leptin基因在团头鲂成鱼各组织和早期发育中的表达分析
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摘要
应用荧光定量PCR方法检测团头鲂leptin及其相关基因—瘦素受体(leptin receptor,leptin-R)和瘦素受体重叠转录产物类似物1基因(leptin receptor overlapping transcript like-1,leprotl-1)在胚胎发育早期及成鱼各组织的表达量。结果显示,3个基因在团头鲂成鱼所有检测组织中均有所表达,表明3个基因在团头鲂机体中具有多重生理功能。早期发育过程中的表达量结果显示leptin-R和leprotl-1基因在胚胎阶段的表达水平变化趋势一致,而leptin在破膜后5d前一直处于较低并呈波动性变化;出膜后12d时leptin-R和leprotl-1基因表达量均达到了峰值,而leptin基因从破膜后第5天开始表达量持续上升,在第15天到达峰值,表明3个基因可能涉及到胚胎发育早期阶段的能量调控。Pearson相关分析显示3个基因在团头鲂胚胎发育早期中的表达均为显著正相关关系(P<0.05)。
In the present study,the expression levels of the leptin and its related genes including leptin receptor(leptin-R)and leptin receptor overlapping transcript like-1(leprotl-1)in Megalobrama amblycephala during embryonic and larval development as well as in various adults' tissues were studied using qRT-PCR.The results showed that the three genes were ubiquitously expressed in all the examined adults' tissues,which suggested their multiple physiology functions in M.amblycephala.During embryonic and larval developmental,the leptin-Rand leprotl-1genes had lower and fluctuant expression levels.Expression levels of the three genes had no differences before 5days post-hatching(dph),which suggested that these genes may be related to the regulation of energy during early developmental stages.Expression levels of both the leptin-Rand leprotl-1genes reached the peak at 12 dph,but the leptin expression increased from 5dph and reached the peak at 15 dph,which indicated that the three genes may participated in the M.amblycephalasex differentiation during larval developmental stages.The correlation coefficients among leptin,leptin-Rand leprotl-1mRNA levels indicated that these three genes were positively correlated during embryonic and larval developmental stages in M.amblycephala.
In the present study,the expression levels of the leptin and its related genes including leptin receptor(leptin-R)and leptin receptor overlapping transcript like-1(leprotl-1)in Megalobrama amblycephala during embryonic and larval development as well as in various adults' tissues were studied using qRT-PCR.The results showed that the three genes were ubiquitously expressed in all the examined adults' tissues,which suggested their multiple physiology functions in M.amblycephala.During embryonic and larval developmental,the leptin-Rand leprotl-1genes had lower and fluctuant expression levels.Expression levels of the three genes had no differences before 5days post-hatching(dph),which suggested that these genes may be related to the regulation of energy during early developmental stages.Expression levels of both the leptin-Rand leprotl-1genes reached the peak at 12 dph,but the leptin expression increased from 5dph and reached the peak at 15 dph,which indicated that the three genes may participated in the M.amblycephalasex differentiation during larval developmental stages.The correlation coefficients among leptin,leptin-Rand leprotl-1mRNA levels indicated that these three genes were positively correlated during embryonic and larval developmental stages in M.amblycephala.
引文
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[23]FANTUZZI G,FAGGIONI R.Leptin in the regulation of immunity,inflammation,and hematopoiesis[J].The Journal of Leukocyte Biology,2000,68(10):437-446.
[24]LAM Q L K,LU L.Role of leptinin immunity[J].Cellular&Molecular Immunology,2007,4(1):1-13.
[25]TROMBLEY S,SCHMITZ M.Leptin in fish:possible role in sexual maturation in male Atlantic salmon[J].Fish Physiology and Biochemistry,2013,39(1):103-106.
[26]MARK R D,QIN L,MASON D K,et al.Leptin expression affects metabolic rate in zebrafish embryos(D.rerio)[J].Frontiers in Physiology,2013,4:160.
[27]ZHANG H X,CHEN H P,ZHANG Y,et al.Molecular cloning,characterization and expression profiles of multiple leptin genes and a leptin receptor gene in orange-spotted grouper(Epinephelus coioides)[J].General and Comparative Endocrinology,2013,181:295-305.
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[2]KUROKAWA T,UJI S,SUZUKI T.Identification of cDNA coding for a homologue to mammalian leptinfrom pufferfish,Takifugu rubripes[J].Peptides,2005,26:745-750.
[3]HUISING M O,GEVEN E J,KRUISWIJK C P,et al.Increased leptinexpression in common carp(Cyprinus carpio)after food intake but not after fasting or feeding to satiation[J].Endocrinology,2006,147:5786-5797.
[4]MURASHITA K,UJI S,YAMAMOTO T,et al.Production of recombinant leptin and its effects on food intake in rainbow trout(Oncorhynchus mykiss)[J].Comparative Biochemistry and Physiology Part B,2008,150(4):377-384.
