Comparison of type 2 inositol 1,4,5-trisphosphate receptor distributionand subcellular Ca2+ release sites that support Ca2+ waves in cultured astrocytes

Carol A. Sheppard, Peter B. Simpson, Alan H. Sharp, Frederick C. Nucifora, Christopher A. Ross, G. David Lange, James T. Russell

Research output: Contribution to journalArticlepeer-review

Abstract

We have examined the mechanisms that underlie Ca2+ wave propagation in cultured cortical astrocytes. Norepinephrine evoked Ca2+ waves in astrocytes that began at discrete initiation loci and propagated throughout the cell by regenerative amplification at a number of cellular sites, as shown by very high Ca2+ release rates at these regions. We have hypothesized previously that domains displaying elevated Ca2+ release kinetics in astrocytes may correspond to sites of high inositol 1,4,5- trisphosphate receptor (InsP3R) density. To examine this possibility, we compared the distribution pattern of endoplasmic reticulum (ER) and InsP3Rs with Ca2+ release kinetics in subcellular regions during propagation of norepinephrine-evoked waves. 3,3'-Dihexyloxacarbocyanine iodide staining revealed that the ER in astrocytes exists as a meshwork of membranes extending throughout the cells, including fine processes. A specific antibody directed against type 2 InsP3Rs (InsP3R2) detected a 260-kDa band in western blotting of astrocyte membranes. Immunocytochemistry using this antibody stained the entire ER system in a punctate, variegated manner. When Ca2+ responses and InsP3R2 immunofluorescence were compared in the same cell, domains of elevated Ca2+ response kinetics (high amplitude and rapid rate of rise) showed significant positive correlation with high local intensity of InsP3R2 staining. It appears, therefore, that specializations in the ER responsible for discrete local Ca2+ release sites that support regenerative wave propagation include increased levels of InsP3R2 expression.

Original languageEnglish (US)
Pages (from-to)2317-2327
Number of pages11
JournalJournal of Neurochemistry
Volume68
Issue number6
DOIs
StatePublished - Jun 1997

Keywords

  • Astrocytes
  • Calcium
  • Endoplasmic reticulum
  • Inositol 1,4,5-trisphosphate
  • Receptors

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Fingerprint Dive into the research topics of 'Comparison of type 2 inositol 1,4,5-trisphosphate receptor distributionand subcellular Ca<sup>2+</sup> release sites that support Ca<sup>2+</sup> waves in cultured astrocytes'. Together they form a unique fingerprint.

Cite this