Matsushige Cassandra, Kitazumi Kaelyn, Beaman Amanda, Miyagi Marissa, Tallquist Michelle D, Yamazaki Yukiko
Reproduction. 2024 Dec 9;169(1). doi: 10.1530/REP-24-0151. Print 2025 Jan 1.
Three-dimensional ovarian tissue culture is a unique model to define the effects of molecules on folliculogenesis. Using this model, we determined that RGD-integrin interaction plays a role in antrum formation and theca cell differentiation.
We recently developed a three-dimensional (3D) ovarian tissue culture system supported by bacterial-derived dextran hydrogel. Arg-Gly-Asp (RGD) is an extracellular matrix-derived triple peptide. Immature ovarian tissues cultured in RGD-modified dextran hydrogel significantly promoted antral follicle growth and oocyte quality compared with those cultured in dextran hydrogel alone. In this study, we examined the mechanism of follicle growth stimulated by RGD treatment in the 3D system. First, we detected that direct contact between RGD-modified dextran hydrogel and ovarian interstitial cells is necessary to promote antral follicle growth. Therefore, we hypothesized that RGD stimulates antral follicle growth through RGD-binding integrin receptors expressed in the interstitial cell mass. Using quantitative PCR (qPCR) and immunochemical staining, we identified that integrins ⍺vβ3 and ⍺v5 are predominantly expressed in the ovarian interstitial compartment. To assess the effect of RGD-integrin interaction on follicle growth, ovarian tissues were cultured with cilengitide (Ci), an inhibitor specific for ⍺vβ3 and ⍺vβ5. Ci treatment suppressed RGD-induced follicle growth and oocyte quality in a dose-dependent manner. When the interstitial cell aggregates were cultured with RGD, cell migration and theca-related gene expression were significantly upregulated. Ci treatment dramatically suppressed these RGD-induced activities. In coculturing the interstitial aggregate and secondary follicles with RGD, migrating cells formed the outermost cell layers around the follicles, like theca layers, which were totally blocked by Ci treatment. In conclusion, our results suggest that RGD stimulates theca cell differentiation in the ovarian interstitial cells through integrins ⍺vβ3 and ⍺v5 to promote antral follicle growth in our 3D system.
三维卵巢组织培养是一种独特的模型，可用于确定分子对卵泡发生的影响。利用该模型，我们确定RGD-整合素相互作用在卵泡腔形成和膜细胞分化中发挥作用。
我们最近开发了一种由细菌衍生的葡聚糖水凝胶支持的三维（3D）卵巢组织培养系统。精氨酸-甘氨酸-天冬氨酸（RGD）是一种源自细胞外基质的三肽。与单独在葡聚糖水凝胶中培养的未成熟卵巢组织相比，在RGD修饰的葡聚糖水凝胶中培养的未成熟卵巢组织显著促进了窦卵泡生长和卵母细胞质量。在本研究中，我们研究了在三维系统中RGD处理刺激卵泡生长的机制。首先，我们检测到RGD修饰的葡聚糖水凝胶与卵巢间质细胞之间的直接接触是促进窦卵泡生长所必需的。因此，我们假设RGD通过间质细胞团中表达的RGD结合整合素受体刺激窦卵泡生长。使用定量PCR（qPCR）和免疫化学染色，我们确定整合素αvβ3和αv5主要在卵巢间质区表达。为了评估RGD-整合素相互作用对卵泡生长的影响，用西仑吉肽（Ci）培养卵巢组织，西仑吉肽是一种对αvβ3和αvβ5特异的抑制剂。Ci处理以剂量依赖性方式抑制RGD诱导的卵泡生长和卵母细胞质量。当间质细胞聚集体与RGD一起培养时，细胞迁移和膜相关基因表达显著上调。Ci处理显著抑制了这些RGD诱导的活性。在用RGD共培养间质聚集体和次级卵泡时，迁移的细胞在卵泡周围形成了最外层细胞层，类似于膜层，而Ci处理完全阻断了这一过程。总之，我们的结果表明，在我们的三维系统中，RGD通过整合素αvβ3和αv5刺激卵巢间质细胞中的膜细胞分化，从而促进窦卵泡生长。
