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Promoter Elements of the Rat Choline Acetyltransferase Gene Allowing Nerve Growth Factor Inducibility in Transfected Primary Cultured Cells
Author(s) -
Bejanin Stephane,
Habert Estelle,
Berrard Sylvie,
Edwards JeanBaptiste Dumas Milne,
Loeffler JeanPhilippe,
Mallet Jacques
Publication year - 1992
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1992.tb11383.x
Subject(s) - chloramphenicol acetyltransferase , choline acetyltransferase , biology , microbiology and biotechnology , transfection , reporter gene , gene , transcription (linguistics) , exon , complementary dna , cholinergic neuron , nerve growth factor , promoter , sp1 transcription factor , gene expression , cholinergic , biochemistry , endocrinology , receptor , linguistics , philosophy
Choline acetyltransferase, the enzyme responsible for the synthesis of acetylcholine, provides a convenient index for cholinergic neurons. Using a previously identified rat cDNA clone, we have isolated several corresponding genomic clones and have characterized a 1,902‐bp fragment that contains part of the first noncoding exon as well as promoter sequences. The promoter activity of this fragment was tested, taking advantage of the recently developed lipopolyamine‐mediated DNA transfer method, which allows transfection of primary neurons. The 1,902‐bp sequence drives the expression of the bacterial chloramphenicol acetyltransferase (CAT) reporter gene in a culture of dissociated cells prepared from the septal area of fetal (embryonic day 17) rats, a structure rich in cholinergic neurons. Moreover, addition of nerve growth factor to the culture increases CAT expression by ∼56‐fold, indicating that our DNA fragment contains sequences required for NGF induction. In addition, it contains consensus sequences for various transcription factors, including those of the basic helix–loop–helix family. Finally, experiments to characterize the transcription start site are presented.