{"refrec":{"BRefID":58151,"RR":"<b>Rees, C.B.; Li, W.</b> (2004). Development and application of a real-time quantitative PCR assay for determining CYP1A transcripts in three genera of salmonids. <i>Aquat. Toxicol. 66(4)</i>: 357-368. <a href=\"http://dx.doi.org/10.1016/j.aquatox.2003.10.004\" target=\"_blank\">http://dx.doi.org/10.1016/j.aquatox.2003.10.004</a>","BEntID":55785,"PublicFlag":1,"CheckedFlag":1,"wosflag":1,"vabbflag":null,"RefStringPartII":". <i>Aquat. Toxicol. 66(4)</i>: 357-368. <a href=\"https://dx.doi.org/10.1016/j.aquatox.2003.10.004\" target=\"_blank\">https://dx.doi.org/10.1016/j.aquatox.2003.10.004</a>","DocTypID":8,"DocType":"Journal article","MarineFlag":1,"FreshFlag":0,"BrackishFlag":0,"TerrestrialFlag":0,"Authorstring":"Rees, C.B.; Li, W.","OrigTitleTranslFlag":0,"Authorstringtrunc":"Rees, C.B.; Li, W.","Englishabstract":"The expression of CYP1A (cytochrome P450 1A) can be induced by a large array of aromatic and organic compounds in teleost fishes. We developed a real-time quantitative PCR assay useful for measuring β-naphthoflavone (BNF) induction of liver CYP1A mRNA in four salmonid species. First, to obtain necessary information for the design of a cRNA standard, full-length CYP1A cDNA sequences were determined for two <i>Salvelinus</i> species, lake trout (<i>S. namaycush</i>) and brook trout (<i>S. fontinalis</i>). Each cDNA was found to share the same characteristics with known CYP1A sequences of Atlantic salmon (<i>Salmo salar</i>) and rainbow trout (<i>Oncorhynchus mykiss</i>): a start codon, conserved heme-binding region, putative poly-adenylation signal, stop codon, relatively long 3'-untranslated region (UTR; >1 kb), and a protein length of 523 amino acid residues. The brook trout and lake trout CYP1A cDNA's were 2636 and 2672 base pairs (bp) in length and shared greater than 97% coding region sequence identity with Atlantic salmon and rainbow trout CYP1A's. Next, using the generated sequence information, we developed a CYP1A-specific real-time quantitative PCR assay. Primers and a fluorescent-labeled probe were designed from a 68 bp region that was found to be conserved among salmonid CYP1A genes. The assay was designed to allow for simultaneous comparison of CYP1A expression among each experimental group. Finally, groups (n=4-8) of hatchery-raised Atlantic salmon, brook trout, lake trout, and rainbow trout were given an intraperitoneal injection of a corn oil control, 25 mg kg<sup>-1</sup> BNF, or 50 mg kg<sup>-1</sup> BNF and sacrificed after 48 h. Liver tissue was collected and CYP1A mRNA levels were estimated. In all species, BNF treated fish showed 1.8-3.0 orders of magnitude higher CYP1A than control fish. The CYP1A induction levels were not different in fish treated with both dosages. Mean base levels of CYP1A expression ranged from 7.24×10<sup>6</sup> (rainbow trout) to 1.05×10<sup>7</sup>  (brook trout) transcripts µg<sup>-1</sup> total RNA. Mean induced levels of CYP1A expression ranged from 1.07×10<sup>8</sup> (lake trout) to 1.05×10<sup>9</sup> (brook trout) trancripts µg<sup>-1</sup> total RNA.","AbstractOtherLang":null,"BibLvlCode":"AS","StandardTitle":"Development and application of a real-time quantitative PCR assay for determining CYP1A transcripts in three genera of salmonids","OrigTitleLangCode":"en","OrigTitleLangCodeExtended":"eng","OrigTitleLangID":15,"DateLastModified":{"date":"2024-12-10 01:33:17.368041","timezone_type":1,"timezone":"+01:00"},"UserAccessRight":null,"UserAccID":null,"AuthorKeywords":"CYP1A; P450; salmon; trout; real-time quantitative PCR; cDNA sequence","OtherDescriptors":"CYP1A; P450; Salmon; Trout; Real-time quantitative PCR; cDNA sequence","Notes":null,"AnaPub":2004,"MonPub":null,"DateUpdate":"2018-05-17","DateCreate":"2004-03-09","SecASFANote":null,"ConfID":null,"PeerRev":1,"VlizCoreFlag":1,"WoScode":"WOS:000189228100002","VABBcode":null,"OpenAcc":1,"DOI":"10.1016/j.aquatox.2003.10.004"},"refs":null,"anarec":{"AnaID":58151,"PubliDate":2004,"Pagination":"357-368","XtraPublOfAnaID":null,"ISBN":null,"Volume":"66","Issue":"4","BRefMon":null,"BRefMonRR":null,"BRefXtra":null,"BRefXtraRR":null,"SerBRefID":42203,"SerRR":"Aquatic Toxicology. 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