甘加型藏绵羊发情周期内输卵管和子宫中MTR1a mRNA的表达
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摘要
旨在研究褪黑素受体(MTR1a)基因在甘加型藏绵羊(Ganga Tibetan ovis aries)输卵管和子宫组织中的表达规律,选取发情周期内正常且未孕的甘加型藏绵羊32只,应用qRT-PCR方法检测输卵管和子宫组织中MTR1a基因的表达差异。结果显示:在整个发情周期中,甘加型藏绵羊输卵管和子宫中均有MTR1a mRNA表达。在发情前期、发情期、发情后期、间情期4个阶段,输卵管组织中MTR1a mRNA的相对表达量分别为0.604±0.253、 0.033±0.031、 0.202±0.151和1.000±0.221,间情期的相对表达量最高,与发情期差异极显著,与其他时期差异显著;子宫组织中MTR1a mRNA的相对表达量分别为0.188±0.086、0.525±0.347、1.878±0.209和1.000±0.094,发情后期的相对表达量最高,与其他3个时期差异极显著。发情周期内MTR1a基因在输卵管和子宫中的表达变化规律表明褪黑素(Melatonin,MT)在动物排卵、受精及早期胚胎发育过程中起调节作用。
We selected 32 normal, unpregnant Ganjia Tibetan sheep during the estrous cycle to study the expression of melatonin(MT) receptor( MTR1 a) gene in the oviduct and uterus of them. MTR1 a gene was detected in the oviduct and uterus by using qRT-PCR. The results showed that MTR1 a mRNA was expressed in the oviduct and uterus tissue of Ganjia Tibetan sheep during the whole estrus cycle. In proestrus, estrus, late estrus and dioestrum stages,the relative expression levels of MTR1 a mRNA in the oviduct tissues were 0.604±0.253, 0.033±0.031, 0.202±0.151 and 1.000±0.221 respectively.The relative expression of MTR1 a mRNA in the dioestrum was the highest and extremely significantly different from that in estrus and significantly different from that those in other two periods. At the same period, the relative expression levels of MTR1 a mRNA in the uterus tissues were 0.188±0.086, 0.525±0.347, 1.878±0.209 and 1.000±0.094 respectively, where the relative expression in the late estrus was the highest, which was extremely significantly different from that in other three stages.During oestrus cycle the dynamic changes of MTR1 a gene in oviduct and uterus tissueshowed that MT plays an important regulatory role in animal ovulation, fertilization and early embryonic development.
We selected 32 normal, unpregnant Ganjia Tibetan sheep during the estrous cycle to study the expression of melatonin(MT) receptor( MTR1 a) gene in the oviduct and uterus of them. MTR1 a gene was detected in the oviduct and uterus by using qRT-PCR. The results showed that MTR1 a mRNA was expressed in the oviduct and uterus tissue of Ganjia Tibetan sheep during the whole estrus cycle. In proestrus, estrus, late estrus and dioestrum stages,the relative expression levels of MTR1 a mRNA in the oviduct tissues were 0.604±0.253, 0.033±0.031, 0.202±0.151 and 1.000±0.221 respectively.The relative expression of MTR1 a mRNA in the dioestrum was the highest and extremely significantly different from that in estrus and significantly different from that those in other two periods. At the same period, the relative expression levels of MTR1 a mRNA in the uterus tissues were 0.188±0.086, 0.525±0.347, 1.878±0.209 and 1.000±0.094 respectively, where the relative expression in the late estrus was the highest, which was extremely significantly different from that in other three stages.During oestrus cycle the dynamic changes of MTR1 a gene in oviduct and uterus tissueshowed that MT plays an important regulatory role in animal ovulation, fertilization and early embryonic development.
引文
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[2] HE C,WANG J,ZHANG Z,et al.Mitochondria synthesize melatonin to ameliorate its function and improve mice oocyte’s quality under in vitro conditions[J].International Journal of Molecular Sciences,2016,17(6): 939.
[3] 孙永明,张燕军,李国华,等.内蒙古绒山羊褪黑激素受体1a基因外显子1克隆及序列分析[J].中国畜牧兽医,2011,38(5):82-84. SUN Y M,ZHANG Y J,LI G H,et al.Cloning and sequence analysis of Exon 1 of melatonin receptor 1a gene in inner mongolia cashmere goat[J].China Animal Husbandry & Veterinary Medicine,2011,38(5):82-84.
[4] 朱根宝,于兰兰,刘亚.褪黑素调控动物生殖内分泌的研究进展[J].动物医学进展,2016,37(3):94-98. ZHU G B,YU L L,LIU Y.Advances in research on melatonin regulating reproductive endocrine in animals[J].Progress in Animal Medicine,2016,37(3):94-98.
