The reproductive lifespan of a woman is determined by the gradual recruitment of quiescent follicles into the growing pool. In humans, ovarian tissue removal from its in vivo environment induces spontaneous activation of resting follicles. Similarly, pharmacological activation of the PI3K/Akt pathway leads to accelerated follicle recruitment, but has been associated with follicular damage. Recent findings demonstrate that everolimus (EVE), an mTORC1 inhibitor, limits primordial follicle activation. However, its potential benefit regarding growing follicle integrity remains unexplored. Ovarian cortical fragments were exposed to ± EVE for 24 h and cultured for an additional 5 days. After 0, 1 and 6 days of culture, fragments were either processed for ultrastructural analysis or subjected to follicular isolation for gene expression and immunofluorescence assessments. Data from transmission electron microscopy showed that growing follicles displayed similar ultrastructural features irrespective of the conditions and maintained close contacts between germinal and stromal compartments. Establishment of intra-follicular communication was confirmed by detection of a gap junction component, Cx43, in both groups throughout culture, whereas transzonal projections, which physically link granulosa cells to oocyte, formed later in EVE-treated follicles. Importantly, levels of GJA1 mRNA, encoding for the Cx43 protein, significantly increased from Day 0 to Day 1 in the EVE group, but not in the control group. Given that EVE-treated follicles were smaller than controls, these findings suggest that EVE might facilitate the establishment of appropriate intercellular communications without impairing follicle ultrastructure. Therefore, mTORC1 inhibitors might represent an attractive tool to delay the culture-induced primordial follicle activation while maintaining follicles in a functionally integrated state.

中文翻译：

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培养的人类早期卵泡暴露于mTORC1抑制剂的超微结构和细胞间接触介导的通讯。

女人的生殖寿命取决于将静止的卵泡逐渐吸收到生长池中。在人类中，从体内环境中去除卵巢组织会诱导静止卵泡的自发激活。同样，PI3K / Akt途径的药理学激活导致卵泡募集加速，但与卵泡损害有关。最近的发现表明，mTORC1抑制剂依维莫司（EVE）限制了原始卵泡的激活。然而，其关于增加卵泡完整性的潜在益处仍未得到开发。将卵巢皮质片段暴露于±EVE 24 h，再培养5天。经过0、1和6天的培养，对片段进行加工以进行超微结构分析或进行滤泡分离以进行基因表达和免疫荧光评估。来自透射电子显微镜的数据显示，无论条件如何，正在生长的卵泡均表现出相似的超微结构特征，并且在生发和基质间室之间保持紧密接触。通过在整个培养过程中在两组中检测到间隙连接成分Cx43来确定小泡内通讯的建立，而将粒细胞与卵母细胞物理连接的跨区投射稍后在EVE处理的卵泡中形成。重要的是，在EVE组中，编码Cx43蛋白的GJA1 mRNA水平从第0天到第1天显着增加，但在对照组中则没有。鉴于经EVE处理的卵泡比对照组小，这些发现表明，EVE可能有助于建立适当的细胞间通讯，而不会损害卵泡超微结构。因此，mTORC1抑制剂可能是一种有吸引力的工具，可以在维持卵泡处于功能整合状态的同时，延迟培养诱导的原始卵泡活化。