| First Author | Takahashi N | Year | 2017 |
| Journal | J Neurochem | Volume | 143 |
| Issue | 6 | Pages | 660-670 |
| PubMed ID | 29049849 | Mgi Jnum | J:254374 |
| Mgi Id | MGI:6102743 | Doi | 10.1111/jnc.14239 |
| Citation | Takahashi N, et al. (2017) Regulatory connection between the expression level of classical protein kinase C and pruning of climbing fibers from cerebellar Purkinje cells. J Neurochem 143(6):660-670 |
| abstractText | Cerebellar Purkinje cells (PCs) express two members of the classical protein kinase C (cPKC) subfamily, namely, PKCalpha and PKCgamma. Previous studies on PKCgamma knockout (KO) mice have revealed a critical role of PKCgamma in the pruning of climbing fibers (CFs) from PCs during development. The question remains as to why only PKCgamma and not PKCalpha is involved in CF synapse elimination from PCs. To address this question, we assessed the expression levels of PKCgamma and PKCalpha in wild-type (WT) and PKCgamma KO PCs using PC-specific quantitative real-time reverse transcription-polymerase chain reaction, western blotting, and immunohistochemical analysis. The results revealed that the vast majority of cPKCs in PCs were PKCgamma, whereas PKCalpha accounted for the remaining minimal fraction. The amount of PKCalpha was not up-regulated in PKCgamma KO PCs. Lentiviral expression of PKCalpha in PKCgamma KO PCs resulted in a 10-times increase in the amount of PKCalpha mRNA in the PKCgamma KO PCs, compared to that in WT PCs. Our quantification showed that the expression levels of cPKC mRNA in PKCgamma KO PCs increased roughly from 1% to 22% of that in WT PCs solely through PKCalpha expression. The up-regulation of PKCalpha in PKCgamma KO PCs significantly rescued the impaired CF synapse elimination. Although both PKCalpha and PKCgamma are capable of pruning supernumerary CF synapses from developing PCs, these results suggest that the expression levels of cPKCs in PKCgamma KO PCs are too low for CF pruning. |
