NGF-induction of the expression of ChAT mRNA in PC12 cells and primary cultures of embryonic rat basal forebrain.
The objective of this study was to examine the role of nerve growth factor (NGF) in regulation of expression of the cholinergic phenotype. NGF was administered to PC12 cells or primary cultures of embryonic (E17) rat basal forebrain for 2 days, then steady-state levels of choline acetyltransferase (ChAT) mRNA was monitored. Expression of ChAT mRNA isoforms was investigated using reverse transcription-polymerase chain reaction (RT-PCR) to amplify different upstream regions of the ChAT transcripts, and Southern blot analysis was used to verify identity of the PCR products. An NGF-induced increase of 1.8- and 1.5-fold in steady-state level of the ChAT transcript containing the M-exon (M-ChAT) was observed in PC12 cells and embryonic rat basal forebrain neurons, respectively. Also, a 2-fold increase in ChAT protein as determined by western blot analysis was associated with an NGF-mediated increase of 1.7-fold in ChAT activity in rat basal forebrain neurons within the same cultures following 4 days of NGF treatment.
Pongrac
J L
JL
Department of Physiology, University of Western Ontario, London, Ontario, Canada.
Rylett
R J
RJ
D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D048429:Cell Size / Q000187:drug effects; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000032:analysis / Q000235:genetics*; D004622:Embryo, Mammalian; D005786:Gene Expression Regulation / Q000187:drug effects*; D008970:Molecular Weight; D009414:Nerve Growth Factors / Q000494:pharmacology*; D016501:Neurites / Q000187:drug effects; D009474:Neurons / Q000166:cytology / Q000201:enzymology; D016716:PC12 Cells; D016133:Polymerase Chain Reaction; D016548:Prosencephalon / Q000166:cytology / Q000201:enzymology*; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis* / Q000187:drug effects*; D051381:Rats; D013997:Time Factors
D009414:Nerve Growth Factors; D012333:RNA, Messenger; D002795:Choline O-Acetyltransferase