111 CLONING AND CHARACTERIZATION OF PIG VASA HOMOLOG GENE AND ITS SPECIFIC EXPRESSION IN GERM CELL LINEAGE

2005 ◽  
Vol 17 (2) ◽  
pp. 206
Author(s):  
G.S. Lee ◽  
S.H. Lee ◽  
H.S. Kim ◽  
E.B. Jeung ◽  
S.K. Kang ◽  
...  
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Germ Line ◽  
Pcr Amplification ◽  
Molecular Probes ◽  
Cloning And Expression ◽  
Soil Nematode ◽  
Rt Pcr ◽  
Specific Expression ◽  
Cell Commitment

All of the vasa homologue genes in C. elegans (Caenorhabditis elegens, a free-living soil nematode), xenopus, zebrafish, mouse, human, chicken, trout, and rat exhibited a germ line-specific expression and are used as specific molecular probes to distinguish the developmental profile of germ cells. In order to determine a useful marker for the research of germ cell commitment and development in pigs, we investigated the cloning and expression profile of porcine vasa homolog gene (Pvh). A Pvh cDNA gene of size 2172 bps (submitted to NCBI gene Bank No. AY626785) was cloned from pig ovary by reverse transcription-polymerase chain reaction (RT-PCR) amplification. The amplification was repeated three times and each RT-PCR product was sequenced. The isolated cDNA had 724 deduced amino acids with significant homology to mouse (85%) or human (91%) vasa. The Pvh sequence presents five copies of the RGG motifs and the DEAD box. By RT-PCR amplification, the expression of Pvh mRNA was restricted to the ovary and testis and was undetectable in somatic tissues including brain, whole blood, heart, lung, kidney, spleen, intestine, and liver. When analyzed by RT-PCR amplification, during pre-implantation embryo development, Pvh was transcribed in oocytes and fertilized 2-cell embryos (no difference in the expression levels between oocytes and fertilized 2-cell embryos), but not in 4-cell, 8-cell, morula and blastocyst stages. Using mouse vasa antibody (kindly donated from Dr. Noce, Japan; tested in porcine cells with porcine oocytes and mouse oocytes as positive control and with porcine brain cells as negative control), immunohistochemical analysis of fetal (Day 100) and adult gonad sections revealed that Pvh protein was specifically expressed in proliferating primordial germ cells (PGC), oocytes and spermatocytes. Interestingly, Pvh protein was not expressed in embryonic germ cells, but it was strongly expressed in freshly isolated PGC. Our results indicate that Pvh gene is specifically transcribed in pig germ cells. This study was supported by grants from the Korean Ministry of Science and Technology (Biodiscovery) and the Biogreen 21-1000520030100000.

scholarly journals Transcription factor AP2 controls cnidarian germ cell induction

Science ◽  
2020 ◽  
Vol 367 (6479) ◽  
pp. 757-762 ◽  
Author(s):  
Timothy Q. DuBuc ◽  
Christine E. Schnitzler ◽  
Eleni Chrysostomou ◽  
Emma T. McMahon ◽  
Febrimarsa ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factor ◽  
Germ Cell ◽  
Cell Fate ◽  
Germ Cells ◽  
Adult Stem Cells ◽  
Germ Line ◽  
Cell Induction ◽  
Cell Commitment ◽  
I Cells

Clonal animals do not sequester a germ line during embryogenesis. Instead, they have adult stem cells that contribute to somatic tissues or gametes. How germ fate is induced in these animals, and whether this process is related to bilaterian embryonic germline induction, is unknown. We show that transcription factor AP2 (Tfap2), a regulator of mammalian germ lines, acts to commit adult stem cells, known as i-cells, to the germ cell fate in the clonal cnidarian Hydractinia symbiolongicarpus. Tfap2 mutants lacked germ cells and gonads. Transplanted wild-type cells rescued gonad development but not germ cell induction in Tfap2 mutants. Forced expression of Tfap2 in i-cells converted them to germ cells. Therefore, Tfap2 is a regulator of germ cell commitment across germ line–sequestering and germ line–nonsequestering animals.

