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Biomed Res Int
2014 Jan 01;2014:654710. doi: 10.1155/2014/654710.
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Regulation of melanopsins and Per1 by α -MSH and melatonin in photosensitive Xenopus laevis melanophores.
Moraes MN
,
dos Santos LR
,
Mezzalira N
,
Poletini MO
,
Castrucci AM
.
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α-MSH and light exert a dispersing effect on pigment granules of Xenopus laevis melanophores; however, the intracellular signaling pathways are different. Melatonin, a hormone that functions as an internal signal of darkness for the organism, has opposite effects, aggregating the melanin granules. Because light functions as an important synchronizing signal for circadian rhythms, we further investigated the effects of both hormones on genes related to the circadian system, namely, Per1 (one of the clock genes) and the melanopsins, Opn4x and Opn4m (photopigments). Per1 showed temporal oscillations, regardless of the presence of melatonin or α-MSH, which slightly inhibited its expression. Melatonin effects on melanopsins depend on the time of application: if applied in the photophase it dramatically decreased Opn4x and Opn4m expressions, and abolished their temporal oscillations, opposite to α-MSH, which increased the melanopsins' expressions. Our results demonstrate that unlike what has been reported for other peripheral clocks and cultured cells, medium changes or hormones do not play a major role in synchronizing the Xenopus melanophore population. This difference is probably due to the fact that X. laevis melanophores possess functional photopigments (melanopsins) that enable these cells to primarily respond to light, which triggers melanin dispersion and modulates gene expression.
Figure 1. Quantitative PCR of Per1 in X. laevis melanophores after 2 medium changes in DD (a); 2 medium changes in LD 12 : 12 (b); 10−9 M α-MSH in DD (c); and 10−9 M melatonin in the photophase of LD 12 : 12 (d). Each bar represents the mean, ± SE, mRNA relative to the noted ZT. (a) Significantly different from (b); (c) significantly different from (d) by one-way ANOVA, Tukey's post-test; ∗ means significantly different from the respective control at the same ZT, by two-way ANOVA and Bonferroni's post-test. n number shown on top of each bar in this and the following figures.
Figure 2. Quantitative PCR of Opn4x in X. laevis melanophores after 2 medium changes in the photophase of LD 12 : 12 (a); 10−9 M melatonin in the photophase of LD 12 : 12 (b); 2 medium changes in the scotophase of LD 12 : 12 (c); 10−9 M melatonin in the scotophase of LD 12 : 12 (d); 2 medium changes in DD (e); 10−9 M α-MSH in DD (f). Each bar represents the mean, ± SE, mRNA relative to the noted ZT. (a) Significantly different from (b); (c) significantly different from (d), except in 2 C where (a) is different from (b) and (c) is different from (a) and (d) by one-way ANOVA and Tukey's post-test; ∗ means significantly different from the respective control at the same ZT, by two-way ANOVA and Bonferroni's post-test.
Figure 3. Quantitative PCR of Opn4m in X. laevis melanophores after to 2 medium changes in the photophase of LD 12 : 12 (a); 10−9 M melatonin in the photophase of LD 12 : 12 (b); 2 medium changes in the scotophase of LD 12 : 12 (c); 10−9 M melatonin in the scotophase of LD 12 : 12 (d); 2 medium changes in DD (e); 10−9 M α-MSH in DD (f). Each bar represents the mean, ± SE, mRNA relative to the noted ZT. (a) Significantly different from (b); (c) significantly different from (d), by one-way ANOVAandTukey's post-test; ∗ means significantly different from the respective control at the same ZT, by two-way ANOVA and Bonferroni's post-test.
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