PUBLICATION
Cloning and characterization of muscarinic receptor genes from the nile tilapia (Oreochromis niloticus)
- Authors
- Seo, J.S., Kim, M.S., Park, E.M., Ahn, S.J., Kim, N.Y., Jung, S.H., Kim, J.W., Lee, H.H., and Chung, J.K.
- ID
- ZDB-PUB-090330-18
- Date
- 2009
- Source
- Molecules and cells 27(3): 383-390 (Journal)
- Registered Authors
- Ahn, Sang Jung
- Keywords
- carbachol, gene cloning, intestinal motility, muscarinic receptor, nile tilapia, Oreochromis niloticus, phopholipase C, real-time PCR
- MeSH Terms
-
- Animals
- Cichlids/genetics*
- Cichlids/metabolism
- Cloning, Molecular
- Muscle, Smooth/chemistry
- Muscle, Smooth/metabolism
- Receptors, Muscarinic/genetics*
- Receptors, Muscarinic/metabolism
- PubMed
- 19326086 Full text @ Mol. Cells
Citation
Seo, J.S., Kim, M.S., Park, E.M., Ahn, S.J., Kim, N.Y., Jung, S.H., Kim, J.W., Lee, H.H., and Chung, J.K. (2009) Cloning and characterization of muscarinic receptor genes from the nile tilapia (Oreochromis niloticus). Molecules and cells. 27(3):383-390.
Abstract
To investigate the regulatory mechanism underlying the contractile response in the intestinal smooth muscle of the nile tilapia (Orechromis niloticus), we used pharmacologic and molecular approaches to identify the muscarinic subreceptors and the intracellular signaling pathways involved in this motility. Myography assays revealed that an M1- and M3-subtype selective antagonist, but not a M2-subtype selective antagonist, inhibited carbachol HCI (CCH)-induced intestinal smooth muscle contraction. In addition, a phospholipase C inhibitor, but not an adenylate cyclase inhibitor, blocked the contractile response to CCH. We also cloned five muscarinic genes (OnM2A, OnM2B, OnM3, OnM5A, and OnM5B) from the nile tilapia. In the phylogenetic analysis and sequence comparison to compare our putative gene products (OnMs) with the sequences obtained from the near complete teleost genomes, we unexpectedly found that the teleost fish have respectively two paralogous genes corresponding to each muscarinic subreceptor, and other teleost fish, except zebrafish, do not possess muscarinic subreceptor M1. In addition, the expression pattern of the nile tilapia muscarinic subreceptor transcripts during CCH-induced intestinal smooth muscle contraction in the proximal intestinal tissue was analyzed by real-time PCR surveys and it was demonstrated that CCH increased the OnMs m RNA expression rapidly and transiently.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping