外源褪黑激素促进绒山羊皮肤毛囊相关基因表达差异的研究
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摘要
旨在通过对非长绒期阿尔巴斯型绒山羊进行褪黑激素埋植,研究褪黑激素对羊绒周期性生长的调控,以期借此延长绒山羊毛囊的兴盛期,提高羊绒产量。将试验羊随机分成两组:埋植褪黑激素的试验组(T)和对照组(C),每组3只(试验组(TI、T2和T3)和对照组(C1、C2和C3)),采集绒山羊的皮肤组织进行组织切片,染色观察埋植褪黑激素对毛囊生长诱导作用。分别抽提两组样本总RNA,逆转录合成相应被标记的cDNA探针,采用Agilent绵羊的8×15K规格全基因组表达谱芯片进行杂交,筛选差异表达基因。利用实时荧光定量PCR技术进行验证。结果表明,从组织学分析可观察到埋植褪黑激素对毛囊生长具有明显的促进作用;芯片数据显示,筛选出差异表达的基因95个,其中61个表达上调、34个下调。GO分析基因数量分布情况:参与分子功能的基因占47.78%,参与生物学过程的基因占33.89%,组成细胞成分的基因占18.33%。与对照组的样本相比,埋植褪黑激素后相关基因的差异性表达涉及毛囊生长及周围皮肤附属物形态发生等生物学过程。这些差异性表达的基因为绒山羊毛囊生长及周期性调控的基因功能研究提供极有价值的参考。
Effects of embedding melatonin for Arbas Cashmere goats in telogen phase on control of cashmere growth cycle were conducted in order to extend anagen phase and increase the cashmere production.Cashmere goats were selected and randomly divided into 2groups,each group with 3goats(the experiment group(T1,T2,T3)and the control group(C1,C2,C3)).Skin tissues ofthe treatment and control groups were taken and tissue sections were observed for the influence of embedding melatonin on the induction of hair follicle growth.The total RNA was extracted from the 2groups of samples and synthesized marked cDNA probes by reverse transcription,and had microarray hybridization on 8×15 K(Agilent)sheep genome-wide expression profile to screen differentially expressed genes.The results were verified by real-time fluorescent quantitative PCR technology.Histological analysis showed that embedding melatonin could obviously promote hair follicle growth.Microarray data showed that there were 95 differently expressed genes,including61up-regulated and 34down-regulated.GO analysis indicated that the genes of 47.78% participated in the molecular function,those of 33.89% were involved in biological processes,and those of 18.33% were associated with the composition of cell.Compared with the control groups,differential expression of related genes in treatment groups were involved in the hair follicle morphogenesis and biological processes of surrounding skin appendages.These differently expressed genes provide valuable reference to research Cashmere goat hair follicle growth and the function of the cyclical controlling genes.
Effects of embedding melatonin for Arbas Cashmere goats in telogen phase on control of cashmere growth cycle were conducted in order to extend anagen phase and increase the cashmere production.Cashmere goats were selected and randomly divided into 2groups,each group with 3goats(the experiment group(T1,T2,T3)and the control group(C1,C2,C3)).Skin tissues ofthe treatment and control groups were taken and tissue sections were observed for the influence of embedding melatonin on the induction of hair follicle growth.The total RNA was extracted from the 2groups of samples and synthesized marked cDNA probes by reverse transcription,and had microarray hybridization on 8×15 K(Agilent)sheep genome-wide expression profile to screen differentially expressed genes.The results were verified by real-time fluorescent quantitative PCR technology.Histological analysis showed that embedding melatonin could obviously promote hair follicle growth.Microarray data showed that there were 95 differently expressed genes,including61up-regulated and 34down-regulated.GO analysis indicated that the genes of 47.78% participated in the molecular function,those of 33.89% were involved in biological processes,and those of 18.33% were associated with the composition of cell.Compared with the control groups,differential expression of related genes in treatment groups were involved in the hair follicle morphogenesis and biological processes of surrounding skin appendages.These differently expressed genes provide valuable reference to research Cashmere goat hair follicle growth and the function of the cyclical controlling genes.
引文
[1]MCGREGOR B A.Nutrition,management and other environmental influences on the quality and production of mohair and cashmere:A review with particular reference to mediterranean and annual temperate climatic zones[J].Small Rumin Res,1998,28(3):199-215.
