Abstract
Many differentiating spermatogenic cells die by apoptosis during the process of mammalian spermatogenesis. However, very few apoptotic spermatogenic cells are detected by histological examination of the testis, probably due to the rapid elimination of dying cells by phagocytosis. Previous in vitro studies showed that Sertoli cells selectively phagocytose dying spermatogenic cells by recognizing the membrane phospholipid phosphatidylserine (PS), which is exposed to the surface of spermatogenic cells during apoptosis. We examined here whether PS-mediated phagocytosis of apoptotic spermatogenic cells occurs in vivo. For this purpose, the PS-binding protein annexin V was microinjected into the seminiferous tubules of normal live mice, and their testes were examined. The injection of annexin V caused no histological changes in the testis, but significantly increased the number of apoptotic spermatogenic cells as assessed by the terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling assay. The number of Sertoli cells did not change in the annexin V-injected testes, and annexin V itself did not induce apoptosis in primary cultured spermatogenic cells. These results indicate that annexin V inhibited the phagocytic clearance of apoptotic spermatogenic cells and suggest that PS-mediated phagocytosis of those cells occurs in vivo. Furthermore, the injection of annexin V into the seminiferous tubules brought about a significant reduction in the number of spermatogenic cells and epididymal sperm in anticancer drug-treated mice. This suggests that the elimination of apoptotic spermatogenic cells is required for the production of sperm.
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Abbreviations
- BSA:
-
bovine serum albumin
- PS:
-
phosphatidylserine
- SR-BI:
-
class B scavenger receptor type I
- TUNEL:
-
terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling
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Acknowledgements
We thank T Ogawa for technical advice and instruction in microinjection, and Y Kawabuchi for tissue preparation. This study was supported by Grants-in-Aid for Scientific Research from Japan Society for the Promotion of Science, a grant from the Mitsubishi Foundation, a grant from the Sumitomo Foundation, and a grant from the Hayashi Memorial Foundation for Female Natural Scientists.
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Maeda, Y., Shiratsuchi, A., Namiki, M. et al. Inhibition of sperm production in mice by annexin V microinjected into seminiferous tubules: possible etiology of phagocytic clearance of apoptotic spermatogenic cells and male infertility. Cell Death Differ 9, 742–749 (2002). https://doi.org/10.1038/sj.cdd.4401046
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DOI: https://doi.org/10.1038/sj.cdd.4401046
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