Regulator of G protein signaling Gb5-R7 is a crucial activator of muscarinic M3 receptor-stimulated insulin secretion

Qiang Wang, Alexey N. Pronin, Konstantin Levay, Joana Almaca, Alessia Fornoni, Diego A Caicedo-Vierkant, Vladlen Z Slepak

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

In pancreatic β cells, muscarinic cholinergic receptor M3 (M3R) stimulates glucose-induced secretion of insulin. Regulator of G protein signaling (RGS) proteins are critical modulators of GPCR activity, yet their role in β cells remains largely unknown. R7 subfamily RGS proteins are stabilized by the G protein subunit Gb5, such that the knockout of the Gnb5 gene results in degradation of all R7 subunits. We found that Gnb5 knockout in mice or in the insulin-secreting MIN6 cell line almost completely eliminates insulinotropic activity of M3R. Moreover, overexpression of Gβ5-RGS7 strongly promotes M3R-stimulated insulin secretion. Examination of this noncanonical mechanism in Gnb5-/- MIN6 cells showed that cAMP, diacylglycerol, or Ca2+ levels were not significantly affected. There was no reduction in the amplitude of freeCa2+ responses in islets from the Gnb5-/- mice, but the frequency of Ca2+ oscillations induced by cholinergic agonist was lowered by more than 30%. Ablation of Gnb5 impaired M3R stimulated phosphorylation of ERK1/2. Stimulation of the ERK pathway in Gnb5-/- cells by epidermal growth factor restored M3R-stimulated insulin release to near normal levels. Identification of the novel role of Gb5-R7 in insulin secretion may lead to a new therapeutic approach for improving pancreatic b-cell function. - Wang, Q., Pronin, A.N.,Levay, K.,Almaca, J., Fornoni, A.,Caicedo, A., Slepak, V.Z. Regulator of G protein signaling Gβ5-R7 is a crucial activator of muscarinic M3 receptor-stimulated insulin secretion.

Original languageEnglish (US)
JournalFASEB Journal
Volume31
Issue number11
DOIs
StatePublished - Jul 1 2017

Fingerprint

GTP-Binding Protein Regulators
Muscarinic M3 Receptors
Insulin
RGS Proteins
Cholinergic Agonists
Gene Knockout Techniques
MAP Kinase Signaling System
Diglycerides
Protein Subunits
Insulin-Secreting Cells
Cholinergic Receptors
Phosphorylation
GTP-Binding Proteins
Epidermal Growth Factor
Knockout Mice
Ablation
Cholinergic Agents
Modulators
Glucose
Cell Line

Keywords

  • CRISPR-Cas9
  • Fluorescent biosensor
  • Islet of Langerhans
  • Pupil sphincter
  • UBO-QIC

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Regulator of G protein signaling Gb5-R7 is a crucial activator of muscarinic M3 receptor-stimulated insulin secretion. / Wang, Qiang; Pronin, Alexey N.; Levay, Konstantin; Almaca, Joana; Fornoni, Alessia; Caicedo-Vierkant, Diego A; Slepak, Vladlen Z.

In: FASEB Journal, Vol. 31, No. 11, 01.07.2017.

Research output: Contribution to journalArticle

@article{53afe5e121aa4d60bf01ba0f6cc968d8,
title = "Regulator of G protein signaling Gb5-R7 is a crucial activator of muscarinic M3 receptor-stimulated insulin secretion",
abstract = "In pancreatic β cells, muscarinic cholinergic receptor M3 (M3R) stimulates glucose-induced secretion of insulin. Regulator of G protein signaling (RGS) proteins are critical modulators of GPCR activity, yet their role in β cells remains largely unknown. R7 subfamily RGS proteins are stabilized by the G protein subunit Gb5, such that the knockout of the Gnb5 gene results in degradation of all R7 subunits. We found that Gnb5 knockout in mice or in the insulin-secreting MIN6 cell line almost completely eliminates insulinotropic activity of M3R. Moreover, overexpression of Gβ5-RGS7 strongly promotes M3R-stimulated insulin secretion. Examination of this noncanonical mechanism in Gnb5-/- MIN6 cells showed that cAMP, diacylglycerol, or Ca2+ levels were not significantly affected. There was no reduction in the amplitude of freeCa2+ responses in islets from the Gnb5-/- mice, but the frequency of Ca2+ oscillations induced by cholinergic agonist was lowered by more than 30{\%}. Ablation of Gnb5 impaired M3R stimulated phosphorylation of ERK1/2. Stimulation of the ERK pathway in Gnb5-/- cells by epidermal growth factor restored M3R-stimulated insulin release to near normal levels. Identification of the novel role of Gb5-R7 in insulin secretion may lead to a new therapeutic approach for improving pancreatic b-cell function. - Wang, Q., Pronin, A.N.,Levay, K.,Almaca, J., Fornoni, A.,Caicedo, A., Slepak, V.Z. Regulator of G protein signaling Gβ5-R7 is a crucial activator of muscarinic M3 receptor-stimulated insulin secretion.",
keywords = "CRISPR-Cas9, Fluorescent biosensor, Islet of Langerhans, Pupil sphincter, UBO-QIC",
author = "Qiang Wang and Pronin, {Alexey N.} and Konstantin Levay and Joana Almaca and Alessia Fornoni and Caicedo-Vierkant, {Diego A} and Slepak, {Vladlen Z}",
year = "2017",
month = "7",
day = "1",
doi = "10.1096/fj.201700197RR",
language = "English (US)",
volume = "31",
journal = "FASEB Journal",
issn = "0892-6638",
publisher = "FASEB",
number = "11",

