Abstract

Sertoli cells are essential for testicular development and spermatogenesis, but the post-transcriptional mechanisms regulating their function in sheep remain incompletely understood. This study investigated the regulatory relationship between miR-132-y and Yes-associated protein 1 (YAP1), a core effector of the Hippo pathway, in primary Sertoli cells isolated from Southdown × Hu F1 sheep. Target prediction and dual-luciferase reporter assays supported a direct interaction between miR-132-y and the YAP1 3′ untranslated region. YAP1 overexpression was associated with increased CCK-8-based cell viability and altered mRNA expression of selected viability-associated, YAP1-related, and Sertoli cell function-associated genes, whereas YAP1 silencing showed opposite trends. Conversely, miR-132-y overexpression reduced YAP1 mRNA abundance and was associated with decreased CCK-8-based cell viability and corresponding transcriptional changes, while miR-132-y inhibition produced the opposite pattern. Rescue experiments showed that ectopic YAP1 expression partially attenuated miR-132-y-associated changes. Overall, these findings provide in vitro, cell-based evidence that miR-132-y targets YAP1 at the transcript level and is associated with viability-related transcriptional responses in sheep Sertoli cells.

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