scholarly journals BMP4 promotes mouse iPS cell differentiation to male germ cells via Smad1/5, Gata4, Id1 and Id2

Reproduction ◽  
2017 ◽  
Vol 153 (2) ◽  
pp. 211-220 ◽  
Author(s):  
Shi Yang ◽  
Qingqing Yuan ◽  
Minghui Niu ◽  
Jingmei Hou ◽  
Zijue Zhu ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Germ Cell ◽  
Germ Cells ◽  
Real Time Pcr ◽  
Molecular Mechanisms ◽  
Ips Cells ◽  
Male Germ Cell ◽  
Western Blots ◽  
Male Germ Cells ◽  
Downstream Effectors

Generation of male germ cells from pluripotent cells could provide male gametes for treating male infertility and offer an ideal model for unveiling molecular mechanisms of spermatogenesis. However, the influence and exact molecular mechanisms, especially downstream effectors of BMP4 signaling pathways, in male germ cell differentiation of the induce pluripotent stem (iPS) cells, remain unknown. This study was designed to explore the role and mechanism of BMP4 signaling in the differentiation of mouse iPS cells to male germ cells. Embryoid body (EB) formation and recombinant BMP4 or Noggin were utilized to evaluate the effect of BMP4 on male germ cell generation from mouse iPS cells. Germ cell-specific genes and proteins as well as the downstream effectors of BMP4 signaling pathway were assessed using real-time PCR and Western blots. We found that BMP4 ligand and its multiple receptors, including BMPR1a, BMPR1b and BMPR2, were expressed in mouse iPS cells. Real-time PCR and Western blots revealed that BMP4 could upregulate the levels of genes and proteins for germ cell markers in iPS cells-derived EBs, whereas Noggin decreased their expression in these cells. Moreover, Smad1/5 phosphorylation, Gata4 transcription and the transcripts of Id1 and Id2 were enhanced by BMP4 but decreased when exposed to Noggin. Collectively, these results suggest that BMP4 promotes the generation of male germ cells from iPS cells via Smad1/5 pathway and the activation of Gata4, Id1 and Id2. This study thus offers novel insights into molecular mechanisms underlying male germ cell development.

scholarly journals EXOC1 regulates cell morphology of spermatogonia and spermatocytes in mice

2020 ◽  
Author(s):  
Yuki Osawa ◽  
Miho Usui ◽  
Yumeno Kuba ◽  
Hoai Thu Le ◽  
Natsuki Mikami ◽  
...  
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Cell Morphology ◽  
Molecular Mechanisms ◽  
Syncytium Formation ◽  
Male Germ Cell ◽  
Cell Morphogenesis ◽  
Male Germ Cells ◽  
Starting Point

AbstractSpermatogenesis requires high regulation of germ cell morphology. The spermatogonia regulates its differentiation state by its own migration. The male germ cells differentiate and mature with the formation of syncytia, failure of forming the appropriate syncytia results in the arrest of spermatogenesis at the spermatocyte stage. However, the detailed molecular mechanisms of male germ cell morphological regulation are unknown. Here, we found that EXOC1 is important for the pseudopod formation of spermatogonia and spermatocyte syncytia in mice. We found that while EXOC1 contributes to the inactivation of Rac1 in the pseudopod formation of spermatogonia, in spermatocyte syncytium formation, EXOC1 and SNAP23 cooperate with STX2. Our results showed that EXOC1 functions in concert with various cell morphology regulators in spermatogenesis. Since EXOC1 is known to bind to several cell morphogenesis factors, this study is expected to be the starting point for the discovery of many morphological regulators of male germ cells.

