Control of Insulin Release by Transient Receptor Potential Melastatin 3 (TRPM3) Ion Channels.

BACKGROUND/AIMS The release of insulin in response to increased levels of glucose in the blood strongly depends on Ca2+ influx into pancreatic beta cells by the opening of voltage-gated Ca2+ channels. Transient Receptor Potential Melastatin 3 proteins build Ca2+ permeable, non-selective cation channels serving as pain sensors of noxious heat in the peripheral nervous system. TRPM3 channels are also strongly expressed in pancreatic beta cells that respond to the TRPM3 agonist pregnenolone sulfate with Ca2+ influx and increased insulin release. Therefore, we hypothesized that in beta cells TRPM3 channels may contribute to pregnenolone sulfate- as well as to glucose-induced insulin release. METHODS We used INS-1 cells as a beta cell model in which we analysed the occurrence of TRPM3 isoformes by immunoprecipitation and western blotting and by cloning of RT-PCR amplified cDNA fragments. We applied pharmacological as well as CRISPR/Cas9-based strategies to analyse the interplay of TRPM3 and voltage-gated Ca2+ channels in imaging experiments (FMP, Fura-2) and electrophysiological recordings. In immunoassays, we examined the contribution of TRPM3 channels to pregnenolone sulfate- and glucose-induced insulin release. To confirm our findings, we generated beta cell-specific Trpm3-deficient mice and compared their glucose clearance with the wild type in glucose tolerance tests. RESULTS TRPM3 channels triggered the activity of voltage-gated Ca2+ channels and both channels together contributed to insulin release after TRPM3 activation. Trpm3-deficient INS-1 cells lacked pregnenolone sulfate-induced Ca2+ signals just like the pregnenolone sulfate-induced insulin release. Both, glucose-induced Ca2+ signals and the glucose-induced insulin release were strongly reduced. Accordingly, Trpm3-deficient mice displayed an impaired decrease of the blood sugar concentration after intraperitoneal or oral administration of glucose. CONCLUSION The present study suggests an important role for TRPM3 channels in the control of glucose-dependent insulin release.

[1]  Xiaotian Liu,et al.  The effect of progesterone and pregnenolone on diabetes status in Chinese rural population: a dose-response analysis from Henan Rural Cohort. , 2019, European journal of endocrinology.

[2]  D. Andersson,et al.  Promiscuous G-Protein-Coupled Receptor Inhibition of Transient Receptor Potential Melastatin 3 Ion Channels by Gβγ Subunits , 2019, The Journal of Neuroscience.

[3]  K. Suhre,et al.  Metabolic and proteomic signatures of hypoglycaemia in type 2 diabetes , 2019, Diabetes, obesity & metabolism.

[4]  T. Voet,et al.  A TRP channel trio mediates acute noxious heat sensing , 2018, Nature.

[5]  Manuela Schmidt,et al.  Anti-nociceptive action of peripheral mu-opioid receptors by G-beta-gamma protein-mediated inhibition of TRPM3 channels , 2017, eLife.

[6]  Tamara Luti Rosenbaum Emir.  The Role of TRP Channels in the Pancreatic Beta-Cell , 2017 .

[7]  F. Marabita,et al.  Expression of Transient Receptor Potential Channels in the Purified Human Pancreatic &bgr;-Cells , 2017, Pancreas.

[8]  S. Barg,et al.  Plasma Membrane Phosphatidylinositol 4,5-Bisphosphate Regulates Ca(2+)-Influx and Insulin Secretion from Pancreatic β Cells. , 2016, Cell chemical biology.

[9]  T. Voets,et al.  Regulation of the transient receptor potential channel TRPM3 by phosphoinositides , 2015, The Journal of general physiology.

[10]  T. Rohacs,et al.  Transient receptor potential melastatin 3 is a phosphoinositide-dependent ion channel , 2015, The Journal of general physiology.

[11]  A. Marchand,et al.  Activation of TRPM3 by a potent synthetic ligand reveals a role in peptide release , 2015, Proceedings of the National Academy of Sciences.

[12]  Feng Zhang,et al.  Genome engineering using CRISPR-Cas9 system. , 2015, Methods in molecular biology.

