共查询到20条相似文献,搜索用时 31 毫秒
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Díaz-López A Rivas C Iniesta P Morán A García-Aranda C Megías D Sánchez-Pernaute A Torres A Díaz-Rubio E Benito M De Juan C 《Experimental cell research》2005,307(1):91-99
We report the characterization of the novel human protein MDGA1 encoded by MDGA1 (MAM domain containing glycosylphosphatidylinositol anchor-1) gene, firstly termed as GPIM. MDGA1 has been mapped to 6p21 and it is expressed in human tissues and tumors. The deduced polypeptide consists of 955 amino acids and exhibits structural features found in different types of cell adhesion molecules (CAMs), such as the presence of both immunoglobulin domains and a MAM domain or the capacity to anchor to the cell membrane by a GPI (glycosylphosphatidylinositol) motif. Our results demonstrate that human MDGA1 (hMDGA1) is localized in the membrane of eukaryotic cells. The protein follows the secretion pathway and finally it is retained in the cell membrane by a GPI anchor, susceptible to be cleavaged by phospholipase C (PI-PLC). Moreover, our results reveal that hMDGA1 is localized specifically into membrane microdomains known as lipid rafts. Finally, as other proteins of the secretory pathway, hMDGA1 undergoes other post-translational modification consisting of N-glycosylation. 相似文献
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Establishment of estrogen receptor 1 (ESR1)‐knockout medaka: ESR1 is dispensable for sexual development and reproduction in medaka,Oryzias latipes 下载免费PDF全文
Saki Tohyama Yukiko Ogino Anke Lange Taijun Myosho Tohru Kobayashi Yu Hirano Gen Yamada Tomomi Sato Norihisa Tatarazako Charles R. Tyler Taisen Iguchi Shinichi Miyagawa 《Development, growth & differentiation》2017,59(6):552-561
Estrogens play fundamental roles in regulating reproductive activities and they act through estrogen receptor (ESR) in all vertebrates. Most vertebrates have two ESR subtypes (ESR1 and ESR2), whereas teleost fish have at least three (Esr1, Esr2a and Esr2b). Intricate functionalization has been suggested among the Esr subtypes, but to date, distinct roles of Esr have been characterized in only a limited number of species. Study of loss‐of‐function in animal models is a powerful tool for application to understanding vertebrate reproductive biology. In the current study, we established esr1 knockout (KO) medaka using a TALEN approach and examined the effects of Esr1 ablation. Unexpectedly, esr1 KO medaka did not show any significant defects in their gonadal development or in their sexual characteristics. Neither male or female esr1 KO medaka exhibited any significant changes in sexual differentiation or reproductive activity compared with wild type controls. Interestingly, however, estrogen‐induced vitellogenin gene expression, an estrogen‐responsive biomarker in fish, was limited in the liver of esr1 KO males. Our findings, in contrast to mammals, indicate that Esr1 is dispensable for normal development and reproduction in medaka. We thus provide an evidence for estrogen receptor functionalization between mammals and fish. Our findings will also benefit interpretation of studies into the toxicological effects of estrogenic chemicals in fish. 相似文献
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Rebecca L. Lowery Yu Zhang Emily A. Kelly Cassandra E. Lamantia Brandon K. Harvey Ania K. Majewska 《Developmental neurobiology》2009,69(10):674-688
Chronic in vivo imaging studies of the brain require a labeling method that is fast, long‐lasting, efficient, nontoxic, and cell‐type specific. Over the last decade, adeno‐associated virus (AAV) has been used to stably express fluorescent proteins in neurons invivo. However, AAV's main limitation for many studies (such as those of neuronal development) is the necessity of second‐strand DNA synthesis, which delays peak transgene expression. The development of double‐stranded AAV (dsAAV) vectors has overcome this limitation, allowing rapid transgene expression. Here, we have injected different serotypes (1, 2, 6, 7, 8, and 9) of a dsAAV vector carrying the green fluorescent protein (GFP) gene into the developing and adult mouse visual cortex