title: Expression of tachykinin receptors (tacr1a and tacr1b) in zebrafish: influence of cocaine and opioid receptors.
abstract: Opioid and tachykinin receptors (TACRs) are closely related in addiction and pain processes. In zebrafish, opioid receptors have been cloned and characterized both biochemically and pharmacologically. However, the tacr1 gene has not yet been described in zebrafish. The aim of this research was to identify the tacr1 gene, study the effects of cocaine on tacr1, and analyze the interaction between tacr1 and opioid receptors. We have identified a duplicate of tacr1 gene in zebrafish, designated as tacr1a and tacr1b. Phylogenetic analyses revealed an alignment of these receptors in the Tacr1 fish cluster, with a clear distinction from other TACR1s of amphibians, birds, and mammals. Our qPCR results showed that tacr1a and tacr1b mRNAs are expressed during embryonic development. Whole-mount in situ hybridization showed tacr1 expression in the CNS and in the peripheral tissues. Cocaine (1.5 μM) induced an upregulation of tacr1a and tacr1b at 24 and 48 h post-fertilization (hpf; except for tacr1a at 48 hpf, which was downregulated). By contrast, HEK-293 cells transfected with tacr1a and tacr1b and exposed to cocaine showed a downregulation of tacr1s. The knockdown of ZfDOR2 and ZfMOR, opioid receptors, induced a down- and upregulation of tacr1a and tacr1b respectively. In conclusion, tacr1a and tacr1b in zebrafish are widely expressed throughout the CNS and peripherally, suggesting a critical role of these tacr1s during embryogenesis. tacr1a and tacr1b mRNA expression is altered by cocaine exposure and by the knockdown of opioid receptors. Thus, zebrafish can provide clues for a better understanding of the relationship between tachykinin and opioid receptors in pain and addiction during embryonic development.
authors.affiliation: Department of Biochemistry and Molecular Biology, Institute of Neuroscience of Castilla y León, University of Salamanca, Salamanca, Spain.
authors.forename: Roger
authors.initials: R
authors.lastname: López-Bellido
authors.forename: Katherine
authors.initials: K
authors.lastname: Barreto-Valer
authors.forename: Raquel E
authors.initials: RE
authors.lastname: Rodríguez
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D003042:Cocaine / Q000494:pharmacology; D005786:Gene Expression Regulation; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011955:Receptors, Drug / Q000235:genetics / Q000378:metabolism; D011957:Receptors, Opioid / Q000235:genetics / Q000378:metabolism; D018039:Receptors, Tachykinin / Q000145:classification / Q000235:genetics*; D016415:Sequence Alignment; D014162:Transfection; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D012333:RNA, Messenger; D011955:Receptors, Drug; D011957:Receptors, Opioid; D018039:Receptors, Tachykinin; C061445:cocaine receptor; D003042:Cocaine