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通过双光子光解笼锁研究 GABA 受体 α 亚基亚型在大鼠小脑颗粒细胞抗分泌因子活性中的作用。

Involvement of GABA receptors containing α subtypes in antisecretory factor activity on rat cerebellar granule cells studied by two-photon uncaging.

机构信息

DIFILAB, Department of Physics, University of Genoa, Genoa, Italy.

Nanoscopy, CHT Erzelli, Istituto Italiano di Tecnologia, Genoa, Italy.

出版信息

Eur J Neurosci. 2022 Sep;56(5):4505-4513. doi: 10.1111/ejn.15775. Epub 2022 Jul 20.

Abstract

The antisecretory factor (AF) is an endogenous protein that counteracts intestinal hypersecretion and various inflammation conditions in vivo. It has been detected in many mammalian tissues and plasma, but its mechanisms of action are largely unknown. To study the pharmacological action of the AF on different GABA receptor populations in cerebellar granule cells, we took advantage of the two-photon uncaging method as this technique allows to stimulate the cell locally in well-identified plasma membrane parts. We compared the electrophysiological response evoked by releasing a caged GABA compound on the soma, the axon initial segment and neurites before and after administering AF-16, a 16 amino acids long peptide obtained from the amino-terminal end of the AF protein. After the treatment with AF-16, we observed peak current increases of varying magnitude depending on the neuronal region. Thus, studying the effects of furosemide and AF-16 on the electrophysiological behaviour of cerebellar granules, we suggest that GABA receptors, containing the α subunit, may be specifically involved in the increase of the peak current by AF, and different receptor subtype distribution may be responsible for differences in this increase on the cell.

摘要

抗分泌因子(AF)是一种内源性蛋白质,可在体内对抗肠道过度分泌和各种炎症状态。它已在许多哺乳动物组织和血浆中被检测到,但它的作用机制在很大程度上尚不清楚。为了研究 AF 对小脑颗粒细胞中不同 GABA 受体群体的药理学作用,我们利用双光子光解方法,因为这种技术允许在细胞局部在明确定义的质膜部分进行刺激。我们比较了在给予 AF-16(一种从 AF 蛋白氨基末端获得的 16 个氨基酸长的肽)前后,在体、轴突起始段和神经突上释放被笼闭的 GABA 化合物引起的电生理反应。在用 AF-16 处理后,我们观察到取决于神经元区域的峰电流的不同幅度的增加。因此,通过研究速尿和 AF-16 对小脑颗粒的电生理行为的影响,我们提出包含α亚基的 GABA 受体可能特异性地参与 AF 引起的峰电流增加,并且不同的受体亚型分布可能导致细胞中这种增加的差异。

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