title: Development of a multiplex gene expression assay for components of the endocrine growth axis in coho salmon.
abstract: This study explores the efficacy of the Quantigene plex (QGP) technology for measuring a panel of endocrine growth-related transcripts in coho salmon, Oncorhynchus kisutch. The QGP technology permits the simultaneous quantification of multiple targeted mRNAs within a single tissue homogenate using sequence-specific probes and requires no reverse transcription (RT) or amplification as is required for RT-quantitative PCR (RT-qPCR). Using liver homogenates from coho salmon under fed and fasted conditions, we compared the detectable fold differences of steady-state mRNA levels between the QGP and probe-based RT-qPCR assays for insulin-like growth factors (igf1 and igf2), insulin-like growth factor binding proteins (igfbp1b, igfbp2a, and igfbp2b), somatolactin receptor (slr), and growth hormone receptors (ghr1 and ghr2). Significant, positive correlations for all genes between the two assays were found. In addition, the relatively low variance of results from the QGP assay suggests that this is a suitable method for a comprehensive analysis of endocrine growth-related transcripts and could potentially be used to develop assays for other gene networks in teleosts.
authors.affiliation: Environmental Physiology Program, Northwest Fisheries Science Center, National Marine Fisheries Service, National Oceanic and Atmospheric Administration, Seattle, WA, United States.
authors.forename: David C
authors.initials: DC
authors.lastname: Metzger
authors.forename: J Adam
authors.initials: JA
authors.lastname: Luckenbach
authors.forename: Jon T
authors.initials: JT
authors.lastname: Dickey
authors.forename: Brian R
authors.initials: BR
authors.lastname: Beckman
mesh_terms: D000818:Animals; D018969:Insulin-Like Growth Factor Binding Proteins / Q000235:genetics; D017684:Oncorhynchus kisutch / Q000235:genetics*; D011986:Receptors, Somatotropin / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D018969:Insulin-Like Growth Factor Binding Proteins; D011986:Receptors, Somatotropin