多疣壁虎胚胎发育过程中角蛋白的表达分析
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摘要
角蛋白包括α角蛋白和β角蛋白两种类型.迄今α角蛋白被发现存在于所有的脊椎动物中,而β角蛋白仅存在于鸟类和爬行类表皮及其附属物中.本研究通过同源搜索比对鉴定了多疣壁虎(Gekko japonicus)的1个β角蛋白基因:G.japonicus claw keratin-like(LOC107121854),暂命名为ge-gprp-13,和1个α角蛋白基因:G.japonicus keratinocyte associated protein 2(KRTCAP2),暂命名为ge-krtcap2.利用荧光定量PCR方法比较分析了多疣壁虎ge-gprp-13和ge-krtcap2基因在其胚胎发育过程中的mRNA表达模式.研究结果表明,在多疣壁虎胚胎发育的不同时期,α角蛋白基因(ge-krtcap2)和β角蛋白基因(ge-gprp-13)的表达模式不同:在趾部发育的初期,α角蛋白基因先表达,而β角蛋白基因主要在趾部发育的后期进行表达,并且在趾部形成的整个过程中,β角蛋白基因的表达量远远高于α角蛋白基因的表达量.
Reptile epidermis contains two types of keratins—α-keratins and β-keratins. Up to date,α-keratins are reported to be present in all vertebrates,while β-keratins are only found in birds and reptiles. In the present study,two keratin genes of Gekko japonicus were identified from the National Center of Biotechnology Information database,one β-keratin gene,G.japonicus claw keratin-like(LOC107121854)tentatively named ge-gprp-13 and one α-keratin gene,G.japonicus keratinocyte associated protein 2(KRTCAP2)tentatively named ge-krtcap2. The mRNA expression of keratin genes during embryonic development of the gecko was studied by using fluorescence quantitative real-time PCR. The results showed that the α-keratin gene(ge-krtcap2)and the β-keratin gene(ge-gprp-13)have different expression patterns in different stages of embryonic development of the gecko. In the early stage of toe development,the α-keratin gene is expressed first,while the β-keratin gene is mainly expressed in the later stage of toe development. And during the formation of the toe,the expression level of the β-keratin gene is much higher than the α-keratin gene.
Reptile epidermis contains two types of keratins—α-keratins and β-keratins. Up to date,α-keratins are reported to be present in all vertebrates,while β-keratins are only found in birds and reptiles. In the present study,two keratin genes of Gekko japonicus were identified from the National Center of Biotechnology Information database,one β-keratin gene,G.japonicus claw keratin-like(LOC107121854)tentatively named ge-gprp-13 and one α-keratin gene,G.japonicus keratinocyte associated protein 2(KRTCAP2)tentatively named ge-krtcap2. The mRNA expression of keratin genes during embryonic development of the gecko was studied by using fluorescence quantitative real-time PCR. The results showed that the α-keratin gene(ge-krtcap2)and the β-keratin gene(ge-gprp-13)have different expression patterns in different stages of embryonic development of the gecko. In the early stage of toe development,the α-keratin gene is expressed first,while the β-keratin gene is mainly expressed in the later stage of toe development. And during the formation of the toe,the expression level of the β-keratin gene is much higher than the α-keratin gene.
引文
[1] ALIBARDI L.Cell biology of adhesive setae in gecko lizards[J].Zoology,2009,112(6):403-424.
[2] SAWYER R H,GLENN T,FRENCH J O,et al.The expression of beta(β)keratins in the epidermal appendages of reptiles and birds[J].American zoologist,2000,40(4):530-539.
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[4] ALIBARDI L.Immunolocalization of specific keratin associated beta-proteins(beta-keratins)in the adhesive setae of Gekko gecko[J].Tissue and cell,2013,45(4):231-240.
[5] GREENWOLD M J,SAWYER R H.Linking the molecular evolution of avian beta(beta)keratins to the evolution of feathers[J].Journal of experimental zoology part b,molecular and developmental evolution,2011,316(8):609-616.
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[7] COULOMBE P A,OMARY M B.‘Hard’and‘soft’principles defining the structure,function and regulation of keratin intermediate filaments[J].Current opinion in cell biology,2002,14(1):110-122.
[8] FUCHS E,CLEVELAND D W.A structural scaffolding of intermediate filaments in health and disease[J].Science,1998,279(5350):514-519.
[9] WYLD J A,BRUSH A H.The molecular heterogeneity and diversity of reptilian keratins[J].Journal of molecular evolution,1979,12(4):331-347.
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[14] 计翔,王培潮,洪卫星.多疣壁虎的繁殖生态研究[J].动物学报,1991(2):185-192.
[15] 赵若男,张黎悦,吴鹏飞,等.多疣壁虎胚胎发育分期的形态学特征[J].动物学杂志,2017,52(6):987-995.
[16] 朱灵君,胡凌君,张永普,等.不同多疣壁虎种群繁殖特征的比较[J].生态学杂志,2009,28(4):692-697.
[17] 许雪峰,计翔.多疣壁虎的雌性繁殖及孵化温度对孵化期和孵出幼体特征的影响[J].生态学杂志,2001(6):8-11.
[18] LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using realtime quantitative PCR and the 2-ΔΔCt Method[J].Methods,2001,25:402-408.
[19] ALIBARDI L,TONI M.Cytochemical and molecular characteristics of the process of cornification during feather morphogenesis[J].Progress in histochemistry and cytochemistry,2008,43(1):1-69.
[20] ALIBARDI L,MICHIELI F,DALLA VALLE L.Low-cysteine alpha-keratins and corneous beta-proteins are initially formed in the regenerating tail epidermis of lizard[J].Journal of morphology,2017,278(1):119-130.
