Identification of a New Theca/Interstitial Cell-Specific Gene and Its Biological Role in Growth of Mouse Ovarian Follicles at the Gonadotropin-Independent Stage
Identification of a New Theca/Interstitial Cell-Specific Gene and Its Biological Role in Growth of Mouse Ovarian Follicles at the Gonadotropin-Independent Stage
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DOI：
10.3389/fendo.2019.00553
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发表时间：
2019-08-14
影响因子：
5.2
通讯作者：
Satake, Honoo
中科院分区：
文献类型：
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作者：
Aoyama, Masato;Shiraishi, Akira;Satake, Honoo
卵泡膜/间质细胞负责卵泡的生长和成熟。然而，对细胞膜/间质细胞特异性基因及其功能知之甚少。在这项研究中，我们通过RNA-seq技术探索了膜膜/间质细胞的转录组，以及膜膜细胞标记物——抗菌素(Aspn)/牙周韧带相关蛋白1 （PLAP-1）的新生物学作用。RNA-seq检测到从3周龄小鼠卵巢分离的膜/间质细胞和颗粒细胞中特异性表达的432和62个基因。基因本体分析表明，这些基因大致可分为四大类：细胞外基质组织相关术语、趋化相关术语、血管生成相关术语和形态发生相关术语。原位杂交表明，新检测到的代表基因Aspn/PLAP-1在卵泡膜细胞外层特异性检测到，与基底层特异性基因Nidgen-1的表达形成对比。有趣的是，一种Aspn/PLAP-1抗体完全阻止了次级卵泡的生长，即不依赖促性腺激素的卵泡发育阶段。此外，转化生长因子- β (tgf - β)触发的信号被Aspn/PLAP-1抗体处理诱导，这与Aspn/PLAP-1对tgf - β的抑制作用是一致的。总之，这些结果表明，卵泡膜细胞根据新的标记基因及其生物学功能被划分为亚群，并提供了Aspn/PLAP-1仅在卵泡膜细胞外层表达的证据，并通过下调典型的tgf - β信号级联在次生卵泡的生长中起关键作用。
Theca/interstitial cells are responsible for the growth and maturation of ovarian follicles. However, little is known about the theca/interstitial cell-specific genes and their functions. In this study, we explored transcriptomes of theca/interstitial cells by RNA-seq, and the novel biological roles of a theca cell marker, asporin (Aspn)/periodontal ligament-associated protein 1 (PLAP-1). RNA-seq detected 432 and 62 genes expressed specifically in theca/interstitial cells and granulosa cells isolated from 3-weeks old mouse ovaries. Gene ontology analysis demonstrated that these genes were largely categorized into four major groups: extracellular matrix organization-related terms, chemotaxis-related terms, the angiogenesis-related terms, and morphogenesis-related terms. in situ hybridization demonstrated that the newly detected representative gene, Aspn/PLAP-1 , was detected specifically in the outer layer of theca cells in contrast with the expression of the basal lamina-specific gene, Nidgen-1. Intriguingly, an Aspn/PLAP-1 antibody completely arrested the growth of secondary follicles that is the gonadotropin-independent follicle developmental stage. Furthermore, transforming growth factor-beta (TGF-beta)-triggered signaling was induced by the Aspn/PLAP-1 antibody treatment, which is consistent with the inhibitory effect of Aspn/PLAP-1 on TGF-beta. Altogether, these results suggest that theca cells are classified into subpopulations on the basis of new marker genes and their biological functions, and provide evidence that Aspn/PLAP-1 is expressed exclusively in the outer layer of theca cells and plays a pivotal role in the growth of secondary follicles via downregulation of the canonical TGF-beta signaling cascade.