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转录组分析揭示了 BmKrüppel 同源物 1 在家蚕卵母细胞发育中的调控网络。

Transcriptomic analysis reveals the regulation network of BmKrüppel homolog 1 in the oocyte development of Bombyx mori.

机构信息

Guangdong Key Laboratory of Insect Developmental Biology and Applied Technology, Guangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Science and Technology & School of Life Sciences, South China Normal University, Guangzhou, China.

出版信息

Insect Sci. 2021 Feb;28(1):47-62. doi: 10.1111/1744-7917.12747. Epub 2020 Apr 13.

Abstract

Krüppel homolog 1 (Kr-h1), a zinc finger transcription factor, is involved in the metamorphosis and adult reproduction of insects. However, the role of Kr-h1 in reproduction of holometabolic insects remains to be elucidated. The regulation network of Kr-h1-associated genes in the reproduction in Bombyx mori was investigated in this study. The higher expression level of BmKr-h1 in the ovaries was detected during the late pupal stage and adults. RNA interference (RNAi)-mediated depletion of BmKr-h1 in the female at day 6 of pupae resulted in abnormal oocytes at 48 h post-double-stranded RNA treatment, which showed less yolk protein deposition and partially transparent chorion. RNA-seq and subsequent differentially expressed transcripts analysis showed that knockdown of BmKr-h1 caused a decrease in the expression of 2882 genes and an increase in the expression of 2565 genes in the oocytes at day 8 of pupae. Totally, 27 genes coding for transcription factors were down-regulated, while six genes coding for other transcription factors were up-regulated. BmKr-h1 bound to the Kr-h1 binding site of the transcription factors AP-1 (activating protein-1) and FOXG1 to increase their messenger RNA transcripts in the BmN cells, respectively. Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses of that positively co-expressed with AP-1 and FOXG1 transcripts showed mainly enrichment in the metabolic-related pathways, the nutrient absorption and the yolk protein absorption processes. These data suggested that BmKr-h1 might directly regulate the metabolic-related pathways, the nutrient absorption and the yolk protein absorption processes or probably through AP-1 and /or FOXG1 to regulate oocyte development.

摘要

Krüppel 同源物 1(Kr-h1)是一种锌指转录因子,参与昆虫的变态和成虫繁殖。然而,Kr-h1 在完全变态昆虫繁殖中的作用仍有待阐明。本研究调查了 BmKr-h1 相关基因在桑蚕生殖中的调控网络。在蛹后期和成虫期检测到 BmKr-h1 在卵巢中的表达水平较高。在第 6 天的蛹期用 RNA 干扰(RNAi)介导的 BmKr-h1 耗尽会导致 48 小时后双链 RNA 处理的异常卵母细胞,表现为卵黄蛋白沉积减少和部分透明卵壳。RNA-seq 及随后的差异表达转录本分析显示,BmKr-h1 的敲低导致蛹期第 8 天的卵母细胞中 2882 个基因的表达减少和 2565 个基因的表达增加。总共 27 个编码转录因子的基因下调,而 6 个编码其他转录因子的基因上调。BmKr-h1 与转录因子 AP-1(激活蛋白-1)和 FOXG1 的 Kr-h1 结合位点结合,分别增加了它们在 BmN 细胞中的信使 RNA 转录物。与 AP-1 和 FOXG1 转录物呈正共表达的基因本体富集和京都基因与基因组百科全书途径分析显示,主要富集在代谢相关途径、营养吸收和卵黄蛋白吸收过程中。这些数据表明,BmKr-h1 可能直接调节代谢相关途径、营养吸收和卵黄蛋白吸收过程,或可能通过 AP-1 和/或 FOXG1 来调节卵母细胞发育。

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