A G s-coupled purinergic receptor boosts Ca 2+ influx and vascular contractility during diabetic hyperglycemia

Elife. 2019 Mar 1;8:e42214. doi: 10.7554/eLife.42214.

Abstract

Elevated glucose increases vascular reactivity by promoting L-type CaV1.2 channel (LTCC) activity by protein kinase A (PKA). Yet, how glucose activates PKA is unknown. We hypothesized that a Gs-coupled P2Y receptor is an upstream activator of PKA mediating LTCC potentiation during diabetic hyperglycemia. Experiments in apyrase-treated cells suggested involvement of a P2Y receptor underlying the glucose effects on LTTCs. Using human tissue, expression for P2Y11, the only Gs-coupled P2Y receptor, was detected in nanometer proximity to CaV1.2 and PKA. FRET-based experiments revealed that the selective P2Y11 agonist NF546 and elevated glucose stimulate cAMP production resulting in enhanced PKA-dependent LTCC activity. These changes were blocked by the selective P2Y11 inhibitor NF340. Comparable results were observed in mouse tissue, suggesting that a P2Y11-like receptor is mediating the glucose response in these cells. These findings established a key role for P2Y11 in regulating PKA-dependent LTCC function and vascular reactivity during diabetic hyperglycemia.

Keywords: arterial tone; biosensors; cell biology; extracellular nucleotides; human; ion channels; molecular biophysics; mouse; structural biology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Vessels / physiopathology*
  • Calcium / metabolism*
  • Calcium Signaling
  • Cyclic AMP-Dependent Protein Kinases / metabolism
  • Hyperglycemia*
  • Mice, Inbred C57BL
  • Muscle Contraction*
  • Receptors, G-Protein-Coupled / metabolism*
  • Receptors, Purinergic / metabolism*

Substances

  • Receptors, G-Protein-Coupled
  • Receptors, Purinergic
  • Cyclic AMP-Dependent Protein Kinases
  • Calcium