Evidence for a role of protein kinase C in FGF signal transduction in the developing chick limb bud

In developing limbs, numerous signaling molecules have been identified but less is know about the mechanisms by which such signals direct patterning. We have explored signal transduction pathways in the chicken limb bud, A cD NA encoding RACK1, a protein that binds and stabilizes activated protein ki nase C (PKC), was isolated in a screen for genes induced by retinoic acid ( RA) in the chick wing bud. Fibroblast growth factor (FGF) also induced RACK 1 and such induction of RACK1 expression was accompanied by a significant a ugmentation in the number of active PKC molecules and an elevation of PKC e nzymatic activity. This suggests that PKCs mediate signal transduction in t he limb bud. Application of chelerythrine, a potent PKC inhibitor, to the p resumptive wing region resulted in buds that did not express sonic hedgehog (Shh) and developed into wings that were severely truncated. This observat ion suggests that the expression of Shh depends on PKCs, Providing ectopic SHH protein, RA or ZPA grafts overcome the effects of blocking PKC with che lerythrine and resulted in a rescue of the wing morphology, Taken together, these findings suggest that the responsiveness of Shh to FGF is mediated, at least in part, by PKCs.