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Development of a Single Cell Ca2+ Imaging System to Study the Role of PKC Substrate B-50 in Neurotransmitter Release and Neurite Outgrowth

  • J. Elands
  • W. H. Gispen
  • W.H.
  • P. N. E. De Graan
Conference paper
Part of the NATO ASI Series book series (volume 52)

Abstract

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.

Keywords

Neurite Outgrowth Transmitter Release Video Microscopy Neuronal Growth Cone Electronic Shutter 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Almers W, Neher E (1985) The calcium signal from fura-2 loaded mast cells depends strongly on the method of dye-loading. FEBS Lett 192: 13–18.PubMedCrossRefGoogle Scholar
  2. Becker PL, Fay FS (1987) Photobleaching of fura-2 and its effect on determination of calcium concentrations. Am J Physiol 253: C613–C618.PubMedGoogle Scholar
  3. Cheek TR, O’Sullivan AJ, Moreton RB, Berridge MJ, Burgoyne RD (1989) Spatial localization of the stimulus-induced rise in cytosolic Ca2+ in bovine adrenal chromaffin cells. Distinct nicotinic and muscarinic patterns. FEBS Lett 247: 429–434.PubMedCrossRefGoogle Scholar
  4. Connor JA (1986) Digital imaging of free calcium changes and of spatial gradients in growing processes in single, mammalian central nervous system cells. Proc Natl Acad Sci USA 83: 6179–6183.PubMedCrossRefGoogle Scholar
  5. Connor JA, Cornwall MC, Williams GH (1987) Spatially resolved cytosolic calcium response to angiotensin II and potassium in rat glomerulosa cells measured by digital imaging techniques. J Biol Chem 262: 2919–2927.PubMedGoogle Scholar
  6. 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
  7. 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
  8. Dekker LV, De Graan PNE, Oestreicher AB, Versteeg DHG, Gispen WH (1989) Inhibition of noradrenaline release by antibodies to B-50 (GAP-43). Nature 342: 74–76.PubMedCrossRefGoogle Scholar
  9. Dekker LV, De Graan PNE, Gispen WH (1991a) Transmitter release: target of regulation by protein kinase C? Progr Brain Res, in press.Google Scholar
  10. 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
  11. Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260: 3440–3450.PubMedGoogle Scholar
  12. Heemskerk FMJ, Schrama LH, Gianotti C, Spierenburg H, Versteeg DHG, De Graan PNE, Gispen WH (1989a) 4-Aminopyridine stimulates B-50/GAP-43 phosphorylation and [3H]-noradrenaline release in rat hippocampal slices. J Neurochem 54: 863–869.CrossRefGoogle Scholar
  13. Heemskerk FMJ, Schrama LH, De Graan PNE, Gispen WH (1990) 4-Aminopyridine stimulates B-50 (GAP-43) phosphorylation in rat synaptosomes. J Mol Neurosci 2: 11–17.PubMedCrossRefGoogle Scholar
  14. 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
  15. Inoue S (1987) Video Microscopy. Plenum Press, NY.Google Scholar
  16. Liu Y, Storm DR (1990) Regulation of free calmodulin levels by neuromodulin: neuron growth and regeneration. Trends Pharmacol Sci 11: 107–111.PubMedCrossRefGoogle Scholar
  17. Nichols RA, Sihra TS, Czernik AJ, Nairn A, Greengard P. (1990) Calcium/calmo-dulin-dependent protein kinase II increases glutamate and noradrenaline release from synaptosomes. Nature 343: 647–651.PubMedCrossRefGoogle Scholar
  18. 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
  19. Skene JHP (1989) Axonal growth-associated proteins. Ann Rev Neurosci 12: 127–156.PubMedCrossRefGoogle Scholar
  20. Tsien RY (1988) Fluoresence measurement and photochemical manipulation of cytosolic free calcium. TINS 11: 419–424.PubMedGoogle Scholar
  21. Tsien RY (1989) Fluorescent indicators of ion concentrations. Meth Cell Biol 30: 127–156.CrossRefGoogle Scholar
  22. Tsien RY, Poenie M (1986) Fluoresence ratio imaging: a new window into intra-cellular ionic signalling. Trends Biochem Sci 11: 450–455.CrossRefGoogle Scholar
  23. Verhage M, Besselsen E, Lopes da Silva FH, Ghijsen WEJM (1988) Evaluation of the Ca2+ concentration in purified nerve tenninals: relationship between Ca2+ homeostasis and synaptosomal preparation. J Neurochem 51: 1667–1674.PubMedCrossRefGoogle Scholar
  24. Williams DA, Fogarty KE, Tsien RY, Fay FS (1985) Calcium gradients in single smooth muscle cells revealed by the digital inmaging microscope using fura-2. Nature 318: 558–561.PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1991

Authors and Affiliations

  • J. Elands
    • 1
  • W. H. Gispen
    • 1
  • W.H.
    • 1
  • P. N. E. De Graan
    • 1
  1. 1.Division of Molecular NeurobiologyRudolf Magnus InstituteUtrechtThe Netherlands

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