by using tapping mode on hydrated samples
(36). The observation that
nanometer-scale islets of MHC class I molecules in the cell membrane are
organized into micrometer-sized “island groups” was extended to
MHC class II (56) and to the
IL-2 receptor α-subunit and the transferrin receptor
(24). Sequential application
of different-sized gold labels targeted to MHC classes I and II also revealed
successively that positive FRET data did not necessarily mean that each
receptor was homo- or heteroassociated in a large population. The degree of
association for MHC class II to MHC class I was 66%, but only 25% of MHC class
I molecules were in the molecular vicinity of MHC class II
(56).Besides establishing the nonrandom distribution of receptors, ImmunoGold
labeling also provided for estimating the average size of higher-order
molecular clusters (or island groups)
(24). These were in the range
of 400–800 nm for various receptors, in good correlation with mean
barrier-free paths reported earlier
13). Confocal laser-scanning
microscopy of fluorescently labeled, live, and fixed cells followed by surface
reconstruction and spatial autocorrelation analysis confirmed the existence of
such receptor clusters with sizes essentially equal to those deduced from
electron microscopy (24).Lipid Rafts: The Functional Equivalent of Receptor Islands?Numerous studies directed at the plasma membrane have provided evidence for
the existence of distinct domains in the submicron range
(24–26,
56–60).
Paralleling these observations, the term “lipid rafts” was coined
based on studies of epithelial cell polarity and gained widespread popularity
in the past years (11).
Biochemical analysis suggested that rafts consist of cholesterol and
sphingolipids in the exoplasmic leaflet of the lipid bilayer and cholesterol
and phospholipids with saturated fatty acids in the endoplasmic leaflet
(61). They are surrounded by
more fluid membrane domains, which are abundant in unsaturated fatty acids
(62). Polyunsaturated
phosphatidylcholines and phosphatidylethanolamines
(63) including ethanolamine
plasmalogens were detected in rafts by quantitative electrospray ionization
and mass spectrometry (64),
probably reflecting a regulatory mechanism that offsets the rigidifying effect
of cholesterol (63). In
addition, 18:1 and 22:6 phosphatidylethanolamines and ethanolamine
plasmalogens are prone to form nonbilayer phases
66), and the association of
soluble proteins (such as G proteins) to membranes depends on the propensity
of reverse hexagonal-phase areas
(67). This observation
highlights the possible role of lipid rafts in signal-transduction processes.
Isolating lipid rafts as detergent-resistant membrane domains, followed by
Western blotting, indicated that rafts indeed are efficient concentrators of
various proteins, many of them active in cell signaling. Among these were
(68), cholesterol-linked and
palmitoylated proteins such as Hedgehog
(69), Src-family kinases and
α-subunits of heterotrimeric G proteins
(70), cytokine receptors
(71), and integrins
(72).The question is how these biochemically identified, detergent-resistant
membrane domains correspond to the observed confinement of membrane proteins.
As Jacobson and Dietrich (73)
pointed out, “... the nature of the in vivo correlate of such
detergent-resistant membranes remains enigmatic. In principle, microscopy
should be able to determine whether the postulated rafts exist.”
Recently, it has been shown that indeed microscopic equivalents of rafts, or,
rather, aggregates of rafts, can be detected by using high-resolution confocal
microscopy and that their disassembly by depletion or in situ
complexation of cholesterol not only destroys their morphology
(24) but also impairs their
signaling capabilities; for example, in T cells, it hinders Stat5 and Stat3
phosphorylation via the IL-2 receptor
(71). Based on the notion that
rafts essentially are membrane units formed from transport vesicles fusing to
the membrane, one would expect their size to be very small
(59). In photonic force
microscopic experiments, it was determined that raft size is <50 nm in
diameter (74), representing
≈3,500 sphingolipid molecules. This indicates that membrane patches
observed in fluorescence microscopy, bearing raft marker proteins and/or
lipids, probably are aggregates of these basic building blocks. In some
instances, these larger aggregates are not observed in resting cells
76) and can be seen only upon
crosslinking the “unit rafts”
(75). In other cases, cells in
their native state present surface patches of submicrometer size, identifiable
as rafts based on their composition
77).Rafts Are Dynamic Structures Reshaped as Function RequiresA few years ago, little was known about the stability and lifespan of lipid
rafts in living cells. It appears now that, compared with the relatively
stable nature of phases in artificial bilayers, lipid rafts in cells are
relatively short-lived. As Edidin points out in a recent review
(78), “... domains are
now thought to be smaller and less stable then they were in 1992.” Lipid
probes with saturated chains on average spend 13 ms in one domain
(79); the average lifetime of
stable domains is found to be on the scale of tens of seconds
(10). Pulsed EPR measurements
indicate a very fast exchange rate between protein-rich and bulk domains in
the membrane, reaching residency times as low as 15 μs
(80). This possibility of
rapidly changing composition and location in the membrane, as well as the
ability to form aggregates of various sizes, easily can account for the
dynamic regulatory role lipid rafts play in various signaling processes. Such
a function has been established for receptor tyrosine kinases
(27) and for immune receptors
such as the TCR (81) and the
IgE receptor (82,
83).The first step in immunoreceptor signaling is represented by
ligand-dependent receptor aggregation, followed by receptor phosphorylation by
tyrosine kinases of the Src family. Lipid rafts have been identified as
platforms wherein signal transduction molecules may interact with the
aggregated immunoreceptors. Multichain immune recognition receptors such as
TCR (84) and FcεRI
(83) use common mechanisms by
which lipid rafts assist in the initiation of signaling.The onset of T cell activation is associated with the formation of the
so-called “immunological synapse”
(85) between T cells and the
cells that they are recognizing. This synapse starts with the initial
clustering of receptors and promotes the centralized accumulation of TCRs that
has been termed the “central supramolecular activating complex”
with a corresponding peripheral ring, consisting of lymphocyte
function-associated antigen 1 (LFA-1) ligands
(86). The surface components
of antigen-presenting cells are also integral to these clusters;
MHC–peptide complexes are found in the central supramolecular activating
complex, whereas intercellular adhesion molecule 1, the LFA-1 counterreceptor,
is concentrated in the peripheral supramolecular activating complex.
