scholarly journals 253.Follicle stimulating-hormone (FSH) withdrawal induces germ cell apoptosis in the mature rat testis in vitro

2004 ◽  
Vol 16 (9) ◽  
pp. 253
Author(s):  
S. M. Degen ◽  
P. G. Stanton ◽  
K. L. Loveland ◽  
S. J. Meachem
Keyword(s):  
Cell Proliferation ◽  
Germ Cell ◽  
Cell Development ◽  
Brdu Incorporation ◽  
Proliferating Cells ◽  
Germ Cell Development ◽  
Sperm Output ◽  
The Impact ◽  
Germ Cell Proliferation

FSH is a key determinant of adult sperm output influencing both Sertoli and germ cell development. The aim of this study was to assess the impact of FSH action on Sertoli and germ cell proliferation and survival in vitro, and to identify FSH-regulated genes that may underpin these responses. Testis fragments from 17-day-old rats were cultured with recombinant human FSH for 2 or 24 h and then labelled with bromodeoxyuridine (BrdU) to identify proliferating cells. The testis fragments were then processed for analysis of cell numbers by stereology, BrdU incorporation by immunohistochemistry, and apoptosis by TUNEL. The TUNEL assay revealed that without FSH, spermatogonial apoptosis was induced to 195% and 179% (P�<�0.05) compared to fragments with FSH after 2 and 24 h, respectively. No difference in apoptosis was observed in spermatocyte or Sertoli cell populations at these time points. No differences in Sertoli or germ cell proliferation were observed with or without FSH. To understand how FSH mediates spermatogonial apoptosis the response of 5 testicular genes of interest was examined. Expression of cyclin D2 (cell cycle, G1-S), N-cadherin (N-Cad; adhesion molecule), Bax (pro-apoptotic), Bcl-w (anti-apoptotic), and stem cell factor (SCF; pro-apoptotic and other functions) was elevated to 151%, 348%, 209%, 258%, and 198%, respectively (all P�<�0.001), in fragments cultured without FSH for 24 h, compared to fragments with FSH. No gene expression differences were observed at 2 h, except for SCF, which was elevated to 135% (P�<�0.01). In conclusion, these studies have examined apoptosis and proliferation activities simultaneously in testis fragments in vitro, and demonstrated that FSH withdrawal induces both spermatogonial apoptosis and expression of testicular genes known to be involved in cell survival. This model will now be used to further investigate FSH-mediation of Sertoli and germ cell development.

scholarly journals Bioactive isoprenoids guide migrating germ cells to the embryonic gonad

2021 ◽  
Author(s):  
Lacy Barton ◽  
Justina Sanny ◽  
Emily P Dawson ◽  
Marcela Nouzova ◽  
Fernando G Noriega ◽  
...  
Keyword(s):  
Cell Migration ◽  
Germ Cell ◽  
Germ Cells ◽  
Primordial Germ Cell ◽  
Cell Development ◽  
Germ Cell Development ◽  
Germ Cell Migration ◽  
The Impact

Germ cells are essential to sexual reproduction. Across the animal kingdom, extracellular isoprenoids, such as retinoic acids (RAs) in vertebrates and juvenile hormones (JHs) in insects, impact the germline lifecycle from meiosis to gametogenesis. Emerging evidence suggests that these bioactive isoprenoids also influence embryonic reproductive development, though the precise functions remain unclear. Here, we investigated the specific molecular pathways by which JHs regulates embryonic germ cell development in Drosophila. With a newly generated in vivo reporter, we find that JH signaling is active in the vicinity of germ cells as they migrate to colonize the somatic gonad. Through a combination of in vivo and in vitro assays, we find that JHs are both necessary and sufficient for primordial germ cell migration through mechanisms independent of canonical nuclear receptor-mediated transcription. These findings reveal that JH is present during Drosophila embryogenesis and that bioactive isoprenoids impact germ cell development earlier than previously appreciated. Interestingly, we find that like JH in Drosophila, RA is sufficient for murine germ cell migration in vitro, suggesting that the impact of bioactive isoprenoids on embryonic germ cell development may be broadly conserved.

