Mimicry and Inhibition of Nerve Growth Factor Effects: Interactions of Staurosporine, Forskolin, and K252a in PC12 Cells and Normal Rat Chromaffin Cells In Vitro

Abstract

The structurally similar compounds staurosporine and K252a are potent inhibitors of protein kinases. K252a has previously been reported to inhibit most or all of the effects of nerve growth factor (NGF) on PC12 pheochromocytoma cells, and staurosporine has been reported both to inhibit and to mimic NGF-induced neurite outgrowth from a PC12 cell subclone in a dose-dependent manner. We have studied the interactions of these agents with each other, with NGF, and with forskolin, an activator of adenylate cyclase, on the parent PC12 cell line and on normal neonatal and adult rat chromaffin cells. Staurosporine alone or in conjunction with forskolin induces outgrowth of short neurites from PC12 cells but does not substitute for NGF in promoting cell survival. It does not abolish NGF-induced neurite outgrowth but does reverse the effects of NGF on catecholamine synthesis. K252a abolishes NGF-induced neurite outgrowth but only partially decreases outgrowth induced by NGF plus forskolin. It does not inhibit neurite outgrowth produced by staurosporine or staurosporine plus forskolin. These findings with PC12 cells suggest that staurosporine might act downstream from K252a and NGF on components of one or more signal transduction pathways by which NGF selectively affects the expression of certain traits. Both neonatal and adult rat chromaffin cells show dramatic flattening and extension of filopodia in response to staurosporine, an observation suggesting that some of the same pathways might remain active in cells that do not exhibit a typical NGF response. Only a small amount of neurite outgrowth is observed, however, and only in neonatal cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

Agonist‐induced desensitization of histamine Hi receptor‐mediated inositol phospholipid hydrolysis in human umbilical vein endothelial cells

“…Consequently, the lack of effect of both staurosporine and the similarly non-selective kinase inhibitor K-252a (1 JAM) (Kase et al, 1987;Ruegg & Burgess, 1989;Tischler et al, 1990) on histamine-induced desensitization supports the data obtained with forskolin and sodium nitroprusside, and appears to rule out a role for either cyclic AMP or cyclic GMP dependent kinases in this process. Interestingly, the selective Ca+/calmodulin dependent kinase II inhibitor, KN-62 (201JM) (Tokumitsu et al, 1990) was also without effect on histamine-induced desensitization.…”

“…Staurosporine is acknowledged to be a potent but rather non-selective inhibitor of protein kinase C (Ruegg & Burgess, 1989) and is in addition a relatively potent inhibitor of protein kinases A and G (Tamaoki et al, 1986;Schachtele et al, 1988;Tischler et al, 1990). Consequently, the lack of effect of both staurosporine and the similarly non-selective kinase inhibitor K-252a (1 JAM) (Kase et al, 1987;Ruegg & Burgess, 1989;Tischler et al, 1990) on histamine-induced desensitization supports the data obtained with forskolin and sodium nitroprusside, and appears to rule out a role for either cyclic AMP or cyclic GMP dependent kinases in this process.…”

Plasmalopsychosine of Human Brain Mimics the Effect of Nerve Growth Factor by Activating Its Receptor Kinase and Mitogen-activated Protein Kinase in PC12 Cells

“…In contrast, high concentrations (50 -200 nM) of staurosporine inhibited MAPK activation by 4,6-PLPS or NGF, but induced neurite outgrowth (of short processes) in the absence of NGF or 4,6-PLPS (data not shown). These results support previous observations that staurosporine by itself induces neurite outgrowth in the absence of NGF at high concentration but inhibits NGF-induced neurite outgrowth at low concentration (24,25).…”

“…No such phosphorylation was detectable in cells preincubated with 100 nM K-252a or staurosporine (Fig. 5), both of which are known tyrosine kinase inhibitors as well as neurotrophic effect blockers (23)(24)(25).…”

Staurosporine-induced neuronal death: multiple mechanisms and methodological implications

“…Low STS concentrations promote neurite outgrowth and survival of other neuronal applications as well: STS (10 ± 50 nM) promotes neurite outgrowth in PC12 cells. 59,60 STS (10 fM ± 10 nM) also promotes the survival of hippocampal, septal, and cortical cells after glucose deprivation 61 and prevents death of hippocampal neurons after amyloid b toxicity or oxidative injury. 62 STS administration in rats also facilitates recovery from deficits induced by basal forebrain lesions 63 and prevents neuronal damage after ischemic injury.…”