Effect of depolarization on the maturation of cerebellar granule cells in culture

Abstract

The effect of depolarization on the maturation of granule cells derived from cerebella of 8-day-old rats can be studied in cultures in chemically defined media because their survival is not dependent on elevated K+ as it is when they are grown in serum-containing media. As an index of maturation, stimulus-coupled transmitter release was examined. This was chosen because it is closely associated with the neuronal phenotype and, in contrast to granule cells grown under depolarizing conditions in serum-containing media, it has not been known whether this property is expressed during the development of serum-free cells in culture. Veratrine-induced release of preloaded D-[3H]aspartate (Asp), an analogue of glutamate (Glu; the transmitter of the granule cells), was not detectable in the serum-free cells at a time (8-12 days in vitro) when this property was fully developed in cells grown in a medium containing serum and 25 mM K+ (reference cultures). This finding may be related to the failure of the expression of voltage-sensitive calcium channels in the serum-free granule cells. However, in comparison with reference cultures, voltage-sensitive 45Ca2+ entry was only transiently retarded in the serum-free cells. Furthermore, in contrast to the exogenous D-[3H]Asp, stimulated release of endogenous Glu was detectable, although it was substantially lower than in the reference cultures. Autoradiographic studies indicated that the failure to elicit evoked release of the exogenous amino acid was due to a severe retardation of the expression of the acidic amino acid carrier in the serum-free granule cells.(ABSTRACT TRUNCATED AT 250 WORDS)