and characterized its expression. We observed labeling of both neurons and astrocytes with serotype‐specific tropism. dsAAV‐GFP labeling showed high levels of neuronal GFP expression as early as 2 days postinjection and as long as a month, surpassing conventional AAV's onset of expression and matching its longevity. Neurons labeled with dsAAV‐GFP appeared structurally and electrophysiologically identical to nonlabeled neurons, suggesting that dsAAV‐GFP is neither cytotoxic nor alters normal neuronal function. We also demonstrated that dsAAV‐labeled cells can be imaged with subcellular resolution in vivo over multiple days. We conclude that dsAAV is an excellent vector for rapid labeling and long‐term in vivo imaging studies of astrocytes and neurons on the single cell level within the developing and adult visual cortex. © 2009 Wiley Periodicals, Inc. Develop Neurobiol 2009 相似文献
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Kenji Murata Sean Degmetich Masato Kinoshita Eriko Shimada 《Development, growth & differentiation》2009,51(2):95-107
The congenital heart disease 5 (CHD5)/tryptophan rich basic protein (WRB) is a protein containing a tryptophan‐rich carboxy‐terminal region, which was discovered in the human fetal heart. In humans, this CHD5/WRB is located between the markers ACTL5‐D21S268 within the Down syndrome (DS) Region‐2 at chromosome 21. Congenital heart disease is commonly linked to DS patients. The functions of this gene product are unknown. To identify the functions of CHD5/WRB in heart formation during embryogenesis, the medaka CHD5 cDNA (mCHD5) was isolated and its gene expression pattern and the localization of its gene product were investigated. The obtained mCHD5 belongs to the CHD5 superfamily, whose members include coiled‐coil proteins. The mCHD5 gene was found to be expressed in the developing heart after stage 28 at which the chamber (ventricle and atrium) differentiation in the heart tube is initiated in the embryo. Its gene product was also detected in the developing heart at embryonic stage 28 and 35. Knocking‐down of mCHD5 function caused severe cardiac disorder, including abnormal chamber differentiation, abnormal looping and ocular abnormality such as Cyclops. Our results provide the mCHD5 gene expression pattern as well as its physiological role during heart formation in a vertebrate model system. 相似文献
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Hideyuki Dekimoto Toshio Terashima Yu Katsuyama 《Development, growth & differentiation》2010,52(2):181-193
Neurons with similar functions including neuronal connectivity and gene expression form discrete condensed structures within the vertebrate brain. This is exemplified within the circuitry formed by the cortical layers and the neuronal nuclei. It is well known that the Reelin protein is required for development of these neuronal structures in rodents and human, but the function of Reelin remains controversial. In this report, we used “layer‐specific markers” of the cerebral cortex to carry out detailed observations of spatial distribution of the neuronal subpopulations in the brain of the Reelin deficient mouse, reeler. We observed a spatially dispersed expression of the markers in the reeler cerebral cortex. These markers are expressed also in other laminated and non‐laminated structures of brain, in which we observed similar abnormal gene expression. Our observations suggest that neurons within the brain structures (such as the layers and the nuclei), which normally exhibit condensed distribution of marker expressions, loosen their segregation or scatter by a lack of Reelin. 相似文献
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Adult neurogenesis in the crayfish brain: Proliferation,migration, and possible origin of precursor cells 总被引:1,自引:0,他引:1
Yi Zhang Silvana Allodi David C. Sandeman Barbara S. Beltz 《Developmental neurobiology》2009,69(7):415-436
