Cloning and functional characterization of a Caenorhabditis elegans muscarinic acetylcholine receptor

J. M. Hwang, D. J. Chang, U. S. Kim, Yong-Seok Lee, Y. S. Park, B. K. Kaang, N. J. Cho

Research output: Contribution to journalArticleResearchpeer-review

39 Citations (Scopus)

Abstract

A cDNA clone encoding a muscarinic acetylcholine receptor (mAChR) has been isolated from the nematode Caenorhabditis elegans. The nematode mAChR, consisted of 585 amino acids, displays a high degree of amino acid sequence homology to other invertebrate and vertebrate mAChRs. Excluding a highly variable middle portion of the third intracellular loop, the C. elegans mAChR shares about 51% amino acid sequence identity with a Drosophila mAChR and 42-44% identity with human ml-m5 mAChR subtypes. Comparison of the cDNA sequence with the corresponding genomic sequence reveals that the C. elegans mAChR gene contains ten introns, eight of them in the coding region. Pharmacological profiles of the C. elegans mAChR expressed in Chinese hamster ovary (CHO) cells were shown to be similar to those of mammalian counterparts, indicating that ligand binding domains of the receptor have been conserved during evolution. When this cloned receptor was expressed in Xenopus oocytes, acetylcholine evoked a transient Cl- current. Furthermore, activation of the receptor with oxotremorine, acetylcholine or carbachol resulted in the stimulation of phosphatidylinositol metabolism in CHO cells, suggesting that the receptor is coupled to phospholipase C activation.

Original languageEnglish
Pages (from-to)415-424
Number of pages10
JournalReceptors and Channels
Volume6
Issue number6
StatePublished - 1 Dec 1999

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Cloning
Caenorhabditis elegans
Muscarinic Receptors
Organism Cloning
Cricetulus
Amino Acids
Acetylcholine
Ovary
Complementary DNA
Chemical activation
Oxotremorine
Amino Acid Sequence Homology
Carbachol
Type C Phospholipases
Invertebrates
Xenopus
Phosphatidylinositols
Metabolism
Introns
Drosophila

Keywords

  • Caenorhabditis elegans
  • Electrophysiology
  • Heterologous gene expression
  • Muscarinic acetylcholine receptor
  • Phosphatidylinositol metabolism

Cite this

Hwang, J. M., Chang, D. J., Kim, U. S., Lee, Y-S., Park, Y. S., Kaang, B. K., & Cho, N. J. (1999). Cloning and functional characterization of a Caenorhabditis elegans muscarinic acetylcholine receptor. Receptors and Channels, 6(6), 415-424.
Hwang, J. M. ; Chang, D. J. ; Kim, U. S. ; Lee, Yong-Seok ; Park, Y. S. ; Kaang, B. K. ; Cho, N. J. / Cloning and functional characterization of a Caenorhabditis elegans muscarinic acetylcholine receptor. In: Receptors and Channels. 1999 ; Vol. 6, No. 6. pp. 415-424.
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abstract = "A cDNA clone encoding a muscarinic acetylcholine receptor (mAChR) has been isolated from the nematode Caenorhabditis elegans. The nematode mAChR, consisted of 585 amino acids, displays a high degree of amino acid sequence homology to other invertebrate and vertebrate mAChRs. Excluding a highly variable middle portion of the third intracellular loop, the C. elegans mAChR shares about 51{\%} amino acid sequence identity with a Drosophila mAChR and 42-44{\%} identity with human ml-m5 mAChR subtypes. Comparison of the cDNA sequence with the corresponding genomic sequence reveals that the C. elegans mAChR gene contains ten introns, eight of them in the coding region. Pharmacological profiles of the C. elegans mAChR expressed in Chinese hamster ovary (CHO) cells were shown to be similar to those of mammalian counterparts, indicating that ligand binding domains of the receptor have been conserved during evolution. When this cloned receptor was expressed in Xenopus oocytes, acetylcholine evoked a transient Cl- current. Furthermore, activation of the receptor with oxotremorine, acetylcholine or carbachol resulted in the stimulation of phosphatidylinositol metabolism in CHO cells, suggesting that the receptor is coupled to phospholipase C activation.",
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Hwang, JM, Chang, DJ, Kim, US, Lee, Y-S, Park, YS, Kaang, BK & Cho, NJ 1999, 'Cloning and functional characterization of a Caenorhabditis elegans muscarinic acetylcholine receptor', Receptors and Channels, vol. 6, no. 6, pp. 415-424.

Cloning and functional characterization of a Caenorhabditis elegans muscarinic acetylcholine receptor. / Hwang, J. M.; Chang, D. J.; Kim, U. S.; Lee, Yong-Seok; Park, Y. S.; Kaang, B. K.; Cho, N. J.

In: Receptors and Channels, Vol. 6, No. 6, 01.12.1999, p. 415-424.

Research output: Contribution to journalArticleResearchpeer-review

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