An efficient method for the long-term and specific expression of exogenous cDNAs in cultured Purkinje neurons.

ABSTRACT: We present a simple and efficient method for expressing cDNAs in Purkinje neurons (PNs) present in heterogeneous mouse cerebellar cultures. The method combines the transfection of freshly dissociated cerebellar cells via nucleofection with the use of novel expression plasmids containing a fragment of the L7 (Pcp2) gene that, within the cerebellum, drives PN-specific expression. The efficiency of PN transfection (determined 13 days post nucleofection) is approximately 70%. Double and triple transfections are routinely achieved at slightly lower efficiencies. Expression in PNs is obvious after one week in culture and still strong after three weeks, by which time these neurons are well-developed. Moreover, high-level expression is restricted almost exclusively to the PNs present in these mixe

SUBMITTER: Wagner W 

PROVIDER: S-EPMC3407467 | biostudies-literature | 2011 Sep

REPOSITORIES: biostudies-literature