3 research outputs found

    Evolutionary analysis of the osteichthyans secretin GPCR family.

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    <p>The maximum likelihood (ML) optimal tree topology is presented and was constructed with MEGA5. ML bootstrap values higher than 50% are indicated at nodes. To facilitate interpretation, PTHR was used as an outgroup based on the proposed models for secretin GPCR family evolution <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053482#pone.0053482-Cardoso1" target="_blank">[7]</a>. The tree supported the identities of lfGHRHR and xGHRHR<sub>2</sub> as the orthologs of mammalian GHRHR and chicken GHRHR<sub>2</sub> respectively. Accession numbers of the sequences used were listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053482#pone.0053482.s009" target="_blank">Table S2</a>.</p

    Functional characterization of lfGHRHR and xGHRHR<sub>2</sub>.

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    <p>Intracellular cAMP accumulation ([cAMP]<sub>i</sub>) in response to 100 nM of GHRH and related peptides on CHO-K1 cells transfected with (A) lfGHRHR and (D) xGHRHR<sub>2</sub> (*** indicates <i>P</i><0.001, ** indicates <i>P</i><0.01, and * indicates <i>P</i><0.05). Effects of GHRH and related peptides on graded concentrations of peptides on [cAMP]<sub>i</sub> (B) lfGHRHR- and (E) xGHRHR<sub>2</sub>-expressing cells. The intracellular calcium mobilization ([Ca<sup>2+</sup>]<sub>i</sub>) assays of (C) lfGHRHR- and (F) xGHRHR<sub>2</sub>-expressing cells. For [cAMP]<sub>i</sub>, values represent mean ± SEM (n = 4). For ([Ca<sup>2+</sup>]<sub>i</sub>, data were expressed in ΔRFU value (maximum changes in the fluorescence signals from baseline) and converted to percentage of the maximum of xGHRH-induced [Ca<sup>2+</sup>]<sub>i</sub> elevation. Results are expressed as mean ± SEM from at least 10 independent experiments, cell number = 20 to 50. Peptide species: h, human; x, <i>X. laevis</i>, zf, zebrafish <i>D. rerio</i>; gf, goldfish <i>C. auratus</i>.</p

    Gene linkage comparisons of GHRHRs in the Osteichthyes lineage.

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    <p>Genes in vicinity of GHRHR were mapped, and syntenic genes were linked by straight lines. Size of the chromosomal region analyzed was given underneath based on the current edition of Ensembl databases. Syntenic genes encoding other secretin GPCR receptors were drawn in grey boxes, and the conserved flanking genes of GHRHR were drawn in closed boxes. (A) Gene environment of GHRHR in the Sarcopterygii lineage represented by human, mouse, lizard, chicken, frog and coelacanth was compared. Despite the syntenic genomic locations of GHRHR and neighbouring genes from human to avians, GHRHR was not located in frog. (B) Gene environment of GHRHR in the Actinopterygii lineage represented by fugu, tetraodon, stickleback, medaka, and zebrafish. Apart from the less conserved genomic region of zebrafish GHRHR, genes in proximity of other teleost GHRHRs were highly syntenic. However, they displayed an entirely different gene environment when compared to the sarcopterygian GHRHRs. (C) Genomic location analysis of xGHRHR characterized in present work and GHRHR<sub>2</sub> in zebrafish and chicken. Gene synteny could neither be identified inter-species nor between the two GHRHR genes in the same species. The figures were not drawn to scale.</p