Markers for Gene Expression in Cultured Cells from the Nervous System

Methods are presented for preparation of extracts from cultured cells from the nervous system and for study of choline acetyltransferase, acetylcholinesterase, glutamate decarboxylase, and catechol O -methyltransferase activities. These enzyme activities are markers that can be used for studying gen...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of biological chemistry 1972-05, Vol.247 (10), p.3159-3169
Main Authors: Wilson, S H, Schrier, B K, Farber, J L, Thompson, E J, Rosenberg, R N, Blume, A J, Nirenberg, M W
Format: Article
Language:English
Subjects:
Acetylcholinesterase - analysis
Acyltransferases - analysis
Animals
Brain - drug effects
Brain - enzymology
Carbon Isotopes
Carboxy-Lyases - analysis
Carboxy-Lyases - antagonists & inhibitors
Catechol O-Methyltransferase - analysis
Cell Line
Cells, Cultured - drug effects
Cells, Cultured - enzymology
Choline
Coenzyme A
Genes
Glutamates
Glycolates - pharmacology
Humans
Methyltransferases - analysis
Mice
Mice, Inbred Strains
Neuroblastoma - enzymology
Neuroglia - drug effects
Neuroglia - enzymology
Rats
S-Adenosylmethionine
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Methods are presented for preparation of extracts from cultured cells from the nervous system and for study of choline acetyltransferase, acetylcholinesterase, glutamate decarboxylase, and catechol O -methyltransferase activities. These enzyme activities are markers that can be used for studying gene expression in neurons. The methods are sufficiently sensitive so that all assays can be performed with protein harvested from one Petri dish. Activities of the marker enzymes were assessed in surface cultures of newborn mouse brain cells, and in glial and nonbrain cell lines. Low activities of choline acetyltransferase, acetylcholinesterase, and glutamate decarboxylase were detected in all the cells tested. All of these activities, and particularly glutamate decarboxylase, were higher in cultured brain cells from newborn animals than in non-neuronal cell lines. Glutamate decarboxylase activity in glial cells and in brain cells was inhibited more than 95% by 1 m m amino-oxyacetic acid.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(19)45227-7