[2]GEBBIE F,FORSYTH I,ARENDT J.Effects of maintaining solstice light and temperature on reproductive activity,coat growth,plasma prolactin and melatonin in goats[J].J Reprod Fertil,1999,116(1):25-33.
[3]IBRAHEEM M,GALBRAITH H,SCAIFE J,et al.Growth of secondary hair follicles of the Cashmere goat in vitro and their response to prolactin and melatonin[J].J Anat,1994,185(Pt 1):135-142.
[4]王潇,尹俊.内蒙古白绒山羊毛囊干细胞巢的间接定位[J].农业生物技术学报,2014,22(3):326-332.WANG X,YIN J.Indirect location of hair follicle stem cell niche in Inner Mongolia White Cashmere goat[J].Journal of Agricultural Biotechnology,2014,22(3):326-332.(in Chinese)
[5]MOELLMANN G,LERNER A B,HENDEE J R.The mechanism of frog skin lightening by acetylcholine[J].Gen Comp Endocrinol,1974,23(1):45-51.
[6]REPPERT S M,WEAVER D R,RIVKEES S A,et al.Putative melatonin receptors in a human biological clock[J].Science,1988,242(4875):78-81.
[7]CONG Y,DENG H,FENG Y,et al.Melatonin implantation from winter solstice could extend the cashmere growth phase effectively[J].Small Rumin Res,2011,99(1):48-53.
[8]TUSHER V G,TIBSHIRANI R,CHU G.Significance analysis of microarrays applied to the ionizing radiation response[J].Proc Natl Acad Sci,2001,98(9):5116-5121.
[9]GHOSH S,SINGH A K,HALDAR C.Seasonal modulation of immunity by melatonin and gonadal steroids in a short day breeder goat capra hircus[J].Theriogenology,2014,82(8):1121-1130.
[10]ZHANG Q,LI J,CHEN Y,et al.Growth and viability of Liaoning Cashmere goat hair follicles during the annual hair follicle cycle[J].Genet Mol Res,2014,13(2):4433-4443.
[11]FOULKES N S,ASSONE-CORSI P,BORJIGIN J,et al.Rhythmic transcription:the molecular basis of circadian melatonin synthesis[J].Trends Neurosci,1997,20(10):487-492.
[12]LANOIX D,BEGHDADI H,LAFOND J,et al.Human placental trophoblasts synthesize melatonin and express its receptors[J].J Pineal Res,2008,45(1):50-60.
[13]LI Y H,ZHANG K,YANG K,et al.Adenovirus-mediated Wnt10boverexpression induces hair follicle regeneration[J].J Invest Dermatol,2013,133(1):42-48.
[14]MYUNG P S,TAKEO M,ITO M,et al.Epithelial Wnt ligand secretion is required for adult hair follicle growth and regeneration[J].J Invest Dermatol,2013,133(1):31-41.
[15]CHEN D,JARRELL A,GUO C,et al.Dermalβ-catenin activity in response to epidermal Wnt ligands is required for fibroblast proliferation and hair follicle initiation[J].Development,2012,139(8):1522-1533.
[16]ALONSO L,FUCHS E.Stem cells in the skin:waste not,Wnt not[J].Genes Dev,2003,17(10):1189-1200.
[17]WANG B,LI H,LIU Y,et al.Expression patterns of WNT/β-CATENIN signaling molecules during human tooth development[J].J Mol Hist,2014,45(5):487-496.
[18]MAEKAWA M,TAKASHIMA N,ARAI Y,et al.Pax6is required for production and maintenance of progenitor cells in postnatal hippocampal neurogenesis[J].Genes Cells,2005,10(10):1001-1014.
[19]ROWE J M,WELSH C,PENA R N,et al.Illuminating role of CYP1A1in skin function[J].J Invest Dermatol,2008,128(7):1866-1868.
[20]MAN X Y,YANG X H,CAI S Q,et al.Expression and localization of vascular endothelial growth factor and vascular endothelial growth factor receptor-2in human epidermal appendages:a comparison study by immunofluorescence[J].Clin Exp Dermatol,2009,34(3):396-401.
[21]WU X J,ZHU J W,JING J,et al.VEGF 165 modulates proliferation,adhesion,migration and differentiation of cultured human outer root sheath cells from central hair follicle epithelium through VEGFR-2activation in vitro[J].J Dermatol Sci,2014,73(2):152-160.