[5] 雷梦媛,狄冉,刘秋月,等.褪黑激素受体1A基因 (MTR1a )的分子遗传学研究进展[J].农业生物技术学报,2015,23(03):388-396. LEI M Y,DI R,LIU Q Y,et al.Progress in molecular genetics of melatonin receptor 1A gene( MTR1a )[J].Journal of Agricultural Biotechnology,2015,23(3):388-396.
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[7] 张前中,包永清,何文,等.甘加羊种质特性[J].中国草食动物,2008,28(3):66-68. ZHANG Q ZH,BAO Y Q ,HE W,et al.Ganjia sheep germplasm characteristics[J].China Herbivores Science,2008,28(3):66-68.
[8] 史军红,何玉琴,葛文博,等.高原甘加型藏绵羊生殖周期垂体FSH β和LH β mRNA表达差异的研究[J].中国兽医科学,2016,46(11):1444-1449. SHI J H,HE Y Q ,GE W B,et al.Study on the difference of FSHβ and LHβ mRNA expression in pituitary gland of Ganjia Tibetan sheep in the plateau[J].Chinese Veterinary Science,2016,46(11):1444-1449.
[9] 李伶俐,林汉华,陈晓娟 ,等.外源性褪黑素水平对雌性性早熟大鼠的影响[J].实用儿科临床杂志,2007,22(20):1539-1541. LI L L,LIN H H,CHEN X J,et al.Effect of exogenous melatonin on female precocious rats[J].Chinese Journal of Applied Clinical Pediatrics,2007,22(20):1539-1541.
[10] 张伟,王世银,甘尚权,等.Kisspeptins/GPR54神经内分泌系统对哺乳动物生殖活动的调控[J].中国细胞生物学学报,2014,36(8):1200-1206. ZHANG W,WANG SH Y,GAN SH Q,et al.Regulation of mammalian reproductive activities by Kisspeptins/GPR54 neuroendocrine system[J].China Journal of Cell Biology,2014,36(8):1200-1206.
[11] 王丽,李泰云,王树迎,等.沂蒙黑山羊发情周期中输卵管促性腺激素受体基因的表达差异研究[J].中国兽医科学,2011,41(8):832-838. WANG L,LI T Y,WANG SH Y,et al.Differential expression of tubal gonadotropin receptor gene in the estrous cycle of Yimeng black goat[J].Chinese Veterinary Science,2011,41(8):832-838.
[12] TABECK-LONCZYNSKA A,MYTYCH J,SOLEK P,et al.Melatonin receptors subtypes (MT1 and MT2) in the uterus masculinus of mature male european bison.Biological and seasonal reproductive role[J].Journal of Physiology and Pharmacology,2018,69(1):67-73.
[13] 霍生东,袁其建,卢建雄,等.褪黑素受体在未妊娠和妊娠牦牛子宫表达的研究[J].畜牧兽医学报,2015,46(2):239-244. HUO SH D,YUAN Q J,LU J X,et al.Study on the expression of melatonin receptor in uterus of nonpregnant and pregnant yak[J].Chinese Journal of Animal and Veterinary Sciences,2015,46(2):239-244.
[14] 陈玉林,张小辉.不同绵羊品种褪黑激素的季节性及昼夜变化规律研究[J].家畜生态学报,2005,26(1):35-38. CHEN Y L,ZHANG X H.Seasonal and day and night changes of melatonin in different sheep breeds[J].Journal of Domestic Animal Ecology,2005,26(1):35-38.
[15] 王凌燕,王树迎,侯衍猛,等.哺乳动物下丘脑-垂体-卵巢轴的研究进展[J].动物医学进展,2005,26(7):8-11. WANG L Y,WANG SH Y,HOU Y M,et al.Progress in research on mammalian hypothalamic-pituitary-ovarian axis[J].Progress in Animal Medicine,2005,26(7): 8-11.
[16] 杨颖,陈黎,卢立志.松果体调控动物季节性繁殖概述[J].农业生物技术学报,2017,25(7):1086-1101. YANG Y,CHEN L,LU L ZH.Overview of pineal body regulating seasonal reproduction of animals[J].Journal of Agricultural Biotechnology,2017,25(7):1086-1101.
[17] 储明星,冯涛,王西筠,等.哺乳动物繁殖季节性相关基因的研究进展[J].中国畜牧兽医,2010,37(6):107-110. CHU M X,FENG T,WANG X J,et al.Advances in research on seasonal genes related to mammalian reproductive[J].China Animal Husbandry & Veterinary Medicine,2010,37(6):107-110.
[18] 朱雪雁,葛闻博,何玉琴,等.甘加藏羊发情周期不同阶段血浆生殖激素分泌规律[J].河南农业科学,2017,46(11):138-143. ZHU X Y,GE W B,HE Y Q,et al.The law of plasma reproductive hormone secretion in different stages of estrus cycle in Ganjia Tibetan sheep[J].Journal of Henan Agricultural Sciences,2017,46(11):138-143.
