Department of Ecology and Evolutionary Biology, University of Toronto, Toronto, ON M5S 3B2, Canada.
Centre for Neural Circuits and Behaviour, Oxford University, Oxford OX1 3SR, UK.
Int J Mol Sci. 2023 Jun 16;24(12):10219. doi: 10.3390/ijms241210219.
The () gene of encodes a cGMP-dependent protein kinase (PKG), which is a major effector of the cGMP signaling pathway involved in the regulation of behaviour and metabolic traits. Despite being well studied at the transcript level, little is known about the gene at the protein level. Here, we provide a detailed characterization of the gene protein (FOR) products and present new tools for their study, including five isoform-specific antibodies and a transgenic strain that carries an HA-labelled allele (). Our results showed that multiple FOR isoforms were expressed in the larval and adult stages of and that the majority of whole-body FOR expression arises from three (P1, P1α, and P3) of eight putative protein isoforms. We found that FOR expression differed between the larval and adult stages and between the dissected larval organs we analyzed, which included the central nervous system (CNS), fat body, carcass, and intestine. Moreover, we showed that the FOR expression differed between two allelic variants of the gene, namely, (sitter) and (rover), that are known to differ in many food-related traits. Together, our in vivo identification of FOR isoforms and the existence of temporal, spatial, and genetic differences in their expression lay the groundwork for determining their functional significance.
() 基因编码一种 cGMP 依赖性蛋白激酶 (PKG)，它是参与调节行为和代谢特征的 cGMP 信号通路的主要效应物。尽管在转录水平上研究得很好，但在蛋白质水平上对 基因知之甚少。在这里，我们详细描述了 基因蛋白 (FOR) 产物，并提供了新的研究工具，包括五种同工型特异性抗体和一种携带 HA 标记的 等位基因 () 的转基因品系。我们的结果表明，多个 FOR 同工型在 和 的幼虫和成虫阶段表达，并且全身 FOR 表达的大部分来自八个假定蛋白同工型中的三个 (P1、P1α 和 P3)。我们发现，FOR 表达在幼虫和成虫阶段以及我们分析的分离幼虫器官（包括中枢神经系统 (CNS)、脂肪体、尸体和肠道）之间存在差异。此外，我们表明， 基因的两个等位基因变体 ()（坐着的）和 （闲逛的）之间的 FOR 表达存在差异，已知它们在许多与食物相关的特征上存在差异。总之，我们在体内鉴定了 FOR 同工型，以及它们表达的时间、空间和遗传差异，为确定它们的功能意义奠定了基础。