Initially, the TCRs are not necessarily engaged in the center, but as
cell–cell interaction develops, they translocate from the periphery into
the center of the synapse (87,
88). According to the serial
triggering model, in this manner, very few peptide-presenting MHC molecules
can activate a high number of TCRs
(89). It appears important in
terms of the interaction that palmitoylation of the membrane-proximal
cysteines of CD4 and the association of CD4 with Lck contribute to the
enrichment of CD4 in lipid rafts
(90) and that, furthermore,
K+ channels also reside in rafts in the molecular vicinity of TCR
(91). The structure of the
CTL–target cell contact is similar to that observed between T cell and
antigen-presenting cell, a ring of adhesion proteins surrounding the inner
signal molecule domain. Lytic granule secretion occurs in a separate domain
within the adhesion ring (92).
As for the spatiotemporal coordination of signaling by the high-affinity IgE
receptor FcεRI, lipid rafts first concentrate Lyn protein kinases while
excluding the FcεRIs from these domains. Upon crosslinking FcεRIs by
the antigen, the receptors rapidly translocate into the lipid rafts followed
by their phosphorylation and the subsequent recruitment of Syk and PLCγ
into these domains (93). The
latter process also involves accumulation of actin cytoskeleton to the active
domains (82).A recent attempt to repeat the classic experiment of Frye and Edidin
(94) with currently available
technologies provided interesting data underlining the dynamism of membrane
domains. Cells labeled with different fluorescent antibodies were fused with
each other. Near-field scanning optical microscopic and parallel FRET studies
revealed that intermixing of micrometer-scale protein clusters started right
after cell fusion, but there was a delay of about 20 min in the intermixing of
nanometer-scale protein associations
(28), which clearly indicates
the hierarchy of protein associations (Fig.
2). Although these experiments corroborate the existence of
protein clusters, they emphasize their dynamism, which may be important for
rapidly reshuffling protein interactions.Fig. 2.Dynamics in the hierarchical association of membrane proteins. Imaging of
nanometer- and micrometer-sized protein clusters give an overview of the
hierarchical association of membrane proteins. Two cell samples previously
labeled with different fluorescent ...Static and Dynamic Factors Organizing Membrane DomainsThe physical and chemical forces giving rise to membrane domains are under
intensive investigation (2,
57). One presumes that several
intracellular and extracellular constraints and forces influence the size and
distribution of these clusters, one of them being the cholesterol content of
the membrane area in question
95), and changing the
cholesterol composition of the cell membrane alters the association pattern
and signaling properties of various molecules
95).Recently, it was proposed that lipids tend to adopt a super-lattice
distribution in fluid-mixed bilayers and distribution of phospholipids in
these structures is determined by the molecular shape and the charge of the
head group (96,
97). These super-lattice
structures do not cover the whole membrane area; rather, they are in dynamic
equilibrium with areas in which lipids are distributed randomly. The presence
of proteins can modify these structures by depleting or attracting certain
lipid species because of similarities or differences in molecular shapes.
However, fatty acid and polar head group composition of phospholipids
(96), as well as the
thermotropic and lamellar- to nonlamellar-phase transitions, are controlled
precisely in a way that overall fluidity is reached below body temperature.