[19] 葛闻博,何玉琴,朱雪雁,等.甘加型藏绵羊发情周期下丘脑GnRH分泌的动态变化规律[J].甘肃农业大学学报,2018,53(4):1-7. GE W B,HE Y Q,ZHU X Y,et al.Dynamic changes of GnRH secretion in hypothalamus during estrus cycle in Ganjia Tibetan sheep[J].Journal of Gansu Agricultural University,2018,53(4):1-7.
[20] CALVO J H,SERRANO M,MARTINEZ-ROYO A,et al.SNP rs403212791 in exon 2 of the MTNR1A gene is associated with reproductive seasonality in the Rasa aragonesa sheep breed[J].Theriogenology,2018,113:63-72.
[21] 王世银,邓双义,杨力伟,等. MTNR1A 基因多态性与绵羊季节性繁殖的相关性分析[J].西北农业学报,2015,24(8):23-30. WANG SH Y,DENG SH Y,YANG L W,et al.Polymorphism of melatonin receptor 1A ( MTNR1A ) and the relationship with seasonal reproduction of sheep[J].Acta Agriculturae Boreali-occidentalis Sinica,2015,24(8):23-30.
[22] MATEESCU R G,LUNSFORD A K,THONNEY M L.Association between melatonin receptor 1A gene polymorphismand reproductive performance in Dorset ewes[J].Journal of Animal Sciences,2009,87(8):2485-2488.
[23] CARCANGIU V,VACCA G M,MURA M C,et al.Relation-ship between MTNR1A melatonin receptor gene polymorphism and seasonal reproduction in different goatbreeds[J].Animal Reproduction Science,2009,110(1/2):71-78.
[2] HE C,WANG J,ZHANG Z,et al.Mitochondria synthesize melatonin to ameliorate its function and improve mice oocyte’s quality under in vitro conditions[J].International Journal of Molecular Sciences,2016,17(6): 939.
[3] 孙永明,张燕军,李国华,等.内蒙古绒山羊褪黑激素受体1a基因外显子1克隆及序列分析[J].中国畜牧兽医,2011,38(5):82-84. SUN Y M,ZHANG Y J,LI G H,et al.Cloning and sequence analysis of Exon 1 of melatonin receptor 1a gene in inner mongolia cashmere goat[J].China Animal Husbandry & Veterinary Medicine,2011,38(5):82-84.
[4] 朱根宝,于兰兰,刘亚.褪黑素调控动物生殖内分泌的研究进展[J].动物医学进展,2016,37(3):94-98. ZHU G B,YU L L,LIU Y.Advances in research on melatonin regulating reproductive endocrine in animals[J].Progress in Animal Medicine,2016,37(3):94-98.
[5] 雷梦媛,狄冉,刘秋月,等.褪黑激素受体1A基因 (MTR1a )的分子遗传学研究进展[J].农业生物技术学报,2015,23(03):388-396. LEI M Y,DI R,LIU Q Y,et al.Progress in molecular genetics of melatonin receptor 1A gene( MTR1a )[J].Journal of Agricultural Biotechnology,2015,23(3):388-396.
[6] 季从亮,储明星,陈国宏.褪黑激素受体基因的研究进展[J].遗传,2003,25(2):221-224. JI C L,CHU M X,CHEN G H.Progress in research on melatonin receptor gene[J].Heredity,2003,25(2):221-224.
[7] 张前中,包永清,何文,等.甘加羊种质特性[J].中国草食动物,2008,28(3):66-68. ZHANG Q ZH,BAO Y Q ,HE W,et al.Ganjia sheep germplasm characteristics[J].China Herbivores Science,2008,28(3):66-68.
[8] 史军红,何玉琴,葛文博,等.高原甘加型藏绵羊生殖周期垂体FSH β和LH β mRNA表达差异的研究[J].中国兽医科学,2016,46(11):1444-1449. SHI J H,HE Y Q ,GE W B,et al.Study on the difference of FSHβ and LHβ mRNA expression in pituitary gland of Ganjia Tibetan sheep in the plateau[J].Chinese Veterinary Science,2016,46(11):1444-1449.
[9] 李伶俐,林汉华,陈晓娟 ,等.外源性褪黑素水平对雌性性早熟大鼠的影响[J].实用儿科临床杂志,2007,22(20):1539-1541. LI L L,LIN H H,CHEN X J,et al.Effect of exogenous melatonin on female precocious rats[J].Chinese Journal of Applied Clinical Pediatrics,2007,22(20):1539-1541.
[10] 张伟,王世银,甘尚权,等.Kisspeptins/GPR54神经内分泌系统对哺乳动物生殖活动的调控[J].中国细胞生物学学报,2014,36(8):1200-1206. ZHANG W,WANG SH Y,GAN SH Q,et al.Regulation of mammalian reproductive activities by Kisspeptins/GPR54 neuroendocrine system[J].China Journal of Cell Biology,2014,36(8):1200-1206.
