Relative expression of clock genes in cultured goldfish liver treated with ghrelin, adenylate cyclase (AC) activator and protein kinase A (PKA) inhibitor.

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Relative expression of clock genes in cultured goldfish liver treated with ghrelin, adenylate cyclase (AC) activator and protein kinase A (PKA) inhibitor for 1 h. Relative expression of clock genes in cultured goldfish liver treated with ghrelin, adenylate cyclase (AC) activator and protein kinase A (PKA) inhibitor for 1 h. The PKA inhibitor H89 (50 or 200 μmol l−1) was added 15 min prior to the addition of the AC activator forskolin (10 μmol l−1) or ghrelin (GRL; 10 nmol l−1). Data obtained by RT-qPCR are shown as means±s.e.m. (n=6, liver aliquots from 6 different fish) in relative units (ΔΔCt method). Asterisks indicate significant inhibition by H89 of forskolin stimulation (interaction P<0.05; two-way ANOVA). Lowercase letters indicate differences among forskolin and H89 groups (SNK test). ¶ indicates significant inhibition by H89 of ghrelin stimulation (interaction P<0.05; two-way ANOVA). Capital letters indicate differences among ghrelin and H89 groups (SNK test). When the two-way ANOVA was significant (P<0.05), but there was no significant interaction between factors, ‡ indicates a significant effect of ghrelin (gper1a, gper2a, gper3, gclock1a) and § indicates a significant effect of H89 (gper3) compared with controls. Aída Sánchez-Bretaño et al. J Exp Biol 2017;220:1295-1306 © 2017. Published by The Company of Biologists Ltd