Molecular architecture of certain phospholipids
(98) and proportion of
nonbilayer-forming lipids (66)
may contribute to this phenomenon. Remarkably, the liquid-ordered- to
liquid-disordered-phase transition temperature of rafts proper is
13–15°C above (99)
the main transition temperature of membranes
(100) because of the high
cholesterol and sphingomyelin content. Under pathological conditions such as
neurodegenerative diseases, this balance is upset.Lipid rafts often are looked at as structures originating solely from
lipid–lipid interactions. One should not, however, overlook the fact
that proteins and protein–lipid interactions could be equally important
in the formation, maintenance, and dynamics of these domains. In fact, one can
imagine the basic unit of a raft as a single protein molecule surrounded by
lipids that are especially suitable as its environment regarding polarity and
steric complementarity. In addition to the classic investigations
(101), where very precise
predictions were given about the membrane-spanning α-helices, recent
data also suggest that the exact composition and nature of α-helical
structures will influence heavily how and in what phospholipid environment
this polypeptide chain can be hosted, if at all
(102). This alone would be
enough to account for protein patterns in cell membranes that are predestined
genetically, as we have predicted decades ago
(32). The primary and
secondary structure of proteins newly synthesized, together with the sorting
capabilities in vesicular transport, can easily determine which proteins will
be embedded into a certain lipid environment and which other proteins will be
their immediate neighbors. For example, it has been demonstrated that the
transmembrane domain of CD40
(103) and influenza
hemagglutinin (104)
determines their partitioning into lipid rafts.On a similar note but looking at the dynamic side, changes in the structure
of proteins [a most evident example being those occurring after their
interaction with other proteins or those generated by changes in membrane
potential (105)], as well as
changes in the constitution or thickness of the various lipid regions, can
easily cause reshuffling of the components of lipid rafts so that the best
steric/energetic stability is achieved again
(106).The strength of interaction between proteins and their immediate lipid
environment is well characterized when lymphoid cells with labeled MHC
molecules are fused (28). MHC
class II molecules from the two cells did not intermix even after 80 min and
maintained their monomeric behavior. It was proposed that the small size of
the microdomain allowed such a strong interaction with the membrane-spanning
parts of the protein molecule that the cohesive forces prevented the fusion of
these microdomains. Interestingly, class II molecules did intermix with class
I molecules of the other cell. MHC class I probably was accommodated in larger
rafts, which could incorporate the small rafts of class II molecules in their
entirety.As for the assembly and maintenance of protein clusters, the
internalization-recycling machinery is one important candidate
“force”: proteins are recruited to sites of endosome formation,
which gives rise to protein associations
108). S—S bonds between
integral membrane proteins were implicated in the case of human killer
cell-activating receptors expressed in the plasma membrane of natural killer
cells as multimeric complexes
(109). Partitioning of
covalently linked, saturated acyl chains into liquid-ordered phase domains is
likely to be an important mechanism for targeting proteins to rafts, whereas
prenylated proteins tend to be excluded from there
(110). Thus, even without
direct protein–protein interactions, preferential accumulation of
certain proteins in a lipid domain may induce homoor heteroassociation as well
as formation of micrometer-sized protein clusters
(Fig. 3).Fig. 3.Association of proteins can be induced by selective accumulation of
proteins in distinct lipid microdomains (a) or by specific
protein–protein interactions (b). (a) The membrane
contains lipid microdomains with distinct lipid compositions. ...Do We Need a New Paradigm?Recent data that do not fit the S-N model can be summarized as follows:
(i) nonrandom codistribution patterns of receptors in the plasma
membrane at different hierarchical levels; (ii) quasipermanent
molecular contacts to cytoskeletal elements and signal-transducing molecules;
(iii) much shorter barrier-free path than expected for unrestricted
diffusion; (iv) domain structure of the lipid components of membranes
has the capacity to segregate or colocalize membrane proteins; (v)
participation of integral membrane proteins in the maintenance of membrane
domains suggests that proteins are as important as structural elements as
lipids; and (vi) dynamic reorganization of protein elements in
membrane domains allows for streamlined cellular responses and is restricted
by protein–lipid and protein–protein interactions.Thus, the straightforward application of the S-N model as a frame of events
is impossible without introducing a new concept. This new concept has the
following attributes emphasizing the colocalization, comobility, and nonrandom
codistribution of a significant number of cell-surface molecules: (i)
the mobility of the cell-surface (transmembrane,
glycosylphosphatidylinositol-anchored, or any other type of) proteins is
restricted by lipid-domain segregation and the length of the free diffusion
pathway covered without bumping into boundaries; (ii) membrane
proteins may colocalize with each other on the 1- to 10-nm scale in a
homologous or heterologous fashion, making the mosaicism of the S-N model
prevalent; (iii) a second hierarchical level of protein clustering
ranging to several hundred nanometers can be observed for many membrane
proteins; (iv) proteins or protein clusters frequently are
accommodated by lipid rafts organized by weak or strong interactions above,
inside, or below the cell membrane; (v) some receptor types (e.g.,
tumor necrosis factor and IL receptors) are in a preassembled supramolecular
formation even in the absence of their physiological ligands and may form a
tighter formation upon ligand binding; (vi) ligand-evoked receptor
aggregations (e.g., epidermal growth factor receptor family) are distinctly
different from preformed oligomers, yet may serve as equally important
amplifying factors of transmembrane signaling; (vii) the smallest
microdomains can be considered modules that accommodate membrane proteins
either alone or in functional oligomers preassembled from subunits, and these
can be the building units of larger signaling domains (such as those formed in
the immunological synapse); (viii) the α-helical
membrane-spanning parts of transmembrane proteins are matched in length and
shape by the aliphatic side chains of lipids constituting the membrane domains
that preferentially accommodate them; (ix) the localization of
proteins in different lipid regions of the plasma membrane can be determined
genetically because the amino acid sequence of the transmembrane domain and
sequence-dependent covalent modifications define the possible, specific
lipid–protein interactions; (x) proteins are likely to have an
equally important role in determining the constituents, structure, and
dynamics of membrane domains; (xi) whereas artificial lipid bilayers
tend to spontaneously form segregated structures, the dynamics and specificity
in the living cell membrane are provided by specific protein–protein and
protein–lipid interactions, as well as the targeted and sensitively
regulated nature of vesicular transport processes; and (xii)
identification of the origin and characteristics of microdomains and receptor
assemblies therein may help us understand the immediate past and future of
cells, their activation state, and reactivity. Such signals may carry
diagnostic, prognostic, or even therapeutic values if these nonrandom receptor
patterns can be linked to diseases affecting the different states and/or
altered genetic material of the cell.In light of the above attributes, we must understand that biological
details are far more complicated than the resolving power of a simple model,
which describes generalized, uniform behavior of molecules in the membrane.
