甘加藏羊发情周期血浆Kisspeptin动态变化和PHPT轴中<i>Kiss1/GPR54</i> 的表达与分布

doi:10.7668/hbnxb.20193072

摘要/Abstract

摘要：

松果体-下丘脑-垂体-甲状腺轴(PHPTA)对动物的繁殖活动有重要的调节作用。为探讨Kiss1/GPR54系统在甘加藏羊PHPT轴的表达及其对繁殖活动的调控,选取24只处于发情周期内健康未孕的甘加藏羊作试验组,6只处于非繁殖季节的甘加藏羊作对照组,运用酶联免疫吸附法检测其血浆Kisspeptin动态变化,用实时荧光定量PCR、蛋白免疫印迹和免疫组织化学方法检测视神经、松果体、下丘脑、垂体和甲状腺组织中Kiss1、GPR54 mRNA及其蛋白的表达与分布。结果显示:血浆Kisspeptin分泌量在发情后期显著高于其他时期(P<0.05),繁殖季节内各时期分泌量均显著高于非繁殖季节(P<0.05)。Kiss1、GPR54 mRNA及其蛋白在视神经、PHPT轴组织中均有表达:视神经Kiss1、GPR54 mRNA相对表达量在发情前期显著高于其余时期;松果体在Kiss1 mRNA及其蛋白乏情期达到最大值,显著高于繁殖周期;下丘脑和甲状腺Kiss1、GPR54 mRNA相对表达量均在发情后期显著升高(P<0.05);垂体组织中二者mRNA的相对表达量在发情期显著高于其他时期。免疫组织化学检测结果显示:在视神经中,Kisspeptin与GPR54分别主要分布在神经胶质细胞核与胞质中;松果体细胞胞质中二者均呈强阳性表达;在下丘脑中,Kisspeptin表达在神经内分泌小细胞和神经胶质细胞中,GPR54表达在神经内分泌小细胞胞质中;在垂体组织中,Kisspeptin和GPR54主要表达在嗜碱性细胞的胞质中;而在甲状腺中,二者主要表达在滤泡细胞胞质中。甘加藏羊发情周期血浆Kisspeptin的动态变化及Kiss1、GPR54在各组织中的差异表达,表明Kiss1/GPR54系统参与调节了甘加藏羊的生殖生理活动。

关键词: 甘加藏羊, 季节性发情, Kiss1/GPR54, 松果体-下丘脑-垂体-甲状腺轴

Abstract:

The Pineal-hypothalamus-pituitary-thyroid axis(PHPTA)could significantly regulate reproductive activities of animals.In order to investigate the expression of Kiss1/GPR54 system on PHPT axis and its regulation on reproductive activities of Tibetan sheep,24 healthy and non-pregnant Tibetan sheep in estrus cycle were selected as experimental group and 6 non-breeding season Tibetan sheep as control group.The dynamic changes of plasma Kisspeptin were detected by ELISA,and the mRNA and protein expression of Kiss1 and GPR54 in optic nerve,pineal body,hypothalamus,pituitary body and thyroid gland were detected by Real-time quantitative PCR,Western Blot and immunohistochemistry.The results showed that the plasma Kisspeptin secretion in late estrus was significantly higher than that in other periods,and that in each period of breeding season was significantly higher than that in non-breeding season.The mRNA and protein of Kiss1 and GPR54 were expressed in optic nerve and PHPT axis.The relative expression of mRNA and protein of Kiss1 and GPR54 in optic nerve were significantly higher in early estrus than in other periods(P<0.05).The mRNA and its protein of Kiss1 of the pineal body reached its maximum value in estrus,significantly higher than that in reproductive cycle.The mRNA relative expression of Kiss1 and GPR54 of hypothalamus and thyroid gland were significantly increased in late estrus(P<0.05).The mRNA relative expression levels of the two in pituitary body were significantly higher in estrus than in other periods.Immunohistochemical results showed that Kisspeptin and GPR54 were mainly distributed in the glial nucleus and cytoplasm,respectively.Both were strongly positive in pineal cell cytoplasm.Kisspeptin was expressed in neuroendocrine small cells and glial cells in hypothalamus,GPR54 was expressed in cytoplasm of neuroendocrine small cells.Kisspeptin and GPR54 were mainly expressed in the cytoplasm of basophils in pituitary body.In thyroid gland,they were mainly expressed in the cytoplasm of follicular cells.The dynamic changes of plasma Kisspeptin and the differential expressions of Kiss1 and GPR54 in different tissues during estrus cycle of Ganjia Tibetan sheep indicated that Kiss1/GPR54 system was involved in regulating reproductive physiological activities of Ganjia Tibetan sheep.

