FGF20及其受体FGFR2和FGFR3在小鼠毛囊形成期的定位及表达
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摘要
旨在研究成纤维细胞生长因子(FGF20)及受体FGFR2和FGFR3在小鼠胚胎期毛囊形成期的表达情况,了解FGF20及受体FGFR2和FG-FR3在小鼠毛囊形成期的作用。采用实时荧光定量PCR、Western blot和免疫组织化学技术检测胚胎期13 d (E13)至18 d (E18)小鼠背部皮肤中FGF20、FGFR2和FGFR3 mRNA及蛋白的差异表达。免疫组化结果显示,FGF20蛋白和FGFR3蛋白E13在表层细胞微弱表达,E14主要表达在表层细胞,且在基底层细胞也有微弱表达,E15主要表达在基板和表层细胞,E16强表达在毛钉与表层细胞,E17和E18主要表达在表层细胞和未成熟的毛囊;FGFR2蛋白在表层细胞、基底层细胞、基板、毛钉、未成熟的毛囊等处均有表达,且E18强表达在表层细胞和未成熟毛囊;RT-PCR及Western blot数据分析结果显示:FGF20 mRNA及蛋白在E16相对表达量最高;FGFR2 mRNA及蛋白在E13相对表达量最低,E18相对表达量最高;FGFR3 mRNA及蛋白表达趋势和FGF20相似。研究结果提示,在毛囊形成期,FGF20可能通过与FGFR3结合从而参与毛囊的诱导和激活,促进毛囊形成并决定其生长方向。FGFR2可能在毛囊形成过程调控表层细胞增殖,促进毛囊形成,诱导毛囊进入第一生长周期。
The aim of this study was to understand the expression patterns of the fibroblast growth factor(FGF) 20 in the embryonic stage of hair follicle formation in mice and the role of FGF20 and its receptors FGFR2,FGFR3 in during formation of hair follicles of mice.The rea-time PCR,Western Blot and immunohistochemistry were adopted to study the differential expressions of FGF20,FGFR2,FGFR3 mRNA and the proteins in the back skin of embryonic mice at 13 d(E13) to 18 d(E18).The results were as follows:1) Immunohistochemistry indicated that the proteins FGF20 and FGFR3 were weakly expressed in the surface cells in E13;in E14 mainly in the surface cells and also weakly in the basal cells;in E15,mainly in the substrate and surface cells;in E16,strongly in the nail and surface cells;and in E17 and E18,mainly in the surface cells and the immature hair follicles.FGFR2 was expressed in the surface cells,basal cells,substrates,nails,and immature hair follicles;and in E18,the most strongly in the surface cells and immature hair follicle.2) RT-PCR and Western Blot showed that the relative expression levels of FGF20 mRNA and the protein was the highest in E16,and that FGFR2 mRNA and the protein had the lowest relative expression in E13,but the highest in E18.The expressed trend of FGFR3 mRNA and the protein was similar to that of FGF20.The present results suggested that during embryonic hair follicle formation,FGF20 might participate in the induction and activation of hair follicles by binding to FGFR3 to promote hair follicle formation and to determine its growth direction.FGFR2 might regulate surface cell proliferation during hair follicle formation,and might promote hair follicle formation and induce hair follicles into the first growth cycle.