217 PROTEIN-INDUCED TRANSDIFFERENTIATION INTO MALE GERM CELL-LIKE LINEAGE FROM CHICKEN FETAL BONE MARROW STEM CELLS

2012 ◽  
Vol 24 (1) ◽  
pp. 220
Author(s):  
J. M. Yoo ◽  
J. J. Park ◽  
K. Gobianand ◽  
J. Y. Ji ◽  
J. S. Kim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Retinoic Acid ◽  
Germ Cell ◽  
Germ Cells ◽  
Cultured Cells ◽  
Male Germ Cell ◽  
Male Germ Cells ◽  
Germ Cell Differentiation ◽  
Transgenic Chickens

Bone marrow (BM)-derived stem cells are capable of transdifferentiation into multilineage cells like muscle, bone, cartilage, fat and nerve cells. In this study, we investigated the capability of mesenchymal stem cells (MSC) derived from BM into germ cell differentiation in the chicken. Chicken MSCs were isolated from BM of day 20 fertilized fetal chicken with Ficoll-Paque Plus. Isolated cells were cultured in advance-DMEM (ADMEM) supplemented with 10% fetal bovine serum and antibiotics. Once confluent, cells were subcultured until five passages. The cultured cells showed fibroblast-like morphology. The cells had positive expressions of Oct4, Sox2 and Nanog. Two induction methods were conducted to examine the ability of transdifferentation into male germ cells. In group 1, MSC were cultured in ADMEM containing retinoic acid and chicken testicular extracts proteins for 10 to 15 days. In group 2, MSC were permeabilized by streptolysin O and treated with chicken testicular protein extracts. In both treatment groups, MSC were cultured in ADMEM containing retinoic acid for 10 to 15 days. We found that chicken MSC had a positive expression of pluripotent proteins such as Oct4, Sox2, Nanog and a small population of chicken MSC seem to transdifferentiate into male germ cell-like cells. These cells expressed early germ cell markers and male germ-cell-specific markers (Dazl, C-kit, Stra8 and DDX4) as analysed by reverse transcription-PCR and immunohistochemistry. These results demonstrated that chicken MSC may differentiate into male germ cells and the same might be used as a potential source of cells for production of transgenic chickens. This study was carried out with the support of Agenda Program (Project No. PJ0064692011), RDA and Republic of Korea.

scholarly journals Different expression and activity of the α1 and α4 isoforms of the Na,K-ATPase during rat male germ cell ontogeny

Reproduction ◽  
2005 ◽  
Vol 130 (5) ◽  
pp. 627-641 ◽  
Author(s):  
K Wagoner ◽  
G Sanchez ◽  
A-N Nguyen ◽  
G C Enders ◽  
G Blanco
Keyword(s):  
Plasma Membrane ◽  
Germ Cell ◽  
Germ Cells ◽  
Cell Biology ◽  
Mrna Level ◽  
Male Gamete ◽  
Male Germ Cell ◽  
Male Germ Cells ◽  
And Function ◽  
Expression And Activity

Two catalytic isoforms of the Na,K-ATPase, α1 and α4, are present in testis. While α1 is ubiquitously expressed in tissues, α4 predominates in male germ cells. Each isoform has distinct enzymatic properties and appears to play specific roles. To gain insight into the relevance of the Na,K-ATPase α isoforms in male germ cell biology, we have studied the expression and activity of α1 and α4 during spermatogenesis and epididymal maturation. This was explored in rat testes at different ages, in isolated spermatogenic cells and in spermatozoa from the caput and caudal regions of the epididymis. Our results show that α1 and α4 undergo differential regulation during development. Whereas α1 exhibits only modest changes, α4 increases with gamete differentiation. The most drastic changes for α4 take place in spermatocytes at the mRNA level, and with the transition of round spermatids into spermatozoa for expression and activity of the protein. No further changes are detected during transit of spermatozoa through the epididymis. In addition, the cellular distribution of α4 is modified with development, being diffusely expressed at the plasma membrane and intracellular compartments of immature cells, finally to localize to the midregion of the spermatozoon flagellum. In contrast, the α1 isoform is evenly present along the plasma membrane of the developing and mature gametes. In conclusion, the Na,K-ATPase α1 and α4 isoforms are functional in diploid, meiotic and haploid male germ cells, α4 being significantly upregulated during spermatogenesis. These results support the importance of α4 in male gamete differentiation and function.