Key words: Ganjia Tibetan sheep, Seasonal estrus, Kiss1/GPR54, Pineal-hypothalamus-pituitary-thyroid axis

杨亚文, 高何璇, 刘莉莉, 包莹莹, 何玉琴, 杨志杰, 陈卫刚, 葛闻博. 甘加藏羊发情周期血浆Kisspeptin动态变化和PHPT轴中Kiss1/GPR54 的表达与分布[J]. 华北农学报, 2022, 37(5): 204-212. doi: 10.7668/hbnxb.20193072.

YANG Yawen, GAO Hexuan, LIU Lili, BAO Yingying, HE Yuqin, YANG Zhijie, CHEN Weigang, GE Wenbo. Dynamic Changes of Plasma Kisspeptin and Expression and Distribution of Kiss1/GPR54 in PHPT Axis of Ganjia Tibetan Sheep During Estrus Cycle[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(5): 204-212. doi: 10.7668/hbnxb.20193072.

http://www.hbnxb.net/CN/Y2022/V37/I5/204

图/表 6

表1 qRT-PCR引物

Tab.1 Primer of qRT-PCR

基因 Gene	引物序列(5' —3') Primer sequence(5' —3')	产物大小/bp Product size	退火温度/℃ Annealing temperature	基因登录号 NCBI reference sequence
Kiss1	F:GCCTGATCCCGGCTCCGA	90	58	NM_001306104.1
	R:CGTAGCGCAGGCCGAAGGAG
GPR54	F:TACATCCAGCAGGTCTCGGTG	71	60	NM_001318077.1
	R:ACGTACCAGCGGTCCACACT
β-actin	F:CCATTGAGCACGGCATTGT	184	60	NM_001009784.3
	R:GCAGGGGTGTTGAAGGTCTC

图1 甘加藏羊发情周期和乏情期血浆中Kisspeptin动态变化 A.发情期(0~36 h)和发情后期(39~72 h);B.间情期(4 dM~13 dN)、发情前期(14 dM~16 dN)和乏情期(F);N.晚;M.早。

Fig.1 Dynamic changes of plasma Kisspeptin in estrus cycle and anestrus of Ganjia Tibetan sheep A.Estrus(0—36 h)and metestrus(39—72 h);B.Diestrus(4 dM—13 dN),proestrus(14 dM—16 dN)and anestrus(F) ;N.Night ;M.Morning.

图2 甘加藏羊发情周期和乏情期视神经、PHPT轴组织中Kiss1、GPR54 mRNA相对表达量同一组织不同时期间进行显著性分析,不同小写字母表示不同时期间差异显著(P<0.05)。图4同。

Fig.2 Optic nerve and PHPT axis tissues in estrus cycle and anestrus of Ganjia Tibetan sheep relative expression levels of Kiss1 and GPR54 mRNA Significance analysis was conducted in different periods of the same organization, and different lowercase letters indicated significant differences in different periods(P<0.05).The same as Fig.4.