The birth of new neurons and their incorporation into functional circuits in the adult brain is a characteristic of many vertebrate and invertebrate organisms, including decapod crustaceans. Precursor cells maintaining life‐long proliferation in the brains of crayfish (Procambarus clarkii, Cherax destructor) and clawed lobsters (Homarus americanus) reside within a specialized niche on the ventral surface of the brain; their daughters migrate to two proliferation zones along a stream formed by processes of the niche precursors. Here they divide again, finally producing interneurons in the olfactory pathway. The present studies in P. clarkii explore (1) differential proliferative activity among the niche precursor cells with growth and aging, (2) morphological characteristics of cells in the niche and migratory streams, and (3) aspects of the cell cycle in this lineage. Morphologically symmetrical divisions of neuronal precursor cells were observed in the niche near where the migratory streams emerge, as well as in the streams and proliferation zones. The nuclei of migrating cells elongate and undergo shape changes consistent with nucleokinetic movement. LIS1, a highly conserved dynein‐binding protein, is expressed in cells in the migratory stream and neurogenic niche, implicating this protein in the translocation of crustacean brain neuronal precursor cells. Symmetrical divisions of the niche precursors and migration of both daughters raised the question of how the niche precursor pool is replenished. We present here preliminary evidence for an association between vascular cells and the niche precursors, which may relate to the life‐long growth and maintenance of the crustacean neurogenic niche. © 2009 Wiley Periodicals, Inc. Develop Neurobiol, 2009 相似文献
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Emilio Casanova Nicole Guetg Réjan Vigot Riad Seddik Marcela Julio‐Pieper Niall P. Hyland John F. Cryan Martin Gassmann Bernhard Bettler 《Genesis (New York, N.Y. : 2000)》2009,47(9):595-602
GABAB receptors are the G‐protein‐coupled receptors for the neurotransmitter γ‐aminobutyric acid (GABA). Receptor subtypes are based on the subunit isoforms GABAB1a and GABAB1b, which combine with GABAB2 subunits to form heteromeric receptors. Here, we used a modified bacterial artificial chromosome (BAC) containing the GABAB1 gene to generate transgenic mice expressing GABAB1a and GABAB1b subunits fused to the enhanced green fluorescence protein (eGFP). We demonstrate that the GABAB1‐eGFP fusion proteins reproduce the cellular expression patterns of endogenous GABAB1 proteins in the brain and in peripheral tissue. Crossing the GABAB1‐eGFP BAC transgene into the GABAB1?/? background restores pre and postsynaptic GABAB functions, showing that the GABAB1‐eGFP fusion proteins substitute for the lack of endogenous GABAB1 proteins. Finally, we demonstrate that the GABAB1‐eGFP fusion proteins replicate the temporal expression patterns of native GABAB receptors in cultured neurons. These transgenic mice therefore provide a validated tool for direct visualization of native GABAB receptors. genesis 47:595–602, 2009. © 2009 Wiley‐Liss, Inc. 相似文献
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Kirischian N McArthur AG Jesuthasan C Krattenmacher B Wilson JY 《Journal of molecular evolution》2011,72(1):56-71
Cytochrome P450 (CYP) proteins compose a highly diverse superfamily found in all domains of life. These proteins are enzymes
involved in metabolism of endogenous and exogenous compounds. In vertebrates, the CYP2 family is one of the largest, most
diverse and plays an important role in mammalian drug metabolism. However, there are more than 20 vertebrate CYP2 subfamilies
with uncertain evolution and fairly discrete subfamily composition within vertebrate classes, hindering extrapolation of knowledge
across subfamilies. To better understand CYP2 diversity, a phylogenetic analysis of 196 CYP2 protein sequences from 16 species
was performed using a maximum likelihood approach and Bayesian inference. The analyses included the CYP2 compliment from human,
fugu, zebrafish, stickleback, medaka, cow, and dog genomes. Additional sequences were included from rabbit, marsupial, platypus,
chicken, frog, and salmonid species. Three CYP2 sequences from the tunicate Ciona intestinalis were utilized as the outgroup. Results indicate a single ancestral vertebrate CYP2 gene and monophyly of all CYP2 subfamilies.