RNA Expression in Male Germ Cells During Spermatogenesis (Male Germ Cell Transcriptome)

2011 ◽  
pp. 107-121 ◽  
Author(s):  
Tin-Lap Lee ◽  
Albert Hoi-Hung Cheung ◽  
Owen M. Rennert ◽  
Wai-Yee Chan
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Male Germ Cell ◽  
Rna Expression ◽  
Male Germ Cells ◽  
Cell Transcriptome

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.

RNA Expression in Male Germ Cells During Spermatogenesis (Male Germ Cell Transcriptome)

2013 ◽  
pp. 105-123
Author(s):  
Tin-Lap Lee ◽  
Albert Hoi-Hung Cheung ◽  
Owen M. Rennert ◽  
Wai-Yee Chan
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Male Germ Cell ◽  
Rna Expression ◽  
Male Germ Cells ◽  
Cell Transcriptome

scholarly journals C-X-C motif chemokine ligand 10 produced by mouse Sertoli cells in response to mumps virus infection induces male germ cell apoptosis

2017 ◽  
Vol 8 (10) ◽  
pp. e3146-e3146 ◽  
Author(s):  
Qian Jiang ◽  
Fei Wang ◽  
Lili Shi ◽  
Xiang Zhao ◽  
Maolei Gong ◽  
...  
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Cell Apoptosis ◽  
Caspase 3 ◽  
Mumps Virus ◽  
Male Germ Cell ◽  
Male Germ Cells ◽  
Germ Cell Apoptosis ◽  
Chemokine Ligand

Abstract Mumps virus (MuV) infection usually results in germ cell degeneration in the testis, which is an etiological factor for male infertility. However, the mechanisms by which MuV infection damages male germ cells remain unclear. The present study showed that C-X-C motif chemokine ligand 10 (CXCL10) is produced by mouse Sertoli cells in response to MuV infection, which induces germ cell apoptosis through the activation of caspase-3. CXC chemokine receptor 3 (CXCR3), a functional receptor of CXCL10, is constitutively expressed in male germ cells. Neutralizing antibodies against CXCR3 and an inhibitor of caspase-3 activation significantly inhibited CXCL10-induced male germ cell apoptosis. Furthermore, the tumor necrosis factor-α (TNF-α) upregulated CXCL10 production in Sertoli cells after MuV infection. The knockout of either CXCL10 or TNF-α reduced germ cell apoptosis in the co-cultures of germ cells and Sertoli cells in response to MuV infection. Local injection of MuV into the testes of mice confirmed the involvement of CXCL10 in germ cell apoptosis in vivo. These results provide novel insights into MuV-induced germ cell apoptosis in the testis.

scholarly journals Cell-fate transition and determination analysis of mouse male germ cells throughout development

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jiexiang Zhao ◽  
Ping Lu ◽  
Cong Wan ◽  
Yaping Huang ◽  
Manman Cui ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Fate ◽  
Germ Cells ◽  
Transitional Cell ◽  
Critical Time ◽  
Mitotic Arrest ◽  
Cell Development ◽  
Male Germ Cell ◽  
Male Germ Cells ◽  
Germ Cell Development

AbstractMammalian male germ cell development is a stepwise cell-fate transition process; however, the full-term developmental profile of male germ cells remains undefined. Here, by interrogating the high-precision transcriptome atlas of 11,598 cells covering 28 critical time-points, we demonstrate that cell-fate transition from mitotic to post-mitotic primordial germ cells is accompanied by transcriptome-scale reconfiguration and a transitional cell state. Notch signaling pathway is essential for initiating mitotic arrest and the maintenance of male germ cells’ identities. Ablation of HELQ induces developmental arrest and abnormal transcriptome reprogramming of male germ cells, indicating the importance of cell cycle regulation for proper cell-fate transition. Finally, systematic human-mouse comparison reveals potential regulators whose deficiency contributed to human male infertility via mitotic arrest regulation. Collectively, our study provides an accurate and comprehensive transcriptome atlas of the male germline cycle and allows for an in-depth understanding of the cell-fate transition and determination underlying male germ cell development.

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