The S-N model is valid, and free diffusion can occur within domain borders,
where molecular interactions do not interfere. This means that the emphasis
must be shifted from the fluidity to the mosaicism of the S-N model. Mosaicism
can restrict free diffusion through one of the following ways: (i)
lipid domain structure, (ii) cytoskeletal or other cytosolic
interactions, or (iii) homo- and heteroassociations with other
integral proteins. These interactions have the capacity to increase the
lifetime of an intermolecular encounter, thereby increasing the possibilities
for bi- or multilateral interactions, which sometimes simply are called
receptor crosstalk. Hence, the overall mobility of molecular elements of the
membrane can be accepted with the above restrictions, making the membrane a
heavily compartmentalized, quasi-two-dimensional structure, which is more
mosaic-like than fluid. In this two-dimensional plane, diffusion,
intermolecular forces, the ever changing membrane potential, and extracellular
influences can dynamically generate and destroy supramolecular structures. We
propose that this new model of the cell membrane be called the dynamically
structured mosaic model.AcknowledgmentsThis work was supported in part by Hungarian Academy of Sciences Grants
OTKA T037831, T034393, TS 040773, TS 044836, T042618, T043061, T043087, and
T043509. G.V. is a Békésy Fellow of the Hungarian Ministry of
Education.NotesAbbreviations: S-N model, Singer–Nicolson fluid mosaic membrane
model; FRET, fluorescence resonance energy transfer; TCR, T cell antigen
receptor.References1. Singer, S. J. & Nicolson, G. L. (1972)
Science 175,
720-731. [PubMed]2. Jacobson, K., Sheets, E. D. & Simson, R. (1995)
1441-1442. [PubMed]3. Sheets, E. D., Simson, R. & Jacobson, K. (1995)
7, 707-714. [PubMed]4. Goding, J. W. & Layton, J. E. (1976) J.
Exp. Med. 144,
857. [PMC free article] [PubMed]5. Damjanovich, S., Gáspár, R., Jr., & Pieri, C.
(1997) Q. Rev. Biophys.
30, 67-106. [PubMed]6. Damjanovich, S., Matkó, J., Mátyus, L., Szabó,
G. J., Szöllősi, J., Pieri, C., Farkas, T. &
Gáspár, R. J. (1998)
Cytometry 33,
225-234. [PubMed]7. Damjanovich, S., Bene, L., Matkó, J., Mátyus, L.,
Krasznai, Z., Szabó, G., Pieri, C., Gáspár, R. &
Szöllősi, J. (1999) Biophys.
Chem. 82,
99-108. [PubMed]8. Kusumi, A., Sako, Y. & Yamamoto, M. (1993)
Biophys. J. 65,
2021-2040. [PMC free article] [PubMed]9. Edidin, M. (1993) J. Cell Sci.
Suppl. 17,
165-169. [PubMed]10. Dietrich, C., Yang, B., Fujiwara, T., Kusumi, A. & Jacobson, K.
(2002) Biophys. J.
274-284. [PMC free article] [PubMed]11. Simons, K. & Ikonen, E. (1997)
569-572. [PubMed]12. Shaikh, S. R., Dumaual, A. C., Jenski, L. J. & Stillwell, W.
(2001) Biochim. Biophys. Acta
317-328. [PubMed]13. Edidin, M., Zuniga, M. C. & Sheetz, M. P. (1994)
3378-3382. [PMC free article] [PubMed]14. Jacobson, K., Elson, E., Koppel, D. & Webb, W.
283-284. [PubMed]15. Anderson, C. M., Georgiou, G. N., Morrison, I. E., Stevenson, G. V.
& Cherry, R. J. (1992) J. Cell Sci.
415-425. [PubMed]16. Simson, R., Sheets, E. D. & Jacobson, K. (1995)
Biophys. J. 69,
989-993. [PMC free article] [PubMed]17. Edidin, M., Kuo, S. C. & Sheetz, M. P. (1991)
1379-1382. [PubMed]18. Sako, Y. & Kusumi, A. (1995) J. Cell
1559-1574. [PMC free article] [PubMed]19. Magde, D., Elson, E. L. & Webb, W. W. (1974)
Biopolymers 13,
29-61. [PubMed]20. Ehrenberg, M. & Rigler, R. (1976) Q.
Rev. Biophys. 9,
69-81. [PubMed]21. Brock, R., Vámosi, G., Vereb, G. & Jovin, T. M.
10123-10128. [PMC free article] [PubMed]22. Schwille, P., Haupts, U., Maiti, S. & Webb, W. W.
(1999) Biophys. J.
