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 Loss of Kallmann syndrome-associated gene WDR11 disrupts primordial germ cell development by affecting canonical and non-canonical Hedgehog signalling

2020 ◽  
Author(s):  
Jiyoung Lee ◽  
Yeonjoo Kim ◽  
Paris Ataliotis ◽  
Hyung-Goo Kim ◽  
Dae-Won Kim ◽  
...  
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Primary Cilia ◽  
Primordial Germ Cell ◽  
Kallmann Syndrome ◽  
Loss Of Function ◽  
Downstream Effectors ◽  
Cell Components ◽  
Underlying Mechanisms ◽  
And Migration

ABSTRACTMutations of WDR11 are associated with Kallmann syndrome (KS) and congenital hypogonadotrophic hypogonadism (CHH), typically caused by defective functions of gonadotrophin-releasing hormone (GnRH) neurones in the brain. We previously reported that Wdr11 knockout mice show profound infertility with significantly fewer germ cells present in the gonads. To understand the underlying mechanisms mediated by WDR11 in these processes, we investigated the effects of Wdr11 deletion on primordial germ cell (PGC) development. Using live-tracking of PGCs and primary co-cultures of genital ridges (GR), we demonstrated that Wdr11-deficient embryos contained reduced numbers of PGCs which had delayed migration due to significantly decreased proliferation and motility. We found primary cilia-dependent canonical Hedgehog (Hh) signalling was required for proliferation of the somatic mesenchymal cells of GR, while primary cilia-independent non-canonical Hh signalling mediated by Ptch2/Gas1 and downstream effectors Src and Creb was required for PGC proliferation and migration, which was disrupted by the loss of function mutations of WDR11. Therefore, canonical and non-canonical Hh signalling are differentially involved in the development of somatic and germ cell components of the gonads, and WDR11 is required for both of these pathways operating in parallel in GR and PGCs, respectively, during normal PGC development. Our study provides a mechanistic link between the development of GnRH neurones and germ cells mediated by WDR11, which may underlie some cases of KS/CHH and ciliopathies.

scholarly journals CRISPR/Cas9-based genetic screen of SCNT-reprogramming resistant genes identifies critical genes for male germ cell development in mice

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Most Sumona Akter ◽  
Masashi Hada ◽  
Daiki Shikata ◽  
Gen Watanabe ◽  
Atsuo Ogura ◽  
...  
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Molecular Mechanisms ◽  
Essential Gene ◽  
Cell Development ◽  
Genetic Screen ◽  
Male Germ Cell ◽  
Specific Expression ◽  
Germ Cell Development ◽  
Resistant Genes

AbstractMale germ cells undergo complex developmental processes eventually producing spermatozoa through spermatogenesis, although the molecular mechanisms remain largely elusive. We have previously identified somatic cell nuclear transfer-reprogramming resistant genes (SRRGs) that are highly enriched for genes essential for spermatogenesis, although many of them remain uncharacterized in knockout (KO) mice. Here, we performed a CRISPR-based genetic screen using C57BL/6N mice for five uncharacterized SRRGs (Cox8c, Cox7b2, Tuba3a/3b, Faiml, and Gm773), together with meiosis essential gene Majin as a control. RT-qPCR analysis of mouse adult tissues revealed that the five selected SRRGs were exclusively expressed in testis. Analysis of single-cell RNA-seq datasets of adult testis revealed stage-specific expression (pre-, mid-, or post-meiotic expression) in testicular germ cells. Examination of testis morphology, histology, and sperm functions in CRISPR-injected KO adult males revealed that Cox7b2, Gm773, and Tuba3a/3b are required for the production of normal spermatozoa. Specifically, Cox7b2 KO mice produced poorly motile infertile spermatozoa, Gm773 KO mice produced motile spermatozoa with limited zona penetration abilities, and Tuba3a/3b KO mice completely lost germ cells at the early postnatal stages. Our genetic screen focusing on SRRGs efficiently identified critical genes for male germ cell development in mice, which also provides insights into human reproductive medicine.

Germ cell-sertoli cell interactions: Regulation by germ cells of the stage-specific expression of CP-2/cathepsin LmRNA by sertoli cells

1995 ◽  
Vol 16 (2) ◽  
pp. 104-113 ◽  
Author(s):  
William W. Wright ◽  
Sonya D. Zabludoff ◽  
Tarja-Leena Penttilä ◽  
Martti Parvinen
Keyword(s):  
Germ Cell ◽  
Sertoli Cell ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Cell Interactions ◽  
Specific Expression

scholarly journals Discrete somatic niches coordinate proliferation and migration of primordial germ cells via Wnt signaling

2016 ◽  
Vol 214 (2) ◽  
pp. 215-229 ◽  
Author(s):  
Andrea V. Cantú ◽  
Svetlana Altshuler-Keylin ◽  
Diana J. Laird
Keyword(s):  
Cell Cycle ◽  
Germ Cell ◽  
Wnt Signaling ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Early Embryo ◽  
Cell Pool ◽  
And Migration ◽  
Selective Mechanism ◽  
Proliferation And Migration