[2]GEBBIE F,FORSYTH I,ARENDT J.Effects of maintaining solstice light and temperature on reproductive activity,coat growth,plasma prolactin and melatonin in goats[J].J Reprod Fertil,1999,116(1):25-33.
[3]IBRAHEEM M,GALBRAITH H,SCAIFE J,et al.Growth of secondary hair follicles of the Cashmere goat in vitro and their response to prolactin and melatonin[J].J Anat,1994,185(Pt 1):135-142.
[4]王潇,尹俊.内蒙古白绒山羊毛囊干细胞巢的间接定位[J].农业生物技术学报,2014,22(3):326-332.WANG X,YIN J.Indirect location of hair follicle stem cell niche in Inner Mongolia White Cashmere goat[J].Journal of Agricultural Biotechnology,2014,22(3):326-332.(in Chinese)
[5]MOELLMANN G,LERNER A B,HENDEE J R.The mechanism of frog skin lightening by acetylcholine[J].Gen Comp Endocrinol,1974,23(1):45-51.
[6]REPPERT S M,WEAVER D R,RIVKEES S A,et al.Putative melatonin receptors in a human biological clock[J].Science,1988,242(4875):78-81.
[7]CONG Y,DENG H,FENG Y,et al.Melatonin implantation from winter solstice could extend the cashmere growth phase effectively[J].Small Rumin Res,2011,99(1):48-53.
[8]TUSHER V G,TIBSHIRANI R,CHU G.Significance analysis of microarrays applied to the ionizing radiation response[J].Proc Natl Acad Sci,2001,98(9):5116-5121.
[9]GHOSH S,SINGH A K,HALDAR C.Seasonal modulation of immunity by melatonin and gonadal steroids in a short day breeder goat capra hircus[J].Theriogenology,2014,82(8):1121-1130.
[10]ZHANG Q,LI J,CHEN Y,et al.Growth and viability of Liaoning Cashmere goat hair follicles during the annual hair follicle cycle[J].Genet Mol Res,2014,13(2):4433-4443.
[11]FOULKES N S,ASSONE-CORSI P,BORJIGIN J,et al.Rhythmic transcription:the molecular basis of circadian melatonin synthesis[J].Trends Neurosci,1997,20(10):487-492.
[12]LANOIX D,BEGHDADI H,LAFOND J,et al.Human placental trophoblasts synthesize melatonin and express its receptors[J].J Pineal Res,2008,45(1):50-60.
[13]LI Y H,ZHANG K,YANG K,et al.Adenovirus-mediated Wnt10boverexpression induces hair follicle regeneration[J].J Invest Dermatol,2013,133(1):42-48.
[14]MYUNG P S,TAKEO M,ITO M,et al.Epithelial Wnt ligand secretion is required for adult hair follicle growth and regeneration[J].J Invest Dermatol,2013,133(1):31-41.
[15]CHEN D,JARRELL A,GUO C,et al.Dermalβ-catenin activity in response to epidermal Wnt ligands is required for fibroblast proliferation and hair follicle initiation[J].Development,2012,139(8):1522-1533.
[16]ALONSO L,FUCHS E.Stem cells in the skin:waste not,Wnt not[J].Genes Dev,2003,17(10):1189-1200.
[17]WANG B,LI H,LIU Y,et al.Expression patterns of WNT/β-CATENIN signaling molecules during human tooth development[J].J Mol Hist,2014,45(5):487-496.
[18]MAEKAWA M,TAKASHIMA N,ARAI Y,et al.Pax6is required for production and maintenance of progenitor cells in postnatal hippocampal neurogenesis[J].Genes Cells,2005,10(10):1001-1014.
[19]ROWE J M,WELSH C,PENA R N,et al.Illuminating role of CYP1A1in skin function[J].J Invest Dermatol,2008,128(7):1866-1868.
[20]MAN X Y,YANG X H,CAI S Q,et al.Expression and localization of vascular endothelial growth factor and vascular endothelial growth factor receptor-2in human epidermal appendages:a comparison study by immunofluorescence[J].Clin Exp Dermatol,2009,34(3):396-401.
[21]WU X J,ZHU J W,JING J,et al.VEGF 165 modulates proliferation,adhesion,migration and differentiation of cultured human outer root sheath cells from central hair follicle epithelium through VEGFR-2activation in vitro[J].J Dermatol Sci,2014,73(2):152-160.