}

TY - JOUR

T1 - Regulator of G protein signaling Gb5-R7 is a crucial activator of muscarinic M3 receptor-stimulated insulin secretion

AU - Wang, Qiang

AU - Pronin, Alexey N.

AU - Levay, Konstantin

AU - Almaca, Joana

AU - Fornoni, Alessia

AU - Caicedo-Vierkant, Diego A

AU - Slepak, Vladlen Z

PY - 2017/7/1

Y1 - 2017/7/1

N2 - In pancreatic β cells, muscarinic cholinergic receptor M3 (M3R) stimulates glucose-induced secretion of insulin. Regulator of G protein signaling (RGS) proteins are critical modulators of GPCR activity, yet their role in β cells remains largely unknown. R7 subfamily RGS proteins are stabilized by the G protein subunit Gb5, such that the knockout of the Gnb5 gene results in degradation of all R7 subunits. We found that Gnb5 knockout in mice or in the insulin-secreting MIN6 cell line almost completely eliminates insulinotropic activity of M3R. Moreover, overexpression of Gβ5-RGS7 strongly promotes M3R-stimulated insulin secretion. Examination of this noncanonical mechanism in Gnb5-/- MIN6 cells showed that cAMP, diacylglycerol, or Ca2+ levels were not significantly affected. There was no reduction in the amplitude of freeCa2+ responses in islets from the Gnb5-/- mice, but the frequency of Ca2+ oscillations induced by cholinergic agonist was lowered by more than 30%. Ablation of Gnb5 impaired M3R stimulated phosphorylation of ERK1/2. Stimulation of the ERK pathway in Gnb5-/- cells by epidermal growth factor restored M3R-stimulated insulin release to near normal levels. Identification of the novel role of Gb5-R7 in insulin secretion may lead to a new therapeutic approach for improving pancreatic b-cell function. - Wang, Q., Pronin, A.N.,Levay, K.,Almaca, J., Fornoni, A.,Caicedo, A., Slepak, V.Z. Regulator of G protein signaling Gβ5-R7 is a crucial activator of muscarinic M3 receptor-stimulated insulin secretion.

AB - In pancreatic β cells, muscarinic cholinergic receptor M3 (M3R) stimulates glucose-induced secretion of insulin. Regulator of G protein signaling (RGS) proteins are critical modulators of GPCR activity, yet their role in β cells remains largely unknown. R7 subfamily RGS proteins are stabilized by the G protein subunit Gb5, such that the knockout of the Gnb5 gene results in degradation of all R7 subunits. We found that Gnb5 knockout in mice or in the insulin-secreting MIN6 cell line almost completely eliminates insulinotropic activity of M3R. Moreover, overexpression of Gβ5-RGS7 strongly promotes M3R-stimulated insulin secretion. Examination of this noncanonical mechanism in Gnb5-/- MIN6 cells showed that cAMP, diacylglycerol, or Ca2+ levels were not significantly affected. There was no reduction in the amplitude of freeCa2+ responses in islets from the Gnb5-/- mice, but the frequency of Ca2+ oscillations induced by cholinergic agonist was lowered by more than 30%. Ablation of Gnb5 impaired M3R stimulated phosphorylation of ERK1/2. Stimulation of the ERK pathway in Gnb5-/- cells by epidermal growth factor restored M3R-stimulated insulin release to near normal levels. Identification of the novel role of Gb5-R7 in insulin secretion may lead to a new therapeutic approach for improving pancreatic b-cell function. - Wang, Q., Pronin, A.N.,Levay, K.,Almaca, J., Fornoni, A.,Caicedo, A., Slepak, V.Z. Regulator of G protein signaling Gβ5-R7 is a crucial activator of muscarinic M3 receptor-stimulated insulin secretion.

KW - CRISPR-Cas9

KW - Fluorescent biosensor

KW - Islet of Langerhans

KW - Pupil sphincter

KW - UBO-QIC

UR - http://www.scopus.com/inward/record.url?scp=85037568019&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85037568019&partnerID=8YFLogxK

U2 - 10.1096/fj.201700197RR

DO - 10.1096/fj.201700197RR

M3 - Article

C2 - 28687610

AN - SCOPUS:85037568019

VL - 31

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 11

ER -