Developmental stage- and spermatogenic cycle-specific expression of transcription factor GATA-1 in mouse Sertoli cells

Development ◽  
1994 ◽  
Vol 120 (7) ◽  
pp. 1759-1766 ◽  
Author(s):  
K. Yomogida ◽  
H. Ohtani ◽  
H. Harigae ◽  
E. Ito ◽  
Y. Nishimune ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Developmental Stage ◽  
Sertoli Cell ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Transcriptional Activation ◽  
Germ Line ◽  
Mouse Testis ◽  
Seminiferous Tubules ◽  
Specific Expression

GATA-1 is an essential factor for the transcriptional activation of erythroid-specific genes, and is also abundantly expressed in a discrete subset of cells bordering the seminiferous epithelium in tubules of the murine testis. In examining normal and germ-line defective mutant mice, we show here that GATA-1 is expressed only in the Sertoli cell lineage in mouse testis. GATA-1 expression in Sertoli cells is induced concomitantly with the first wave of spermatogenesis, and GATA-1-positive cells are uniformly distributed among all tubules during prepubertal testis development. However, the number of GATA-1-positive cells declines thereafter and were found only in the peripheral zone of seminiferous tubules in stages VII, VIII and IX of spermatogenesis in the adult mouse testis. In contrast, virtually every Sertoli cell in mutant W/Wv, jsd/jsd or cryptorchid mice (all of which lack significant numbers of germ cells) expresses GATA-1, thus showing that the expression of this transcription factor is negatively controlled by the maturing germ cells. These observations suggest that transcription factor GATA-1 is a developmental stage- and spermatogenic cycle-specific regulator of gene expression in Sertoli cells.

scholarly journals Serotonergic regulation of distention-induced ATP release from the urothelium

2016 ◽  
Vol 310 (7) ◽  
pp. F646-F655 ◽  
Author(s):  
Kazumasa Matsumoto-Miyai ◽  
Erika Yamada ◽  
Eriko Shinzawa ◽  
Yoshihisa Koyama ◽  
Shoichi Shimada ◽  
...  
Keyword(s):  
Urinary Bladder ◽  
Tryptophan Hydroxylase ◽  
Atp Release ◽  
Pcr Amplification ◽  
Inhibitory Effect ◽  
Visceral Sensation ◽  
Rt Pcr ◽  
Specific Expression ◽  
Bladder Distention ◽  
Gr 127935

Serotonin [5-hydroxytryptamine (5-HT)] is involved in both motor and sensory functions in hollow organs, especially in the gastrointestinal tract. However, the involvement of 5-HT in visceral sensation of the urinary bladder remains unknown. Because distention-induced ATP release from the urothelium plays an essential role in visceral sensation of the urinary bladder, we investigated the regulation of urothelial ATP release by the 5-HT signaling system. RT-PCR and immunohistochemical analyses of the urothelium revealed specific expression of 5-HT1D and 5-HT4 receptors. The addition of 5-HT did not affect urothelial ATP release without bladder distention, but it significantly reduced distention-induced ATP release by physiological pressure during urine storage (5 cmH2O). The inhibitory effect of 5-HT on distention-elicited ATP release was blocked by preincubation with the 5-HT1B/1D antagonist GR-127935 but not by the 5-HT4 antagonist SB-204070. mRNA encoding tryptophan hydroxylase 1 was detected in the urinary bladder by nested RT-PCR amplification, and l-tryptophan or the selective serotonin reuptake inhibitor citalopram also inhibited ATP release, indicating that 5-HT is endogenously synthesized and released in the urinary bladder. The addition of GR-127935 significantly enhanced the distention-elicited ATP release 40 min after distention, whereas SB-204070 reduced the amount of ATP release 20 min after distention. These data suggest that 5-HT4 facilitates the distention-induced ATP release at an earlier stage, whereas 5-HT1D inhibits ATP release at a later stage. The net inhibitory effect of 5-HT indicates that the action of 5-HT on the urothelium is mediated predominantly by 5-HT1D.

