Department of Biological Sciences, University of Wisconsin, Milwaukee, Wisconsin 53211, USA.
Dev Neurobiol. 2010 Nov;70(13):884-96. doi: 10.1002/dneu.20830.
A precise organization of gene expression is required for developing neural and muscular systems. Steroid hormones can control the expression of genes that are critical for development. In this study we test the hypothesis that the steroid hormone ecdysone regulates gene expression of the voltage-gated calcium-activated potassium ion channel, Slowpoke or KCNMA1. Late in adult development of the tobacco hawkmoth Manduca sexta, slowpoke (msslo) levels increased contributing to the maturation of the dorsal longitudinal flight muscles (DLMs) and CNS. We show that critical components of ecdysteroid gene regulation were present during upreglation of msslo in late adult DLM and CNS development. Ecdysteroid receptor complex heterodimeric partner proteins, the ecdysteroid receptor (EcR) and ultraspiracle (USP), and the ecdysone-induced early gene, msE75B, were expressed at key developmental time points, suggesting that ecdysteroids direct aspects of gene expression in the DLMs during these late developmental stages. We provide evidence that ecdysteroids suppress msslo transcription in the DLMs; when titers decline msslo transcript levels increase. These results are consistent with msslo being a downstream gene in an ecdysteroid-mediated gene cascade during DLM development. We also show that the ecdysteroids regulate msslo transcript levels in the developing CNS. These results will contribute to our understanding of how the spatiotemporal regulation of slowpoke transcription contributes to tailoring cell excitability to the differing physiological and behavioral demands during development.
精确的基因表达调控对于神经系统和肌肉系统的发育至关重要。类固醇激素可以控制对发育至关重要的基因的表达。在这项研究中,我们检验了这样一个假设,即类固醇激素蜕皮激素调节电压门控钙激活钾离子通道Slowpoke 或 KCNMA1 的基因表达。在烟草天蛾 Manduca sexta 的成虫后期发育过程中,Slowpoke(msslo)的水平增加,有助于背纵飞行肌(DLM)和中枢神经系统的成熟。我们表明,在成年后期 DLM 和中枢神经系统发育过程中 msslo 的上调过程中,存在蜕皮激素基因调控的关键组成部分。蜕皮激素受体复合物异二聚体伴侣蛋白,蜕皮激素受体(EcR)和超螺内酯(USP),以及蜕皮激素诱导的早期基因 msE75B,在关键的发育时间点表达,表明蜕皮激素在这些晚期发育阶段指导 DLM 中的基因表达。我们提供的证据表明,蜕皮激素在 DLM 中抑制 msslo 的转录;当浓度下降时,msslo 的转录水平增加。这些结果与 msslo 是 DLM 发育过程中蜕皮激素介导的基因级联中的下游基因一致。我们还表明,蜕皮激素调节发育中的中枢神经系统中的 msslo 转录水平。这些结果将有助于我们理解 Slowpoke 转录的时空调控如何使细胞兴奋性适应发育过程中不同的生理和行为需求。