Research Papers: Immunology:
B7-H3 promoted proliferation of mouse spermatogonial stem cells via the PI3K signaling pathway
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Xuedong Wei1,*, Kai Li1,2,*, Guangbo Zhang3, Yuhua Huang1, Jinxing Lv1, Miao Li1, Lun Zhao1, Caibin Fan2, Jinxian Pu1, Jianquan Hou1 and Hexing Yuan1
1 Department of Urology, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, People’s Republic of China
2 Department of Urology, Suzhou Municipal Hospital, Suzhou, Jiangsu, People’s Republic of China
3 Department of Clinical Immunology Laboratory, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, People’s Republic of China
* These authors have contributed equally to this work
Hexing Yuan, email:
Jianquan Hou, email:
Keywords: B7-H3; testis; mouse spermatogonial stem cell; proliferation; PI3K pathway; Immunology
Received: May 18, 2017 Accepted: December 06, 2017 Published: December 20, 2017
Objective: We found seminal B7-H3 was associated with human sperm concentration. However, the mechanism is unclear. The purpose of this study was to investigate the expression of B7-H3 in mouse testis and determine the effects of B7-H3 on the proliferation of mouse spermatogonial stem cells (SSCs) and the underlying mechanisms.
Methods: B7-H3 expression in the testis of mice at different ages (3 weeks, 8 weeks, 4 months and 9 months) was detected by western blot and immunohistochemistry. CCK-8 were used to measure mouse SSCs proliferation after incubation with different concentrations of B7-H3 for 1-72 h in vitro. Flow cytometry was used to analyze the cell cycle of mouse SSCs after incubation with different concentrations of B7-H3 for 48 and 72 h. The signaling pathways involved were assessed by western blot.
Results: Four-month-old mice had the highest expression of B7-H3 in the testis, while 3-week-old mice had the lowest expression of B7-H3. B7-H3 was predominantly detected on the membrane and in the cytoplasm of Sertoli cells. Furthermore, B7-H3 promoted mouse SSCs proliferation and increased the percentage of cells in S+G2/M phase in a time- and dose-dependent manner in vitro. These effects were inhibited by LY294002, indicating the involvement of the phosphoinositide 3-kinase signaling pathway.
Conclusions: The expression of B7-H3 in mouse testis, especially Sertoli cells, was associated with mouse age. In vitro, B7-H3 promoted the proliferation and accelerated the cell cycle of mouse SSCs via the PI3K pathway, indicating a critical role of B7-H3 expressed by Sertoli cells in mouse spermatogenesis.
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