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. 2013 Sep;10(3):343–352. doi: 10.1089/zeb.2012.0841

FIG. 4.

FIG. 4.

Response to alpha-melanocyte-stimulating hormone (α-MSH) in the Alamar Blue assay. One (B) or three (A, C–F) fish/well were incubated in 4 mM sodium bicarbonate with 1% Alamar Blue. Fluorescence of the solution was measured at time 0 (4 days postfertilization; dpf) and 24 h later (5 dpf). Data are reported as the relative change in fluorescence (mean±SEM). (A) α-MSH response in wild-type AB fish. B–F show that α-MSH (10 μM) induced a metabolic rate increase in fish lacking 1 or multiple melanocortin receptors. (B) α-MSH response in melanocortin 4-receptor (mc4r), wild-type (+/+), heterozygous (+/−), and knockout (−/−) fish. (C) α-MSH response in fish treated with 0.4, 0.8, or 1.6 ng/embryo of a morpholino targeted to knockdown melanocortin 3-receptor (mc3r). (D) α-MSH response in fish treated with 0.8, 1.6, or 2.4 ng/embryo of a morpholino targeted to knockdown melanocortin 5-receptor b (mc5rb). (E) α-MSH response in mc4r −/− fish treated with 1.6 ng/embryo of a morpholino targeted to knockdown mc3r. (F) α-MSH response in mc4r −/− fish treated with 3.2 ng/embryo standard control morpholino, 1.6 ng/embryo mc5rb morpholino+1.6 ng/embryo control morpholino, or 1.6 ng/embryo mc5rb morpholino and 1.6 ng/embryo mc3r morpholino. Numbers within each bar indicate the number of samples (n). a,bColumns with differing superscripts differ significantly (p<0.05). *Indicates difference from 0 α-MSH dose (p<0.05). †Indicates difference from no morpholino injection (p<0.05).