[5]GORISSEN M,BERNIER N J,NABUURS S B.Two divergent leptin paralogues in zebrafish(Danio rerio)that originate early in teleostean evolution[J].Endocrinology,2009,201(3):329-339.
[6]LI G G,LIANG X F,XIE Q L.Gene structure,recombinant expression and functional characterization of grass carp leptin[J].General and Comparative Endocrinology,2010,166(1):117-127.
[7]GONG Y,LUO Z,QING L Z,et al.Characterization and tissue distribution of leptin,leptin receptor and leptin receptor overlapping transcript genes in yellow catfish Pelteobagrus fulvidraco[J].General and Comparative Endocrinology,2013,182:1-6.
[8]TANG C H,LU D Y,YANG R S,et al.Leptin-induced IL-6production is mediat ed by leptin receptor,insulin receptor substrate-1,phosphatidylinositol 3-kinase,Akt,NF-kappaB,and p300pathway in microglia[J].Immunology,2007,179(2):1292-1302.
[9]KUROKAWA T,MURASHITA K,SUZUKI T,et al.Genomic characterization and tissue distribution of leptinR and leptinR overlapping transcript genes in the pufferfish Takifugu rubripes[J].General and Comparative Endocrinology,2008,158:108-114.
[10]KUROKAWA T,MURASHITA T.Genomic characterization of multiple leptin genes and a leptin receptor gene in the Japanese medaka,Oryzias latipes[J].General and Comparative Endocrinology,2009,161(2):229-237.
[11]BAILLEUL B,AKERBLOM I,STROSBERG A D.The leptin receptor promoter controls expression of a second distinct protein[J].Nucleic Acids Research,1997,25:2752-2758.
[12]STRAUSBERG R L,FEINGOLD E A,GROUSE L H,et al.Generation and initial analysis of more than 15000full-length human and mouse cDNA sequences[J].Proc Natl AcadSci USA,2002,99(26):16899-16903.
[13]李光照,梁旭方,陈亮,等.鳜鱼肥胖基因的克隆与组织表达[J].暨南大学学报(自然科学与医学版),2010,31(3):318-323,330.
[14]MARNIX G,GERT F.Leptin in teleostean fish,towards the origins of leptin[J].Physiology,2014,61/62:200-206.
[15]曾聪,曹小娟,高泽霞,等.团头鲂生长性状的遗传力和育种值估计[J].华中农业大学学报,2014,33(2):89-95.
[16]易少奎,高泽霞,赵鸿昊,等.团头鲂生长相关基因在不同发育时期生长轴组织的表达分析[J].水生生物学报,2015,39(1):101-109.
[17]高泽霞,王卫民,蒋恩明,等.团头鲂种质资源及遗传改良研究进展[J].华中农业大学学报,2014,33(3):138-144.
[18]KENNETH J L,THOMAS D S.Analysis of relative gene expression data using real-time quantitative PCR and the2-△△CT method[J].Methods,2001,25(4):402-408.
[19]LONDRAVILLE R L,DUVALL C S.Murine leptininjections increase intracellular fatty acid-binding protein in green sunfish(Lepomis cyanellus)[J].General and Comparative Endocrinology,2002,129(1):56-62.
[20]VOLKOFF H,EYKELBOSH A J,PETER R E.Role of leptin in the control of feeding of goldfish(Carassius auratus):interactions with cholecystokinin,neuropeptide Y and orexin A,and modulation by fasting[J].Brain Research,2003,962(1/2):90-109.
[21]李观贵.草鱼瘦素的基因结构、重组表达与功能研究[D].广州:暨南大学图书馆,2010.
[22]唐永凯,俞菊华,徐跑,等.吉富罗非鱼leptin基因的克隆及其表达[J].动物学杂志,2012,47(3):89-94.
[23]FANTUZZI G,FAGGIONI R.Leptin in the regulation of immunity,inflammation,and hematopoiesis[J].The Journal of Leukocyte Biology,2000,68(10):437-446.
[24]LAM Q L K,LU L.Role of leptinin immunity[J].Cellular&Molecular Immunology,2007,4(1):1-13.
[25]TROMBLEY S,SCHMITZ M.Leptin in fish:possible role in sexual maturation in male Atlantic salmon[J].Fish Physiology and Biochemistry,2013,39(1):103-106.
[26]MARK R D,QIN L,MASON D K,et al.Leptin expression affects metabolic rate in zebrafish embryos(D.rerio)[J].Frontiers in Physiology,2013,4:160.
[27]ZHANG H X,CHEN H P,ZHANG Y,et al.Molecular cloning,characterization and expression profiles of multiple leptin genes and a leptin receptor gene in orange-spotted grouper(Epinephelus coioides)[J].General and Comparative Endocrinology,2013,181:295-305.
[28]PRASHANT B,LEAH K L.Discrimination begins in the womb:evidence of sex-selective prenatal investments[J].Human Resources,2013,48:71-113.