Glucose increases intracellular free Ca(2+) in tanycytes via ATP released through connexin 43 hemichannels

Glia. 2012 Jan;60(1):53-68. doi: 10.1002/glia.21246. Epub 2011 Oct 10.

Abstract

The ventromedial hypothalamus is involved in regulating feeding and satiety behavior, and its neurons interact with specialized ependymal-glial cells, termed tanycytes. The latter express glucose-sensing proteins, including glucose transporter 2, glucokinase, and ATP-sensitive K(+) (K(ATP) ) channels, suggesting their involvement in hypothalamic glucosensing. Here, the transduction mechanism involved in the glucose-induced rise of intracellular free Ca(2+) concentration ([Ca(2+) ](i) ) in cultured β-tanycytes was examined. Fura-2AM time-lapse fluorescence images revealed that glucose increases the intracellular Ca(2+) signal in a concentration-dependent manner. Glucose transportation, primarily via glucose transporters, and metabolism via anaerobic glycolysis increased connexin 43 (Cx43) hemichannel activity, evaluated by ethidium uptake and whole cell patch clamp recordings, through a K(ATP) channel-dependent pathway. Consequently, ATP export to the extracellular milieu was enhanced, resulting in activation of purinergic P2Y(1) receptors followed by inositol trisphosphate receptor activation and Ca(2+) release from intracellular stores. The present study identifies the mechanism by which glucose increases [Ca(2+) ](i) in tanycytes. It also establishes that Cx43 hemichannels can be rapidly activated under physiological conditions by the sequential activation of glucosensing proteins in normal tanycytes.

Publication types

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

MeSH terms

  • Adenosine Triphosphate / metabolism*
  • Animals
  • Animals, Newborn
  • Calcium / metabolism*
  • Calcium Signaling / drug effects
  • Calcium Signaling / physiology
  • Cations / metabolism
  • Cells, Cultured
  • Connexin 43 / antagonists & inhibitors
  • Connexin 43 / metabolism*
  • Cytochalasin B / pharmacology
  • Dose-Response Relationship, Drug
  • Enzyme Inhibitors / pharmacology
  • Glucokinase / metabolism
  • Glucose / metabolism
  • Glucose / pharmacology*
  • Glutamate Plasma Membrane Transport Proteins / metabolism
  • Hypothalamus / cytology
  • Intracellular Fluid / metabolism*
  • Ki-67 Antigen / metabolism
  • Male
  • Membrane Potentials / drug effects
  • Membrane Potentials / physiology
  • Microscopy, Confocal
  • Nerve Tissue Proteins / metabolism
  • Neuroglia / cytology
  • Neuroglia / drug effects*
  • Patch-Clamp Techniques
  • Probenecid / pharmacology
  • Rats
  • Rats, Sprague-Dawley
  • von Willebrand Factor / metabolism

Substances

  • Cations
  • Connexin 43
  • Enzyme Inhibitors
  • Glutamate Plasma Membrane Transport Proteins
  • Ki-67 Antigen
  • Nerve Tissue Proteins
  • von Willebrand Factor
  • Cytochalasin B
  • Adenosine Triphosphate
  • Glucokinase
  • Glucose
  • Probenecid
  • Calcium