Expression of the ammonia transporter proteins Rh B glycoprotein and Rh C glycoprotein in the intestinal tract

2005 ◽  
Vol 288 (5) ◽  
pp. G1036-G1047 ◽  
Author(s):  
Mary E. Handlogten ◽  
Seong-Pyo Hong ◽  
Li Zhang ◽  
Allen W. Vander ◽  
Marshall L. Steinbaum ◽  
...  
Keyword(s):  
Gastrointestinal Tract ◽  
Epithelial Cells ◽  
Intestinal Tract ◽  
Pcr Amplification ◽  
Immunoblot Analysis ◽  
Rt Pcr ◽  
Specific Expression ◽  
Ammonia Metabolism ◽  
Transporter Family ◽  
Ammonia Transport

Ammonia metabolism is important in multiple aspects of gastrointestinal physiology, but the mechanisms of ammonia transport in the gastrointestinal tract remain incompletely defined. The present study examines expression of the ammonia transporter family members Rh B glycoprotein (RhBG) and Rh C glycoprotein (RhCG) in the mouse gastrointestinal tract. Real-time RT-PCR amplification and immunoblot analysis identified mRNA and protein for both RhBG and RhCG were expressed in stomach, duodenum, jejunum, ileum, and colon. Immunohistochemistry showed organ and cell-specific expression of both RhBG and RhCG. In the stomach, both RhBG and RhCG were expressed in the fundus and forestomach, but not in the antrum. In the forestomach, RhBG was expressed by all nucleated squamous epithelial cells, whereas RhCG was expressed only in the stratum germinativum. In the fundus, RhBG and RhCG immunoreactivity was present in zymogenic cells but not in parietal or mucous cells. Furthermore, zymogenic cell RhBG and RhCG expression was polarized, with apical RhCG and basolateral RhBG immunoreactivity. In the duodenum, jejunum, ileum, and colon, RhBG and RhCG immunoreactivity was present in villous, but not in mucous or crypt cells. Similar to the fundic zymogenic cell, RhBG and RhCG expression in villous epithelial cells was polarized when apical RhCG and basolateral RhBG immunoreactivity was present. Thus the ammonia transporting proteins RhBG and RhCG exhibit cell-specific, axially heterogeneous, and polarized expression in the intestinal tract suggesting they function cooperatively to mediate gastrointestinal tract ammonia transport.

The fog-3 gene and regulation of cell fate in the germ line of Caenorhabditis elegans.

Genetics ◽  
1995 ◽  
Vol 139 (2) ◽  
pp. 561-577 ◽  
Author(s):  
R E Ellis ◽  
J Kimble
Keyword(s):  
Caenorhabditis Elegans ◽  
Germ Cell ◽  
Sexual Identity ◽  
Cell Fate ◽  
Germ Cells ◽  
Regulatory Network ◽  
Germ Line ◽  
The Other ◽  
Nematode Caenorhabditis Elegans ◽  
Inhibitory Interactions

Abstract In the nematode Caenorhabditis elegans, germ cells normally adopt one of three fates: mitosis, spermatogenesis or oogenesis. We have identified and characterized the gene fog-3, which is required for germ cells to differentiate as sperm rather than as oocytes. Analysis of double mutants suggests that fog-3 is absolutely required for spermatogenesis and acts at the end of the regulatory hierarchy controlling sex determination for the germ line. By contrast, mutations in fog-3 do not alter the sexual identity of other tissues. We also have characterized the null phenotype of fog-1, another gene required for spermatogenesis; we demonstrate that it too controls the sexual identity of germ cells but not of other tissues. Finally, we have studied the interaction of these two fog genes with gld-1, a gene required for germ cells to undergo oogenesis rather than mitosis. On the basis of these results, we propose that germ-cell fate might be controlled by a set of inhibitory interactions among genes that specify one of three fates: mitosis, spermatogenesis or oogenesis. Such a regulatory network would link the adoption of one germ-cell fate to the suppression of the other two.

scholarly journals Germline Specific Expression of a vasa Homologue Gene in the Viviparous Fish Black Rockfish (Sebastes schlegelii) and Functional Analysis of the vasa 3′ Untranslated Region

2020 ◽  
Vol 8 ◽  
Author(s):  
Li Zhou ◽  
Xueying Wang ◽  
Shuran Du ◽  
Yanfeng Wang ◽  
Haixia Zhao ◽  
...  
Keyword(s):  
Germ Cell ◽  
Marine Fish ◽  
Germ Cells ◽  
Untranslated Region ◽  
Cell Manipulation ◽  
Marine Medaka ◽  
Specific Expression ◽  
Black Rockfish ◽  
Sebastes Schlegelii ◽  
And Migration

