伸展摇蚊(Chironomus tentans)转录组测序及胰岛素信号通路关键基因序列分析

doi:10.19741/j.issn.1673-4831.2023.0132

摘要/Abstract

摘要： 为了探索基于胰岛素信号通路的无脊椎动物内分泌干扰性基因标志物,以伸展摇蚊(Chironomus tentans)为模式生物,在转录组测序基础上识别胰岛素信号通路中的关键基因,并针对InR、Pdk、Akt、FoxO等上游基因进行了cDNA全长扩增和氨基酸序列的多物种比对。基于该研究获取的cDNA全长序列,伸展摇蚊InR、Pdk、Akt、FoxO基因可分别编码含1 436、522、520、470个氨基酸的蛋白序列,序列同源性比对、系统发育树分析结果验证了其进化保守性。伸展摇蚊Ⅰ~Ⅳ龄幼虫、蛹、雌性成虫、雄性成虫中基因相对表达量的实时荧光定量PCR(qRT-PCR)检测结果表明,InR、Pdk、Akt、FoxO在伸展摇蚊体内的表达趋势相似。Ⅰ龄和Ⅳ龄幼虫的基因表达峰值提示胰岛素信号在Ⅰ龄和Ⅳ龄幼虫中诱导了较快的生长发育速率;雌性成虫InR、Pdk、Akt、FoxO的表达显著高于雄性,这可能与胰岛素信号参与雌性生殖系统发育有关。

关键词: 胰岛素信号通路, 转录组学, 序列分析, qRT-PCR

Abstract: To explore the endocrine disrupting gene markers of invertebrates based on the insulin signal pathway, this study selected Chironomus tentans as the model organism, identified the key genes in the insulin signal pathway on the basis of transcriptome sequencing, and carried out the full-length amplification of cDNA and the multi-species comparison of amino acid sequences for the upstream genes such as InR, Pdk, Akt and FoxO. Based on the cDNA full-length sequence, the InR, Pdk, Akt and FoxO genes of Chironomus tentans can encode protein sequences containing 1 436, 522, 520 and 470 amino acids, respectively. The sequence homology comparison and phylogenetic tree analysis results verified its evolutionary conservatism. The qRT-PCR results of the relative expression of genes in Ⅰ-Ⅳ instar larvae, pupae, female adults and male adults show that the expression trend of InR, Pdk, Akt and FoxO in C. tentans was similar. The peaks of gene expression in I and IV instar indicate that insulin signaling induces faster growth and development rates in I and IV instar larvae. The expression of InR, Pdk, Akt, and FoxO in female adults was significantly higher than that in males, which may be related to the involvement of insulin signaling in the development of the female reproductive system.

Key words: insulin signaling pathway, transcriptomics, sequential analysis, qRT-PCR

中图分类号:

X171.5

张成林, 古文, 汪贞, 石利利, 王蕾, 杨家新. 伸展摇蚊(Chironomus tentans)转录组测序及胰岛素信号通路关键基因序列分析[J]. 生态与农村环境学报, 2023, 39(12): 1646-1656.

ZHANG Cheng-lin, GU Wen, WANG Zhen, SHI Li-li, WANG Lei, YANG Jia-xin. Transcriptome Sequencing and Sequence Analysis of Key Genes in the Insulin Signaling Pathway of Chironomus tentans[J]. Journal of Ecology and Rural Environment, 2023, 39(12): 1646-1656.

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