Regulation of chick dorsal root ganglion growth cone filopodia by protein kinase C

Growth cone filopodia function both as structural and sensory devices during neuronal pathfinding and their presence is important for correct growth cone navigation. It is assumed that a growth cone can adjust the area of the environment it can explore by regulating the length and number of its filopodial sensors, and in several cell types, these parameters are controlled by the intracellular calcium concentration ([Ca(2+)](i)). In the present report, we address the question whether [Ca(2+)](i) is a general regulator of growth cone filopodia, or whether different cell types utilize different second-messenger systems for this purpose. We show that increasing [Ca(2+)](i) in growth cones of chick dorsal root ganglion (DRG) neurons does not affect average filopodial length in this cell type, suggesting that this parameter is not controlled by [Ca(2+)](i) in chick DRG neurons. Further, we demonstrate that the second-messenger protein kinase C (PKC) is involved in the regulation of filopodial length in chick DRG neurons. Activation of PKC with the phorbol ester, phorbol myristate-13-acetate (PMA), caused filopodial shortening, whereas inhibition of PKC with either bisindolylmaleimide I or calphostin C caused a significant elongation of filopodia. Although the pathway through which PKC mediates its effect on growth cone filopodia in chick DRG neurons remains to be identified, our results indicate that filopodial regulation by [Ca(2+)](i), though clearly important in several other neuronal cell types in vitro, appears to be less important in chick DRG neurons. Rather, we find that in chick DRG neurons, filopodial parameters are controlled by PKC.