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Antisecretory Factor Modulates GABAA Receptor Activity in Neurons

Journal article
Authors V. Bazzurro
E. Gatta
A. Cupello
Stefan Lange
M. Robello
Published in Journal of Molecular Neuroscience
Volume 64
Issue 2
Pages 312-320
ISSN 0895-8696
Publication year 2018
Published at Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
Pages 312-320
Language en
Links https://doi.org/10.1007/s12031-017-...
Keywords Antisecretory factor, Central nervous system, Cerebellar granule cells, Enteric nervous system, GABAA receptors, Patch clamp, 4 aminobutyric acid A receptor, amino acid, antisecretory factor 16 peptide, neurotransmitter, unclassified drug, amino acid sequence, animal cell, animal cell culture, Article, cerebellum granular layer, chloride current, electrophysiological procedures, enteropathy, immunocytochemistry, immunofluorescence, intestinal hypersecretion, nerve cell membrane, nonhuman, rat, steady state, whole cell patch clamp
Subject categories Cell Biology

Abstract

The antisecretory factor is an endogenous protein found in all mammalian tissues investigated so far. It acts by counteracting intestinal hypersecretion and various forms of inflammation, but the detailed mechanism of antisecretory factor (AF) action is unknown. We tested neuronal GABAA receptors by means of AF-16, a potent AF peptide derived from amino acids 36–51 from the NH2 part of AF. Cultured rat cerebellar granule cells were used, and the effects on the GABA-mediated chloride currents were determined by whole-cell patch clamp. Both the neurotransmitter GABA and AF-16 were added by perfusion of the experimental system. A 3-min AF-16 preincubation was more efficacious than 30 s in significantly elevating the rapidly desensitizing GABA-activated chloride current. No effect was found on the tonic, slowly desensitizing current. The GABA-activated current increase by AF-16 demonstrated a low k of 41 pM with a maximal increase of 37% persisting for some minutes after AF washout, independent from GABA concentration. This indicates an effect on the maximal stimulation (E%Max) excluding an altered affinity between GABA and its receptor. An immunocytochemical fluorescence approach with anti γ2 subunit antibodies demonstrated an increased expression of GABAA receptors. Thus, both the electrophysiological and the immunofluorescence approach indicate an increased appearance of GABAA receptors on the neuronal membrane. The rationale of the experiments was to test the effect of AF on a defined neuronal population of GABAA receptors. The implications of the results on the impact of AF on the enteric nervous system or on brain function are discussed. © 2018, Springer Science+Business Media, LLC, part of Springer Nature.

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