Abstract
The incretin hormone, glucagon-like peptide-1 (GLP-1) is released from intestinal L-cells following food ingestion. Its secretion is triggered by a range of nutrients, including fats, carbohydrates and proteins. We reported previously that Na(+)-dependent glutamine uptake triggered electrical activity and GLP-1 release from the L-cell model line GLUTag. However, whereas alanine also triggered membrane depolarization and GLP-1 secretion, the response was Na+ independent. A range of alanine analogues, including d-alanine, beta-alanine, glycine and l-serine, but not d-serine, triggered similar depolarizing currents and elevation of intracellular [Ca2+], a sensitivity profile suggesting the involvement of glycine receptors. In support of this idea, glycine-induced currents and GLP-1 release were blocked by strychnine, and currents showed a 58.5 mV shift in reversal potential per 10-fold change in [Cl-], consistent with the activation of a Cl(-)-selective current. GABA, an agonist of related Cl- channels, also triggered Cl- currents and secretion, which were sensitive to picrotoxin. GABA-triggered [Ca2+]i increments were abolished by bicuculline and partially impaired by (1,2,5,6-tetrahydropyridine-4-yl)methylphosphinic acid (TPMPA), suggesting the involvement of both GABA(A) and GABA(C) receptors. Expression of GABA(A), GABA(C) and glycine receptor subunits was confirmed by RT-PCR. Glycine-triggered GLP-1 secretion was impaired by bumetanide but not bendrofluazide, suggesting that a high intracellular [Cl-] maintained by Na(+)-K(+)-2Cl- cotransporters is necessary for the depolarizing response to glycine receptor ligands. Our results suggest that GABA and glycine stimulate electrical activity and GLP-1 release from GLUTag cells by ligand-gated ion channel activation, a mechanism that might be important in responses to endogenous ligands from the enteric nervous system or dietary sources.
MeSH Terms
Action Potentials/drug effects
Animals
Calcium/metabolism
Cell Line, Tumor
Chlorides/metabolism
Dose-Response Relationship, Drug
GABA Antagonists/pharmacology
Glucagon-Like Peptide 1/metabolism
Glycine/pharmacology
Ion Channel Gating/drug effects
Mice
Neurotransmitter Agents/pharmacology
RNA, Messenger/metabolism
Receptors, GABA/drug effects,genetics,metabolism
Receptors, GABA-A/drug effects,genetics,metabolism
Receptors, Glycine/drug effects,genetics,metabolism
Sodium Potassium Chloride Symporter Inhibitors/pharmacology
Sodium-Potassium-Chloride Symporters/drug effects,metabolism
Strychnine/pharmacology
gamma-Aminobutyric Acid/pharmacology
Chemicals
Chlorides
GABA Antagonists
GABA-C receptor
Neurotransmitter Agents
RNA, Messenger
Receptors, GABA
Receptors, GABA-A
Receptors, Glycine
Sodium Potassium Chloride Symporter Inhibitors
Sodium-Potassium-Chloride Symporters
gamma-Aminobutyric Acid
Glucagon-Like Peptide 1
Strychnine
Calcium
Glycine
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Gameiro A
Wellcome Trust/MRC Building, Addenbrooke's Hospital, Hills Road, Cambridge CB2 2XY, UK.
Reimann F
Habib A M
O'Malley D
Williams L
Simpson A K
Gribble F M
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