Germ cells play a key role in gonad development. As precursors, primordial germ cells (PGCs) are particularly important for germline formation. However, the origination and migration patterns of PGCs are poorly studied in marine fish, especially for viviparous economic species. The vasa gene has been widely used as a germ cell marker to identify a germline because vasa RNA is a component of germ plasm. In this study, we described the expression pattern of black rockfish (Sebastes schlegelii) vasa (Ssvas) in gonadal formation and development by in situ hybridization. The results showed that Ssvas failed in localization at the cleavage furrows until the late gastrula stage, when PGCs appeared and migrated to the genital ridge and formed elongated gonadal primordia at 10 days after birth. This study firstly revealed the PGCs origination and migration characteristics in viviparous marine fish. Furthermore, we microinjected chimeric mRNA containing EGFP and the 3′untranslated region (3′UTR) of Ssvas into zebrafish (Danio rerio) and marine medaka (Oryzias melastigma) fertilized eggs for tracing PGCs. We found that, although Sebastes schlegelii lacked early localization, similar to red seabream (Pagrus major) and marine medaka, only the 3′UTR of Ssvas vasa 3′UTR of black rockfish was able to label both zebrafish and marine medaka PGCs. In comparison with other three Euteleostei species, besides some basal motifs, black rockfish had three specific motifs of M10, M12, and M19 just presented in zebrafish, which might play an important role in labeling zebrafish PGCs. These results will promote germ cell manipulation technology development and facilitate artificial reproduction regulation in aquaculture.

scholarly journals Quantification of healthy and atretic germ cells and follicles in the developing and post-natal ovary of the South American plains vizcacha, Lagostomus maximus: evidence of continuous rise of the germinal reserve

Reproduction ◽  
2014 ◽  
Vol 147 (2) ◽  
pp. 199-209 ◽  
Author(s):  
P I F Inserra ◽  
N P Leopardo ◽  
M A Willis ◽  
A L Freysselinard ◽  
A D Vitullo
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Germ Line ◽  
Cell Number ◽  
South American ◽  
The South ◽  
Foetal Life ◽  
Female Germ Line ◽  
Lagostomus Maximus ◽  
Mitotic Proliferation

The female germ line in mammals is subjected to massive cell death that eliminates 60–85% of the germinal reserve by birth and continues from birth to adulthood until the exhaustion of the germinal pool. Germ cell demise occurs mainly through apoptosis by means of a biased expression in favour of pro-apoptotic members of theBCL2gene family. By contrast, the South American plains vizcacha,Lagostomus maximus, exhibits sustained expression of the anti-apoptoticBCL2gene throughout gestation and a low incidence of germ cell apoptosis. This led to the proposal that, in the absence of death mechanisms other than apoptosis, the female germ line should increase continuously from foetal life until after birth. In this study, we quantified all healthy germ cells and follicles in the ovaries ofL. maximusfrom early foetal life to day 60 after birth using unbiased stereological methods and detected apoptosis by labelling with TUNEL assay. The healthy germ cell population increased continuously from early-developing ovary reaching a 50 times higher population number by the end of gestation. TUNEL-positive germ cells were <0.5% of the germ cell number, except at mid-gestation (3.62%). Mitotic proliferation, entrance into prophase I stage and primordial follicle formation occurred as overlapping processes from early pregnancy to birth. Germ cell number remained constant in early post-natal life, but a remnant population of non-follicular VASA- and PCNA-positive germ cells still persisted at post-natal day 60.L. maximusis the first mammal so far described in which female germ line develops in the absence of constitutive massive germ cell elimination.Free Spanish abstractSpanish translation of this abstract is freely available athttp://www.reproduction-online.org/content/147/2/199/suppl/DC1

scholarly journals AZFa candidate gene UTY and its X homologue UTX are expressed in human germ cells

2021 ◽  
Author(s):  
Peter H Vogt ◽  
Jutta Zimmer ◽  
Ulrike Bender ◽  
Thomas Strowitzki
Keyword(s):  
Germ Cell ◽  
Candidate Gene ◽  
Y Chromosome ◽  
Germ Cells ◽  
Protein Interactions ◽  
Germ Line ◽  
Primordial Germ Cells ◽  
Basal Membrane ◽  
Specific Protein ◽  
Disorders Of Sexual Development

