title: Evidence that Ca2+ within the microdomain of the L-type voltage gated Ca2+ channel activates ERK in MIN6 cells in response to glucagon-like peptide-1.
abstract: Glucagon like peptide-1 (GLP-1) is released from intestinal L-cells in response to nutrient ingestion and acts upon pancreatic β-cells potentiating glucose-stimulated insulin secretion and stimulating β-cell proliferation, differentiation, survival and gene transcription. These effects are mediated through the activation of multiple signal transduction pathways including the extracellular regulated kinase (ERK) pathway. We have previously reported that GLP-1 activates ERK through a mechanism dependent upon the influx of extracellular Ca(2+) through L-type voltage gated Ca(2+) channels (VGCC). However, the mechanism by which L-type VGCCs couple to the ERK signalling pathway in pancreatic β-cells is poorly understood. In this report, we characterise the relationship between L-type VGCC mediated changes in intracellular Ca(2+) concentration ([Ca(2+)](i)) and the activation of ERK, and demonstrate that the sustained activation of ERK (up to 30 min) in response to GLP-1 requires the continual activation of the L-type VGCC yet does not require a sustained increase in global [Ca(2+)](i) or Ca(2+) efflux from the endoplasmic reticulum. Moreover, sustained elevation of [Ca(2+)](i) induced by ionomycin is insufficient to stimulate the prolonged activation of ERK. Using the cell permeant Ca(2+) chelators, EGTA-AM and BAPTA-AM, to determine the spatial dynamics of L-type VGCC-dependent Ca(2+) signalling to ERK, we provide evidence that a sustained increase in Ca(2+) within the microdomain of the L-type VGCC is sufficient for signalling to ERK and that this plays an important role in GLP-1- stimulated ERK activation.
authors.affiliation: Department of Cell Physiology and Pharmacology, Henry Wellcome Building, University of Leicester, Leicester, United Kingdom.
authors.forename: Joanne
authors.initials: J
authors.lastname: Selway
authors.forename: Roberto
authors.initials: R
authors.lastname: Rigatti
authors.forename: Nina
authors.initials: N
authors.lastname: Storey
authors.forename: Jing
authors.initials: J
authors.lastname: Lu
authors.forename: Gary B
authors.initials: GB
authors.lastname: Willars
authors.forename: Terence P
authors.initials: TP
authors.lastname: Herbert
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020746:Calcium Channels, L-Type / Q000378:metabolism*; D002460:Cell Line; D004789:Enzyme Activation / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D052216:Glucagon-Like Peptide 1 / Q000378:metabolism* / Q000494:pharmacology; D051379:Mice; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D020746:Calcium Channels, L-Type; D052216:Glucagon-Like Peptide 1; D048049:Extracellular Signal-Regulated MAP Kinases; D002118:Calcium