Abstract：In order to reveal the genetic diversity and tissue expression pattern of Parathyroid hormone-related peptide gene (PTHrP) in Gaojiao chickens (Gallus), Gaojiao chickens in Guizhou province were used in the present study, Aijiao chickens and Baiyi black chickens as controls. Its polymorphic loci were detected with DNA pool, Single Strand Conformation Polymorphism (SSCP) and DNA sequencing techniques, and its differential expression levels in heart, liver, breast muscl and leg muscle were analyzed by using real-time quantitative PCR technology. The results showed that the C→T point mutation was detected in 209bp of exon 3 of PTHrP gene of Gaojiao chickens，which was a silent mutation, hree genotypes (CC, CT and TT) were found.The relationship between PTHrP gene and shank length in Gaojiao chickens was analyzed. The C209T polymorphism wad significantly associate with shank length of Gaojiao chicken, and the individuals with CT genotype had significant shank length than CC and TT (P＜0.05). Thefitness test indicated that Gaojiao chickens at the mutation locus were Hardy-weinberg equilibrium(P＞0.05), and itpresented moderate polymorphism (PIC=0.350).. The RQ-PCR detection showed that the expression level of PTHrP gene was the highest in leg muscle of Gaojiao chickens, and significant higher than other tissues (P＜0.05). In addition, compare with Aijiao chicken and Baiyi black chicken,the expression levels of various tissues of Gaojiao chickens were higher than that of Baiyi black chickens (P＜0.05)，but lower than Aijiao chickens (P＜0.05). The results indicate that polymorphisms of PTHrP gene are significantly correlated with shank length traits of Gaojiao chickens, and individuals with CT genotype can be used as molecular genetic marker to select shank length of Gaojiao chicken for reproduction traits, and the expression levels of PTHrP has an influence on chickens' shank length.
[1] Moseley JM, Kubota M, Diefenbach-Jagger H, et al.Parathyroid hormone-related protein purified from a human lung cancer cell line [J].Proc Natl Acad sci, 1987, 84(14):5048-5052.
[2] Hock JM; Fonseca J; Gunness-Hey M.Comparison of the anabolic effects of synthetic parathyroid hormone-related protein(PTHrP) 1-34 and PTH 1-34 on bone in rats[J].Endocrinology, 1989, 125(4):2022-2027
[3]李瑛, 王金泉.甲状旁腺激素相关蛋白的新认识[J].肾脏病与透析肾移植杂志, 2003, 12(4):362-366
[4] Gujral, A, Burton, DW, Terkeltaub, R, Deftos, LJ.Parathyroid hormone-related protein induces interleukin 8 production by prostate cancer cells via a novel intracrine mechanism not mediated by its classical nuclear localization sequence[J].Cancer Research, ,2001, 61:2282-2288.
[5]李明辉, 郭风劲.骨骺生长板-信号轴及相关因子调节机制研究进展[J].国际骨科学杂志, 2007, 28(6):348-350
[6]郭敏, 张婧, 鹿蕾等.实验性单侧前牙反牙合修复体对大鼠髁突软骨中甲状旁腺激素相关蛋白和受体-表达的影响[J].华西口腔医学杂志, 2013, 31(2):122-126
[7]黄生高, 钟孝欢, 王会欣等.牵张力作用下甲状旁腺激素相关蛋白对人成骨样细胞增殖的影响[J].华西口腔医学杂志, 2009, 27(3):344-347
[8]段炳南, 邓锡云, 黄河等.体外骨对甲状旁腺激素相关蛋白-的双相反应[J].重庆医科大学学报, 2009, 34(2):148-151
[9]Kobayashi T, Chung U, Schipani E, et al.PTHrP and Indian hedgehog control differentiation of growth plate chondrocytes at multiple steps[J].Development, 2002, 129(12):2977-2986
[10] Lanske, Beate, Amling, Michael, Neff, Lynn等.Ablation of the PTHrP gene or the PTH/PTHrP receptor gene leads to distinct abnormalities in bone development. [J].J Clin Invest, 1999, 104(4):399.-407
[11]熊红珍, 徐会勇, 李松等.甲状旁腺激素相关蛋白在髁突软骨与生长板软骨发育早期的表达变化[J].广东牙病防治, 2012, 20(8):409-412
[12]马荣生, 周振雷, 侯加法等.信号轴对软骨内成骨的调控作用[J].畜牧与兽医, 2013, 45(7):96-99
[13]周坚, 贺林.软骨生长板的发育遗传学研究进展[J].国际遗传学杂志, 2008, 31(4):267-271
[14] Shawn P Grogan, Tsaiwei Olee, Koji Hiraoka, Martin K Lotz.Repression of Chondrogenesis Notch Signaling Proteins Hes-1 and Hey-1 Bind N-box Domains in the Col2a1 Enhancer Site[J].Arthritis & Rheumatism, 2008, 58(9):2754-2763
[15] 林飞太.PTHrP和Notch双信号通路调控骨骺干细胞增殖的研究[D].厦门大学, 2011.
