Albino Apostichopus japonicus occur both in the wild and in captivity. The offspring of albino A. japonicus also suffer from albinism. The formation of melanin in the melanocytes is dependant on microphthalmia-associa...Albino Apostichopus japonicus occur both in the wild and in captivity. The offspring of albino A. japonicus also suffer from albinism. The formation of melanin in the melanocytes is dependant on microphthalmia-associated transcription factor (MITF). To investigate the role of MITF in controlling albinism, we cloned the full-length MITF cDNA from A. japonicus and compared MITF mRNA expression in albino and normal A. japonicus. In addition, we used light and electron microscopy to compare histological samples of normal and albino A. japonicus. The body wall of albino adults was characterized by significantly lower levels of MITF expression and lower numbers of epidermal melanocytes, which also contained less melanin. In albino juvenile offspring, MITF expression levels were significantly lower 32 d after fertilization and there were fewer, and less developed, epidermal melanocytes. Thus, we conclude that albino A. japonicus have fewer melanocytes and a reduced ability to synthesize melanin, likely because of lower expression of MITF.展开更多
Microphthalmia-associated transcription factor(MITF)is an essential transactivator in melanin synthesis.To characterize the role of MITF in the pearl mussel Hyriopsis cumingii,the MITF homolog of H.cumingii was isolat...Microphthalmia-associated transcription factor(MITF)is an essential transactivator in melanin synthesis.To characterize the role of MITF in the pearl mussel Hyriopsis cumingii,the MITF homolog of H.cumingii was isolated.The full-length HcMitf cDNA consisted of a 1332-bp with an open reading frame that encode for a 443 amino acid protein that contain a conserved basic helix-loop-helix zipper domain.The HcMitf was found to widespread tissue distribution but expression was higher in purple mussels than in white mussels,mostly in mantle,liver,kidney,gill,and foot with the exception of the adductor mussel.HcMitf and its downstream gene tyrosinase(HcTyr)were highly expressed at the nacre deposition stage after implantation of mantle tissue to produce pearls.Using RNA interference,the expression of HcMitf was reduced by 78%(P<0.01)and expression of HcTyr was also significantly suppressed and consequently total melanin content was decreased(P<0.05).The results suggest that HcMitf plays an important role in melanin synthesis,nacre formation and shell pigmentation in the H.cumingii.展开更多
The present study investigates the expression of microphthalmia-associated transcription factor(MITF) and its contribution to the melanin deposition in Liancheng white ducks.Nested PCR was used to clone the MITF gene ...The present study investigates the expression of microphthalmia-associated transcription factor(MITF) and its contribution to the melanin deposition in Liancheng white ducks.Nested PCR was used to clone the MITF gene sequence from the skin tissue of female Liancheng white ducks.Ultraviolet spectrophotometry was used to detect the melanin deposition.MITF mRNA expression and melanin deposition in different tissues and organs were detected and their correlation was analyzed.The MITF gene(GenBank number: MG516570) was 1 323 bp in length,contains a complete CDS region(34-1 323 bp) and codes 429 amino acids with 100% homology to the MITF of Anas platyrhynchos and over 95% homology to those of Gallus gallus and Coturnix japonica.Genetic evolution analysis reveals a close relationship of Liancheng white ducks with A.platyrhynchos,and also to lesser extents with Anser cygnoides,silky fowl and G.gallus,as well as Sus scrofa,Ovis aries and other mammals.Real-time quantitative PCR(qPCR) analysis demonstrated that MITF was expressed in skin,gizzard,liver,kidney and muscle,and of these tissues,its expression was the highest in the skin tissue(skin>gizzard>liver>kidney>muscle).Ultraviolet spectrophotometry showed that melanin deposition was positively correlated with the MITF expression level in these five tissues and organs(P<0.05).Together,these results demonstrated a tissue-specific pattern of MITF expression and a positive correlation between MITF expression and melanin deposition,indicating that MITF expression may contribute to the melanin deposition in Liancheng white ducks.展开更多
基金Supported by the National Natural Science Foundation of China(No. 40976089)the National Marine Public Welfare Research Project(No. 200805069)+1 种基金the National Science and Technology Support Program of China (No. 2011BAD13B02)the Breeding Project of Shandong Province (China)
文摘Albino Apostichopus japonicus occur both in the wild and in captivity. The offspring of albino A. japonicus also suffer from albinism. The formation of melanin in the melanocytes is dependant on microphthalmia-associated transcription factor (MITF). To investigate the role of MITF in controlling albinism, we cloned the full-length MITF cDNA from A. japonicus and compared MITF mRNA expression in albino and normal A. japonicus. In addition, we used light and electron microscopy to compare histological samples of normal and albino A. japonicus. The body wall of albino adults was characterized by significantly lower levels of MITF expression and lower numbers of epidermal melanocytes, which also contained less melanin. In albino juvenile offspring, MITF expression levels were significantly lower 32 d after fertilization and there were fewer, and less developed, epidermal melanocytes. Thus, we conclude that albino A. japonicus have fewer melanocytes and a reduced ability to synthesize melanin, likely because of lower expression of MITF.
