锦鲤早期发育过程中色素代谢相关酶的活性和基因表达变化

Changes of pigment-related enzyme activity and gene expression at early developmental stage of koi carp

  • 摘要: 通过显微观察、酶联免疫吸附及荧光定量PCR等方法,探究了红白、红黑锦鲤(Cyprinus carpio var. koi) 子一代 (F1)早期体色发育过程及不同发育时期色素代谢相关酶(酪氨酸酶、胱氨酸酶)的活性和基因(agoutimc1rmitftyr)的表达变化。结果表明,锦鲤出膜后呈淡黄色半透明,黑色素细胞在出膜8 d后开始出现。胱氨酸酶活性随锦鲤发育呈升高趋势,出膜7 d后显著升高(P<0.05);7日龄仔鱼前,红黑组与红白组间无显著差异(P>0.05);出膜7 d后红黑组比同时期红白组显著升高(P<0.05)。酪氨酸酶在锦鲤发育过程中也呈升高趋势,但略有起伏,眼色素积累期及出膜23 d后酪氨酸酶活性均比相邻各期高(P<0.05);组间比较发现发育各期红黑组均比红白组高(P<0.05)。表明酪氨酸酶、胱氨酸酶活性变化与锦鲤色素细胞发育存在密切关联。agoutimc1rmitftyr 4个基因均在原肠期与神经胚期时表达最高(P<0.01),随后均呈下降趋势,表明这4个基因可能在胚胎发育早期的体色形成过程中发挥着重要作用。

     

    Abstract: We investigated the changes of tyrosinase and cystinase activities, as well as the expressions of tyr, mitf, agouti and mc1r mRNAs at the early developmental stage of Kohaku and Hi-Utsuri koi carp (Cyprinus carpio) by using histological observation, enzyme-linked immunoassay and quantitative reverse transcription-polymerase chain reaction methods. The results show that the hatched carps became light yellow and translucent. The melanocytes occurred on 8 dph (day-post-hatching). The activity of cystinase increased with the koi carp development, and no significant difference was observed between Kohaku and Hi-Utsuri groups before 7 dph (P>0.05), while Hi-Utsuri group showed significantly higher enzyme activity than Kohaku group at the same stages after 7 dph (P<0.05). Tyrosinase activity showed an up-regulation trend with slight fluctuation during the development of koi carp, and peaked at the ocular pigment stage and on 23 dph (P<0.05). The enzyme activity was significantly higher in Hi-Utsuri group than in Kohaku group at each developmental stage (P<0.05), which indicates that tyrosinase and cystinase activities were closely associated with pigment cell development of koi carp. The expressions of agouti, mc1r, mitf and tyr mRNAs peaked in gastrula and blastula periods (P<0.01), then decreased. Thus, agouti, mc1r, mitf and tyr mRNAs might play an important role in the formation of body colors in early embryonic development.

     

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