Abstract
Experimental autoimmune orchitis (EAO) is a chronic inflammatory disorder that causes progressive spermatogenic impairment. EAO is characterized by high intratesticular levels of nitric oxide (NO) and tumor necrosis factor alpha (TNFα) causing germ cell apoptosis and Sertoli cell dysfunction. However, the impact of this inflammatory milieu on the spermatogenic wave is unknown. Therefore, we studied the effect of inflammation on spermatogonia and preleptotene spermatocyte cell cycle progression in an EAO context and through the intratesticular DETA-NO and TNFα injection in the normal rat testes. In EAO, premeiotic germ cell proliferation is limited as a consequence of the undifferentiated spermatogonia (CD9+) cell cycle arrest in G2/M and the reduced number of differentiated spermatogonia (c-kit+) and preleptotene spermatocytes that enter in the meiotic S-phase. Although inflammation disrupts spermatogenesis in EAO, it is maintained in some seminiferous tubules at XIV and VII–VIII stages of the epithelial cell cycle, thereby guaranteeing sperm production. We found that DETA-NO (2 mM) injected in normal testes arrests spermatogonia and preleptotene spermatocyte cell cycle; this effect reduces the number of proliferative spermatogonia and the number of preleptotene spermatocytes in meiosis S-phase (36 h after). The temporal inhibition of spermatogonia clonal amplification delayed progression of the spermatogenic wave (5 days after) finally altering spermatogenesis. TNFα (0.5 and 1 µg) exposure did not affect premeiotic germ cell cycle or spermatogenic wave. Our results show that in EAO the inflammatory microenvironment altered spermatogenesis kinetics through premeiotic germ cell cycle arrest and that NO is a sufficient factor contributing to this phenomenon.
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Acknowledgements
We thank the Instituto Nacional de Microbiología “A. Malbrán”, División Vacunas Bacterianas for their generous gift of Bordetella pertussis, and M. Imsem, and C. García for their technical assistance.
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Universidad de Buenos Aires (UBA 20020150100104BA) and the Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT PICT 0497).
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María Eugenia Ferreiro, Cinthia Soledad Méndez, and Leilane Glienke: investigation, methodology, formal analysis; Maria Jimena Ferraris and Patricia Verónica Jacobo: conceptualization, review and editing; Cristian Marcelo Sobarzo: visualization; Daniel Pisera and Livia Lustig: supervision, review and editing; Maria Susana Theas: conceptualization supervision, validation, resources, original draft.
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The use of rats followed the National Institutes of Health (NIH) guidelines for care and use of experimental animals, and ethical approval was granted by local the committee: CICUAL-Facultad de Medicina, Universidad de Buenos Aires [RES (CD): 2029/2019].
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Maria Eugenia Ferreiro and Cinthia Soledad Méndez are co-author.
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Ferreiro, M.E., Méndez, C.S., Glienke, L. et al. Unraveling the effect of the inflammatory microenvironment in spermatogenesis progression. Cell Tissue Res 392, 581–604 (2023). https://doi.org/10.1007/s00441-022-03703-z
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DOI: https://doi.org/10.1007/s00441-022-03703-z