2251-2265. [PMC free article] [PubMed]23. Elson, E. L. (2001) Traffic
2, 789-796. [PubMed]24. Vereb, G., Matkó, J., Vámosi, G., Ibrahim, S. M.,
Magyar, E., Varga, S., Szöllősi, J., Jenei, A.,
Gáspár, R., Jr., Waldmann, T. A., et al.
6013-6018. [PMC free article] [PubMed]25. Vereb, G., Meyer, C. K. & Jovin, T. M. (1997) in
Interacting Protein Domains: Their Role in Signal and Energy
Transduction, NATO ASI series, ed. Heilmeyer, L. M. G., Jr.
(Springer, New York), Vol. H102, pp.
49-52.26. Hwang, J., Gheber, L. A., Margolis, L. & Edidin, M.
(1998) Biophys. J.
2184-2190. [PMC free article] [PubMed]27. Nagy, P., Jenei, A., Kirsch, A. K., Szöllősi, J.,
Damjanovich, S. & Jovin, T. M. (1999) J. Cell
1733-1741. [PubMed]28. Nagy, P., Mátyus, L., Jenei, A., Panyi, G., Varga, S.,
Matkó, J., Szöllősi, J., Gáspár, R., Jovin,
T. M. & Damjanovich, S. (2001) J. Cell
4063-4071. [PubMed]29. Edidin, M. (2001) Traffic
2, 797-803. [PubMed]30. Edidin, M. J. (1988) Immunol.
Today 9,
218-219. [PubMed]31. Cherry, R. J., Smith, P. R., Morrison, I. E. & Fernandez, N.
(1998) FEBS Lett.
430, 88-91. [PubMed]32. Damjanovich, S., Somogyi, B. & Trón, L.
(1981) in Advances in Physiological
Sciences, Neural Communication and Control, eds. Székely,
G., Lábos, E. & Damjanovich, S. (Pergamon, Oxford), Vol.
30, pp. 9-21.33. Damjanovich, S., Tron, L., Szöllősi, J., Zidovetzki,
R., Vaz, W. L., Regateiro, F., Arndt-Jovin, D. J. & Jovin, T. M.
(1983) Proc. Natl. Acad. Sci. USA
5985-5989. [PMC free article] [PubMed]34. Damjanovich, S., Szöllősi, J. & Trón, L.
(1992) Immunol. Today
A12-A15. [PubMed]35. Szöllősi, J. & Damjanovich, S. (1994)
in Mobility and Proximity in Biological Membranes,
eds. Damjanovich, S. & Edidin, M. (CRC, Boca Raton, FL), pp.
49-108.36. Damjanovich, S., Vereb, G., Schaper, A., Jenei, A., Matkó,
J., Starink, J. P., Fox, G. Q., Arndt-Jovin, D. J. & Jovin, T. M.
(1995) Proc. Natl. Acad. Sci. USA
1122-1126. [PMC free article] [PubMed]37. Damjanovich, S., Bene, L., Matkó, J., Alileche, A., Goldman,
C. K., Sharrow, S. & Waldmann, T. A. (1997) Proc.
13134-13139. [PMC free article] [PubMed]38. Fernandez, S. M. & Berlin, R. D. (1976)
Nature 264,
411-415. [PubMed]39. Chan, S. S., Arndt-Jovin, D. J. & Jovin, T. M.
(1979) J. Histochem. Cytochem.
27, 56-64. [PubMed]40. Szöllősi, J., Damjanovich, S., Mulhern, S. A. &
Tron, L. (1987) Prog. Biophys. Mol. Biol.
49, 65-87. [PubMed]41. Jovin, T. M. & Arndt-Jovin, D. J. (1989) in
Cell Structure and Function by
Microspectrofluorimetry, eds. Kohen, E. & Hirschberg, J. G.
(Academic, San Diego), pp. 99-117.42. Szöllősi, J., Damjanovich, S., Balázs, M., Nagy,
P., Tron, L., Fulwyler, M. J. & Brodsky, F. M. (1989)
208-213. [PubMed]43. Szöllősi, J., Damjanovich, S., Goldman, C. K.,
Fulwyler, M. J., Aszalós, A. A., Goldstein, G., Rao, P., Talle, M. A.
& Waldmann, T. A. (1987) Proc. Natl. Acad. Sci.
7246-7250. [PMC free article] [PubMed]44. Edidin, M., Aszalós, A., Damjanovich, S. & Waldmann, T.
A. (1988) J. Immunol.
1206-1210. [PubMed]45. Burton, J., Goldman, C. K., Rao, P., Moos, M. & Waldmann, T. A.
(1990) Proc. Natl. Acad. Sci. USA
7329-7333. [PMC free article] [PubMed]46. Mátyus, L., Bene, L., Heyligen, H., Raus, J. &
Damjanovich, S. (1995) Immunol. Lett.
203-208. [PubMed]47. Mittler, R. S., Goldman, S. J., Spitalny, G. L. & Burakoff, S.
J. (1989) Proc. Natl. Acad. Sci. USA
8531-8535. [PMC free article] [PubMed]48. Lee, P. U. & Kranz, D. M. (2003) Mol.
Immunol. 39,
687-695. [PubMed]49. De la Hera, A., Muller, U., Olsson, C., Isaaz, S. &
Tunnacliffe, A. J. (1991) J. Exp. Med.
