title: Gene-specific disruption of endocannabinoid receptor 1 (cnr1a) by ethanol probably leads to the development of fetal alcohol spectrum disorder (FASD) phenotypes in Japanese rice fish (Oryzias latipes) embryogenesis.
abstract: The present study was designed to investigate the probable roles played by cannabinoid (CB) receptors in fetal alcohol spectrum disorder (FASD) induction in Japanese rice fish (Oryzias latipes). Searching of public databases (GenBank, Ensembl) indicated that the Japanese rice fish genome includes three human ortholog CB receptor genes (cnr1a, cnr1b and cnr2). Quantitative real-time PCR (qPCR) and whole mount in situ hybridization (WMISH) techniques were used to analyze the expression of these cnr genes during Japanese rice fish embryogenesis and also in response to developmental ethanol exposure. qPCR analyses showed that the expression of all three CB receptor genes were developmentally regulated and only cnr2 showed maternal expression. The mRNA concentrations of these genes were found to be enhanced after 3 dpf and attained maximal levels either prior to or after hatching. WMISH technique indicated that all three cnr genes were expressed in the head region of hatchlings. During development, ethanol selectively attenuated the expression of cnr1a mRNA only. Blocking of cnr1a mRNA by CB1 receptor antagonists rimonabant (10-20 μM) or AM251 (0.2-1 μM) 0-2 dpf were unable to induce any FASD-related phenotypic features in embryos or in hatchlings. However, continuous exposure of the embryos (0-6 dpf) to AM251 (1 μM) was able to reduce the hatching efficiency of the embryos. Our data indicated that in Japanese rice fish, ethanol disrupted the expression of only cnr1a in a concentration-dependent manner that induced delay in hatching and might be responsible for the development of FASD phenotypes.
authors.affiliation: National Center for Natural Product Research, University of Mississippi, University, MS, USA; Department of BioMolecular Sciences, Division of Pharmacology, University of Mississippi, University, MS, USA. Electronic address: asok@olemiss.edu.
authors.forename: Asok K
authors.initials: AK
authors.lastname: Dasmahapatra
authors.affiliation: National Center for Natural Product Research, University of Mississippi, University, MS, USA.
authors.forename: Ikhlas A
authors.initials: IA
authors.lastname: Khan
mesh_terms: D000818:Animals; D004625:Embryo, Nonmammalian / Q000002:abnormalities / Q000187:drug effects*; D047108:Embryonic Development / Q000187:drug effects; D000431:Ethanol / Q000633:toxicity*; D063647:Fetal Alcohol Spectrum Disorders; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D009990:Oryzias / Q000196:embryology*; D043882:Receptors, Cannabinoid / Q000235:genetics / Q000378:metabolism*
chemical_list: D043882:Receptors, Cannabinoid; D000431:Ethanol
keywords: Alcohol; Development; Endocannabinoid receptors; Fetal alcohol spectrum disorder; Japanese rice fish