The aim of this study was to understand the expression patterns of the fibroblast growth factor(FGF) 20 in the embryonic stage of hair follicle formation in mice and the role of FGF20 and its receptors FGFR2,FGFR3 in during formation of hair follicles of mice.The rea-time PCR,Western Blot and immunohistochemistry were adopted to study the differential expressions of FGF20,FGFR2,FGFR3 mRNA and the proteins in the back skin of embryonic mice at 13 d(E13) to 18 d(E18).The results were as follows:1) Immunohistochemistry indicated that the proteins FGF20 and FGFR3 were weakly expressed in the surface cells in E13;in E14 mainly in the surface cells and also weakly in the basal cells;in E15,mainly in the substrate and surface cells;in E16,strongly in the nail and surface cells;and in E17 and E18,mainly in the surface cells and the immature hair follicles.FGFR2 was expressed in the surface cells,basal cells,substrates,nails,and immature hair follicles;and in E18,the most strongly in the surface cells and immature hair follicle.2) RT-PCR and Western Blot showed that the relative expression levels of FGF20 mRNA and the protein was the highest in E16,and that FGFR2 mRNA and the protein had the lowest relative expression in E13,but the highest in E18.The expressed trend of FGFR3 mRNA and the protein was similar to that of FGF20.The present results suggested that during embryonic hair follicle formation,FGF20 might participate in the induction and activation of hair follicles by binding to FGFR3 to promote hair follicle formation and to determine its growth direction.FGFR2 might regulate surface cell proliferation during hair follicle formation,and might promote hair follicle formation and induce hair follicles into the first growth cycle.
引文
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[16]程笳琪,于秀菊,高淑媛,等.FGF家族对鼠类毛发生长作用的研究[J].黑龙江畜牧兽医,2016,9(17):215-217.
[17]Chiu H C,Chang C H,Chen J S,et al.Human hair follicle dermal papilla cell,dermal sheath cell and interstitial dermal fibroblast characteristics[J].Journal of the Formosan Medical Association,1996,95(9):667-674.
[18]杨卫兵,郝飞.毛乳头细胞在毛囊形态学发生和生长调控中的作用[J].国外医学:皮肤性病学分册,2003,29(5):312-314.
[19]Jahoda C A B,Oliver R F,Reynolds A J,et al.Trans species hair growth induction by human hair follicle dermal papillae[J].Experimental Dermatology,2010,10(4):229-237.
[20]Richardson G D,Bazzi H,Fantauzzo K A,et al.KGF and EGF signalling block hair follicle induction and promote interfollicular epidermal fate in developing mouse skin[J].Development,2009,136(13):2153-2164.
[21]孙晓艳,付小兵,伊海英,等.碱性成纤维细胞生长因子诱导表皮细胞去分化的初步研究[J].第三军医大学学报,2008,30(21):2003-2007.
[22]王博,王春生,安铁洙.小鼠皮肤及其毛囊早期发育的组织学观察[J].中国实验动物学报,2007,15(6):413-415.
[23]Abe Y,Tanaka N.Roles of the Hedgehog Signaling Pathway in Epidermal and Hair Follicle Development,Homeostasis,and Cancer[J].Journal of Developmental Biology,2017,5(4):12.
[24]Itoh N.The Fgf families in humans,mice,and zebrafish:their evolutional processes and roles in development,metabolism,and disease[J].Biologica&Pharmaceutical Bulletin,2007,30(10):1819-1825.
[25]Ohuchi H,Hori Y,Yamasaki M,et al.FGF10 acts as a major ligand for FGF receptor 2 IIIb in mouse multi-organ development[J].Biochemical&Biophysical Research Communications,2000,277(3):643-649.
[26]伊海英,孙晓艳,付小兵,等.胎儿皮肤发育中成纤维细胞生长因子受体-2表达的免疫组织化学研究[J].第三军医大学学报,2008,30(21):2008-2010.
[27]Petiot A,Conti F J,Grose R,et al.A crucial role for Fgfr2-b signalling in epidermal development and hair follicle patterning[J].Development,2003,(22):5493-5010.
[28]Rosenquist T A,Martin G R.Fibroblast growth factor signalling in the hair growth cycle:expression of the fibroblast growth factor receptor and ligand genes in the murine hair follicle[J].Developmental Dynamics An Official Publication of the American Association of Anatomists,1996,205(4):379-86.
[29]单慧,劳力民.成纤维细胞生长因子受体及其在皮肤的表达[J].浙江临床医学,2007,9(12):1687-1688.
