Cloning, characterization and expression of the D2 dopamine receptor from the tilapia pituitary

B Levavi-Sivan; Joseph Aizen; A Avitan

doi:10.1016/j.mce.2005.03.010

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Cloning, characterization and expression of the D2 dopamine receptor from the tilapia pituitary

Journal article

Peer reviewed

Cloning, characterization and expression of the D2 dopamine receptor from the tilapia pituitary

B Levavi-Sivan, Joseph Aizen and A Avitan

Molecular and Cellular Endocrinology, Vol.236(1-2), pp.17-30

2005

DOI: https://doi.org/10.1016/j.mce.2005.03.010

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Abstract

Biological Sciences

Dopamine receptor

Estradiol

cAMP

adenylate cyclase

steroid feedback

tilapia

mullet

catfish

pituitary

A full-length cDNA encoding a dopamine receptor (DA-R) was obtained from the pituitary of tilapia (ta). This cDNA encodes a protein of 469 amino acids that exhibits the typical arrangement of GPCR. The taDA-R shows high similarity to the DA-Rs of mullet and fugu, and over 70% similarity to Xenopus, mouse and turkey D2 DA-Rs. Northern blot analysis revealed transcript for a single transcript in the pituitary, of approximately 3 kb. In a Southern analysis, the tilapia probe recognized specific bands in the genomic DNA of both mullet and catfish, suggesting high similarity between the corresponding genes. Phylogenetic analysis clearly aligned the taDA-D2-R with all vertebrate D2-like receptor sequences cloned to date, and it was therefore designated taDA-D2-R. taDA-D2-R was transiently expressed in COS-7 cells together with the reporter construct CRE-luciferase. Addition of the specific D2 dopamine agonists quinpirole or bromocriptine, in the presence of forskolin, led to a dose-dependent decrease in forskolin-induced cAMP levels. Both agonists yielded the same maximal inhibition (around 40%). However, the potency of taDA-D2-R for bromocriptine was higher than for quinpirole. As established for mammalian D2-like receptors, stimulation of the taDA-D2-R with quinpirole triggers pertussis-toxin-sensitive Gi/o-mediated, but not Gs-mediated signaling. In contrast to mammals, PCR analysis gave no evidence of alternative splicing in taDA-D2-R. Pharmacological and genetic manipulation of the taDA-D2-R should enable us to better define its physiological role and to further explore the usefulness of fish as a model system for understanding dopaminergic function in higher organisms.

Details

Title: Cloning, characterization and expression of the D2 dopamine receptor from the tilapia pituitary
Authors: B Levavi-Sivan (Author) - Hebrew University of Jerusalem, Israel
Joseph Aizen (Author) - Hebrew University of Jerusalem, Israel
A Avitan (Author) - Hebrew University of Jerusalem, Israel
Publication details: Molecular and Cellular Endocrinology, Vol.236(1-2), pp.17-30
Publisher: Elsevier Ireland Ltd.
Date published: 2005
DOI: 10.1016/j.mce.2005.03.010
ISSN: 0303-7207
Organisation Unit: University of the Sunshine Coast, Queensland; GeneCology Research Centre - Legacy; Centre for Bioinnovation
Language: English
Record Identifier: 99448781002621
Output Type: Journal article

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