金边鲤皮肤黑色素合成及转运相关基因表达分析

Skin melanin synthesis of Jinbian carp and expression analysis of transport related genes

【目的】研究影响金边鲤皮肤色素沉着的关键基因及信号通路,揭示其皮肤颜色差异形成的原因,为金边鲤良种培育提供候选基因信息。【方法】基于突变型金边鲤黄色皮肤(Y组)和黑色皮肤(B组)及野生型金边鲤黑色皮肤(W组)的转录组测序数据,筛选分析获得部分共有差异表达基因,利用实时荧光定量PCR检测差异表达基因和黑色素合成相关基因的表达情况,并分析各基因间的相关性。【结果】依据转录组数据筛选获得ACT3、MYH2、TPM1、TPM2和TPM3共5个来源于心肌肾上腺素信号通路及与细胞动力相关的基因;除ACT3基因外,其余4个基因(MYH2、TPM1、TPM2和TPM3)在Y组皮肤中的相对表达量均显著低于B组和W组(P<0.05,下同),与转录组分析的差异基因表达趋势一致。MC-1R、TYR、TYRP1和TYRP2等4个黑色素合成相关基因在金边鲤皮肤中的表达情况为:B组皮肤中MC-1R基因的相对表达量显著高于Y组和W组;TYR、TYRP1和TYRP2基因在3组皮肤中的相对表达量排序均为W组>B组>Y组,其中W组的相对表达量显著高于B组和Y组。相关性分析结果显示,MC-1R和TYR基因与ACT3、MYH2、TPM1、TPM2和TPM3基因呈显著或极显著(P<0.01)正相关,而TYRP1和TYRP2基因与这5个差异表达基因均无显著相关性(P>0.05)。【结论】ACT3、MYH2、TPM1、TPM2和TPM3基因及其所在的心肌肾上腺素信号通路可能影响黑色素合成与运动,且与黑色素合成相关基因存在相关性,可作为金边鲤体色变异研究中的标记基因。

【Objective】The key genes and signaling pathways affecting skin pigmentation of Jinbian carp were studied,to reveal the causes of skin color variation mechanism and provide candidate gene information for further cultivating new varieties of Jinbian carp.【Method】Part of differentially co-expressed genes were screened based on the transcriptome sequencing data of yellow skin(group Y)and black skin(group B)in mutant type,and black skin(group W)in wild type of Jinbian carp,thenreal-time fluorescence quantitative PCR was used to detect the expressions of co-differentially expressed genes and genes related to melanin synthesis,and the correlations among the genes were analyzed.【Result】A total of five genes,including ACT3,MYH2,TPM1,TPM2 and TPM3,were derived from the adrenergic signaling in cardiomyocytes pathways and associated with cell power were screened according to the transcriptome data. Apart from ACT3,the mRNA expression levels of the other fourgenes(MYH2,TPM1,TPM2 and TPM3)in group Y were all significantly lower than group B and group W(P<0.05,the same below),which was consistent with the trend of the result in transcriptome analysis. The mRNA expression levels of four genesrelating to melanin synthetic,including MC-1 R,TYR,TYRP1 and TYRP2 were detected,which showed that the relative expression of MC-1 R gene in group B was significantly higher than group Y and group W. The expression of TYR,TYRP1 and TYRP2 in the skin of the three groups showed an order of group W>group B>group Y,in which group W was significantly higher than group B and group Y. Correlation analysis showed that the mRNA expressions levels of MC-1 R and TYR gene were significantor extremely(P<0.01)positively correlated with ACT3,MYH2,TPM1,TPM2 and TPM3 genes,while TYRP1 and TYRP2 had no significant correlation with the fivegenes(P>0.05).【conclusion】ACT3,MYH2,TPM1,TPM2 and TPM3 genes and their adrenergic signaling in cardiomyocytes pathways may affect the synthesis and movement of melanin,which may be related to the skin color variation of the Jinbian carp. These genes can be used as marker genes in the study of skin color variation of Jinbian carp.