[30]Takenaka H,Yasuno H,Kishimoto S.Immunolocalization of fibroblast growth factor receptors in normal and wounded human skin[J].Journal of Dermatological Science,2002,294(7):331-338.
[31]Katoh M.Cancer genomics and genetics of FGFR2(Review)[J].International Journal of Oncology,2008,33(2):233-7.
[32]赵央,田海山,李校堃,等.成纤维细胞生长因子20研究进展[J].中国生物工程杂志,2015,35(8):103-108.
[33]Jeffers M,Shimkets R,Prayaga S,et al.Identification of a novel human fibroblast growth factor and characterization of its role in oncogenesis[J].Cancer Research,2001,61(7):3131-3138.
[34]Thisse B,Thisse C.Functions and regulations of fibroblast growth factor signaling during embryonic development[J].Developmental Biology,2005,287(2):390-402.
[35]Mena L D,Cardo L F,Coto E,et al.FGF20 rs12720208 SNP and microRNA-433 variation:no association with Parkinson’s disease in Spanish patients[J].Neuroscience Letters,2010,479(1):22-25.
[36]程笳琪.FGF7亚家族对小鼠毛囊形成期和发育作用的研究[D].太谷:山西农业大学,2017.
[2]Rogers G E.Hair follicle differentiation and regulation[J].International Journal of Developmental Biology,2004,48(2-3):163-170.
[3]Enshellseijffers D,Lindon C,Kashiwagi M.β-catenin activity in the dermal papilla regulates morphogenesis and regeneration of hair[J].Developmental Cell,2010,18(4):633-642.
[4]Stecksén Blicks C,Falk K C,Schmittegenolf M,et al.Hair shaft structures in EDAR induced ectodermal dysplasia[J].BMCMedical Genetics,2015,16(1):1-6.
[5]Genander M,Cook P J,Ramskold D.BMP Signaling and Its p S-MAD1/5 Target Genes Differentially Regulate Hair Follicle Stem Cell Lineages[J].Cell Stem Cell,2014,15(5):619-633.
[6]Itoh N,Ornitz D M.Evolution of the Fgf and Fgfr gene families[J].Trends in Genetics,2004,20(11):563-569.
[7]韩萨茹拉,高爱琴,李金泉,等.成纤维细胞生长因子(FGF)研究进展[J].安徽农业科学,2009,37(7):3008-3010.
[8]Huh SH,Narhi K,Lindfors P H,et al.Fgf20 governs formation of primary and secondary dermal condensations in developing hair follicles[J].Genes&Development,2013,27(4):450-458.
[9]Su N,Jin M,Chen L.Role of FGF/FGFR signaling in skeletal development and homeostasis:learning from mouse models[J].Bone Res,2014,2(1):9-32.
[10]Muenke M,Schell U.Fibroblast-growth-factor receptor mutations in human skeletal disorders[J].Trends Genet,1995,11(8):308-313.
[11]苏钰涵,杜华,牛广明,等.成纤维细胞生长因子的信号通路[J].中国组织工程研究,2016,20(15):2255-2264.
[12]李殿启,杨甲梅,殷正丰.FGF-20:成纤维细胞生长因子家族新成员[J].生命的化学,2006,26(5):445-448.
[13]李国强,纪影畅,李宇.毛囊形态发生的分子机制[J].国外医学:皮肤性病学分册,2004,30(1):38-40.
[14]Paus R,Muller-Rover S,Van D V C,et al.A comprehensive guide for the recognition and classification of distinct stages of hair follicle morphogenesis[J].Journal of Investigative Dermatology,1999,113(4):523-532.
[15]魏泓.医学实验动物学[M].成都:四川科学技术出版社,1998:423-427.
[16]程笳琪,于秀菊,高淑媛,等.FGF家族对鼠类毛发生长作用的研究[J].黑龙江畜牧兽医,2016,9(17):215-217.
