Isolation and characterization of sexual dimorphism genes expressed in chicken embryonic gonads
Isolation and characterization of sexual dimorphism genes expressed in chicken embryonic gonads
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鸡胚胎性腺中表达的性二态性基因的分离和表征
DOI：
10.1093/abbs/gmp012
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发表时间：
2009-04-01
影响因子：
3.7
通讯作者：
Gong, Yanzhang
中科院分区：
文献类型：
--
作者：
Feng, Yanping;Peng, Xiuli;Gong, Yanzhang
鸡的双能性胚胎性腺分化为一对睾丸或一个卵巢，但与性腺性别分化相关的基因尚不清楚，性别决定基因也不清楚。为了进一步研究鸡性别分化的相关基因，本研究采用抑制性消减杂交技术分离了鸡E3.5-E6期性腺的性别差异表达基因。利用斑点杂交技术，共筛选到88个基因的152个cDNA克隆（41个来自F-M文库，47个来自M-F文库）。这些基因主要位于大染色体（1-5）上，其中5个位于性染色体（1个在W，4个在Z）上，编码属于酶、DNA缔合、RNA缔合和结构蛋白的4种主要分子类别。将获得的cDNA序列与鸡EST数据库中的cDNA序列进行比较，发现F-M文库中的32个基因和M-F文库中的16个基因在两个已报道的胚胎性腺cDNA文库中具有同源性。对8个表观遗传和转录调控基因的实时荧光定量PCR分析表明，CDK 2AP 1、SMARCE 1、SAP 18、SUDS 3和PQBP 1在性腺发育早期（E4）出现了显著的性别差异表达。在性别差异表达基因功能比较的基础上，分析了ATP 5A 1 W、CDK 2AP 1、线粒体转录本等重要性别差异表达基因的功能。总之，分离基因的特征将提供有价值的线索，以确定潜在的候选人参与鸡性别分化和性腺发育的遗传机制。
In chicken, the bipotential embryonic gonad differentiates into either a pair of testes or an ovary, but few genes that underlying the gonadal sex differentiation have been identified and the sex-determination gene is still unknown. To identify more genes involved in chicken sex differentiation, we employed suppression subtractive hybridization to isolate differentially expressed genes between sexes from chicken gonads during a period of E3.5-E6. A total of 152 cDNA clones corresponding to 88 genes (41 from F-M library and 47 from M-F library) were screened using dot-blot analysis. These genes are located mainly on the macrochromosomes (1-5) with five in the sex chromosomes (one in W and four in Z), encoding four dominating molecular categories belonging to enzyme, DNA association, RNA association, and structural protein. Comparing the obtained cDNA sequences with those in chicken EST database, it showed that cDNAs of 32 genes from F-M library and 16 from M-F library have homologs in two reported embryonic gonad cDNA libraries. Quantitative real-time PCR analysis of eight genes involved in epigenetic and transcription regulation showed significantly different expression between sexes of CDK2AP1, SMARCE1, SAP18, SUDS3, and PQBP1 appeared at the early stage in gonad development (E4). Based on the functional comparison of sexual differentially expressed genes, the roles of some putatively important genes including ATP5A1W, CDK2AP1, mitochondrial transcripts, etc. have been analyzed. In conclusion, characterization of isolated genes would provide valuable clues to identify potential candidates involved in genetic mechanisms of chicken sex differentiation and gonad development.