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Analysis of mechanism of changes in function and size of golden hamster testis with special reference to TGF-β signal transduction

特别关注TGF-β信号转导的金黄地鼠睾丸功能和大小变化机制分析

基本信息

批准号：
11460137
负责人：
HAYASHI Yoshihiro
金额：
$ 8.38万
依托单位：
THE UNIVERSITY OF TOKYO
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11460137/
关键词：
testis short photoperiod spermatogonia spermatocyte mouse hamster Smad2 Smad3 Smadファミリー

项目摘要

First, during prespermatogenesis when the testis prominently increases in size, the reproliferation and relocation of gonocytes were examined by BrdU method and electron microscopy. Gonocytes began to move from 17.5dpc and relocated on the basement membrane from 18.5dpc. BrdU-labeled gonocytes were first detected on 1.5dpp. They increased in number abruptly from 2.5dpp and then gradually increased. Secondly, histology and lectin-binding patterns in the testes of two different-body sized bats (Java fruit bat, average b.w. : 567g ; Japanese lesser horseshoe bat, average b.w. : 7g) were investigated. Both testicular morphology and lectin-bindig patterns were almost similar between these two species. Thirdly, the mouse Smad3 cDNA including the open reading frame was cloned from the mouse brain using RACE technique, and its expression pattern was analyzed by northern blot. The predicted amino acid sequences of mouse Smad3 showed a high homology with human Smad3 (99.3%) and mouse Smad2 (85.4%). Northern blot analysis revealed that Smad3 was highly expressed in brain and ovary. In situ hybridization revealed that Smad3 was detected in pyramid cells of hippocampus, granular cells of cerebral cortex, and granulosa cells of ovary. Next, the expression of Smad2 and Smad3 mRNA was examined under the influence of long and/or short photoperiod in hamsters. In situ hybridization detected both Smad2 and Smad3 mRNA in spermatogonia and spermatocytes in both photoperiods. Northern blots showed that Smad2 mRNA was detected at all stages in both photoperiods, whereas Smad3 mRNA was expressed in a short photoperiod. The photoperiodic condition would change the balance between Smad2 and Smad3 transcripts. Moreover, the localization of Smad2 and Smad3 protein was examined in both photoperiods. The Smad2 and Smad3 proteins were localized in spermatocyte cytoplasm in a long photoperiod. They accumulated in spermatocyte nucleus in a short photoperiod.

首先，在精子发生前，当睾丸明显增大时，用BrdU法和电子显微镜观察生殖细胞的再增殖和移位。性细胞从17.5dpc开始运动，18.5dpc开始迁移到基底膜。在1.5dpp时，首次检测到BrdU标记的生殖细胞。它们的数量从2.5dpp突然增加，然后逐渐增加。其次，对两种不同体型的蝙蝠(爪哇果蝠，平均体重)的睾丸组织学和凝集素结合模式进行了研究。：567g；日本小马蹄蝠，平均体长。：7g)。这两个物种的睾丸形态和凝集素结合模式几乎是相似的。第三，利用RACE技术从小鼠脑中克隆了含开放阅读框的小鼠Smad3基因，并用Northern印迹分析了其表达模式。预测的小鼠Smad3氨基酸序列与人Smad3(99.3%)和小鼠Smad2(85.4%)有很高的同源性。Northern印迹分析表明，Smad3在脑和卵巢中高表达。原位杂交结果显示，Smad3在海马区锥体细胞、大脑皮层颗粒细胞和卵巢颗粒细胞中均有表达。其次，研究了长光周期和短光周期对仓鼠Smad2和Smad3基因表达的影响。原位杂交检测到Smad2和Smad3在两个光周期的精原细胞和精母细胞中均有表达。Northern杂交结果表明，Smad2在两个光周期的各个时期都有表达，而Smad3在短光周期中表达。光周期条件会改变Smad2和Smad3转录本之间的平衡。此外，还检测了Smad2和Smad3蛋白在两个光周期中的定位。Smad2和Smad3蛋白在较长的光周期中定位于精母细胞胞浆。在短光周期内，它们聚集在精母细胞核中。