图3 甘加藏羊发情周期和乏情期视神经、PHPT轴组织Western Blot A.视神经;B.松果体;C.下丘脑;D.垂体;E.甲状腺。

Fig.3 Western Blot results of optic nerve and PHPT axis tissues of Ganjia Tibetan sheep during estrus and anestrus A.Optic nerve;B.Pineal body;C.Hypothalamus;D.Pituitary body;E.Thyroid gland.

图4 甘加藏羊发情周期和乏情期视神经、PHPT轴组织中Kisspeptin、GPR54蛋白相对表达量

Fig.4 Optic nerve and PHPT axis tissues in estrus cycle and anestrus of Ganjia Tibetan sheep relative expression levels of Kisspeptin and GPR54 proteins

图5 甘加藏羊视神经和PHPT轴组织中Kisspeptin、GPR54免疫阳性产物分布(IHC) 1.神经胶质细胞;2.松果体细胞;3.神经细胞;4.神经细胞核;5.嗜酸性细胞;6.嗜碱性细胞;7.嫌色细胞;8.滤泡上皮细胞;9.滤泡胶质;10.滤泡旁细胞。

Fig.5 Distribution of Kisspeptin and GPR54 immunopositive products(IHC)in optic nerve and PHPT axis tissues of Ganjia Tibetan sheep 1.Glial cell;2.Pineal body cell;3.Neuron;4.Nerve nucleus;5.Eosinophilic cell;6.Basophilic cell ; 7.Chromophobe;8.Follicular epithelial cell;9.Follicular glial cell;10.Parafollicular cell.

