摘要
为了明确17α,20β双羟孕酮(DHP)及其核受体调控促性腺激素释放激素mRNA在雄性斑马鱼脑组织中的表达机制,采用实时荧光定量PCR方法、类固醇激素活体和离体暴露方法进行试验。结果表明,野生型雄性斑马鱼活体暴露于100 nmol/L DHP水溶液,伴随暴露时间的延长,gnrh2和gnrh3 mRNA表达量呈逐渐升高趋势,24 h出现表达最高峰;野生型雄性斑马鱼活体分别暴露于10和100 nmol/L DHP水溶液24 h。与对照组相比,100 nmol/L DHP试验组gnrh2和gnrh3 mRNA表达量升高,差异显著(P<0.05),而10 nmol/L DHP试验组无显著变化(P>0.05);野生型雄性斑马鱼活体暴露于100 nmol/L DHP与100 nmol/L RU486混合水溶液24 h,试验组gnrh2和gnrh3 mRNA表达量与对照组相比无显著变化(P>0.05);野生型和孕酮核受体敲除型(pgr-/-)雄性斑马鱼活体分别暴露于100 nmol/L DHP水溶液24 h,与野生型试验组相比pgr-/-型试验组gnrh2和gnrh3 mRNA表达量显著降低(P<0.05);野生型和pgr-/-型雄性斑马鱼脑组织离体暴露于含100 nmol/L DHP的L-15培养液24 h,与野生型试验组相比pgr-/-型试验组gnrh2和gnrh3 mRNA表达量显著降低(P<0.05)。由此可见,DHP与核受体结合可直接调控雄性斑马鱼脑组织中促性腺激素释放激素gnrh2和gnrh3 mRNA表达。
In order to clarify that 17α,20βdihydroxyproterone(DHP)and its nuclear receptor regulated expression mechanism of gonadotropin-releasing hormone(GnRH)mRNA in male zebrafish,real-time PCR,steroid hormone exposure in vivo and in vitro were used in this test.The results showed that it took 24 h for gnrh2 and 24 h for gnrh3 to respond significantly(P<0.05).24 h 100 nmol/L DHP exposure significantly increased the expression of gnrh2 and gnrh3 mRNA(P<0.05).With and without DHP in vivo treatment,compared with wild type zebrafish treatment group,mRNA expressions of gnrh2 and gnrh3 were significantly decreased in pgr-/-zebrafish treatment group(P<0.05),while that in wild-type zebrafish and pgr-/-zebrafish brain tissue were all exposed to L-15 culture medium containing 100 nmol/L DHP for 24 h,compared with wild-type zebrafish treatment group,mRNA expressions of gnrh2 and gnrh3 were significantly decreased in pgr-/-zebrafish treatment group(P<0.05).DHP and its nuclear receptors could regulate mRNA expressions of gonadotropin-releasing hormone in male zebrafish.
作者
方敏敏
王翠丽
FANG Min-min;WANG Cui-li(Institute of Geography and Oceanography,Nanning Normal University,Nanning,Guangxi 530000;Key Laboratory of Environment Change and Resources Use in Beibu Gulf,Nanning Normal University,Nanning,Guangxi 530000)
出处
《安徽农业科学》
CAS
2019年第23期99-102,共4页
Journal of Anhui Agricultural Sciences
基金
广西自然科学基金项目(2018GXNSFBA281137)
北部湾环境演变与资源利用教育部重点实验室—广西地表过程与智能模拟重点实验室(南宁师范大学)联合开放基金项目(GTEU-KLOP-X1702)
2018年自治区级大学生创新创业训练计划项目(201810603295)
关键词
斑马鱼
17α
20β双羟孕酮
孕酮核受体
促性腺激素释放激素
Zebrafish
17α
20β-dihydroxy-4-pregnen-3-one
Nuclear progesterone receptor
Gonadotropin-releasing hormone
