Long-term effects of early postnatal stress on Sertoli cells

Abstract

Sertoli cells are somatic cells in testis essential for spermatogenesis, that support the development, maturation, and differentiation of germ cells. Sertoli cells are metabolically highly active and physiologically regulated by external signals, particularly factors in the blood stream. In disease conditions, circulating pathological signals may affect Sertoli cells and consequentially, alter germ cells and fertility. While the effects of stress on reproductive cells have been well studied, how Sertoli cells respond to stress remains poorly characterized. We used a mouse model of early postnatal stress to assess the effects of stress on Sertoli cells. We developed an improved strategy based on intracellular stainings and obtained enriched preparations of Sertoli cells from exposed males. We show that adult Sertoli cells have impaired electron transport chain (ETC) pathways and that several components of ETC complexes particularly complex I, III, and IV are persistently affected. We identify serum as potential mediator of the effects of stress on Sertoli cells by showing that it can recapitulate ETC alterations in primary cells. These results highlight Sertoli cells as cellular targets of stress in early life that can keep a trace of exposure until adulthood.

What are Sertoli cells? Historical, methodological, and functional aspects

“…Our recent work proposed to use intracellular staining and FACS parameters to distinguish Sertoli cells from other testicular cells. 113 After enzymatic digestion, we suggest to stain the testicular cell suspension with Hoechst, to identify ploidy and with MitoTracker, indicating mitochondrial activity. As Sertoli cells are rather large, 114 are diploid due to mitotic arrest in adulthood, 4 and are metabolically highly active, 115 they can be distinguished by their FACS forward scatter and the intensity of Hoechst and MitoTracker staining.…”

“…As Sertoli cells are rather large, 114 are diploid due to mitotic arrest in adulthood, 4 and are metabolically highly active, 115 they can be distinguished by their FACS forward scatter and the intensity of Hoechst and MitoTracker staining. 113 Such staining may be transferable to other species, and therefore be less restrictive than transgenic lines which are mostly in mice. While improving enrichment strategies, one also has to consider that Sertoli cells go through various "stages of seminiferous tubules" (modes A and B illustrated in Figure 2).…”

“…However, transgenic lines are mostly available for rodents but not for other species, which restricts their applicability. Our recent work proposed to use intracellular staining and FACS parameters to distinguish Sertoli cells from other testicular cells 113 . After enzymatic digestion, we suggest to stain the testicular cell suspension with Hoechst, to identify ploidy and with MitoTracker, indicating mitochondrial activity.…”

“…After enzymatic digestion, we suggest to stain the testicular cell suspension with Hoechst, to identify ploidy and with MitoTracker, indicating mitochondrial activity. As Sertoli cells are rather large, 114 are diploid due to mitotic arrest in adulthood, 4 and are metabolically highly active, 115 they can be distinguished by their FACS forward scatter and the intensity of Hoechst and MitoTracker staining 113 . Such staining may be transferable to other species, and therefore be less restrictive than transgenic lines which are mostly in mice.…”