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通过逆转录-定量聚合酶链反应对斑马鱼(Danio rerio Hamilton)中枢神经系统中γ-氨基丁酸信号系统的表征。

Characterization of the γ-aminobutyric acid signaling system in the zebrafish (Danio rerio Hamilton) central nervous system by reverse transcription-quantitative polymerase chain reaction.

作者信息

Cocco Arianna, Rönnberg A M Carolina, Jin Zhe, André Gonçalo Igreja, Vossen Laura E, Bhandage Amol K, Thörnqvist Per-Ove, Birnir Bryndis, Winberg Svante

机构信息

Department of Neuroscience, Biomedical Centre, Uppsala University, P.O. Box 593, 751 24 Uppsala, Sweden.

出版信息

Neuroscience. 2017 Feb 20;343:300-321. doi: 10.1016/j.neuroscience.2016.07.018. Epub 2016 Jul 21.

DOI:10.1016/j.neuroscience.2016.07.018
PMID:27453477
Abstract

In the vertebrate brain, inhibition is largely mediated by γ-aminobutyric acid (GABA). This neurotransmitter comprises a signaling machinery of GABA, GABA receptors, transporters, glutamate decarboxylases (gads) and 4-aminobutyrate aminotransferase (abat), and associated proteins. Chloride is intimately related to GABA receptor conductance, GABA uptake, and GADs activity. The response of target neurons to GABA stimuli is shaped by chloride-cation co-transporters (CCCs), which strictly control Cl gradient across plasma membranes. This research profiled the expression of forty genes involved in GABA signaling in the zebrafish (Danio rerio) brain, grouped brain regions and retinas. Primer pairs were developed for reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The mRNA levels of the zebrafish GABA system share similarities with that of mammals, and confirm previous studies in non-mammalian species. Proposed GABA receptors are αβγ, αβδ, αβγ, αβδ, αβγ, αβδ, αβγ and αβδ. Regional brain differences were documented. Retinal hetero- or homomeric ρ-composed GABA receptors could exist, accompanying αβγ, αβδ, αβγ, αβδ. Expression patterns of α and α were opposite, with the former being more abundant in retinas, the latter in brains. Given the stoichiometry αβγ, α- or α-containing receptors likely have different regulatory mechanisms. Different gene isoforms could originate after the rounds of genome duplication during teleost evolution. This research depicts that one isoform is generally more abundantly expressed than the other. Such observations also apply to GABA receptors, GABA transporters, GABA-related enzymes, CCCs and GABA receptor-associated proteins, whose presence further strengthens the proof of a GABA system in zebrafish.

摘要

在脊椎动物大脑中,抑制作用主要由γ-氨基丁酸(GABA)介导。这种神经递质包括GABA、GABA受体、转运体、谷氨酸脱羧酶(GADs)和4-氨基丁酸转氨酶(ABAT)以及相关蛋白的信号传导机制。氯离子与GABA受体电导、GABA摄取和GADs活性密切相关。靶神经元对GABA刺激的反应由氯离子-阳离子共转运体(CCC)塑造,其严格控制跨质膜的氯离子梯度。本研究分析了斑马鱼(Danio rerio)大脑、分组脑区和视网膜中参与GABA信号传导的40个基因的表达情况。开发了用于逆转录定量聚合酶链反应(RT-qPCR)的引物对。斑马鱼GABA系统的mRNA水平与哺乳动物相似,并证实了先前在非哺乳动物物种中的研究。推测的GABA受体为αβγ、αβδ、αβγ、αβδ、αβγ、αβδ、αβγ和αβδ。记录了脑区差异。视网膜中可能存在由αβγ、αβδ、αβγ、αβδ伴随的异聚或同聚ρ组成的GABA受体。α和α的表达模式相反,前者在视网膜中更丰富,后者在大脑中更丰富。鉴于化学计量比为αβγ,含α或α的受体可能具有不同的调节机制。不同的基因亚型可能源于硬骨鱼进化过程中的基因组复制轮次。本研究表明,一种亚型通常比另一种更丰富地表达。这些观察结果也适用于GABA受体、GABA转运体、GABA相关酶、CCC和GABA受体相关蛋白,它们的存在进一步加强了斑马鱼中存在GABA系统的证据。

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