Antisense GAP-43 inhibits the evoked release of dopamine from PC12 cells

Kathryn J. Ivins, Kim Neve, Daniel J. Feller, Seth A. Fidel, Rachael L. Neve

Research output: Contribution to journalArticle

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Abstract

To investigate the role of the neuronal growth-associated protein GAP-43 (neuromodulin, B-50, F1. P-57) in neurotransmitter release, we transfected PC12 cells with a recombinant expression vector coding for antisense human GAP-43 cRNA. Two stable transfectants, designated AS1 and AS2. were selected that had integrated the recombinant sequence and expressed antisense GAP-43 RNA. Immunoblot analysis of proteins from AS1 and AS2 cells indicated that the level of GAP-43 in these cell lines was reduced. In the presence of extracellular calcium, a depolarizing concentration of K+ (56 mM) evoked dopamine release from control cells, but not from AS1 and AS2 cells. Similarly, the calcium ionophore A23187 evoked dopamine release from control cells, but was ineffective in stimulating dopamine release from AS I and AS2 cells. The antisense transfectants, as well as the control cells, contained appreciable quantities of dopamine and secretory granules with a normal appearance. Because the expression of antisense GAP-43 RNA in PC 12 cells leads to a decrease in GAP-43 expression and to the loss of evoked dopamine release, these results provide evidence of a role for GAP-43 in calcium-dependent neurotransmitter release.

Original languageEnglish (US)
Pages (from-to)626-633
Number of pages8
JournalJournal of Neurochemistry
Volume60
Issue number2
StatePublished - 1993

Fingerprint

GAP-43 Protein
PC12 Cells
Dopamine
Neurotransmitter Agents
RNA
Calcium
Complementary RNA
Calcium Ionophores
Nerve Growth Factors
Calcimycin
Secretory Vesicles
Cells
Cell Line

Keywords

  • Antisense RNA
  • Dopamine release
  • GAP-43
  • PC12 cells
  • Transfectant

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Ivins, K. J., Neve, K., Feller, D. J., Fidel, S. A., & Neve, R. L. (1993). Antisense GAP-43 inhibits the evoked release of dopamine from PC12 cells. Journal of Neurochemistry, 60(2), 626-633.

Antisense GAP-43 inhibits the evoked release of dopamine from PC12 cells. / Ivins, Kathryn J.; Neve, Kim; Feller, Daniel J.; Fidel, Seth A.; Neve, Rachael L.

In: Journal of Neurochemistry, Vol. 60, No. 2, 1993, p. 626-633.

Research output: Contribution to journalArticle

Ivins, KJ, Neve, K, Feller, DJ, Fidel, SA & Neve, RL 1993, 'Antisense GAP-43 inhibits the evoked release of dopamine from PC12 cells', Journal of Neurochemistry, vol. 60, no. 2, pp. 626-633.
Ivins, Kathryn J. ; Neve, Kim ; Feller, Daniel J. ; Fidel, Seth A. ; Neve, Rachael L. / Antisense GAP-43 inhibits the evoked release of dopamine from PC12 cells. In: Journal of Neurochemistry. 1993 ; Vol. 60, No. 2. pp. 626-633.
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