[11] 王丽,李泰云,王树迎,等.沂蒙黑山羊发情周期中输卵管促性腺激素受体基因的表达差异研究[J].中国兽医科学,2011,41(8):832-838. WANG L,LI T Y,WANG SH Y,et al.Differential expression of tubal gonadotropin receptor gene in the estrous cycle of Yimeng black goat[J].Chinese Veterinary Science,2011,41(8):832-838.
[12] TABECK-LONCZYNSKA A,MYTYCH J,SOLEK P,et al.Melatonin receptors subtypes (MT1 and MT2) in the uterus masculinus of mature male european bison.Biological and seasonal reproductive role[J].Journal of Physiology and Pharmacology,2018,69(1):67-73.
[13] 霍生东,袁其建,卢建雄,等.褪黑素受体在未妊娠和妊娠牦牛子宫表达的研究[J].畜牧兽医学报,2015,46(2):239-244. HUO SH D,YUAN Q J,LU J X,et al.Study on the expression of melatonin receptor in uterus of nonpregnant and pregnant yak[J].Chinese Journal of Animal and Veterinary Sciences,2015,46(2):239-244.
[14] 陈玉林,张小辉.不同绵羊品种褪黑激素的季节性及昼夜变化规律研究[J].家畜生态学报,2005,26(1):35-38. CHEN Y L,ZHANG X H.Seasonal and day and night changes of melatonin in different sheep breeds[J].Journal of Domestic Animal Ecology,2005,26(1):35-38.
[15] 王凌燕,王树迎,侯衍猛,等.哺乳动物下丘脑-垂体-卵巢轴的研究进展[J].动物医学进展,2005,26(7):8-11. WANG L Y,WANG SH Y,HOU Y M,et al.Progress in research on mammalian hypothalamic-pituitary-ovarian axis[J].Progress in Animal Medicine,2005,26(7): 8-11.
[16] 杨颖,陈黎,卢立志.松果体调控动物季节性繁殖概述[J].农业生物技术学报,2017,25(7):1086-1101. YANG Y,CHEN L,LU L ZH.Overview of pineal body regulating seasonal reproduction of animals[J].Journal of Agricultural Biotechnology,2017,25(7):1086-1101.
[17] 储明星,冯涛,王西筠,等.哺乳动物繁殖季节性相关基因的研究进展[J].中国畜牧兽医,2010,37(6):107-110. CHU M X,FENG T,WANG X J,et al.Advances in research on seasonal genes related to mammalian reproductive[J].China Animal Husbandry & Veterinary Medicine,2010,37(6):107-110.
[18] 朱雪雁,葛闻博,何玉琴,等.甘加藏羊发情周期不同阶段血浆生殖激素分泌规律[J].河南农业科学,2017,46(11):138-143. ZHU X Y,GE W B,HE Y Q,et al.The law of plasma reproductive hormone secretion in different stages of estrus cycle in Ganjia Tibetan sheep[J].Journal of Henan Agricultural Sciences,2017,46(11):138-143.
[19] 葛闻博,何玉琴,朱雪雁,等.甘加型藏绵羊发情周期下丘脑GnRH分泌的动态变化规律[J].甘肃农业大学学报,2018,53(4):1-7. GE W B,HE Y Q,ZHU X Y,et al.Dynamic changes of GnRH secretion in hypothalamus during estrus cycle in Ganjia Tibetan sheep[J].Journal of Gansu Agricultural University,2018,53(4):1-7.
[20] CALVO J H,SERRANO M,MARTINEZ-ROYO A,et al.SNP rs403212791 in exon 2 of the MTNR1A gene is associated with reproductive seasonality in the Rasa aragonesa sheep breed[J].Theriogenology,2018,113:63-72.
[21] 王世银,邓双义,杨力伟,等. MTNR1A 基因多态性与绵羊季节性繁殖的相关性分析[J].西北农业学报,2015,24(8):23-30. WANG SH Y,DENG SH Y,YANG L W,et al.Polymorphism of melatonin receptor 1A ( MTNR1A ) and the relationship with seasonal reproduction of sheep[J].Acta Agriculturae Boreali-occidentalis Sinica,2015,24(8):23-30.
[22] MATEESCU R G,LUNSFORD A K,THONNEY M L.Association between melatonin receptor 1A gene polymorphismand reproductive performance in Dorset ewes[J].Journal of Animal Sciences,2009,87(8):2485-2488.
[23] CARCANGIU V,VACCA G M,MURA M C,et al.Relation-ship between MTNR1A melatonin receptor gene polymorphism and seasonal reproduction in different goatbreeds[J].Animal Reproduction Science,2009,110(1/2):71-78.