173, 7-17. [PMC free article] [PubMed]50. Szöllősi, J., Horejsi, V., Bene, L., Angelisova, P.
& Damjanovich, S. (1996) J. Immunol.
2939-2946. [PubMed]51. Chan, F. K., Chun, H. J., Zheng, L., Siegel, R. M., Bui, K. L.
& Lenardo, M. J. (2000) Science
2351-2354. [PubMed]52. Siegel, R. M., Frederiksen, J. K., Zacharias, D. A., Chan, F. K.,
Johnson, M., Lynch, D., Tsien, R. Y. & Lenardo, M. J. (2000)
2354-2357. [PubMed]53. Tagaya, Y., Burton, J. D., Miyamoto, Y. & Waldmann, T. A.
4928-4939. [PMC free article] [PubMed]54. Waldmann, T. A. & Tagaya, Y. (1999)
19-49. [PubMed]55. Waldmann, T. A. (2003) Annu. Rev.
Immunol. 21,
1-27. [PubMed]56. Jenei, A., Varga, S., Bene, L., Mátyus, L., Bodnar, A.,
Bacso, Z., Pieri, C., Gáspár, R., Jr., Farkas, T. &
Damjanovich, S. (1997) Proc. Natl. Acad. Sci.
7269-7274. [PMC free article] [PubMed]57. Edidin, M. (1997) Curr. Opin. Struct.
528-532. [PubMed]58. Kenworthy, A. K. & Edidin, M. (1998) J.
69-84. [PMC free article] [PubMed]59. Varma, R. & Mayor, S. (1998)
798-801. [PubMed]60. Horejsi, V. (2002) Trends
Immunol. 23,
562-564. [PubMed]61. Fridriksson, E. K., Shipkova, P. A., Sheets, E. D., Holowka, D.,
Baird, B. & McLafferty, F. W. (1999)
Biochemistry 38,
8056-8063. [PubMed]62. Schroeder, R., London, E. & Brown, D. (1994)
12130-12134. [PMC free article] [PubMed]63. Blom, T. S., Koivusalo, M., Kuismanen, E., Kostiainen, R.,
Somerharju, P. & Ikonen, E. (2001)
14635-14644. [PubMed]64. Pike, L. J., Han, X., Chung, K. N. & Gross, R. W.
2075-2088. [PubMed]65. Giorgione, J., Epand, R. M., Buda, C. & Farkas, T.
9767-9770. [PMC free article] [PubMed]66. Lohner, K. (1996) Chem. Phys.
Lipids 81,
167-184. [PubMed]67. Escriba, P. V., Ozaita, A., Ribas, C., Miralles, A., Fodor, E.,
Farkas, T. & Garcia-Sevilla, J. A. (1997) Proc.
11375-11380. [PMC free article] [PubMed]68. Horejsi, V., Cebecauer, M., Cerny, J., Brdicka, T., Angelisova, P.
& Drbal, K. (1998) Immunol. Lett.
63, 63-73. [PubMed]69. Rietveld, A., Neutz, S., Simons, K. & Eaton, S.
12049-12054. [PubMed]70. Resh, M. D. (1999) Biochim. Biophys.
Acta 1451,
1-16. [PubMed]71. Matkó, J., Bodnar, A., Vereb, G., Bene, L., Vámosi,
G., Szentesi, G., Szöllősi, J., Gáspár, R., Horejsi,
V., Waldmann, T. A., et al. (2002) Eur. J.
Biochem. 269,
1199-1208. [PubMed]72. Baron, W., Decker, L., Colognato, H. & ffrench-Constant, C.
151-155. [PubMed]73. Jacobson, K. & Dietrich, C. (1999)
Trends Cell Biol. 9,
87-91. [PubMed]74. Pralle, A., Keller, P., Florin, E. L., Simons, K. & Horber, J.
K. (2000) J. Cell Biol.
997-1008. [PMC free article] [PubMed]75. Harder, T., Scheiffele, P., Verkade, P. & Simons, K.
929-942. [PMC free article] [PubMed]76. Kenworthy, A. K., Petranova, N. & Edidin, M.
(2000) Mol. Biol. Cell
1645-1655. [PMC free article] [PubMed]77. Pyenta, P. S., Holowka, D. & Baird, B. (2001)
2120-2132. [PMC free article] [PubMed]78. Edidin, M. (2001) Trends Cell
492-496. [PubMed]79. Schutz, G. J., Kada, G., Pastushenko, V. P. & Schindler, H.
892-901. [PMC free article] [PubMed]80. Kawasaki, K., Yin, J. J., Subczynski, W. K., Hyde, J. S. &
Kusumi, A. (2001) Biophys. J.
738-748. [PMC free article] [PubMed]81. Langlet, C., Bernard, A. M., Drevot, P. & He, H. T.
(2000) Curr. Opin. Immunol.
250-255. [PubMed]82. Holowka, D., Sheets, E. D. & Baird, B. (2000)
1009-1019. [PubMed]83. Wofsy, C., Vonakis, B. M., Metzger, H. & Goldstein, B.