[13]  M. Boutros,et al.  E-CRISP: fast CRISPR target site identification , 2014, Nature Methods.

[14]  Jin-Soo Kim,et al.  Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleases , 2014, Bioinform..

[15]  M. Schaefer,et al.  Flavanones That Selectively Inhibit TRPM3 Attenuate Thermal Nociception In Vivo , 2013, Molecular Pharmacology.

[16]  S. Philipp,et al.  Citrus fruit and fabacea secondary metabolites potently and selectively block TRPM3 , 2013, British journal of pharmacology.

[17]  A. Beck,et al.  Membrane Potential Measurements of Isolated Neurons Using a Voltage-Sensitive Dye , 2013, PloS one.

[18]  Lenka Gryčová,et al.  PtdIns(4,5)P2 interacts with CaM binding domains on TRPM3 N-terminus , 2012, Channels.

[19]  Stefanie Mannebach,et al.  Alternative Splicing of a Protein Domain Indispensable for Function of Transient Receptor Potential Melastatin 3 (TRPM3) Ion Channels* , 2012, The Journal of Biological Chemistry.

[20]  Bernd Nilius,et al.  TRPM3 Is a Nociceptor Channel Involved in the Detection of Noxious Heat , 2011, Neuron.

[21]  C. Harteneck,et al.  Fenamates as TRP channel blockers: mefenamic acid selectively blocks TRPM3 , 2011, British journal of pharmacology.

[22]  M. Tominaga,et al.  Lack of TRPM2 Impaired Insulin Secretion and Glucose Metabolisms in Mice , 2010, Diabetes.

[23]  David J Beech,et al.  Pregnenolone Sulphate- and Cholesterol-Regulated TRPM3 Channels Coupled to Vascular Smooth Muscle Secretion and Contraction , 2010, Circulation research.

[24]  S. Philipp,et al.  TRPM3 channels provide a regulated influx pathway for zinc in pancreatic beta cells , 2010, Pflügers Archiv - European Journal of Physiology.

[25]  K. Lemaire,et al.  Loss of high-frequency glucose-induced Ca2+ oscillations in pancreatic islets correlates with impaired glucose tolerance in Trpm5−/− mice , 2010, Proceedings of the National Academy of Sciences.

[26]  J. Bogan,et al.  Cholesterol Regulates Glucose-stimulated Insulin Secretion through Phosphatidylinositol 4,5-Bisphosphate* , 2009, The Journal of Biological Chemistry.

[27]  W. Han,et al.  Role of type Ialpha phosphatidylinositol-4-phosphate 5-kinase in insulin secretion, glucose metabolism, and membrane potential in INS-1 beta-cells. , 2009, Endocrinology.

[28]  Veit Flockerzi,et al.  Alternative Splicing Switches the Divalent Cation Selectivity of TRPM3 Channels* , 2005, Journal of Biological Chemistry.

[29]  Andrea L. Szymczak,et al.  Development of 2A peptide-based strategies in the design of multicistronic vectors , 2005, Expert opinion on biological therapy.

[30]  V. Flockerzi,et al.  TRPC3 Mediates T-cell Receptor-dependent Calcium Entry in Human T-lymphocytes* , 2003, Journal of Biological Chemistry.

[31]  G. Schultz,et al.  Molecular and Functional Characterization of the Melastatin-related Cation Channel TRPM3* , 2003, Journal of Biological Chemistry.

[32]  Shujian Wu,et al.  Expression and Characterization of Human Transient Receptor Potential Melastatin 3 (hTRPM3)* , 2003, Journal of Biological Chemistry.

[33]  Shujian Wu,et al.  Expression and Characterization of Human Transient Receptor Potential Melastatin 3 (hTRPM3)* , 2003, Journal of Biological Chemistry.

[34]  F. Brunicardi,et al.  Development of a transgenic mouse model using rat insulin promoter to drive the expression of CRE recombinase in a tissue-specific manner , 1999, International journal of pancreatology : official journal of the International Association of Pancreatology.

[35]  F. Alt,et al.  Efficient in vivo manipulation of mouse genomic sequences at the zygote stage. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[36]  C B Wollheim,et al.  Establishment of 2-mercaptoethanol-dependent differentiated insulin-secreting cell lines. , 1992, Endocrinology.