[17]Chiu H C,Chang C H,Chen J S,et al.Human hair follicle dermal papilla cell,dermal sheath cell and interstitial dermal fibroblast characteristics[J].Journal of the Formosan Medical Association,1996,95(9):667-674.
[18]杨卫兵,郝飞.毛乳头细胞在毛囊形态学发生和生长调控中的作用[J].国外医学:皮肤性病学分册,2003,29(5):312-314.
[19]Jahoda C A B,Oliver R F,Reynolds A J,et al.Trans species hair growth induction by human hair follicle dermal papillae[J].Experimental Dermatology,2010,10(4):229-237.
[20]Richardson G D,Bazzi H,Fantauzzo K A,et al.KGF and EGF signalling block hair follicle induction and promote interfollicular epidermal fate in developing mouse skin[J].Development,2009,136(13):2153-2164.
[21]孙晓艳,付小兵,伊海英,等.碱性成纤维细胞生长因子诱导表皮细胞去分化的初步研究[J].第三军医大学学报,2008,30(21):2003-2007.
[22]王博,王春生,安铁洙.小鼠皮肤及其毛囊早期发育的组织学观察[J].中国实验动物学报,2007,15(6):413-415.
[23]Abe Y,Tanaka N.Roles of the Hedgehog Signaling Pathway in Epidermal and Hair Follicle Development,Homeostasis,and Cancer[J].Journal of Developmental Biology,2017,5(4):12.
[24]Itoh N.The Fgf families in humans,mice,and zebrafish:their evolutional processes and roles in development,metabolism,and disease[J].Biologica&Pharmaceutical Bulletin,2007,30(10):1819-1825.
[25]Ohuchi H,Hori Y,Yamasaki M,et al.FGF10 acts as a major ligand for FGF receptor 2 IIIb in mouse multi-organ development[J].Biochemical&Biophysical Research Communications,2000,277(3):643-649.
[26]伊海英,孙晓艳,付小兵,等.胎儿皮肤发育中成纤维细胞生长因子受体-2表达的免疫组织化学研究[J].第三军医大学学报,2008,30(21):2008-2010.
[27]Petiot A,Conti F J,Grose R,et al.A crucial role for Fgfr2-b signalling in epidermal development and hair follicle patterning[J].Development,2003,(22):5493-5010.
[28]Rosenquist T A,Martin G R.Fibroblast growth factor signalling in the hair growth cycle:expression of the fibroblast growth factor receptor and ligand genes in the murine hair follicle[J].Developmental Dynamics An Official Publication of the American Association of Anatomists,1996,205(4):379-86.
[29]单慧,劳力民.成纤维细胞生长因子受体及其在皮肤的表达[J].浙江临床医学,2007,9(12):1687-1688.
[30]Takenaka H,Yasuno H,Kishimoto S.Immunolocalization of fibroblast growth factor receptors in normal and wounded human skin[J].Journal of Dermatological Science,2002,294(7):331-338.
[31]Katoh M.Cancer genomics and genetics of FGFR2(Review)[J].International Journal of Oncology,2008,33(2):233-7.
[32]赵央,田海山,李校堃,等.成纤维细胞生长因子20研究进展[J].中国生物工程杂志,2015,35(8):103-108.
[33]Jeffers M,Shimkets R,Prayaga S,et al.Identification of a novel human fibroblast growth factor and characterization of its role in oncogenesis[J].Cancer Research,2001,61(7):3131-3138.
[34]Thisse B,Thisse C.Functions and regulations of fibroblast growth factor signaling during embryonic development[J].Developmental Biology,2005,287(2):390-402.
[35]Mena L D,Cardo L F,Coto E,et al.FGF20 rs12720208 SNP and microRNA-433 variation:no association with Parkinson’s disease in Spanish patients[J].Neuroscience Letters,2010,479(1):22-25.
[36]程笳琪.FGF7亚家族对小鼠毛囊形成期和发育作用的研究[D].太谷:山西农业大学,2017.
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