Mechanisms of intercellular calcium signaling in glial cells studied with dantrolene and thapsigargin.
Mechanical stimulation of a single cell in a primary mixed glial cell culture induced a wave of increased intracellular calcium concentration ([Ca2+]i) that was communicated to surrounding cells. Following propagation of the Ca2+ wave, many cells showed asynchronous oscillations in [Ca2+]i. Dantrolene sodium (10 microM) inhibited the increase in [Ca2+]i associated with this Ca2+ wave by 60-80%, and prevented subsequent Ca2+ oscillations. Despite the markedly decreased magnitude of the increase in [Ca2+]i, the rate of propagation and the extent of communication of the Ca2+ wave were similar to those prior to the addition of dantrolene. Thapsigargin (10 nM to 1 microM) induced an initial increase in [Ca2+]i ranging from 100 nM to 500 nM in all cells that was followed by a recovery of [Ca2+]i to near resting levels in most cells. Transient exposure to thapsigargin for 2 min irreversibly blocked communication of Ca2+ wave from the stimulated cell to adjacent cells. Glutamate (50 microM) induced an initial increase in [Ca2+]i in most cells that was followed by sustained oscillations in [Ca2+]i in some cells. Dantrolene (10 microM) inhibited this initial [Ca2+]i increase caused by glutamate by 65-90% and abolished subsequent oscillations. Thapsigargin (10 nM to 1 micron) abolished the response to glutamate in over 99% of cells. These results suggest that while both dantrolene and thapsigargin inhibit intracellular Ca2+ release, only thapsigargin affects the mechanism that mediates intercellular communication of Ca2+ waves. These findings are consistent with the hypothesis that inositol trisphosphate (IP3) mediates the propagation of Ca2+ waves whereas Ca(2+)-induced Ca2+ release amplifies Ca2+ waves and generates subsequent Ca2+ oscillations.
Charles
A C
AC
Department of Neurology, UCLA School of Medicine 90024.
Dirksen
E R
ER
Merrill
J E
JE
Sanderson
M J
MJ
D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000502:physiology; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D003620:Dantrolene / Q000494:pharmacology*; D005971:Glutamates / Q000494:pharmacology; D018698:Glutamic Acid; D007700:Kinetics; D009457:Neuroglia / Q000187:drug effects / Q000502:physiology*; D010812:Physical Stimulation; D051381:Rats; D012433:Ryanodine / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013050:Spectrometry, Fluorescence; D013729:Terpenes / Q000494:pharmacology*; D019284:Thapsigargin
D005971:Glutamates; D013729:Terpenes; D012433:Ryanodine; D002110:Caffeine; D018698:Glutamic Acid; D019284:Thapsigargin; D000252:Calcium-Transporting ATPases; D003620:Dantrolene; D002118:Calcium