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
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
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
黑色素合成是极其复杂的过程,其中涉及多种基因和miRNAs的参与。miR-186-5p在绵羊不同毛色皮肤中差异表达,说明其可能与毛色形成有关,生物信息学预测和绵羊不同毛色皮肤中miR-186-5p和Mitf的差异表达间接说明二者可能存在靶向关系,双荧...黑色素合成是极其复杂的过程,其中涉及多种基因和miRNAs的参与。miR-186-5p在绵羊不同毛色皮肤中差异表达,说明其可能与毛色形成有关,生物信息学预测和绵羊不同毛色皮肤中miR-186-5p和Mitf的差异表达间接说明二者可能存在靶向关系,双荧光报告直接证实了二者的靶向关系。为了证实miR-186-5p在黑色素形成中的作用,本研究构建miR-186-5p真核表达载体,并转染绵羊黑素细胞。结果显示,miR-186-5p通过与Mitf mRNAs的3′UTR结合抑制Mitf的表达和翻译。MITF是Tyr基因家族的调节因子。实时荧光聚合酶链反应(quantitative real time polymerase chain reaction, qRT-PCR)和Western免疫印迹结果表明,过表达miR-186-5p引起的MITF下调抑制了TYR、TYRP1和TYRP2的表达。其中,mRNA表达水平分别下降34%、66%和52%;蛋白质表达水平分别下降32%、6%和46%。分光光度检测结果显示,黑色素含量下降59%倍。上述结果表明,miR-186-5p通过抑制靶基因Mitf的表达,抑制了绵羊黑素细胞内黑色素的生成。展开更多
Vitiligo results in an autoimmune disorder destructing skin pigment cells,melanocytes(Mcs).This study aimed to investigate whether Astragaloside IV(AIV)could efficiently induce differentiation of bone marrow mesenchym...Vitiligo results in an autoimmune disorder destructing skin pigment cells,melanocytes(Mcs).This study aimed to investigate whether Astragaloside IV(AIV)could efficiently induce differentiation of bone marrow mesenchymal stem cells(BMMSCs)into Mcs.BMMSCs were induced and differentiated into Mcs with 0.1,0.2,and 0.4 mg/L AIV during 150-day.Morphologic changes of differentiated cells were observed.Levels of some melanocytic specific genes(TRP-1,TRP-2,MART-1,Mitf)were measured with quantitative polymerase chain reaction(qPCR)at 90,120,and 150 days of induction.After 90-day induction,the differentiated cells with 0.4 mg/L AIV demonstrated the typical morphology of Mcs,positive 3,4 dihydroxyphenylalanine staining,and positive staining of TRP-1,TRP-2,MART-1,and Mitf.After 90-and 120-days’induction with 0.4 mg/L AIV,TRP-1 expression was significantly elevated(p<0.01),and TRP-2 expression was significantly increased in 0.4 mg/L AIV-treated group compared to negative control(p<0.01),0.1 mg/L(p<0.01),and 0.2 mg/L(p<0.01)AIV-treated groups.Moreover,MART-1 expression was significantly up-regulated in 0.4 mg/L AIV-treated group compared to negative control,but without difference compared to 0.1 mg/L(p>0.05)and 0.2 mg/L(p>0.05)AIV-treated groups.During 90 to 150-day induction,there were no significant differences for Mitf levels between AIV-treated groups and negative control(p>0.05).In conclusion,90-day induction with 0.4 mg/L AIV up-regulated TRP-1,TRP-2,and MART-1 expression,indicating that AIV can efficiently induce Mcs differentiation from BMMSCs.These results provide experimental and theoretic evidence for AIV application in clinical vitiligo repigmentation treatment.展开更多
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.
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.
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.
文摘黑色素合成是极其复杂的过程,其中涉及多种基因和miRNAs的参与。miR-186-5p在绵羊不同毛色皮肤中差异表达,说明其可能与毛色形成有关,生物信息学预测和绵羊不同毛色皮肤中miR-186-5p和Mitf的差异表达间接说明二者可能存在靶向关系,双荧光报告直接证实了二者的靶向关系。为了证实miR-186-5p在黑色素形成中的作用,本研究构建miR-186-5p真核表达载体,并转染绵羊黑素细胞。结果显示,miR-186-5p通过与Mitf mRNAs的3′UTR结合抑制Mitf的表达和翻译。MITF是Tyr基因家族的调节因子。实时荧光聚合酶链反应(quantitative real time polymerase chain reaction, qRT-PCR)和Western免疫印迹结果表明,过表达miR-186-5p引起的MITF下调抑制了TYR、TYRP1和TYRP2的表达。其中,mRNA表达水平分别下降34%、66%和52%;蛋白质表达水平分别下降32%、6%和46%。分光光度检测结果显示,黑色素含量下降59%倍。上述结果表明,miR-186-5p通过抑制靶基因Mitf的表达,抑制了绵羊黑素细胞内黑色素的生成。
基金the National Natural Science Foundation of China(Grant No.81703140).
文摘Vitiligo results in an autoimmune disorder destructing skin pigment cells,melanocytes(Mcs).This study aimed to investigate whether Astragaloside IV(AIV)could efficiently induce differentiation of bone marrow mesenchymal stem cells(BMMSCs)into Mcs.BMMSCs were induced and differentiated into Mcs with 0.1,0.2,and 0.4 mg/L AIV during 150-day.Morphologic changes of differentiated cells were observed.Levels of some melanocytic specific genes(TRP-1,TRP-2,MART-1,Mitf)were measured with quantitative polymerase chain reaction(qPCR)at 90,120,and 150 days of induction.After 90-day induction,the differentiated cells with 0.4 mg/L AIV demonstrated the typical morphology of Mcs,positive 3,4 dihydroxyphenylalanine staining,and positive staining of TRP-1,TRP-2,MART-1,and Mitf.After 90-and 120-days’induction with 0.4 mg/L AIV,TRP-1 expression was significantly elevated(p<0.01),and TRP-2 expression was significantly increased in 0.4 mg/L AIV-treated group compared to negative control(p<0.01),0.1 mg/L(p<0.01),and 0.2 mg/L(p<0.01)AIV-treated groups.Moreover,MART-1 expression was significantly up-regulated in 0.4 mg/L AIV-treated group compared to negative control,but without difference compared to 0.1 mg/L(p>0.05)and 0.2 mg/L(p>0.05)AIV-treated groups.During 90 to 150-day induction,there were no significant differences for Mitf levels between AIV-treated groups and negative control(p>0.05).In conclusion,90-day induction with 0.4 mg/L AIV up-regulated TRP-1,TRP-2,and MART-1 expression,indicating that AIV can efficiently induce Mcs differentiation from BMMSCs.These results provide experimental and theoretic evidence for AIV application in clinical vitiligo repigmentation treatment.
