Search

[Objectives] This study investigates the capability of exosome secretion by Sertoli cells and its influence on the self-renewal and differentiation status of porcine spermatogonial stem cells(pSSC) using an in vitro model. [Methods] The two-step enzyme digestion method was employed to isolate Sertoli cells and pSSC, and an in vitro co-culture model of Sertoli cells and pSSC was established. Suppression of exosome secretion by Sertoli cells was achieved to examine the effects on pSSC self-renewal and differentiation markers, thereby studying the impact of Sertoli cell-derived exosome on the fate of pSSC. [Results] The isolated Sertoli cells exhibited a Wilm tumor 1(WT1) positive rate of (87.1±0.02)% and an intorgrin β-1(ITGB1) positive rate of (94.2±0.01)%. The isolated pSSC showed a promyelocytic lenkaemia zine finger protein (PLZF) positive rate of (80.6±0.02) %. Presence of exosome secreted by Sertoli cells was confirmed through Western blot analysis and immunofluorescence assays. Treatment with 20 µmol L-1 GW4869 effectively inhibited exosome secretion without compromising Sertoli cells. Co-culture of pSSC with Sertoli cells treated with the inhibitor revealed significant differences in pSSC status compared to the control group. RT-PCR results showed a significant increase in the expression of the PLZF gene, while expression levels of receptor tyrosine kinase gene (KIT), thymus cell antigen 1 gene (THYI), and dead box 4 gene (DDX4) were significantly reduced. Western blot analysis indicated that levels of protein gene product 9.5(PGP9.5) protein expression elevated and DDX4 protein expression level decreased in pSSC co-cultured with inhibitor-treated Sertoli cells. [Conclusions] Exosomes secreted by Sertoli cells had a regulatory role in both self-renewal and differentiation of pSSC in vitro. This finding implied that exosome represented an important means through which Sertoli cells influence the fate of spermatogonial stem cells, offering novel insights for enhancing boar reproductive performance and fertility improvement. [ABSTRACT FROM AUTHOR]

[Objectives]The purpose : of this study is to investigate the effect of zearalcnone( ZKAJon lactic acid production in Sertoli cells (SC). [Methods]The SC of Vi istar rats were cultured in vitro as test materials. 0(control), 0.1,1,10,20,30 μ.mol·L-1 ZEA were used to detect the cytotoxicity of ZKA against SC by I.DH release assay kit. The lactic acid test (lactic acid kit) was used to observe the effect of ZKA on lactic acid production in SC. The pyruvate test was performed to observe the effect of ZKA on pyruvate production in SC. The Western blot was used to detect the expression of lactic acid prodi jcing relative proteins. The immunofluorescence assay was performed aiming at detecting the key prot ein I.DH of lactic acid produc tion in SC. [Results] With the increase of ZKA concentration, the cytotoxicity of ZKA on SC gradually increased( P<0.01), and the content of lactic acid inside and outside the cell, and tl: ic concentration of pyruvate in the cell showed a downward trend (P< 0.05 or P<0.01 ). The expressions of lactic acid production-related proteins GLUT1 and I.DH exposed to ZKA at 1,10,20,30 μ.mol·L group and above significantly decreased (P<0.01), and the expression of MCT4 at the 30μmol·L-1 ZKA group significantly decreased ( P<0.05). The immunofluorescence assay showed that the fluorescence intensitv of I.DH decreased with the increase of the concentration. Compared with the control group, the protein expressions in the eells exposed to ZEA at 10,20,30 (μmol·L-1 groups, and above significantly decreased (f<0.01). [ Conclusions ] The above results indicate ZEA can inhibit the producti on of lactic acid in SC, and interfere with the energy supply of SC to germ eells, whieh damage* > the mal e reproductive function. [ABSTRACT FROM AUTHOR]