Cellular distribution and activation by phorbol myristate acetate (PMA) of classical (alpha, betaI, betaII,gamma), novel (delta, epsilon, theta, eta), and atypical (zeta, iota) protein kinase C (PKC) isoforms were studied in cultured rat neonatal microglial and astroglial cells by Western blot analysis. Among the classical isoforms, only betaII was expressed in microglia and astrocytes in the same abundance. The expression of betaI in microglia was less abundant, while PKCalpha was not detectable in this cell type. PKCgamma was absent in both cell populations. A different pattern of expression was also found for novel and atypical isoenzymes: Both cell types expressed delta, theta, eta, zeta, and iota isoforms, but PKCepsilon was absent in microglia and the expression of PKCzeta and PKCiota in these cells was low compared to astrocytes. The pattern of PKC distribution in cytosolic and particulate fractions as well as activation by short (10 min) and prolonged (4 hr) PMA treatment in both cell types were similar. On the whole, in comparison with astrocytes, PKC in microglial cells was less expressed, both in terms of number of isoforms and level of expression. The microglial profile of PKC isoforms differed from that of rat peritoneal macrophages, which did express PKCalpha. Preliminary evidence suggests that the ability of PMA to enhance cyclic AMP responses in astrocytes, but not in microglia, is related to the different pattern of expression of PKCalpha and PKCepsilon in the two cell types.