Sequential gonadotropin treatment of immature mice leads to amplification of transforming growth factor β action, via upregulation of receptor-type 1, Smad 2 and 4, and downregulation of Smad 6
Sequential gonadotropin treatment of immature mice leads to amplification of transforming growth factor β action, via upregulation of receptor-type 1, Smad 2 and 4, and downregulation of Smad 6
复制标题
DOI：
10.1095/biolreprod.103.021162
复制
发表时间：
2004-03-01
影响因子：
3.6
通讯作者：
Gougeon, A
中科院分区：
文献类型：
--
作者：
Guéripel, X;Benahmed, M;Gougeon, A
本研究旨在建立细胞定位和表达的转化生长因子0（TGF β）信号通路组件，包括TGF β 1和β 2; TGF β受体I型（TbetaRI）和II型（TbetaRII）;和Smads 2，3，4，和6在促性腺激素诱导的卵泡成熟和排卵的小鼠卵巢。未成熟的21日龄小鼠依次接受重组人FSH（每天5 IU，持续3天）和hCG（在出生后第24天）治疗一次。免疫组织化学实验显示，TGF β 1染色颗粒细胞（GC）和卵泡膜细胞（TIC）以及在卵母细胞，而TGF β 2主要定位于卵母细胞和GC。在TIC中观察到TbetaRI和-RII的强免疫染色，并且在较小程度上在GC中观察到。而卵母细胞没有表现出任何染色的TbetaRII，其TbetaRI免疫染色是强的。在卵母细胞、GC和黄体细胞中检测到Smads，在TIC中检测到少量Smads; Smad 4的免疫染色最强。Western blotting和逆转录-聚合酶链反应（RT-PCR）分析表明，促性腺激素处理后，卵巢组织匀浆中TGF β 2、TbetaRI、Smad 2和Smad 4 mRNA表达水平升高，而Smad 6 mRNA表达水平降低。总之，这些结果表明，在暴露于促性腺激素序贯治疗的未成熟小鼠模型中，FSH和LH通过增加TGF β 2、TbetaRI、刺激性Smad 2和常见Smad 4表达来增加TGF β信号传导系统的表达，这伴随着抑制性Smad 6表达的降低。
The present study was designed to establish the cellular localization and expression of transforming growth factor 0 (TGFbeta) signaling pathway components, including TGFbeta1 and beta2; TGFbeta receptors type I (TbetaRI) and II (TbetaRII); and Smads 2, 3, 4, and 6 during gonadotropin-induced follicular maturation and ovulation in the mouse ovary. Immature 21-day-old mice were sequentially treated with recombinant human FSH, 5 IU daily for 3 days, and hCG once at Day 24 of life. Immunohistochemical experiments revealed a TGFbeta1 staining in granulosa cells (GC) and theca interna cells (TIC) as well as in oocytes, whereas that of TGFbeta2 was mainly localized in oocytes and GC. Strong immunostaining for both TbetaRI and -RII was observed in the TIC and, to a lesser extent, in GC. Whereas oocytes did not exhibit any staining for TbetaRII, their TbetaRI immunostaining was strong. Smads were detected in oocytes, GC, and luteal cells and in a lesser amount in TIC; the immunostaining for Smad 4 was the strongest. Western blotting and reverse transcription-polymerase chain reaction analyses indicated that, in response to gonadotropins, TGFbeta2, TbetaRI, Smad 2 and Smad 4 mRNA add protein levels increased, while those of Smad 6 decreased in ovarian homogenates. In conclusion, these results show that, in a model of immature mouse exposed to a sequential gonadotropin treatment, FSH and LH increased the expression of the TGFbeta signaling system through the increase of TGFbeta2, TbetaRI, stimulatory Smad 2, and common Smad 4 expression, which occurred concomitantly with a decrease of the inhibitory Smad 6 expression.