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通过与 kisspeptin 处理相关的比较转录组分析鉴定黑龙江鲟(Acipenser schrenckii)大脑下丘脑 - 垂体 - 性腺轴中的基因。

Identification of genes in the hypothalamus-pituitary-gonad axis in the brain of Amur sturgeons (Acipenser schrenckii) by comparative transcriptome analysis in relation to kisspeptin treatment.

作者信息

Jin Shubo, Sun Dajiang, Xi Qingkai, Dong Xiaoli, Song Dan, Fu Hongtuo, Zhang Ying

机构信息

Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, PR China.

Heilongjiang River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Haebin, PR China.

出版信息

Gene. 2016 Dec 20;595(1):53-61. doi: 10.1016/j.gene.2016.09.026. Epub 2016 Sep 26.

DOI:10.1016/j.gene.2016.09.026
PMID:27688073
Abstract

Kisspeptin plays an important role in the reproduction and onset of puberty in vertebrates through stimulation of gonadotropin-releasing hormone (GnRH). However, the mechanisms whereby kisspeptin-related genes regulate sexual differentiation in teleosts are poorly understood. We aimed to study the relationship between the hypothalamus-pituitary-gonad (HPG) axis and sexual differentiation in relation to kisspeptin in the sturgeon Acipenser schrenckii. We performed comparative transcriptomic analysis of the brains of sturgeons treated with KISS1-10 during the gonadal sex-differentiation-sensitive period (170-210days post-hatching (dph)) using an Illumina sequencing platform. We also analyzed mRNA expression levels of genes in the HPG axis using real-time quantitative polymerase chain reaction, and measured estradiol-17β (E) and testosterone (T) levels in the brain and gonads using radioimmunological methods. A total of 75,960 and 74,907 unigenes were produced from Kisspeptin-treated and physiological saline-treated fish, respectively, among which 47,891 genes were matched to the non-redundant nr database. Potential genes and their functions were identified by GO (32,435), KEGG (37,619), and COG analyses (18,502). A total of 3169 unigenes were differentially expressed between transcriptomes in KISS1-10- and saline-injected fish, including 300 up-regulated and 2869 down-regulated unigenes. Gene expression levels of KISS1, G protein-coupled receptor-54, GnRH, androgen receptor, estrogen receptor, and Cyp19a in the brain and gonad were significantly affected by KISS1-10 treatment. KISS1-10 injection also significantly increased brain levels of E and T, compared with controls. These results support important roles for KISS1 in the regulation of the HPG axis, and in sex differentiation and reproduction in the Amur sturgeon.

摘要

亲吻素通过刺激促性腺激素释放激素(GnRH),在脊椎动物的生殖和青春期启动过程中发挥重要作用。然而,亲吻素相关基因调控硬骨鱼性别分化的机制尚不清楚。我们旨在研究施氏鲟下丘脑-垂体-性腺(HPG)轴与亲吻素相关的性别分化之间的关系。我们使用Illumina测序平台,对性腺性别分化敏感期(孵化后170-210天(dph))接受KISS1-10处理的施氏鲟大脑进行了比较转录组分析。我们还使用实时定量聚合酶链反应分析了HPG轴中基因的mRNA表达水平,并使用放射免疫法测量了大脑和性腺中雌二醇-17β(E)和睾酮(T)的水平。接受亲吻素处理和生理盐水处理的鱼分别产生了总共75,960个和74,907个单基因,其中47,891个基因与非冗余nr数据库匹配。通过GO(32,435)、KEGG(37,619)和COG分析(18,502)鉴定了潜在基因及其功能。在注射KISS1-10和生理盐水的鱼的转录组之间,共有3169个单基因差异表达,包括300个上调和2869个下调的单基因。KISS1-10处理显著影响了大脑和性腺中KISS1、G蛋白偶联受体-54、GnRH、雄激素受体、雌激素受体和Cyp19a的基因表达水平。与对照组相比,注射KISS1-10还显著提高了大脑中E和T的水平。这些结果支持了KISS1在调节HPG轴以及在施氏鲟性别分化和生殖中的重要作用。

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