Bovine chromaffin cells: Studies on the biosynthesis of phospholipids in chromaffin granules

The protein kinase inhibitor staurosporine inhibited, and often abolished, activity-dependent destaining of frog motor nerve terminals that had been preloaded with the fluorescent dye FM1-43. Staurosporine did not, however, block synaptic transmission; staurosporine treated muscles twitched in response to nerve stimulation, and the amplitudes of evoked end plate potentials were reduced only slightly, and in some cases not at all. The blockade of FM1-43 destaining was not reversed by washing, although treatment with black widow spider venom caused complete destaining. Nerve terminal pretreated with staurosporine could subsequently be stained with FM1-43 (and then destained by black widow spider venom). Thus, staurosporine blocked destaining but not staining of nerve terminals. Staurosporine treatment had little effect on the ultra-structure of resting terminals, the main difference we noted being a somewhat closer packing of synaptic vesicles after exposure to staurosporine. However, staurosporine blocked completely the ultrastructural changes produced by prolonged nerve stimulation, such as depletion of synaptic vesicles, appearance of intraterminal cisternae, and the uptake of horseradish peroxidase. The effects of staurosporine were not mimicked by KN-62, H7, calmidozolium, or trifluoroperazine. These and other observations are consistent with, but do not prove the hypothesis that, after exposure to staurosporine, the exocytotic fusion pore behaved like a valve, letting FM1-43 in, but not out, as if staurosporine interfered with the postexocytotic collapse of synaptic vesicles into the surface membrane.

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Staurosporine blocks evoked release of FM1-43 but not acetylcholine from frog motor nerve terminals

Henkel

Betz

1995

J. Neurosci.

116

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Bovine chromaffin cells: Studies on the biosynthesis of phospholipids in chromaffin granules

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Staurosporine blocks evoked release of FM1-43 but not acetylcholine from frog motor nerve terminals

Staurosporine blocks evoked release of FM1-43 but not acetylcholine from frog motor nerve terminals

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