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Characterization of fish activin

鱼激活素的表征

基本信息

批准号：
09306013
负责人：
TAKASHIMA Fumio
金额：
$ 15.62万
依托单位：
Tokyo University of Fisheries
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

We cloned cDNAs of rainbow trout activin βA and βB, inhibin A, medaka activinβA and βB, and carp activinβA and βB. The mature protein coding regions of these cDNA have high homologies to those of human activin and inhibin. The genes of rainbow trout activinβA and inhibin A were also cloned. The structure of both genes βA and inhibin A have two exons mad one intron. The expression of rainbow trout activin genes was examined by RT-PCR. The major expression tissue of rainbow trout activin was ovary and brain at the mRNA level, and the major expression subunit of rainbow trout was βB subunit.Distribution of inhibin α and activin βA and βB in different ovarian and testis compartments were studied in rainbow trout by in situ hybridization with complementary RNA probes. In testes, inhibin α-and activin βA and βB were only expressed in the testicular interstitia between the seminal lobules, where Sertori cells and Leydig cells are distributed. In all the ovaries sampled every month period, theca cell layers of follicles had the strong reactions of cRNA probes hybridizing against inhibin α and activin βA and βB. In the regressive oocyte found in the July-sampled ovary, the positive reaction appeared.Distribution of activin A and its seasonal changes in the gonads of rainbow trout were immunohistochemically investigated using an heterologous antibody against recombinant human activin A. There were clear seasonal changes in activin A-ir in the gonads. In the testis, activin A-ir was detected during the degeneration of spermatozoa at the end of the spawning season. In the ovary, activin A-ir increased from gonadal recrudescence to ovulation, but disappeared completely after ovulation.

我们克隆了虹鱼激活素βA和βB，抑制素A，青竹激活素βA和βB，鲤鱼激活素βA和βB的基因，这些基因的成熟蛋白编码区与人激活素和抑制素有很高的同源性。此外，还克隆了虹鱼激活素βA和抑制素A基因。βA和抑制素A的结构都有两个外显子和一个内含子。用RT-PCR方法检测虹鱼激活素基因的表达。在基因水平上，虹鱼激活素的主要表达组织是卵巢和脑，其主要表达亚基是βB亚基。用互补α探针原位杂交的方法，研究了抑制素β和激活素βA和激活素B在虹鱼卵巢和睾丸不同组织中的分布。在睾丸中，抑制素α-和激活素βA和βB仅在精小叶之间的睾丸间质中表达，该间质中分布着Sertori细胞和Leydig细胞。在每月采集的所有卵巢中，卵泡膜细胞层均有抑制素α与激活素βA和βB杂交的cRNA探针的强烈反应。在7月采集的卵巢退化的卵母细胞中，出现阳性反应。用抗重组人激活素A的异源抗体免疫组织化学方法研究了激活素A在虹鲑性腺的分布及其季节变化。在精巢中，在产卵季末精子退化过程中检测到激活素A-ir。在卵巢中，激活素A-ir在性腺复发至排卵期间逐渐增加，但在排卵后完全消失。