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Synergistic inhibition of csal1 and csal3 in granulosa cell proliferation and steroidogenesis of hen ovarian prehierarchical development.

Spalt-like 1 (SALL1) and spalt-like 3 (SALL3) are multi-zinc finger transcription factors that play an essential role in regulating developmental processes and organogenesis in many species. However, the functional role of SALL1 (csal1) and SALL3 (csal3) in chicken prehierarchical follicle (PF) development is unknown. This study aimed to explore the potential role and mechanism of csal1 and csal3 in granulosa cells (GCs) proliferation, differentiation, and follicle selection within the PFs of hen ovary. Our data demonstrated that the csal1 and csal3 transcriptions were highly expressed in GCs of the various-sized PFs and their proteins were mainly localized in the cytoplasm of granulosa cells and oocytes of the examined PFs as well as in the ovarian stroma and epithelium. It initially revealed that both csal1 and csal3 may be involved in chicken PF development via a translocation mechanism. Furthermore, our results showed an abundance of CCND1, Bcat, StAR, CYP11A1, and FSHR mRNA in GCs and the proliferation levels of GCs from the PFs were significantly increased by siRNA-mediated knockdown of csal1 or/and csal3. Conversely, the overexpression of csal1 or/and csal3 in the GCs led to a remarkably decreased mRNA expression of the five genes correlating with an decline in cell proliferation levels. Moreover, csal1 and csal3 togather exert a much stronger effect on the regulation than any of csal1 or csal3. These results indicated that csal1 and csal3 play synergistic inhibitory roles on GC proliferation, differentiation, and steroidogenesis during PF development in vitro. The current data provides a basis for further investigation of molecular mechanisms of csal1 and csal3 in controlling the PF development and growth of hen ovary in vivo.

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