A subpopulation of cerebellar granule neurons in culture expresses a functional mGluR1 metabotropic glutamate receptor: effect of depolarizing growing conditions.
Homogeneous neuronal cultures of cerebellar granule neurons express different levels of metabotropic glutamate receptor mGluR1 mRNA depending on the concentration of KCl present during the culture period. We have studied the effect of KCl on mGluR1 expression at the single neuron level by measuring: i) the effect of mGluR agonist 1S,3R-aminocyclopentane-1,3-dicarboxylic acid, tACPD, on intracellular free calcium concentration, [Ca2+]i; ii) the immunocytochemical quantitation of mGluR1 alpha protein. tACPD-induced increases in cytoplasmic free Ca2+ were pertussis toxin-sensitive. The number of tACPD-responding and mGluR1 alpha-positive neurons was higher in cultures grown in the presence of 15 mM than 25 mM KCl, but it never exceeded 50% of the cells. The measurement of basal intracellular free Ca2+ under the respective growing condition revealed a bigger subpopulation of cells with high basal Ca2+ concentration in neurons maintained in 25 mM KCl respect to 15 mM KCl. Hence, in primary cultures of cerebellar granule neurons, depolarization (possibly via Ca(2+)-regulated mechanisms) modulates the expression of a functional mGluR1 only in a subpopulation of cells and thus may account for a functional heterogeneity of morphologically similar cells.
Milani
D
D
Fidia Research Laboratories, Abano Terme, Italy.
Candeo
P
P
Favaron
M
M
Blackstone
C D
CD
Manev
H
H
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002455:Cell Division; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D003515:Cycloleucine / Q000031:analogs & derivatives / Q000494:pharmacology; D007424:Intracellular Fluid / Q000378:metabolism; D008564:Membrane Potentials; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D037342:Pertussis Toxin; D011189:Potassium Chloride / Q000494:pharmacology; D051381:Rats; D018094:Receptors, Metabotropic Glutamate / Q000187:drug effects / Q000378:metabolism*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D018094:Receptors, Metabotropic Glutamate; D010566:Virulence Factors, Bordetella; D003515:Cycloleucine; C036361:1-amino-1,3-dicarboxycyclopentane; D011189:Potassium Chloride; D037342:Pertussis Toxin; D002118:Calcium