[21] ALIBARDI L,YHOMPSON M B.Fine structure of the developing epidermis in the embryo of the American alligator(Alligator mississippiensis,Crocodilia,Reptilia)[J].Journal of anatomy,2001,198(Pt 3):265-282.
[22] ALIBARDI L,YHOMPSON M B.Keratinization and ultrastructure of the epidermis of late embryonic stages in the alligator(Alligator mississippiensis)[J].Journal of anatomy,2002,201(1):71-84.
[23] WU P,NG C S,YAN J,et al.Topographical mapping of alpha- and beta-keratins on developing chicken skin integuments:functional interaction and evolutionary perspectives[J].Proceedings of the national academy sciences of the United States of America,2015,112(49):E6770-9.
[24] DALLA V L,NARDI A,TOFFOLO,et al.Cloning and characterization of scale beta-keratins in the differentiating epidermis of geckoes show they are glycine-proline-serine-rich proteins with a central motif homologous to avian beta-keratins[J].Developmental dynamics,2007,236(2):374-388.
[2] SAWYER R H,GLENN T,FRENCH J O,et al.The expression of beta(β)keratins in the epidermal appendages of reptiles and birds[J].American zoologist,2000,40(4):530-539.
[3] ALIBARDI L,SAWYER R H.Immunocytochemical analysis of beta keratins in the epidermis of chelonians,lepidosaurians,and archosaurians[J].The journal of experimental zoology,2002,293(1):27-38.
[4] ALIBARDI L.Immunolocalization of specific keratin associated beta-proteins(beta-keratins)in the adhesive setae of Gekko gecko[J].Tissue and cell,2013,45(4):231-240.
[5] GREENWOLD M J,SAWYER R H.Linking the molecular evolution of avian beta(beta)keratins to the evolution of feathers[J].Journal of experimental zoology part b,molecular and developmental evolution,2011,316(8):609-616.
[6] STEINERT P M.Structure,function,and dynamics of keratin intermediate filaments[J].The journal of investigative dermatology,1993,100(6):729-734.
[7] COULOMBE P A,OMARY M B.‘Hard’and‘soft’principles defining the structure,function and regulation of keratin intermediate filaments[J].Current opinion in cell biology,2002,14(1):110-122.
[8] FUCHS E,CLEVELAND D W.A structural scaffolding of intermediate filaments in health and disease[J].Science,1998,279(5350):514-519.
[9] WYLD J A,BRUSH A H.The molecular heterogeneity and diversity of reptilian keratins[J].Journal of molecular evolution,1979,12(4):331-347.
[10] ALIBARDI L.Ultrastructural autoradiographic and immunocytochemical analysis of setae formation and keratinization in the digital pads of the gecko Hemidactylus turcicus(Gekkonidae,Reptilia)[J].Tissue and cell,2003,35(4):288-296.
[11] ALIBARDI L,TONI M.Cytochemical,biochemical and molecular aspects of the process of keratinization in the epidermis of reptilian scales[J].Progress in histochemistry and cytochemistry,2006,40(2):73-134.
[12] 计翔,王培潮.多疣壁虎卵的孵化特征[J].动物学研究,1991(1):28-72.
[13] 计翔,王培潮.温度对多疣壁虎摄食量和同化效率的影响[J].杭州师范学院学报(自然科学版),1990(6):90-94.
[14] 计翔,王培潮,洪卫星.多疣壁虎的繁殖生态研究[J].动物学报,1991(2):185-192.
[15] 赵若男,张黎悦,吴鹏飞,等.多疣壁虎胚胎发育分期的形态学特征[J].动物学杂志,2017,52(6):987-995.
[16] 朱灵君,胡凌君,张永普,等.不同多疣壁虎种群繁殖特征的比较[J].生态学杂志,2009,28(4):692-697.
[17] 许雪峰,计翔.多疣壁虎的雌性繁殖及孵化温度对孵化期和孵出幼体特征的影响[J].生态学杂志,2001(6):8-11.
[18] LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using realtime quantitative PCR and the 2-ΔΔCt Method[J].Methods,2001,25:402-408.
[19] ALIBARDI L,TONI M.Cytochemical and molecular characteristics of the process of cornification during feather morphogenesis[J].Progress in histochemistry and cytochemistry,2008,43(1):1-69.
[20] ALIBARDI L,MICHIELI F,DALLA VALLE L.Low-cysteine alpha-keratins and corneous beta-proteins are initially formed in the regenerating tail epidermis of lizard[J].Journal of morphology,2017,278(1):119-130.
[21] ALIBARDI L,YHOMPSON M B.Fine structure of the developing epidermis in the embryo of the American alligator(Alligator mississippiensis,Crocodilia,Reptilia)[J].Journal of anatomy,2001,198(Pt 3):265-282.
[22] ALIBARDI L,YHOMPSON M B.Keratinization and ultrastructure of the epidermis of late embryonic stages in the alligator(Alligator mississippiensis)[J].Journal of anatomy,2002,201(1):71-84.
[23] WU P,NG C S,YAN J,et al.Topographical mapping of alpha- and beta-keratins on developing chicken skin integuments:functional interaction and evolutionary perspectives[J].Proceedings of the national academy sciences of the United States of America,2015,112(49):E6770-9.
[24] DALLA V L,NARDI A,TOFFOLO,et al.Cloning and characterization of scale beta-keratins in the differentiating epidermis of geckoes show they are glycine-proline-serine-rich proteins with a central motif homologous to avian beta-keratins[J].Developmental dynamics,2007,236(2):374-388.