摘要: |
为研究体外培养的小鼠卵巢颗粒细胞Bmp8a和Bmp8b基因的表达情况,用10 IU PMSG 皮下注射21~23日龄健康雌性ICR小白鼠,48 h后刺破卵泡获取颗粒细胞,置入10% FBS的DMEM/F12中体外培养5 d,用Trizol提取总RNA, RT-PCR验证后切取目的DNA片段进行测序。RT-PCR测序结果表明体外培养的小鼠卵巢颗粒细胞可扩增BMP-8a transcript variant 2 和BMP-8b的mRNA序列,不扩增BMP-8a transcript variant 1的mRNA序列。结果显示体外培养的小鼠卵巢颗粒细胞可表达BMP-8a transcript variant 2和Bmp8b基因。 |
关键词: 小鼠 卵巢颗粒细胞 Bmp8a Bmp8b |
DOI:10.11841/j.issn.1007-4333.2013.03.016 |
投稿时间:2012-09-11 |
基金项目:国家转基因重大专项资助(2011ZX08008-003) |
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Expressions of Bmp8a and Bmp8b in culturedmouse granulosa cells |
WANG Jian-qi, Selvaraj J.NIMAL, CAO Wen-guang
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(Institute of Animal Sciences and Veterinary Medicine, Chinese Academy of Agricultural Sciences, Beijing 100193, China) |
Abstract: |
This study was to analyze the expressions of Bmp8a and Bmp8b in cultured mouse granulosa cells,for further understanding of the mechanism of follicular development.Twenty-one to twenty-three day-old female mice were hypodermically injected with 10IU of PMSG.After 48 hours,the bilateral ovaries were removed and the granulosa cells were collected by piercing the ovarian follicles.The granulosa cells were seeded in cell culture plates in DMEM/F12 supplemented with 10% FBS.After 5 days,the cells were collected from the wells and total RNA was extracted.The expressions of Bmp8a and Bmp8b were analyzed by RT-PCR and confirmed by sequencing.RT-PCR sequencing results showed that in vitro cultured mouse granulosa cells generated BMP-8a transcript variant 2 and BMP-8b mRNA.And BMP-8a transcript variant 1 mRNA was found to be absent.Our results showed that in vitro cultured mouse granulosa cells could express BMP-8a transcript variant 2 and Bmp8b. |
Key words: mice granulosa cells Bmp8a Bmp8b |