Inheritance depends on the expansion of a small number of primordial germ cells (PGCs) in the early embryo. Proliferation of mammalian PGCs is concurrent with their movement through changing microenvironments; however, mechanisms coordinating these conflicting processes remain unclear. Here, we find that PGC proliferation varies by location rather than embryonic age. Ror2 and Wnt5a mutants with mislocalized PGCs corroborate the microenvironmental regulation of the cell cycle, except in the hindgut, where Wnt5a is highly expressed. Molecular and genetic evidence suggests that Wnt5a acts via Ror2 to suppress β-catenin–dependent Wnt signaling in PGCs and limit their proliferation in specific locations, which we validate by overactivating β-catenin in PGCs. Our results suggest that the balance between expansion and movement of migratory PGCs is fine-tuned in different niches by the opposing β-catenin–dependent and Ror2-mediated pathways through Wnt5a. This could serve as a selective mechanism to favor early and efficient migrators with clonal dominance in the ensuing germ cell pool while penalizing stragglers.

Primordial germ cell migration in Drosophila melanogaster is controlled by somatic tissue

Development ◽  
1994 ◽  
Vol 120 (1) ◽  
pp. 83-89 ◽  
Author(s):  
M.K. Jaglarz ◽  
K.R. Howard
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Characteristic Time ◽  
Primordial Germ Cells ◽  
Primordial Germ Cell ◽  
Somatic Tissue ◽  
Migratory Behavior ◽  
Somatic Tissues ◽  
Germ Cell Migration ◽  
And Migration

In Drosophila, as in many other organisms, primordial germ cells show invasive and migratory behavior moving from their site of origin to the somatic component of the gonad. At a characteristic time in development, the primordial germ cells pass across the primordium of the gut and migrate on its outer surface toward the mesoderm, where they eventually associate with the somatic tissues of the gonad. Here we demonstrate that the exit and migration are specific behaviors of the primordial germ cells and that they are controlled by the somatic tissue of the embryo rather than by a germ cell autonomous clock. Using mutations, we show that these controlling somatic events probably occur in the tissue of the gut primordium itself.

Migration of Drosophila germ cells: analysis using enhancer trap lines

Development ◽  
1993 ◽  
Vol 119 (Supplement) ◽  
pp. 213-218 ◽  
Author(s):  
Ken Howard ◽  
Mariusz Jaglarz ◽  
Nian Zhang ◽  
Jaymini Shah ◽  
Rahul Warrior
Keyword(s):  
Cell Migration ◽  
Germ Cell ◽  
Germ Cells ◽  
Early Phase ◽  
Enhancer Trap ◽  
Somatic Gonad ◽  
Germ Cell Migration ◽  
And Migration ◽  
Enhancer Trap Lines

Cell migration is a common feature of development. In order to understand more about the factors that control these movements we have embarked on further analysis of the migration of Drosophila germ cells. This process involves passage of the germ cells across the gut primordium and migration toward the mesoderm where the somatic gonad forms. We are particularly interested in the early phase of this migration when the germ cells interact with the amnioproctodeal invagination, the developing gut, before entering into association with the mesoderm. We will summarize the results of our and other studies of these events before describing a number of enhancer trap lines which show expression in the amnioproctodeal invagination during the early phase of germ cell migration. These reveal more about the complexity of this tissue and suggest this tissue is capable of guiding the early phase of germ cell migration.

scholarly journals Pou5f1 and Nanog Are Reliable Germ Cell-Specific Genes in Gonad of a Protogynous Hermaphroditic Fish, Orange-Spotted Grouper (Epinephelus coioides)

Genes ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 79
Author(s):  
Chaoyue Zhong ◽  
Meifeng Liu ◽  
Yuhao Tao ◽  
Xi Wu ◽  
Yang Yang ◽  
...  
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Cell Development ◽  
Marker Genes ◽  
Epinephelus Coioides ◽  
Cell Marker ◽  
Specific Expression ◽  
Germ Cell Development ◽  
Somatic Tissues

Pluripotency markers Pou5f1 and Nanog are core transcription factors regulating early embryonic development and maintaining the pluripotency and self-renewal of stem cells. Pou5f1 and Nanog also play important roles in germ cell development and gametogenesis. In this study, Pou5f1 (EcPou5f1) and Nanog (EcNanog) were cloned from orange-spotted grouper, Epinephelus coioides. The full-length cDNAs of EcPou5f1 and EcNanog were 2790 and 1820 bp, and encoded 475 and 432 amino acids, respectively. EcPou5f1 exhibited a specific expression in gonads, whereas EcNanog was expressed highly in gonads and weakly in some somatic tissues. In situ hybridization analyses showed that the mRNA signals of EcNanog and EcPou5f1 were exclusively restricted to germ cells in gonads. Likewise, immunohistofluorescence staining revealed that EcNanog protein was limited to germ cells. Moreover, both EcPou5f1 and EcNanog mRNAs were discovered to be co-localized with Vasa mRNA, a well-known germ cell maker, in male and female germ cells. These results implied that EcPou5f1 and EcNanog could be also regarded as reliable germ cell marker genes. Therefore, the findings of this study would pave the way for elucidating the mechanism whereby EcPou5f1 and EcNanog regulate germ cell development and gametogenesis in grouper fish, and even in other protogynous hermaphroditic species.

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.

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