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Journal of Neuroscience, Vol 15, 7158-7167, Copyright © 1995 by Society for Neuroscience
Intraneuronal delivery of protein kinase C pseudosubstrate leads to growth cone collapse
L Theodore, D Derossi, G Chassaing, B Llirbat, M Kubes, P Jordan, H Chneiweiss, P Godement and A Prochiantz
URA 1414, Centre National de la Recherche Scientifique, Ecole Normale Superieure, Paris, France.
Axonal navigation during development requires that cues present in the
extracellular environment be capable of modifying the structure of the cone
in a dynamic way. Protein kinase C (PKC) has long been suspected to be one
of the multiple molecular relays present in the terminal structure of the
developing axon and involved in the transduction of extracellular signals.
The latter proposal is, however, based on the use of drugs or of protocols
leading to pleiotropic and often nonspecific effects. In the present study,
we have taken advantage of the discovery of a peptide capable of
translocating across biological membranes and to accumulate in the
cytoplasm and nucleus of cells in culture, to internalize a highly specific
peptidic inhibitor of PKC. We demonstrate that linking the two peptides
(vector and PKC inhibitor) allows the internalization of the latter in live
cells, specifically inhibits PKC and provokes a rapid modification of
growth cone morphology. This set of data thus establishes that a peptidic
inhibitor of PKC activity, once internalized, provokes a change in growth
cone morphology, reminiscent of the collapse phenotype. In addition, the
present study describes a new efficient and harmless way to introduce
pharmacologically active substances in neural cells in culture.
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