Inhibition of protein kinase C prevents Purkinje cell death but does not affect axonal regeneration.
In organotypic cultures, mouse Purkinje cells regenerate their axons from embryonic day 18 (E18) to postnatal day 0 (P0), die of apoptosis between P1 and P7, and survive but do not regenerate at P10. This particular behavior of Purkinje cells did not allow us to find out when the developmental switch between regeneration and lack of regeneration occurs. This work was undertaken to suppress Purkinje cell apoptosis and to investigate whether the same molecules that prevent apoptosis could also influence axonal growth, regeneration, or both. We show that brain-derived neurotrophic factor, neurotrophin 3, and insulin-like growth factor I have marginal effects on P3 Purkinje cell death. The use of Gö6976 [a protein kinase C (PKC) inhibitor] or a transgenic mouse line, in which a pseudosubstrate PKC inhibitor has been specifically targeted to Purkinje cells, prevents the massive Purkinje cell death in P3 organotypic cultures. In addition, Gö6976 promotes axotomized Purkinje cell survival up to P7. Thus, the inhibition of PKC activity is able to prevent Purkinje cell apoptosis in organotypic cultures. Furthermore, Gö6976 increases the outgrowth of dendrites and axon collateralization, as shown after gene gun enhanced green fluorescent protein transfection. In contrast, PKC inhibitors do not influence the axonal regenerative capability of Purkinje cell during development; the latter decreases between E18 and P7 after the same time course in control and Gö6976-treated slices. Thus, because inhibition of PKC prevents Purkinje cell death but does not affect axonal regeneration, these two events (cell death and axonal regeneration) seem to be differentially regulated.
Ghoumari
Abdel M
AM
Institut National de la Santé et de la Recherche Médicale Unité 106, Hôpital de la Salpêtrière, 75651 Paris Cedex 13, France.
Wehrlé
Rosine
R
De Zeeuw
Chris I
CI
Sotelo
Constantino
C
Dusart
Isabelle
I
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001369:Axons / Q000187:drug effects* / Q000502:physiology; D019771:Axotomy; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology; D002227:Carbazoles / Q000494:pharmacology; D002454:Cell Differentiation / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D002531:Cerebellum / Q000166:cytology / Q000196:embryology; D003712:Dendrites / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D018507:Gene Expression Regulation, Developmental; D066298:In Vitro Techniques; D007211:Indoles / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D051379:Mice; D008822:Mice, Transgenic; D009416:Nerve Regeneration / Q000187:drug effects* / Q000502:physiology; D020933:Neurotrophin 3 / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D011689:Purkinje Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D013997:Time Factors
D019208:Brain-Derived Neurotrophic Factor; D002227:Carbazoles; D004791:Enzyme Inhibitors; D007211:Indoles; D020933:Neurotrophin 3; C081021:Go 6976; D007334:Insulin-Like Growth Factor I; D011493:Protein Kinase C