Two subfamilies (CYP2R and CYP2U) pre-date vertebrate diversification, allowing direct comparison across vertebrate classes,
while all other subfamilies originated during vertebrate diversification, often within specific vertebrate lineages. Analysis
of site-specific evolution indicates that some substrate recognition sites (SRS) previously proposed for CYP genes do not
have elevated rates of evolution, suggesting that these regions of the protein are not necessarily important in recognition
of CYP2 substrates. Type II functional divergence analysis identified multiple residues in the active site of CYP2F, CYP2A,
and CYP2B proteins that have undergone radical biochemical changes and may be functionally important. 相似文献
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Nishitsuji K Horie T Ichinose A Sasakura Y Yasuo H Kusakabe TG 《Development, growth & differentiation》2012,54(2):177-186
The tunicate Ciona intestinalis larva has a simple central nervous system (CNS), consisting of fewer than 400 cells, which is homologous to the vertebrate CNS. Recent studies have revealed neuronal types and networks in the larval CNS of C. intestinalis, yet their cell lineage and the molecular mechanism by which particular types of neurons are specified and differentiate remain poorly understood. Here, we report cell lineage origin and a cis‐regulatory module for the anterior caudal inhibitory neurons (ACINs), a putative component of the central pattern generator regulating swimming locomotion. The vesicular GABA/glycine transporter gene Ci‐VGAT, a specific marker for GABAergic/glycinergic neurons, is expressed in distinct sets of neurons, including ACINs of the tail nerve cord and others in the brain vesicle and motor ganglion. Comparative genomics analysis between C. intestinalis and Ciona savignyi and functional analysis in vivo identified the cis‐regulatory module responsible for Ci‐VGAT expression in ACINs. Our cell lineage analyses inferred that ACINs derive from A11.116 cells, which have been thought to solely give rise to glial ependymal cells of the lateral wall of the nerve cord. The present findings will provide a solid basis for future studies addressing the molecular mechanism underlying specification of ACINs, which play a critical role in controlling larval locomotion. 相似文献
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Nicholas W. Plummer Teresa L. Squire Sudha Srinivasan Elizabeth Huang Jon S. Zawistowski Hiroaki Matsunami Laura P. Hale Douglas A. Marchuk 《Mammalian genome》2006,17(2):119-128
Cerebral cavernous malformations are vascular defects of the central nervous system consisting of clusters of dilated vessels
that are subject to frequent hemorrhaging. The genes mutated in three forms of autosomal dominant cerebral cavernous malformations
have been cloned, but it remains unclear which cell type is ultimately responsible for the lesion. In this article we describe
mice with a gene trap insertion in the Ccm2 gene. Consistent with the human phenotype, heterozygous animals develop cerebral vascular malformations, although penetrance
is low. β-galactosidase activity in heterozygous brain and in situ hybridization in wild-type brain revealed Ccm2 expression in neurons and choroid plexus but not in vascular endothelium of small vessels in the brain. The expression pattern
of Ccm2 is similar to that of the Ccm1 gene and its interacting protein ICAP1 (Itgb1bp1). These data suggest that cerebral cavernous malformations arise as a result of defects in the neural parenchyma surrounding
the vascular endothelial cells in the brain.
