Interaction Between Allopregnanolone and Amiloride Binding Sites on the GABA_A Receptor.

IF 1.8 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Cell Biochemistry and Biophysics Pub Date : 2024-12-28 DOI:10.1007/s12013-024-01654-6

Julia V Bukanova, Rodion V Kondratenko, Elena I Solntseva

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引用次数: 0

Abstract

Allopregnanolone (Allo) is a positive allosteric modulator of the GABA_A receptor, and amiloride (Ami) is a competitive antagonist of the GABA_A receptor. The purpose of this work was to study the combined effect of Allo and Ami on functional activity of GABA_A receptor. The GABA-induced chloride current (I_GABA) was measured in isolated Purkinje cells of rat cerebellum using the patch-clamp technique and a system of fast application. Our results indicate that Allo suppresses the inhibitory effect of Ami on I_GABA, the IC₅₀ value of Ami concentration-response curve was increased from 164 to 547 µM (P < 0.001) in the presence of Allo. Next, GABA concentration-response curves (EC₅₀ = 5.8 µM) were constructed in the presence of Allo (EC₅₀ = 1.2 µM), Ami (EC₅₀ = 25.5 µM), and the combination of Allo+Ami (EC₅₀ = 3.2 µM). Changes in EC₅₀ values as a percentage relative to the control were calculated. The blocking effect of Ami is reduced in the presence of Allo (340% vs 150%, P < 0.01) and the potentiating effect of Allo does not change in the presence of Ami (78% vs 87%, P > 0.05). The results suggest that there is an allosteric relationship between the Allo and Ami binding sites on GABA_A receptor that operates in one direction, from Allo sites to Ami site, but not vice versa.

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GABAA受体上异孕酮与阿米洛利结合位点的相互作用。

异孕酮（Allo）是GABAA受体的正变构调节剂，阿米洛利（Ami）是GABAA受体的竞争性拮抗剂。本研究旨在研究Allo和Ami对GABAA受体功能活性的联合影响。采用膜片钳技术和快速应用系统对大鼠小脑浦肯野细胞gaba诱导的氯化电流（IGABA）进行了测定。结果表明，Allo抑制了Ami对IGABA的抑制作用，在Allo （EC50 = 1.2µM）、Ami （EC50 = 25.5µM）和Allo+Ami （EC50 = 3.2µM）存在的情况下，Ami浓度响应曲线的IC50值从164µM增加到547µM （p50 = 5.8µM）。计算EC50值相对于对照组的百分比变化。Allo存在时Ami的阻断作用降低（340% vs 150%, P < 0.05）。结果表明，GABAA受体上的Allo和Ami结合位点之间存在一种单向的变构关系，即从Allo位点到Ami位点，而不是相反。

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

Cell Biochemistry and Biophysics 生物-生化与分子生物学

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

7.5 months

期刊介绍： Cell Biochemistry and Biophysics (CBB) aims to publish papers on the nature of the biochemical and biophysical mechanisms underlying the structure, control and function of cellular systems The reports should be within the framework of modern biochemistry and chemistry, biophysics and cell physiology, physics and engineering, molecular and structural biology. The relationship between molecular structure and function under investigation is emphasized. Examples of subject areas that CBB publishes are: · biochemical and biophysical aspects of cell structure and function; · interactions of cells and their molecular/macromolecular constituents; · innovative developments in genetic and biomolecular engineering; · computer-based analysis of tissues, cells, cell networks, organelles, and molecular/macromolecular assemblies; · photometric, spectroscopic, microscopic, mechanical, and electrical methodologies/techniques in analytical cytology, cytometry and innovative instrument design For articles that focus on computational aspects, authors should be clear about which docking and molecular dynamics algorithms or software packages are being used as well as details on the system parameterization, simulations conditions etc. In addition, docking calculations (virtual screening, QSAR, etc.) should be validated either by experimental studies or one or more reliable theoretical cross-validation methods.