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Evidence that Protein Kinase C Activities Involved in Regulating Neurite Growth Are Localized to Distal Neurites
Author(s) -
Campenot Robert B.,
Draker Dwight D.,
Senger Donna L.
Publication year - 1994
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1994.63030868.x
Subject(s) - neurite , calphostin c , protein kinase c , chelerythrine , microbiology and biotechnology , growth cone , biology , phosphorylation , chemistry , biochemistry , in vitro , axon
Previously, we observed that long‐term treatment of distal nerve fibers of rat sympathetic neurons in compartmented cultures with phorbol 12‐myristate 13‐acetate (PMA) caused a reduction in the rate of neurite elongation by >50%. In the present report we show that protein kinase C (PKC) activity could be measured in extracts of distal neurites by an assay of the Ca 2+ ‐dependent phosphorylation of a PKC‐specific octapeptide substrate. We found that local application of 1 µ M PMA for 24 h to distal neurites caused nearly complete down‐regulation of Ca 2+ ‐dependent PKC activity measured in this manner. We determined that the inhibition of neurite elongation by PMA was mediated by local mechanisms in the neurites because local application of PMA to center compartments containing cell bodies and proximal neurites did not inhibit the rate of elongation of distal neurites. We then investigated the effects of the recently available PKC inhibitors, calphostin C and chelerythrine, finding that, like PMA, these inhibited the growth of distal neurites when applied locally to them, and had no effect when applied to cell bodies and proximal neurites. However, the inhibition of neurite growth by calphostin C occurred at a concentration far below its IC 50 value for protein kinase inhibition, and both calphostin C and chelerythrine inhibited distal neurite growth even in neurons pretreated with PMA. Thus, it appears that these agents do not all inhibit neurite growth through the same mechanisms. Although the PKC activities involved in neurite elongation in sympathetic neurons have not been precisely defined, these data presented in this study indicate that protein kinases localized to growth cones play a complex and important role in regulating axonal growth.