Home LiteratureArticle Details
PMID: 8464921 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

An 83-nucleotide promoter of the acetylcholine receptor epsilon-subunit gene confers preferential synaptic expression in mouse muscle.

Duclert A, Savatier N, Changeux JP

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
References (31)
31 references, click to expand
  1. Two adjacent MyoD1-binding sites regulate expression of the acetylcholine receptor alpha-subunit gene.
    Nature. 1990 May 24;345(6273):353-5 PMID: 2342565
  2. Direct gene transfer into mouse muscle in vivo.
    Science. 1990 Mar 23;247(4949 Pt 1):1465-8 PMID: 1690918
  3. Paired MyoD-binding sites regulate myosin light chain gene expression.
    Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1242-6 PMID: 1847512
  4. An acetylcholine receptor alpha-subunit promoter conferring preferential synaptic expression in muscle of transgenic mice.
    EMBO J. 1991 Mar;10(3):625-32 PMID: 1900467
  5. Nerve-dependent modulation of acetylcholine receptor epsilon-subunit gene expression.
    J Neurosci. 1991 May;11(5):1291-9 PMID: 2027048
  6. Functional adult acetylcholine receptor develops independently of motor innervation in Sol 8 mouse muscle cell line.
    EMBO J. 1991 Sep;10(9):2411-8 PMID: 1868829
  7. Different mechanisms regulate muscle-specific AChR gamma- and epsilon-subunit gene expression.
    EMBO J. 1991 Oct;10(10):2957-64 PMID: 1655408
  8. Induction of adult-type nicotinic acetylcholine receptor gene expression in noninnervated regenerating muscle.
    Neuron. 1991 Oct;7(4):649-58 PMID: 1931052
  9. A developmental and tissue-specific enhancer in the mouse skeletal muscle acetylcholine receptor alpha-subunit gene regulated by myogenic factors.
    J Biol Chem. 1991 Nov 25;266(33):22588-96 PMID: 1658001
  10. Compartmentalization of acetylcholine receptor gene expression during development of the neuromuscular junction.
    Cold Spring Harb Symp Quant Biol. 1990;55:381-96 PMID: 2132828
  11. Selective expression of an acetylcholine receptor-lacZ transgene in synaptic nuclei of adult muscle fibers.
    Development. 1991 Dec;113(4):1181-91 PMID: 1811935
  12. Spatial restriction of AChR gene expression to subsynaptic nuclei.
    Development. 1992 Mar;114(3):545-53 PMID: 1618127
  13. Regulation of an acetylcholine receptor LacZ transgene by muscle innervation.
    Neuroreport. 1992 Nov;3(11):973-6 PMID: 1482767
  14. Host-range mutants of adenovirus type 5 defective for growth in HeLa cells.
    Virology. 1977 Mar;77(1):319-29 PMID: 841862
  15. In vitro differentiation of satellite cells isolated from normal and dystrophic mammalian muscles. A comparison with embryonic myogenic cells.
    Cell Differ. 1980 Dec;9(6):357-68 PMID: 7438216
  16. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells.
    Mol Cell Biol. 1982 Sep;2(9):1044-51 PMID: 6960240
  17. A short amino acid sequence able to specify nuclear location.
    Cell. 1984 Dec;39(3 Pt 2):499-509 PMID: 6096007
  18. Concentration of acetylcholine receptor mRNA in synaptic regions of adult muscle fibres.
    Nature. 1985 Sep 5-11;317(6032):66-8 PMID: 3839905
  19. Transcriptional regulation of the muscle creatine kinase gene and regulated expression in transfected mouse myoblasts.
    Mol Cell Biol. 1986 Aug;6(8):2855-64 PMID: 3785216
  20. A 5'-flanking region of the chicken acetylcholine receptor alpha-subunit gene confers tissue specificity and developmental control of expression in transfected cells.
    Mol Cell Biol. 1987 Feb;7(2):951-5 PMID: 3821734
  21. The muscle creatine kinase gene is regulated by multiple upstream elements, including a muscle-specific enhancer.
    Mol Cell Biol. 1988 Jan;8(1):62-70 PMID: 3336366
  22. Detection of the nicotinic acetylcholine receptor alpha-subunit mRNA by in situ hybridization at neuromuscular junctions of 15-day-old chick striated muscles.
    EMBO J. 1988 Mar;7(3):603-9 PMID: 3396533
  23. Firefly luciferase luminescence assays using scintillation counters for quantitation in transfected mammalian cells.
    Anal Biochem. 1988 Jun;171(2):404-8 PMID: 3407940
  24. Isolation and characterization of the mouse acetylcholine receptor delta subunit gene: identification of a 148-bp cis-acting region that confers myotube-specific expression.
    J Cell Biol. 1988 Dec;107(6 Pt 1):2271-9 PMID: 3198687
  25. Developmental regulation of five subunit specific mRNAs encoding acetylcholine receptor subtypes in rat muscle.
    FEBS Lett. 1989 Jan 2;242(2):419-24 PMID: 2914620
  26. MyoD is a sequence-specific DNA binding protein requiring a region of myc homology to bind to the muscle creatine kinase enhancer.
    Cell. 1989 Sep 8;58(5):823-31 PMID: 2550138
  27. Muscle-specific gene expression controlled by a regulatory element lacking a MyoD1-binding site.
    Nature. 1989 Oct 26;341(6244):716-20 PMID: 2797207
  28. A cell type-specific enhancer drives expression of the chick muscle acetylcholine receptor alpha-subunit gene.
    Neuron. 1988 Aug;1(6):527-34 PMID: 3272179
  29. Spatial and temporal expression of acetylcholine receptor RNAs in innervated and denervated rat soleus muscle.
    Neuron. 1989 Aug;3(2):219-28 PMID: 2483113
  30. Imprinting of acetylcholine receptor messenger RNA accumulation in mammalian neuromuscular synapses.
    Nature. 1990 Apr 5;344(6266):544-7 PMID: 2320126
  31. The myoD gene family: nodal point during specification of the muscle cell lineage.
    Science. 1991 Feb 15;251(4995):761-6 PMID: 1846704
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1993-04-01
Pages
3043-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC46233
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]