Aim： To determine the effectiveness of the ski 1, sk9 and ski 1 TNUA5 Sertoli cell lines in binding germ cells in vitro. Methods： The immortalized Sertoli cell lines sk9, ski 1 and ski 1 TNUA5 were used in co-cultur...Aim： To determine the effectiveness of the ski 1, sk9 and ski 1 TNUA5 Sertoli cell lines in binding germ cells in vitro. Methods： The immortalized Sertoli cell lines sk9, ski 1 and ski 1 TNUA5 were used in co-culture experiments with germ cells in media with or without reproductive hormones and incubated for 44 h at 32℃. The number of germ cells bound to Sertoli cells was then determined and statistically analyzed. Western blot analysis and reverse transcriptasepolymerase chain reaction （RT-PCR） studies were employed to investigate the presence of cell adhesion proteins and follicle stimulating hormone （FSH） receptor, respectively. Results： No statistical difference between the number of bound step-8 spermatids and bound pre-step 8 spermatids on Sertoli cells from any of the cell lines existed. After the addition of germ cells, Sertoli ceils showed more lipid accumulation in their cytoplasm, indicating active phagocytosis. Western blot analysis in the ski I TNUA5 line indicated the expression of N-cadherin. FSH-only and testosterone-only treatments increased N-cadherin expression, regardless of germ cell addition. The addition of germ cells to the ski l TNUA5 Sertoli cells increased the expression of espin, as did the addition of FSH with germ cells. RT-PCR studies of the ski I TNUA5 cells indicated that the mRNA for FSH receptor decreased with successive passages. Conclusion： In vitro binding between isolated germ cells and sk9, skll or skll TNUA5 Sertoli cells is not feasible, and therefore these cell lines are not useful for the in vitro investigation of Sertoli-germ cell interactions and primary Sertoli cell isolates must still be used.展开更多
Heat stress is an important influence on the male reproductive organs.Therefore,the effects of heat stress on genes or pathways related to the reproductive system of male mice were experimentally explored in this pape...Heat stress is an important influence on the male reproductive organs.Therefore,the effects of heat stress on genes or pathways related to the reproductive system of male mice were experimentally explored in this paper to further determine the effects of heat stimulation on mammals.Herein,models of heat-exposed mouse testicular tissue and heatexcited cells were successfully established.Many scorched vesicles were found after heat excitation of testis supporting cells,testicular mesenchymal(TM4)cells.Western blot,in situ terminal deoxynucleotide transferase dUTP Nick end labeling(TUNEL)and transmission electron microscopy showed that membrane rupture,mitochondrial damage and autophagic vesicles occurred in TM4 cells after thermal excitation.The N-segment fragment of the associated protein shear was increased,and the TUNEL result was positive.In conclusion,thermal excitation induced apoptosis and scorch death in TM4 cells.Thus,the Hippo pathway and apoptosis-related pathway were significantly enriched after heat stimulation in mouse testis,and the scorch death effect in TM4 cells was induced by heat excitation.展开更多
The phosphoinositide-3-kinase/Akt(PI3K/AKT)signaling pathway is crucial for Sertoli cell development and completing spermatogenesis.Its main role is to promote proliferation and inhibit apoptosis.Many factors activate...The phosphoinositide-3-kinase/Akt(PI3K/AKT)signaling pathway is crucial for Sertoli cell development and completing spermatogenesis.Its main role is to promote proliferation and inhibit apoptosis.Many factors activate the PI3K/AKT pathway,like hormones,such as follicle stimulating hormone(FSH),androgen,estrogen,insulin to name a few.Many of these factors have receptors inside or on the surface of Sertoli cells(SCs).This review summarizes how these hormones directly regulate the PI3K/AKT signaling pathway in SCs,which in turn affects SC proliferation and differentiation.Further,hormone-mediated PI3K/AKT signaling also stimulates SC secretion,which is essential for germ cell development,suggesting an indirect role of PI3K/AKT signaling during spermatogenesis.These functions include promoting spermatogonia proliferation and differentiation,meiosis of spermatocytes,sperm maturation,and their release.This review also provides potential hints for clinically treating male infertility issues like cryptorchidism and Sertoli cell-only syndrome.展开更多