8615-8620. [PMC free article] [PubMed]84. Bini, L., Pacini, S., Liberatori, S., Valensin, S., Pellegrini, M.,
Raggiaschi, R., Pallini, V. & Baldari, C. T. (2003)
Biochem. J. 369,
301-309. [PMC free article] [PubMed]85. Paul, W. E. & Seder, R. A. (1994)
241-251. [PubMed]86. Krummel, M. F. & Davis, M. M. (2002)
66-74. [PubMed]87. Dustin, M. L. (2002) J. Clin.
Invest. 109,
155-160. [PMC free article] [PubMed]88. Hiltbold, E. M., Poloso, N. J. & Roche, P. A.
(2003) J. Immunol.
1329-1338. [PubMed]89. Valitutti, S., Muller, S., Cella, M., Padovan, E. &
Lanzavecchia, A. (1995) Nature
148-151. [PubMed]90. Fragoso, R., Ren, D., Zhang, X., Su, M. W., Burakoff, S. J. &
Jin, Y. J. (2003) J. Immunol.
913-921. [PubMed]91. Panyi, G., Bagdány, M., Bodnar, A., Vámosi, G.,
Szentesi, G., Jenei, A., Mátyus, L., Varga, S., Waldmann, T. A.,
Gáspár, R., et al. (2003) Proc.
2592-2597. [PMC free article] [PubMed]92. Stinchcombe, J. C., Bossi, G., Booth, S. & Griffiths, G. M.
751-761. [PubMed]93. Kovarova, M., Tolar, P., Arudchandran, R., Draberova, L., Rivera,
J. & Draber, P. (2001) Mol. Cell.
8318-8328. [PMC free article] [PubMed]94. Frye, L. D. & Edidin, M. (1970) J. Cell
319-335. [PubMed]95. Rothberg, K. G., Ying, Y. S., Kamen, B. A. & Anderson, R. G.
(1990) J. Cell Biol.
2931-2938. [PMC free article] [PubMed]96. Somerharju, P., Virtanen, J. A. & Cheng, K. H.
(1999) Biochim. Biophys. Acta
32-48. [PubMed]97. Virtanen, J. A., Cheng, K. H. & Somerharju, P.
4964-4969. [PMC free article] [PubMed]98. Farkas, T., Kitajka, K., Fodor, E., Csengeri, I., Lahdes, E., Yeo,
Y. K., Krasznai, Z. & Halver, J. E. (2000) Proc.
6362-6366. [PMC free article] [PubMed]99. Gousset, K., Wolkers, W. F., Tsvetkova, N. M., Oliver, A. E.,
117-128. [PubMed]100. Tablin, F., Oliver, A. E., Walker, N. J., Crowe, L. M. & Crowe,
J. H. (1996) J. Cell. Physiol.
305-313. [PubMed]101. Hartmann, E., Rapoport, T. A. & Lodish, H. F.
(1989) Proc. Natl. Acad. Sci. USA
5786-5790. [PMC free article] [PubMed]102. Lewis, R. N., Zhang, Y. P., Hodges, R. S., Subczynski, W. K.,
Kusumi, A., Flach, C. R., Mendelsohn, R. & McElhaney, R. N.
12103-12111. [PubMed]103. Bock, J. & Gulbins, E. (2003) FEBS
Lett. 534,
169-174. [PubMed]104. Scheiffele, P., Roth, M. G. & Simons, K. (1997)
5501-5508. [PMC free article] [PubMed]105. Fromherz, P. (1988) Proc. Natl. Acad. Sci.
6353-6357. [PMC free article] [PubMed]106. Bene, L., Szöllősi, J., Balázs, M.,
Mátyus, L., Gáspár, R., Ameloot, M., Dale, R. E. &
Damjanovich, S. (1997) Cytometry
353-357. [PubMed]107. Gheber, L. A. & Edidin, M. (1999)
Biophys. J. 77,
3163-3175. [PMC free article] [PubMed]108. Tang, Q. & Edidin, M. (2001) Biophys.
J. 81,
196-203. [PMC free article] [PubMed]109. Olcese, L., Cambiaggi, A., Semenzato, G., Bottino, C., Moretta, A.
& Vivier, E. (1997) J. Immunol.
5083-5086. [PubMed]110. Melkonian, K. A., Ostermeyer, A. G., Chen, J. Z., Roth, M. G. &
Brown, D. A. (1999) J. Biol. Chem.