参考文献 34

[1]	包莹莹. 甘加型藏羊发情周期血浆E₂动态变化规律及ERs在HPOA中的表达与分布[D]. 兰州: 甘肃农业大学, 2021,doi: 10.27025/d.cnki.ggsnu.2021.000135. doi: 10.27025/d.cnki.ggsnu.2021.000135
	Bao Y Y. The dynamic variation of E₂ in plsama and the distribution and expression of ERs in HPOA of Ganjia Tibetan sheep during estrus cycle[D]. Lanzhou: Gansu Agricultural University, 2021,
[2]	郎多勇, 李发弟, 郎侠, 张新慧. 甘加型藏羊种质特性研究[J]. 畜牧兽医杂志, 2010, 29(3):31-34.doi: 10.3969/j.issn.1004-6704.2010.03.011. doi: 10.3969/j.issn.1004-6704.2010.03.011
	Lang D Y, Li F D, Lang X, Zhang X H. Study on species property of Ganjia Tibetan sheep[J]. Journal of Animal Science and Veterinary Medicine, 2010, 29(3):31-34.
[3]	薛海龙, 赵蒙, 常彤, 许保增. 光周期调控动物繁殖的信号通路研究进展[J]. 特产研究, 2020, 42(2):76-78.doi: 10.16720/j.cnki.tcyj.2020.02.013. doi: 10.16720/j.cnki.tcyj.2020.02.013
	Xue H L, Zhao M, Chang T, Xu B Z. Advances in research on signal pathways related with regulating animal reproduction by photoperiod[J]. Special Wild Economic Animal and Plant Research, 2020, 42(2):76-78.
[4]	黄冬维, 储明星. 动物季节性繁殖分子调控机理研究进展[J]. 遗传, 2011, 33(7):695-706.doi: 10.3724/SP.J.1005.2011.00695. doi: 10.3724/SP.J.1005.2011.00695
	Huang D W, Chu M X. Research progress in molecular mechanism of animal seasonal reproduction[J]. Journal of Distance Education, 2011, 33(7):695-706.
[5]	Lee J H, Miele M E, Hicks D J, Phillips K K, Trent J M, Weissman B E, Welch D R. KiSS-1,a novel human malignant melanoma metastasis-suppressor gene[J]. Journal of the National Cancer Institute, 1996, 88(23):1731-1737.doi: 10.1093/jnci/88.23.1731. doi: 10.1093/jnci/88.23.1731 pmid: 8944003
[6]	王建发, 张旭, 徐丹丹, 李启涛, 张义爽, 柳巨雄. Kisspeptin/GPR54系统功能研究进展[J]. 中国畜牧杂志, 2015, 51(19):85-90.doi: 10.3969/j.issn.0258-7033.2015.19.019. doi: 10.3969/j.issn.0258-7033.2015.19.019
	Wang J F, Zhang X, Xu D D, Li Q T, Zhang Y S, Liu J X. Recent advance in into the physiological roles of kisspeptin/GPR54 system[J]. Chinese Journal of Animal Science, 2015, 51(19):85-90.
[7]	Harter C J L, Kavanagh G S, Smith J T. The role of kisspeptin neurons in reproduction and metabolism[J]. The Journal of Endocrinology, 2018, 238(3):R173-R183.doi: 10.1530/JOE-18-0108. doi: 10.1530/JOE-18-0108 URL
[8]	张伟, 王世银, 甘尚权, 石国庆. 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.doi: 10.11844/cjcb.2014.08.0074. doi: 10.11844/cjcb.2014.08.0074
	Zhang W, Wang S Y, Gan S Q, Shi G Q. Kisspeptins/GPR54 system and reproduction regulation in mammal[J]. Chinese Journal of Cell Biology, 2014, 36(8):1200-1206.
[9]	冯涛, 韦尚丽, 许晓玲, 白佳桦, 肖银霞, 郑琛, 田见晖, 刘彦. Kisspeptin和N-氨甲酰谷氨酸对GT1-7细胞GnRH分泌的影响[J]. 生物学杂志, 2019, 36(6):47-50,58.doi: 10.3969/j.issn.2095-1736.2019.06.047. doi: 10.3969/j.issn.2095-1736.2019.06.047
	Feng T, Wei S L, Xu X L, Bai J H, Xiao Y X, Zheng C, Tian J H, Liu Y. Effects of kisspeptin and N-carbamylgulamate on GnRH secretion in GT1-7 cells[J]. Journal of Microbiology, 2019, 36(6):47-50,58.
[10]	冯涛, 储明星, 王西筠, 张新珍, 刘雪云, 乔念民, 郭志勇, 刘新朋. 哺乳动物繁殖节律相关基因的研究进展[J]. 中国畜牧兽医, 2010, 37(7):89-92.
	Feng T, Chu M X, Wang X J, Zhang X Z, Liu X Y, Qiao N M, Guo Z Y, Liu X P. Advances on genes with reproductive rhythm in mammals[J]. China Animal Husbandry & Veterinary Medicine, 2010, 37(7):89-92.