文摘Aim： To determine the effectiveness of the ski 1, sk9 and ski 1 TNUA5 Sertoli cell lines in binding germ cells in vitro. Methods： The immortalized Sertoli cell lines sk9, ski 1 and ski 1 TNUA5 were used in co-culture experiments with germ cells in media with or without reproductive hormones and incubated for 44 h at 32℃. The number of germ cells bound to Sertoli cells was then determined and statistically analyzed. Western blot analysis and reverse transcriptasepolymerase chain reaction （RT-PCR） studies were employed to investigate the presence of cell adhesion proteins and follicle stimulating hormone （FSH） receptor, respectively. Results： No statistical difference between the number of bound step-8 spermatids and bound pre-step 8 spermatids on Sertoli cells from any of the cell lines existed. After the addition of germ cells, Sertoli ceils showed more lipid accumulation in their cytoplasm, indicating active phagocytosis. Western blot analysis in the ski I TNUA5 line indicated the expression of N-cadherin. FSH-only and testosterone-only treatments increased N-cadherin expression, regardless of germ cell addition. The addition of germ cells to the ski l TNUA5 Sertoli cells increased the expression of espin, as did the addition of FSH with germ cells. RT-PCR studies of the ski I TNUA5 cells indicated that the mRNA for FSH receptor decreased with successive passages. Conclusion： In vitro binding between isolated germ cells and sk9, skll or skll TNUA5 Sertoli cells is not feasible, and therefore these cell lines are not useful for the in vitro investigation of Sertoli-germ cell interactions and primary Sertoli cell isolates must still be used.
基金This research was funded by the Anhui Provincial Key Research and Development Project of the People’s Republic of China(202104j07020016)the 512 Talent Cultivation Plan of Middle-aged Backbone Teachers of Bengbu Medical College of China(by51201207)the Climbing Plan of Natural Science General Project of Bengbu Medical College(China)(2021bypd003).
文摘Heat stress is an important influence on the male reproductive organs.Therefore,the effects of heat stress on genes or pathways related to the reproductive system of male mice were experimentally explored in this paper to further determine the effects of heat stimulation on mammals.Herein,models of heat-exposed mouse testicular tissue and heatexcited cells were successfully established.Many scorched vesicles were found after heat excitation of testis supporting cells,testicular mesenchymal(TM4)cells.Western blot,in situ terminal deoxynucleotide transferase dUTP Nick end labeling(TUNEL)and transmission electron microscopy showed that membrane rupture,mitochondrial damage and autophagic vesicles occurred in TM4 cells after thermal excitation.The N-segment fragment of the associated protein shear was increased,and the TUNEL result was positive.In conclusion,thermal excitation induced apoptosis and scorch death in TM4 cells.Thus,the Hippo pathway and apoptosis-related pathway were significantly enriched after heat stimulation in mouse testis,and the scorch death effect in TM4 cells was induced by heat excitation.
基金supported in part by the National Natural Science Foundation of China(Nos.32270555 and 32072954).
文摘The phosphoinositide-3-kinase/Akt(PI3K/AKT)signaling pathway is crucial for Sertoli cell development and completing spermatogenesis.Its main role is to promote proliferation and inhibit apoptosis.Many factors activate the PI3K/AKT pathway,like hormones,such as follicle stimulating hormone(FSH),androgen,estrogen,insulin to name a few.Many of these factors have receptors inside or on the surface of Sertoli cells(SCs).This review summarizes how these hormones directly regulate the PI3K/AKT signaling pathway in SCs,which in turn affects SC proliferation and differentiation.Further,hormone-mediated PI3K/AKT signaling also stimulates SC secretion,which is essential for germ cell development,suggesting an indirect role of PI3K/AKT signaling during spermatogenesis.These functions include promoting spermatogonia proliferation and differentiation,meiosis of spermatocytes,sperm maturation,and their release.This review also provides potential hints for clinically treating male infertility issues like cryptorchidism and Sertoli cell-only syndrome.
