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  Assembly of an adult type acetylcholine receptor in a mouse cell line transfected with rat muscle ϵ-subunit DNA

Criado, M., Koenen, M., & Sakmann, B. (1990). Assembly of an adult type acetylcholine receptor in a mouse cell line transfected with rat muscle ϵ-subunit DNA. FEBS Letters, 270(1-2), 95-99. doi:10.1016/0014-5793(90)81242-G.

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 Creators:
Criado, Manuel, Author
Koenen, Michael1, 2, 3, 4, Author              
Sakmann, Bert4, Author              
Affiliations:
1Molecular anatomy of the neuromuscular junction, Max Planck Institute for Medical Research, Max Planck Society, ou_1497727              
2Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497704              
3Working Group Witzemann / Koenen, Max Planck Institute for Medical Research, Max Planck Society, ou_1497748              
4Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497701              

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Free keywords: Acetylcholine receptor; Muscle development; Transfection
 Abstract: The mouse muscle cell line BC3H-1 expresses an acetylcholine receptor (AChR) composed of α-,β-, and δ-subunits [1]. The functional characteristics of this AChR are comparable to the non-synaptic AChR subtype in mouse muscle [2,3]. To investigate the role of the ϵ-subunit, which is believed to replace the γ-subunit in forming the adult AChR subtype [4], BC3H-1 cells were stably transfected with cDNA encoding the rat muscle AChR ϵ-subunit. Expression of this cDNA was under the control of a heat shock promoter, and the plasmid carried the neomycin resistance gene for selection. Several clones were isolated that had integrated the plasmid DNA in a stable form and produced ϵ-subunit specific RNA after heat induction. Single-channel current recording from cells which contained abundant ϵ-subunit mRNA identified a novel AChR channel having a larger conductance than the native AChR in these cells. These results suggest that the rat muscle ϵ-subunit may assemble with mouse muscle α-, β- and δ-subunits to form a mouse-rat hybrid AChR with properties similar to that of end-plate channels in the mature mammalian neuromuscular synapse. The novel AChR channel appears in the surface membrane within a few hours following the rise in ϵ-subunit mRNA. Thus, the notion that replacement of the γ-subunit by the ϵ-subunit during development is the result of the postnatal rise in the level of ϵ-subunit specific mRNA is further supported.

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Language(s): eng - English
 Dates: 1990-07-241990-06-252001-10-291990-09-17
 Publication Status: Published in print
 Pages: 9
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1016/0014-5793(90)81242-G
 Degree: -

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Title: FEBS Letters
  Other : FEBS Lett.
Source Genre: Journal
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Publ. Info: Amsterdam : Elsevier
Pages: - Volume / Issue: 270 (1-2) Sequence Number: - Start / End Page: 95 - 99 Identifier: ISSN: 0014-5793
CoNE: https://pure.mpg.de/cone/journals/resource/954925399501