基金Work was supported by the earmarked fund for Modern Agro-industry Technology Research System(CARS-49)the National Natural Science Foundation of China(31672654)the Project of Shanghai Engineering and Technology Center for Promoting Ability(16DZ2281200).
文摘Microphthalmia-associated transcription factor(MITF)is an essential transactivator in melanin synthesis.To characterize the role of MITF in the pearl mussel Hyriopsis cumingii,the MITF homolog of H.cumingii was isolated.The full-length HcMitf cDNA consisted of a 1332-bp with an open reading frame that encode for a 443 amino acid protein that contain a conserved basic helix-loop-helix zipper domain.The HcMitf was found to widespread tissue distribution but expression was higher in purple mussels than in white mussels,mostly in mantle,liver,kidney,gill,and foot with the exception of the adductor mussel.HcMitf and its downstream gene tyrosinase(HcTyr)were highly expressed at the nacre deposition stage after implantation of mantle tissue to produce pearls.Using RNA interference,the expression of HcMitf was reduced by 78%(P<0.01)and expression of HcTyr was also significantly suppressed and consequently total melanin content was decreased(P<0.05).The results suggest that HcMitf plays an important role in melanin synthesis,nacre formation and shell pigmentation in the H.cumingii.
文摘以小干扰RNA(siRNA)技术抑制恶性黑素瘤细胞株A375的黑素皮质素受体1(melanocortin-1-receptor,MC1R)、小眼球相关转录因子(microphthalmia-associated transcription factor,MITF)基因表达,从而抑制细胞黑色素的生成。通过RT-PCR检测MC1R、MITF、酪氨酸酶(tyrosinase,TYR)、酪氨酸酶相关蛋白l(tyrosinase related protein-1,TRP-1)基因mRNA水平的变化,通过测定黑色素含量的变化检测蛋白水平的改变。并用台盼蓝活细胞计数法检测其对细胞增殖的影响。经RT-PCR检测,特异性siRNA作用的靶基因显著下调,黑色素含量明显降低。细胞增殖的变化说明,脂质体包裹的siRNA对A375细胞的毒性远小于熊果苷等化学合成药物。结果显示,利用siRNA对A375细胞中靶基因的调控,有效的抑制了黑色素的生成,siRNA技术与传统的调控黑色素的方法(化学合成物——褪色剂)比较,具有用量小、抑制效率高、细胞毒性低等优点。同时,为进一步研究黑色素形成通路中各基因之间的的关系提供了新思路。
基金supported by the National Natural Science Foundation of China (31372474)the National Guidance Project for Local Science and Technology Development,China (2018L3004)+3 种基金the National and FNU Training Program of Innovation and Enterpreneurship for Undergraduates, China (201910394023, CXXL2019268 and CXXL2019274)the Fujian Province Public Welfare Scientific Project, China (018R1023-2)the Fujian Development and Reform Commission “Five News” Project of China (fjfgw201806)the Young Talent Innovation Funding of Fujian Academy of Agricultural Sciences of China (YC2017-7 and MYQJ2015-5)
文摘The present study investigates the expression of microphthalmia-associated transcription factor(MITF) and its contribution to the melanin deposition in Liancheng white ducks.Nested PCR was used to clone the MITF gene sequence from the skin tissue of female Liancheng white ducks.Ultraviolet spectrophotometry was used to detect the melanin deposition.MITF mRNA expression and melanin deposition in different tissues and organs were detected and their correlation was analyzed.The MITF gene(GenBank number: MG516570) was 1 323 bp in length,contains a complete CDS region(34-1 323 bp) and codes 429 amino acids with 100% homology to the MITF of Anas platyrhynchos and over 95% homology to those of Gallus gallus and Coturnix japonica.Genetic evolution analysis reveals a close relationship of Liancheng white ducks with A.platyrhynchos,and also to lesser extents with Anser cygnoides,silky fowl and G.gallus,as well as Sus scrofa,Ovis aries and other mammals.Real-time quantitative PCR(qPCR) analysis demonstrated that MITF was expressed in skin,gizzard,liver,kidney and muscle,and of these tissues,its expression was the highest in the skin tissue(skin>gizzard>liver>kidney>muscle).Ultraviolet spectrophotometry showed that melanin deposition was positively correlated with the MITF expression level in these five tissues and organs(P<0.05).Together,these results demonstrated a tissue-specific pattern of MITF expression and a positive correlation between MITF expression and melanin deposition,indicating that MITF expression may contribute to the melanin deposition in Liancheng white ducks.
