Development of a Single Cell Ca2+ Imaging System to Study the Role of PKC Substrate B-50 in Neurotransmitter Release and Neurite Outgrowth
One of the well-characterized substrates of protein kinase C (PKC) in neurons is the protein B-50. B-50 is a nervous tissue-specific substrate of PKC associated with the cytosolic face of the presynaptic membrane (see De Graan et al., 1991). Protein B-50 is identical to the growth-associated protein GAP-43, the calmodulin (CaM)-binding protein neuromodulin, and protein F1, which is implicated in long-term potentiation (for a review see Skene, 1989). B-50 is one of the abundant proteins in the neuronal growth cone (Skene, 1989) and has been implicated in signal transduction and the mechanism of neurite outgrowth during development and differentiation (Skene, 1989). In a series of studies we have shown that the degree of PKC-mediated phosphorylation of B-50 in hippocampal slices and synaptosomes is correlated with transmitter release (Dekker et al., 1991a; Heemskerk et al., 1989,1990). Based on these correlative studies and the fact that phorbol esters which stimulate PKC enhance neurotransmitter release (see Dekker et al., 1991a), we have suggested that PKC-mediated B-50 phosphorylation may be involved in the regulation of neurotransmitter release.
KeywordsNeurite Outgrowth Transmitter Release Video Microscopy Neuronal Growth Cone Electronic Shutter
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- De Graan PNE, Oestreicher AB, De Wit M, Kroef M, Schrama LH, Gispen WH (1990) Evidence for the binding of calmodulin to endogenous B-50 (GAP-43) in native synaptosomal plasma membranes. J Neurochem 55: in press.Google Scholar
- De Graan PNE, Schrama LH, Oestreicher AB, Schotman P, Gispen WH (1991) Protein kinase C substrate B-50 (GAP-43) and neurotransmitter release. Progr Brain Res, in press.Google Scholar
- Dekker LV, De Graan PNE, Gispen WH (1991a) Transmitter release: target of regulation by protein kinase C? Progr Brain Res, in press.Google Scholar
- Dekker LV, De Graan PNE, Pijnappel P, Oestreicher AB, Gispen WH (1991b) Nor-adrenaline release from streptolysin-O-permeated rat cortical synaptosomes. J Neurochem, in press.Google Scholar
- Heemskerk FMJ, Schrama LH, Ghijsen WEHM, De Graan PNE, Lopes da Silva FH, Gis-pen WH (1991) Presynaptic mechanism of action of 4-aminopyridine: changes in [Ca2+]i and its relationship to B-50 (GAP-43) phosphorylation. J Neurochem, in press.Google Scholar
- Inoue S (1987) Video Microscopy. Plenum Press, NY.Google Scholar
- Schmidt D, Papadopoulos MT, Crooke ST, Stassen FL (1988) Potential role of protein kinase C in the regulation of vasopressin (V1) receptors of vascular smooth muscle cells (A10) Faseb J 2: A624.Google Scholar