Abstract
The expression of the acetylcholine receptor epsilon-subunit gene is restricted to the endplate of adult muscle fibers. We have started to study the regulatory elements of the epsilon-subunit gene promoter that are important for its synaptic expression. We used, for this purpose, a rapid method of in vivo expression after DNA injection into the muscle tissue [Wolff, J. A., Malone, R. W., Williams, P., Chong, W., Acsadi, G., Jani, A. & Felgner, P. L. (1990) Science 247, 1465-1468]. Our results show that a construction containing 83 nucleotides upstream from the transcription start site is sufficient to obtain preferential endplate expression. Moreover, mutation of a MyoD binding site located around position-70 does not alter this synaptic expression. We also studied the expression of this promoter in vitro in muscle primary cultures and showed the presence of a positive element between positions -122 and -83. Comparison of in vivo and in vitro results reveals that the elements important for in vivo localization at the synapse and in vitro expression in cultured muscle cells may differ.
MeSH Terms
Animals
Base Sequence
Cells, Cultured
Cloning, Molecular
DNA/genetics
Gene Expression
Genomic Library
Luciferases/genetics,metabolism
Macromolecular Substances
Mice
Mice, Inbred DBA
Molecular Sequence Data
Muscles/innervation,physiology
Mutagenesis, Site-Directed
Oligodeoxyribonucleotides
Promoter Regions, Genetic
Receptors, Cholinergic/genetics
Recombinant Fusion Proteins/metabolism
Restriction Mapping
Sequence Deletion
Synapses/physiology
Transfection
Chemicals
Macromolecular Substances
Oligodeoxyribonucleotides
Receptors, Cholinergic
Recombinant Fusion Proteins
DNA
Luciferases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Duclert A
Centre National de la Recherche Scientifique 0210, Département des Biotechnologies, Institut Pasteur, Paris, France.
Savatier N
Changeux J P
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