[16] 金睿, 叶方, 朱明贵等.普定高脚鸡资源现状分析[J].现代农业科技, 2011, 331-331, 337., 10:311-311,377
[17]李蓓, 何小龙, 芒来.蒙古马突触融合蛋白基因多态性及在不同毛色皮肤组织中的表达分析[J].畜牧兽医学报, 2011, 42(11):1632-1637
[18]杨丽华, 白东义, 高雅罕, 等.蒙古马运动相关基因的表达量差异研究[J].畜牧兽医学报, 2013, 44(12):1900-1906
[19]付玉婷, 熊利峰, 于世宾等.实验性咬合紊乱对大鼠髁突软骨中及其受体表达的影响[J].口腔医学研究, 2010, 26(3):305-308
[20]Lozano D, de Castro LF, Dapia S et al.Role of parathyroid hormone-related protein in the decreased osteoblast function in diabetes-related osteopenia[J].Endocrinology, 2009, 150(5):2027-2035
[21]YAO S; PAN F; WISE G E.Chronological gene expression of parathyroid hormone-related protein(PTHrP) in the stellate reticulum of the rat:implications for tooth eruption[J].Archives of Oral Biology, 2007, 52(03):228-232
[22] Rotllant J, Worthington GP, Fuentes J, et al.Determination of tissue and plasma concentrations of PTHRP in fish: development and validation of a radioimmunoassay using a teleost 1-34 N-terminal petide[J].General and Comparative Endocrinology, 2003, 133:146-153
[23] 张存芳.黄牛PTHRP、GHRH和GHRHR基因SNP及其与生长性状关联分析[D].西北农林科技大学, 2008.
[24]郑惠玲, 杨振宇, 祝珍珍等.山羊基因的多克隆抗体制备和组织表达图谱分析[J].畜牧兽医学报, 2011, 42(2):177-181
[25]Zhao Q, Brauer PR, Xiao L et al.Expression of parathyroid hormone-related peptide (PthrP) and its receptor (PTH1R) during the histogenesis of cartilage and bone in the chicken mandibular process[J].Journal of Anatomy, 2002, 201(pt 2):137-151
[26]Ben, Bassat-S, Genina, O, Lavelin, I et al.Parathyroid receptor gene expression by epiphyseal growth plates in rickets and tibial dyschondroplasia[J].Molecular and Cellular Endocrinology, 1999, 149(12):185-195
[27] 王健.锰缺乏对肉仔鸡胫骨生长板软骨细胞发育相关因子的影响[D].山东农业大学, 2012.
[28]曹娟, 陈彬, 傅筑荫等.贵州兴义矮脚鸡遗传方式研究[J].中国家禽, 2010, 33(2):23-25
[29]Landauer W, Dunn L C.Studies on the Creeper Fowl [J].J Genet, 1930, 23:397-413