Nicholas W. Plummer, Teresa L. Squire and Sudha Srinivasan contributed equally to this work. 相似文献
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Ingo Braasch Daniel Liedtke Jean‐Nicolas Volff Manfred Schartl 《Pigment cell & melanoma research》2009,22(6):839-850
The function of the tyrosinase‐related protein 1 (Tyrp1) has not yet been investigated in vertebrates basal to tetrapods. Teleost fishes have two duplicates of the tyrp1 gene. Here, we show that the teleost tyrp1 duplicates have distributed the ancestral gene expression in the retinal pigment epithelium (RPE) and melanophores in a species‐specific manner. In medaka embryos, tyrp1a expression is found in the RPE and in melanophores while tyrp1b is only expressed in melanophores. In zebrafish embryos, expression of tyrp1 paralogs overlaps in the RPE and in melanophores. Knockdown of each zebrafish tyrp1 duplicate alone does not show pigmentary defects, but simultaneous knockdown of both tyrp1 genes results in the formation of brown instead of black eumelanin accompanied by severe melanosome defects. Our study suggests that the brown melanosome color in Tyrp1‐deficient vertebrates is an effect of altered eumelanin synthesis. Black eumelanin formation essentially relies on the presence of Tyrp1 and some of its function is most likely conserved from the common ancestor of bony vertebrates. 相似文献
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Gare Hoon Yeo Felicia S. H. Cheah Christoph Winkler Ethylin Wang Jabs Byrappa Venkatesh Samuel S. Chong 《Development genes and evolution》2009,219(6):289-300
Four members of the twist gene family (twist1a, 1b, 2, and 3) are found in the zebrafish, and they are thought to have arisen through three rounds of gene duplication, two of which occurred
prior to the tetrapod-fish split. Phylogenetic analysis groups most of the vertebrate Twist1 peptides into clade I, except
for the Twist1b proteins of the acanthopterygian fish (medaka, pufferfish, stickleback), which clustered within clade III.
Paralogies and orthologies among the zebrafish, medaka, and human twist genes were determined using comparative synteny analysis of the chromosomal regions flanking these genes. Comparative nucleotide
substitution analyses also revealed a faster rate of nucleotide mutation/substitution in the acanthopterygian twist1b compared to the zebrafish twist1b, thus accounting for their anomalous phylogenetic clustering. We also observed minimal expression overlap among the four
twist genes, suggesting that despite their significant peptide similarity, their regulatory controls have diverged considerably,
with minimal functional redundancy between them.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
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Michael A. Plesha Ting‐Kuo Huang Abhaya M. Dandekar Bryce W. Falk Karen A. McDonald 《Biotechnology progress》2009,25(3):722-734
Use of transient expression for the rapid, large‐scale production of recombinant proteins in plants requires optimization of existing methods to facilitate scale‐up of the process. We have demonstrated that the techniques used for agroinfiltration and induction greatly impact transient production levels of heterologous protein. A Cucumber mosaic virus inducible viral amplicon (CMViva) expression system was used to transiently produce recombinant alpha‐1‐antitrypsin (rAAT) by co‐infiltrating harvested Nicotiana benthamiana leaves with two Agrobacterium tumefaciens strains, one containing the CMViva expression cassette carrying the AAT gene and the other containing a binary vector carrying the gene silencing suppressor p19. Harvested leaves were both infiltrated and induced by either pressure or vacuum infiltration. Using the vacuum technique for both processes, maximum levels of functional and total rAAT were elevated by (190 ± 8.7)% and (290 ± 7.5)%, respectively, over levels achieved when using the pressure technique for both processes. The bioprocessing conditions for vacuum infiltration and induction were optimized and resulted in maximum rAAT production when using an A. tumefaciens concentration at OD600 of 0.5 and a 0.25‐min vacuum infiltration, and multiple 1‐min vacuum inductions further increased production 25% and resulted in maximum levels of functional and total rAAT at (2.6 ± 0.09)% and (4.1 ± 0.29)% of the total soluble protein, respectively, or (90 ± 1.7) and (140 ± 10) mg per kg fresh weight leaf tissue at 6 days post‐induction. Use of harvested plant tissue with vacuum infiltration and induction demonstrates a bioprocessing route that is fully amenable to scale‐up. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 相似文献
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Teruhiro Okuyama Yasuko Isoe Masahito Hoki Yuji Suehiro Genki Yamagishi Kiyoshi Naruse Masato Kinoshita Yasuhiro Kamei Atushi Shimizu Takeo Kubo Hideaki Takeuchi 《PloS one》2013,8(6)