长链非编码RNA lncPrep+96kb在Notch信号调控颗粒细胞分化中的作用研究

Notch通路是非常保守的信号传导途径，我们前期报道其通过调控颗粒细胞增殖影响卵泡发育；目前预实验发现，Notch通路抑制剂显著上调颗粒细胞FSHR水平，过表达Notch2则抑制FSHR表达。IncPrept+96kb是最新报道颗粒细胞高表达的长链非编码RNA,但作用尚不清楚。我们发现激活Notch通路显著上调了IncPrept+96kb表达；RNA免疫共沉淀发现lncPrept+96kb与调控FSHR的转录因子(USF1和USF2)直接结合。因此我们假设lncPrept+96kb在Notch调控颗粒细胞FSHR中起重要作用。本课题将通过Notch通路敲除小鼠模型和颗粒细胞培养观察Notch通路对颗粒细胞分化和lncPrep+96kb的影响，观察lncPrep+96kb在Notch通路调控FSHR中的作用及机理。该研究有助于阐明Notch调控颗粒细胞分化的机制，为卵泡发育机理提供实验依据。

Notch signaling defines an evolutionary ancient cell interaction. In our previous study, we reported that Notch signaling was involved in follicle growth through regulating granulosa cell proliferation. In present study, we found that blocking of Notch signaling upregulated FSHR expression and overexpression of Notch2 inhibited the expression of FSHR. LncPrept+96kb is one new long non-coding RNA, which is highly expressed in granulosa cells. We found that activation of Notch signaling increased its expression in granulosa cells. RNA immunoprecipitation enriched lncPrept+96kb binding protein: USF1 and USF2, which are two important transcription factors of regulating FSHR transcription. Based on these results, we hypothesize that lncPrept+96kb is involved in FSHR expression regulated by Notch signaling. To test the hypothesis, we will employ Notch signaling knockout mice and granulosa cell culture in vitro to investigate the influence of Notch signaling on FSHR and lncPrept+96kb expression and explore the function and mechanism of lncPrept+96kb on FSHR expression regulated by Notch signaling. Completion of this study will be instrumental to understand the mechanism of Notch signaling on granulosa cell differentiation and provide novel insight for follicle development.