孕酮处理对蛋鸡蛋壳钙化过程中CaBP-d28k基因表达的影响
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摘要
实验旨在研究孕酮对蛋鸡CaBP-d28k基因表达的影响。对产蛋高峰期蛋鸡进行孕酮和孕酮受体拮抗剂(RU-486)处理,利用实时荧光定量PCR技术(QRT-PCR),测定在蛋壳形成过程中输卵管子宫部钙结合蛋白(CaBP-d28k)、孕酮受体(PGR)的表达量;同时对血液中钙离子和孕酮浓度变化进行测定。结果表明:孕酮处理后,血液中钙离子浓度显著降低(P<0.05),输卵管子宫部CaBP-d28k表达量也显著降低(P<0.05);孕酮受体拮抗剂处理后,血液中钙离子浓度显著升高(P<0.05),输卵管子宫部CaBP-d28k表达量极显著升高(P<0.01)。结果提示,在蛋壳钙化过程中,孕酮对CaBP-d28k表达和钙离子转运起负调控作用。
This study was aimed to study the effect of progesterone on CaBP-d28k gene expression during the process of eggshell calcification in laying hens.In the present study,the hens were treated with progesterone or progesterone receptor antagonist(RU-486)during the peaking period of egg-laying.Real-time PCR was used to detect the expression levels of CaBP-d28k and PGR mRNA in the uterus of eggshell formation.Meanwhile,differential levels of serum calcium ion(Ca2+)and progesterone concentrations were also detected.Results showed that serum Ca2+ concentrations in test group,which treated by progesterone,was significantly lower than that in control group( P<0.05),and significantly lower expression levels of CaBP-d28k mRNA in the uterus was also detected in test group,when compared with the controls(P<0.05).Whereas,in group which treated by progesterone receptor antagonist,the serum Ca2+concentrations was significantly higher(P<0.05)than that in control group,and very significantly higher expression levels of CaBP-d28k mRNA in uterus occurred in this group(P<0.01),when compared with controls.It is indicated that progesterone has negative regulation on expression of CaBP-d28k and Ca2+ transport during eggshell calcification process in hens.
This study was aimed to study the effect of progesterone on CaBP-d28k gene expression during the process of eggshell calcification in laying hens.In the present study,the hens were treated with progesterone or progesterone receptor antagonist(RU-486)during the peaking period of egg-laying.Real-time PCR was used to detect the expression levels of CaBP-d28k and PGR mRNA in the uterus of eggshell formation.Meanwhile,differential levels of serum calcium ion(Ca2+)and progesterone concentrations were also detected.Results showed that serum Ca2+ concentrations in test group,which treated by progesterone,was significantly lower than that in control group( P<0.05),and significantly lower expression levels of CaBP-d28k mRNA in the uterus was also detected in test group,when compared with the controls(P<0.05).Whereas,in group which treated by progesterone receptor antagonist,the serum Ca2+concentrations was significantly higher(P<0.05)than that in control group,and very significantly higher expression levels of CaBP-d28k mRNA in uterus occurred in this group(P<0.01),when compared with controls.It is indicated that progesterone has negative regulation on expression of CaBP-d28k and Ca2+ transport during eggshell calcification process in hens.
引文
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[13]许多,朱伟杰,王自能.钙结合蛋白Calbindin-d28k在雌性(女)性生殖中的作用[J].生殖与避孕,2005,25(11):686-689.
[14]Luu K,Nie G Y,Salamonsen L A.Endometrial calbindins are critical for embryo implantation:Evidence form in vivo use of morpholino antisense oligonucleotides[J].Proc Natl Acad Sci USA,2004,101(21):8028-8033.
[15]Yang H,Kim T H,Lee H H,et al.Expression of calbindin-D28k and its regulation by estrogen in he human endometrium during the menstrual cycle[J].Reprod Biol Endocrinol,2011,9:28.
[16]Strunker T,Goodwin N,Brenker C,et al.The CatSper channel mediates progesterone-induced Ca2+influx in human sperm[J].Nature,2011,471(7338):382-386.
[2]李治学,魏丽娜,章世元.鸡蛋壳质量与结构关系的研究[J].中国畜牧杂志,2008,44(1):35-39.
[3]Rekawiecki R,Kowalik M K,Kotwica J.Nuclear progesterone receptor isoforms and their functions in the female reproductive tract[J].Pol J Vet Sci,2011,14(1):149-158.
[4]Krzysik-Walker S M,Ocón-Grove O M,Maddineni S B,et al.Identification of calcitonin expression in the chicken ovary:influence of follicular maturation and ovarian steroids[J].Biol Reprod,2007,77(10):626-635.
[5]Ding Y Q,Zhu L J,Bagchi M K,et al.Progesterone stimulates calcitonin gene expression in the uterus during implantation[J].Endocrinology,2001,135:2265-2274.
[6]Bar A,Vax E,Hunziker W,et al.The role of gonadal hormones in gene expression of Calbindin(Mr 28,000)in the laying hen[J].Gen Comp Endocr,1996,103(1):115-122.
[7]Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCTmethod[J].Methods,2001,25(4):402-408.
[8]陈桂勇.甾体激素核受体与辅助调节因子对基因表达的协同调控[J].国外医学计划生育分册,2004,23(2):103-105.
[9]Leonhardt S A,Boonyaratanakornkit V,Edwards D P.Progesterone receptor transcription and non-transcription signaling mechanisms[J].Steroids,2003,68(10-13):761-770.
[10]张宝云,狄冉,储明星,等.孕酮受体基因的研究进展[J].遗传,2008,30(12):1536-1544.
[11]康岳华,张宝云,王凭青,等.孕酮受体介导哺乳动物雌性生殖活动的分子机理[J].遗传,2012,34(10):1223-1232.
[12]姚佳.孕酮与干扰素_对体外培养的牛子宫内膜细胞基因组表达谱的影响[D].雅安:四川农业大学,2011.
[13]许多,朱伟杰,王自能.钙结合蛋白Calbindin-d28k在雌性(女)性生殖中的作用[J].生殖与避孕,2005,25(11):686-689.
[14]Luu K,Nie G Y,Salamonsen L A.Endometrial calbindins are critical for embryo implantation:Evidence form in vivo use of morpholino antisense oligonucleotides[J].Proc Natl Acad Sci USA,2004,101(21):8028-8033.
[15]Yang H,Kim T H,Lee H H,et al.Expression of calbindin-D28k and its regulation by estrogen in he human endometrium during the menstrual cycle[J].Reprod Biol Endocrinol,2011,9:28.
[16]Strunker T,Goodwin N,Brenker C,et al.The CatSper channel mediates progesterone-induced Ca2+influx in human sperm[J].Nature,2011,471(7338):382-386.