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. 1995 Jan;114(2):433–441. doi: 10.1111/j.1476-5381.1995.tb13245.x

Necessity of protein kinase C activity for maintenance of acetylcholine receptor function at snake twitch fibre endplates.

J C Hardwick 1, R L Parsons 1
PMCID: PMC1510267  PMID: 7533618

Abstract

1. The extent of recovery of endplate sensitivity following a 5 or 10 min exposure to carbachol was determined from measurements of miniature endplate current (m.e.p.c.) amplitudes in voltage-clamped snake twitch fibre endplates. M.e.p.c. amplitude recovery was dependent on the carbachol concentration (0.27-5.4 mM) and duration of application. Staurosporine pretreatment (0.5 microM for approximately 15 min) further decreased the extent of m.e.p.c. amplitude recovery. 2. The decrease in m.e.p.c. amplitude at control endplates exposed to high concentrations of agonist (5.4 mM carbachol for 10 min) was due to an apparent decrease in postsynaptic receptor density, not to a change in the conductance of the acetylcholine (ACh)-activated channels. 3. Pretreatment with either 1 microM lavendustin A or 50 microM KN-62 had no effect on m.e.p.c. amplitude recovery, whereas pretreatment with either 0.5 microM staurosporine, 50 microM sphingosine, or 0.5 microM calphostin C significantly reduced m.e.p.c. amplitude recovery following carbachol exposure. 4. Sphingosine and staurosporine produced a concentration-dependent decrease in the extent of m.e.p.c. amplitude recovery, but had no effect on m.e.p.c. characteristics in the absence of carbachol. In addition, this decrease in m.e.p.c. amplitude was not due to the presence of a subpopulation of small amplitude m.e.p.cs. 5. Prolonged treatment (18-20 h) of muscles with 200 nM phorbol 12-myristate 13-acetate (PMA), to down regulate protein kinase C, resulted in a significant reduction in m.e.p.c. amplitudes following exposure to carbachol. Conversely, treatment with 200 nM 4 alpha PMA, an inactive analogue, had no effect on m.e.p.c. amplitude recovery.(ABSTRACT TRUNCATED AT 250 WORDS)

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Selected References

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