[11]	Martins Trevisan C, Naslavsky M S, Monfardini F, Wang J, Zatz M, Peluso C, Pellegrino R, Mafra F, Hakonarson H, Ferreira F M, Nakaya H, Christofolini D M, Montagna E, Crandall K A, Barbosa C P, Bianco B. Variants in the kisspeptin-GnRH pathway modulate the hormonal profile and reproductive outcomes[J]. DNA and Cell Biology, 2020, 39(6):1012-1022.doi: 10.1089/dna.2019.5165. doi: 10.1089/dna.2019.5165 pmid: 32352843
[12]	张明泉. 下丘脑-垂体-甲状腺轴细胞分子机制的季节变化与中医肾本质研究的探讨[D]. 北京: 北京中医药大学, 2007.
	Zhang M Q. Seasonal variation of molecular mechanism of hypothalamic-pituitary-thyroid axis and study of renal essence in Traditional Chinese medicine[D]. Beijing: Beijing University of Chinese Medicine, 2007.
[13]	闫哲, 杨斯琪, 黄慧. 甲状腺激素及自身抗体对女性生殖功能的影响[J]. 西部医学, 2021, 33(10):1557-1560.doi: 10.3969/j.issn.1672-3511.2021.10.032. doi: 10.3969/j.issn.1672-3511.2021.10.032
	Yan Z, Yang S Q, Huang H. Thyroid hormone,autoantibodies and female reproduction[J]. Medical Journal of West China, 2021, 33(10):1557-1560.
[14]	陈逗逗, 刘坤钰, 郑旭琴. 甲状腺疾病对生殖障碍的影响[J]. 中国现代医学杂志, 2020, 30(16):55-58.doi: 10.3969/j.issn.1005-8982.2020.16.010. doi: 10.3969/j.issn.1005-8982.2020.16.010
	Chen D D, Liu K Y, Zheng X Q. The influence of thyroid diseases on reproductive disorders[J]. China Journal of Modern Medicine, 2020, 30(16):55-58.
[15]	陈卫刚. 甘加型藏羊发情周期下丘脑—垂体—卵巢轴kisspeptin/GPR54的表达与分布[D]. 兰州: 甘肃农业大学, 2020.
	Chen W G. Expression and distribution of kisspeptin/GPR54 in the hypothalamic-pituitary-ovary axis of Ganjia Tibetan sheep during the estrous cycle[D]. Lanzhou: Gansu Agricultural University, 2020.
[16]	Luo Q Q, Li W H, Li M, Zhang X Z, Zhang H Q. Leptin/leptinR-kisspeptin/kiss1r-GnRH pathway reacting to regulate puberty onset during negative energy balance[J]. Life Sciences, 2016, 153:207-212.doi: 10.1016/j.lfs.2016.03.048. doi: 10.1016/j.lfs.2016.03.048 pmid: 27212704
[17]	葛闻博, 何玉琴, 朱雪雁, 史军红, 华道才让, 韩志磊, 王文芳. 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.doi: 10.13432/j.cnki.jgsau.2018.04.001. doi: 10.13432/j.cnki.jgsau.2018.04.001
	Ge W B, He Y Q, Zhu X Y, Shi J H, Huadao C R, Han Z L, Wang W F. Dynamic secretion changes of GnRH on plasma in oestrous cycle of Ganjia Tibetan sheep[J]. Journal of Gansu Agricultural University, 2018, 53(4):1-7.
[18]	史军红, 何玉琴, 葛文博, 华道才让, 韩志磊, 王文芳, 朱雪雁. 高原甘加藏羊发情周期垂体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.doi: 10.16656/j.issn.1673-4696.2016.11.017. doi: 10.16656/j.issn.1673-4696.2016.11.017
	Shi J H, He Y Q, Ge W B, Huadao C R, Han Z L, Wang W F, Zhu X Y. Expression of FSH β mRNA and LH β mRNA in the pituitary of estrous cycle of Ganjia Tibetan sheep[J]. Chinese Veterinary Science, 2016, 46(11):1444-1449.
[19]	Jayasena C N, Abbara A, Comninos A N, Nijher G M K, Christopoulos G, Narayanaswamy S, Izzi-Engbeaya C, Sridharan M, Mason A J, Warwick J, Ashby D, Ghatei M A, Bloom S R, Carby A, Trew G H, Dhillo W S. Kisspeptin-54 triggers egg maturation in women undergoing in vitro fertilization[J]. The Journal of Clinical Investigation, 2014, 124(8):3667-3677.doi: 10.1172/JCI75730. doi: 10.1172/JCI75730 URL
[20]	安雪姣, 赵生国, 潘章源, 文禹粱, 田志龙, 蔡原, 储明星. 绵羊KISS1基因组织表达及其多态性与产羔数关联分析[J]. 中国畜牧杂志, 2019, 55(6):45-50.doi: 10.19556/j.0258-7033.2019-06-045. doi: 10.19556/j.0258-7033.2019-06-045