Bradykinin stimulates prepubertal rat germ cell proliferation in vitro

1998 ◽  
Vol 40 (3) ◽  
pp. 173-178 ◽  
Author(s):  
Nina Atanassova ◽  
Ludmila Kancheva ◽  
Boris Somlev
Keyword(s):  
Cell Proliferation ◽  
Germ Cell ◽  
Proliferation In Vitro ◽  
Germ Cell Proliferation

scholarly journals Modelling germ cell development in vitro

2008 ◽  
Vol 14 (9) ◽  
pp. 501-511 ◽  
Author(s):  
A. J. Childs ◽  
P. T.K. Saunders ◽  
R. A. Anderson
Keyword(s):  
Germ Cell ◽  
Cell Development ◽  
Germ Cell Development ◽  
Development In Vitro

scholarly journals In Situ Analysis of Cellular Proliferation in Canine, Feline and Equine Tumors by Immunohistochemistry: A Comparison of Bromodeoxyuridine, Proliferating Cell Nuclear Antigen, and Interchromatin-Associated Antigen Immunostaining Techniques

1994 ◽  
Vol 6 (4) ◽  
pp. 453-457 ◽  
Author(s):  
Alain Pierre Théon ◽  
Loretta Metzger ◽  
Stephen Griffey
Keyword(s):  
Cell Proliferation ◽  
Proliferating Cell Nuclear Antigen ◽  
Cellular Proliferation ◽  
Nuclear Antigen ◽  
Brdu Incorporation ◽  
Immunohistochemical Detection ◽  
Proliferating Cells ◽  
Proliferating Cell

Cell proliferation in canine, feline, and equine tumors was evaluated using immunohistochemical detection of in vitro 5–bromodeoxyuridine (BrdU) incorporation, proliferating cell nuclear antigen (PCNA), and interchromatin-associated antigen (p105). Ten tumors in each species were analyzed. The tumor proliferative fraction (PF) was defined as the percentage of labeled nuclei for 5,000 tumor nuclei counted. Immunoreactivity was observed with all techniques in all species. A good correlation was observed between the proliferative fractions measured with the BrdU (PFBrdU) and PCNA (PFPCNA) techniques ( rs = 0.523, P = 0.0026). There was no correlation between the PFs measured with the BrdU (PFBrdU) and p105 (PFP105) techniques. Using the median values obtained from the different approaches as cutoff points to define slowly and rapidly proliferating tumors, there was an 80% agreement ( P = 0.009) between PFBrdU and PFPCNA and no agreement between PFBrdU and PFP105 The results of this study indicate that both BrdU and PCNA labeling methods can be used reliably for identifying proliferating cells in animal tumors. In addition, PCNA could be used to replace the BrdU method to assess tumor proliferative fraction because it does not require pretreatment of tissues.

VEGF/VEGFR2 Signaling Regulates Germ Cell Proliferation in vitro and Promotes Mouse Testicular Regeneration in vivo

2016 ◽  
Vol 201 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Ruhui Tian ◽  
Shi Yang ◽  
Yong Zhu ◽  
Shasha Zou ◽  
Peng Li ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Germ Cell ◽  
Germ Cells ◽  
Seminiferous Tubules ◽  
Testicular Development ◽  
Male Germ Cells ◽  
Vegfr2 Signaling ◽  
Proliferation In Vitro ◽  
Germ Cell Proliferation

Vascular endothelial growth factor (VEGF) plays fundamental roles in testicular development; however, its function on testicular regeneration remains unknown. The objective of this study was to explore the roles VEGF/VEGFR2 signaling plays in mouse germ cells and in mouse testicular regeneration. VEGF and the VEGFR2 antagonist SU5416 were added to culture medium to evaluate their effects on spermatogonial stem cell line (C18-4 cells) proliferation. Testicular cells obtained from newborn male ICR mice were grafted into the dorsal region of male BALB/c nude mice. VEGF and SU5416 were injected into the graft sites to assess the effects of the VEGF and VEGFR2 signaling pathways on testicular reconstitution. The grafts were analyzed after 8 weeks. We found that VEGF promoted C18-4 proliferation in vitro, indicating its role in germ cell survival. HE staining revealed that seminiferous tubules were reconstituted and male germ cells from spermatogonia to spermatids could be observed in testis-like tissues 8 weeks after grafting. A few advantaged male germ cells, including spermatocytes and spermatids, were found in SU5416-treated grafts. Moreover, VEGF enhanced the expression of genes specific for male germ cells and vascularization in 8-week grafts, whereas SU5416 decreased the expression of these genes. SU5416-treated grafts had a lower expression of MVH and CD31, indicating that blockade of VEGF/VEGFR2 signaling reduces the efficiency of seminiferous tubule reconstitution. Collectively, these data suggest that VEGF/VEGFR2 signaling regulates germ cell proliferation and promotes testicular regeneration via direct action on germ cells and the enhancement of vascularization.

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