3910-3917. [PubMed]Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences
Formats:Article | PubReader | ePub (beta) | PDF (413K)Related citations in PubMedThe Fluid-Mosaic Model of Membrane Structure: Still relevant to understanding the structure, function and dynamics of biological membranes after more than 40years.[Biochim Biophys Acta. 2013]Nicolson GL. Biochim Biophys Acta. 2013 Nov 1; . Epub 2013 Nov 1.Is a fluid-mosaic model of biological membranes fully relevant? Studies on lipid organization in model and biological membranes.[Cell Mol Biol Lett. 2003]Wiśniewska A, Draus J, Subczynski WK. Cell Mol Biol Lett. 2003; 8(1):147-59. Dynamic organizing principles of the plasma membrane that regulate signal transduction: commemorating the fortieth anniversary of Singer and Nicolson's fluid-mosaic model.[Annu Rev Cell Dev Biol. 2012]Kusumi A, Fujiwara TK, Chadda R, Xie M, Tsunoyama TA, Kalay Z, Kasai RS, Suzuki KG. Annu Rev Cell Dev Biol. 2012; 28:215-50. Epub 2012 Aug 16.Supramolecular receptor structures in the plasma membrane of lymphocytes revealed by flow cytometric energy transfer, scanning force- and transmission electron-microscopic analyses.[Cytometry. 1998]Damjanovich S, Matkó J, Mátyus L, Szabó G Jr, Szöllosi J, Pieri JC, Farkas T, Gáspár R Jr. Cytometry. 1998 Oct 1; 33(2):225-33. Dynamics in the plasma membrane: how to combine fluidity and order.[EMBO J. 2006]Marguet D, Lenne PF, Rigneault H, He HT. EMBO J. 2006 Aug 9; 25(15):3446-57. Epub 2006 Jun 22.See reviews...See all...Cited by other articles in PMCMonte Carlo simulations of plasma membrane corral-induced EGFR clustering[Journal of biotechnology. 2011]Costa MN, Radhakrishnan K, Edwards JS. Journal of biotechnology. 2011 Feb 10; 151(3)10.1016/j.jbiotec.2010.12.009Melanoma Biomolecules: Independently Identified but Functionally Intertwined[Frontiers in Oncology. ]Dye DE, Medic S, Ziman M, Coombe DR. Frontiers in Oncology. 3252Monolayer curvature stabilizes nanoscale raft domains in mixed lipid bilayers[Proceedings of the National Academy of Scie...]Meinhardt S, Vink RL, Schmid F. Proceedings of the National Academy of Sciences of the United States of America. 2013 Mar 19; 110(12)4476-4481Non-Enzymatic Modification of Aminophospholipids by Carbonyl-Amine Reactions[International Journal of Molecular Sciences...]Naudí A, Jové M, Ayala V, Cabré R, Portero-Otín M, Pamplona R. International Journal of Molecular Sciences. 14(2)3285-3313Identification of Cell-Surface Molecular Interactions under Living Conditions by Using the Enzyme-Mediated Activation of Radical Sources (EMARS) Method[Sensors (Basel, Switzerland). ]Honke K, Kotani N. Sensors (Basel, Switzerland). 12(12)16037-16045See all...LinksMedGenMedGenRelated information in MedGenPubMedPubMedPubMed citations for these articlesTaxonomyTaxonomyRelated taxonomy entryTaxonomy TreeTaxonomy TreeRecent ActivityClearTurn OffTurn OnDynamic, yet structured: The cell membrane three decades after the
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Railroad Maps, 1828 to 1900, Available Online, 1880/1889, Coal Mines and Mining, Ontario, Middle West, Canada, Maps | Library of Congress
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Résumé - Opinion summary Polygamy and the Rights of Women
P. 1Résumé - Opinion summary Polygamy and the Rights of WomenRésumé - Opinion summary Polygamy and the Rights of WomenRatings: (0)|Views: 1|Likes: 1Published by NinthCircleOfHellRésumé - Opinion summary Polygamy and the Rights of WomenRésumé - Opinion summary Polygamy and the Rights of WomenMore info:Categories:Types, ResearchPublished by: NinthCircleOfHell on Dec 13, 2012Copyright:Attribution Non-commercialAvailability:Read on Scribd mobile: iPhone, iPad and Android.download as PDF, TXT or read online from ScribdFlag for inappropriate content|Add to collectionSee moreSee lesshttps://www.scribd.com/doc/116619571/Resume-Opinion-summary-Polygamy-and-the-Rights-of-Women12/13/2012pdftextoriginal You're Reading a Free Preview
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”Unaffordable” exotics have their place in the M4/3 system: Micro Four Thirds Talk Forum: Digital Photography Review
MOD Tom Caldwell • Forum Pro • Posts: 38,465
I think that this is fair comment but people like to read about Ferraris in magazines without complaining that they cannot afford one in their wildest dreams. My analogy is that if complaints are made about “unaffordable” lenses made by any manufacturer (Be it Sony, Canon, Nikon, Olympus, even Panasonic) then it is much the same issue.
We should simply bathe ourselves in the wonder that such lenses can be made and offered for sale and understand that our “f2.8” is quite a lot cheaper, more versatile, and quite good enough for our purposes.
So what is a stop and a half, or even a stop if we wish to carp, for a whole lot more money to get something unique and cutting edge for the M4/3 system between friends?
We are not forced to buy these exotic lenses any more than we are forced to buy these cutting edge lenses when their purpose is more cutting edge than we might ever need.
In Australia this is known as the “tall poppy syndrome” where those that might show ostentatious wealth need cutting down to “everyday” size.
Some people even take great delight in demonstrating that their mobile phone camera can take “just as good images” as our relatively humble M4/3 cameras
So instead of complaining that “unaffordable”, heavy, and equivalence-challenged lenses are made just think “extraordinary what M4/3 can offer ... but not for me” and pass on very glad that M4/3 already offers about 100 lenses in all sizes and price ranges and surely if i get to own 10% of them in time I will be very well equipped indeed.
So the “unaffordable exotics” are there to show those that like to own the very best gear that they need not leave M4/3 to get their hands on the cutting edge lenses that most “normal” people would never make themselves afford.

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