Novel tools for studying the fish growth hormone axis in vivo.

IF 3.1 2区 医学 Q1 ENDOCRINOLOGY & METABOLISM
Genevieve L Fernandes, Deodatta S Gajbhiye, Yaara Y Columbus-Shenkar, Matan Golan
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引用次数: 0

Abstract

Growth hormone (Gh) is the main regulator of fish growth, making it vital to understand how Gh is controlled. However, studying Gh regulation has been limited by the lack of suitable fish models. We identified conserved regulatory elements in tilapia gh and cloned a functional promoter. By using this promoter, we generated reporter transgenic fish lines of zebrafish and tilapia with labeled somatotrophs. Our in vitro and in vivo investigation revealed that the first intron of the tilapia gh contains functional enhancer elements crucial for transgene expression. Tilapia transgenic lines expressing the calcium-sensitive protein GCaMP6s allow the visualization of calcium activity in somatotrophs within intact pituitary glands. Network plots derived from the cell coactivation revealed distinct patterns of activity, showing cell groups with synchronized behavior, suggesting the presence of functional clusters within the somatotroph population. Furthermore, our zebrafish reporter lines revealed higher spontaneous calcium activity of Gh cells in juveniles compared with adults, suggesting increased somatotroph activity in rapidly growing young fish. Overall, our study demonstrates the functionality of tilapia and zebrafish Gh transgenic lines for investigating Gh cell activity in vivo. Combined with previously established methodologies, these transgenic lines serve as valuable tools for studying the regulation and release of Gh in the largest group of living vertebrates.NEW & NOTEWORTHY This study describes the first transgenic reporter tools for real-time imaging of calcium activity and reporter gene expression in fish growth hormone cells. By identifying critical enhancer elements in gh, we successfully targeted somatotroph cells and performed functional analysis of their behavior within the physiological context. These tools could provide valuable insights into growth hormone (Gh) regulation, with important implications in aquaculture and our understanding of the evolution of vertebrate endocrine systems.

研究鱼类体内生长激素轴的新工具。
生长激素(Gh)是鱼类生长的主要调节剂,因此了解Gh是如何控制的至关重要。然而,由于缺乏合适的鱼类模型,对Gh调控的研究受到了限制。我们鉴定了罗非鱼基因中保守的调控元件,并克隆了一个功能性启动子。利用该启动子,我们获得了带有生长营养标记的斑马鱼和罗非鱼的报告基因转基因鱼系。体外和体内研究表明,罗非鱼的第一个内含子含有对-转基因表达至关重要的功能增强元件。表达钙敏感蛋白GCaMP6s的罗非鱼转基因品系可以可视化显示完整垂体内生长因子中的钙活性。来源于细胞共激活的网络图揭示了不同的活动模式,显示细胞群具有同步的行为,表明在生长缺陷群体中存在功能集群。此外,我们的斑马鱼报告细胞系显示,与成年鱼相比,幼鱼Gh细胞的自发钙活性更高,这表明快速生长的幼鱼的生长营养活性增加。总的来说,我们的研究证明了罗非鱼和斑马鱼Gh转基因系在体内研究Gh细胞活性的功能。结合先前建立的方法,这些转基因系为研究最大的活脊椎动物群体中Gh的调节和释放提供了有价值的工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
9.80
自引率
0.00%
发文量
98
审稿时长
1 months
期刊介绍: The American Journal of Physiology-Endocrinology and Metabolism publishes original, mechanistic studies on the physiology of endocrine and metabolic systems. Physiological, cellular, and molecular studies in whole animals or humans will be considered. Specific themes include, but are not limited to, mechanisms of hormone and growth factor action; hormonal and nutritional regulation of metabolism, inflammation, microbiome and energy balance; integrative organ cross talk; paracrine and autocrine control of endocrine cells; function and activation of hormone receptors; endocrine or metabolic control of channels, transporters, and membrane function; temporal analysis of hormone secretion and metabolism; and mathematical/kinetic modeling of metabolism. Novel molecular, immunological, or biophysical studies of hormone action are also welcome.
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