The Ubiquitous Transcribed Y (UTY) AZFa candidate gene on the human Y chromosome and its paralog on the X chromosome, UTX, encode a histone lysine demethylase removing chromatin H3K27 methylation marks at genes transcriptional start sites for activation. Both proteins harbour the conserved Jumonji C (JmjC) domain, functional in chromatin metabolism, and an extended N-terminal tetratrico peptide repeat (TPR) block involved in specific protein-interactions. Specific antisera for human UTY and UTX proteins were developed to distinguish expression of both proteins in human germ cells by immunohistochemical experiments on appropriate tissue sections. In the male germ line, UTY was expressed in the fraction of A spermatogonia located at the basal membrane probably including spermatogonia stem cells. UTX expression was more spread in all spermatogonia and in early spermatids. In female germ line, UTX expression was found in the primordial germ cells of the ovary. UTY was also expressed during fetal male germ cell development, whereas UTX expression was visible only at distinct gestation weeks. Based on these results and the conserved neighboured location of UTY and DDX3Y in Yq11 found in mammals of distinct lineages, we conclude that UTY –like DDX3Y- is part of the Azoospermia factor a (AZFa) locus functioning in human spermatogonia to support the balance of their proliferation-differentiation rate before meiosis. Comparable UTY and DDX3Y expression was also found in gonadoblastoma and dysgerminoma cells found in germ cell nests of the dysgenetic gonads of individuals with disorders of sexual development and a Y chromosome in karyotype (DSD-XY). This confirms that AZFa overlaps with GBY, the Gonadoblastoma susceptibility Y locus, and includes the UTY gene.

scholarly journals The C. elegans gonadal sheath Sh1 cells associate selectively with a differentiating germ cell population in the proliferative zone

2021 ◽  
Author(s):  
Xin Li ◽  
Noor Singh ◽  
Camille Miller ◽  
India Washington ◽  
Bintou Sosseh ◽  
...  
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Germ Line ◽  
Fluorescent Proteins ◽  
Temperature Sensitive ◽  
Variable Expression ◽  
C Elegans ◽  
Proliferative Zone ◽  
Adult Hermaphrodite ◽  
Sheath Cells

The C. elegans adult hermaphrodite germ line is surrounded by a thin tube formed by somatic sheath cells that support germ cells as they mature from the stem-like mitotic state through meiosis, gametogenesis and ovulation. Recently, we discovered that the distal-most Sh1 sheath cells associate with mitotic germ cells as they exit the niche. Here we report that these distal sheath-associated germ cells differentiate first in animals with temperature-sensitive mutations affecting germ cell state, and stem-like germ cells are maintained distal to the Sh1 boundary. We analyze several markers of the distal sheath, which is best visualized with endogenously tagged membrane proteins, as overexpressed fluorescent proteins fail to localize to distal membrane processes and can cause gonad morphology defects. However, such reagents with highly variable expression can be used to determine the relative positions of the two Sh1 cells, one of which often extends further distal than the other.

scholarly journals Functional Recovery of the Germ Line Following Splicing Collapse

2021 ◽  
Author(s):  
Wei Cao ◽  
Christopher Tran ◽  
Stuart K Archer ◽  
Sandeep Gopal ◽  
Roger Pocock
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Rna Splicing ◽  
Messenger Rna ◽  
Germ Line ◽  
Intron Retention ◽  
Mrna Splicing ◽  
C Elegans ◽  
Mrna Transcripts ◽  
Line Following

Splicing introns from precursor-messenger RNA (pre-mRNA) transcripts is essential for translating functional proteins. Here, we report that the previously uncharacterized Caenorhabditis elegans protein MOG-7, acts as a pre-mRNA splicing factor. Depleting MOG-7 from the C. elegans germ line causes intron retention in the majority of germline-expressed genes, impeding the germ cell cycle, and causing defects in nuclear morphology, germ cell identity and sterility. Despite the deleterious consequences caused by MOG-7 loss, the adult germ line can functionally recover to produce viable and fertile progeny when MOG-7 is restored. Germline recovery is dependent on a burst of apoptosis that likely clears defective germ cells, and viable gametes generated from the proliferation of germ cells in the progenitor zone. Together, these findings reveal that MOG-7 is essential for germ cell development, and that the germ line is able to functionally recover after a collapse in RNA splicing.

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