	An X J, Zhao S G, Pan Z Y, Wen Y L, Tian Z L, Cai Y, Chu M X. Tissue expression and polymorphism of sheep KISS1 gene and their association with litter size[J]. Chinese Journal of Animal Science, 2019, 55(6):45-50.
[21]	Higo S, Iijima N, Ozawa H. Characterisation of Kiss1r(Gpr54)-expressing neurones in the arcuate nucleus of the female rat hypothalamus[J]. Journal of Neuroendocrinology, 2017, 29(2). doi: 10.1111/jne.12452. doi: 10.1111/jne.12452
[22]	Tovar S, Vázquez M J, Navarro V M, Fernández-Fernández R, Castellano J M, Vigo E, Roa J, Casanueva F F, Aguilar E, Pinilla L, Dieguez C, Tena-Sempere M. Effects of single or repeated intravenous administration of kisspeptin upon dynamic LH secretion in conscious male rats[J]. Endocrinology, 2006, 147(6):2696-2704.doi: 10.1210/en.2005-1397. doi: 10.1210/en.2005-1397 pmid: 16513831
[23]	Lents C A, Heidorn N L, Barb R, Ford J J. Central and peripheral administration of kisspeptin activates gonadotropin but not somatotropin secretion in prepubertal gilts[J]. Society for Reproduction and Fertility, 2018, 135(6):879-887.doi: 10.1530/rep-07-0502. doi: 10.1530/rep-07-0502
[24]	Wang X S, Tang J. A new reproduction hormone modulation system:Kisspeptin/GPR54[J]. Foreign Medical Sciences, 2006, 25(4):197-199.
[25]	Funes S, Hedrick J A, Vassileva G, Markowitz L, Abbondanzo S, Golovko A, Yang S J, Monsma F J, Gustafson E L. The KiSS-1 receptor GPR54 is essential for the development of the murine reproductive system[J]. Biochemical and Biophysical Research Communications, 2003, 312(4):1357-1363.doi: 10.1016/j.bbrc.2003.11.066. doi: 10.1016/j.bbrc.2003.11.066 pmid: 14652023
[26]	Tena-Sempere M. GPR54 and kisspeptin in reproduction[J]. Human Reproduction Update, 2006, 12(5):631-639.doi: 10.1093/humupd/dml023. doi: 10.1093/humupd/dml023 pmid: 16731583
[27]	Um H N, Han J M, Hwang J I, Hong S I, Vaudry H, Seong J Y. Molecular coevolution of kisspeptins and their receptors from fish to mammals[J]. Annals of the New York Academy of Sciences, 2010, 1200:67-74.doi: 10.1111/j.1749-6632.2010.05508.x. doi: 10.1111/j.1749-6632.2010.05508.x pmid: 20633134
[28]	Tsutsui K, Bentley G E, Kriegsfeld L J, Osugi T, Seong J Y, Vaudry H. Discovery and evolutionary history of gonadotrophin-inhibitory hormone and kisspeptin:New key neuropeptides controlling reproduction[J]. Journal of Neuroendocrinology, 2010, 22(7):716-727.doi: 10.1111/j.1365-2826.2010.02018.x. doi: 10.1111/j.1365-2826.2010.02018.x pmid: 20456604
[29]	Smith J T, Coolen L M, Kriegsfeld L J, Sari I P, Jaafarzadehshirazi M R, Maltby M, Bateman K, Goodman R L, Tilbrook A J, Ubuka T, Bentley G E, Clarke I J, Lehman M N. Variation in kisspeptin and RFamide-related peptide(RFRP)expression and terminal connections to gonadotropin-releasing hormone neurons in the brain:a novel medium for seasonal breeding in the sheep[J]. Endocrinology, 2008, 149(11):5770-5782.doi: 10.1210/en.2008-0581. doi: 10.1210/en.2008-0581 URL
[30]	王军, 孙蕾, 常喆. KiSS-1基因与生殖相关的表达及其调节[J]. 中国农学通报, 2011, 27(20):11-13.
	Wang J, Sun L, Chang Z. Expression and its regulation of KiSS-1 gene related with reproduction[J]. Chinese Agricultural Science Bulletin, 2011, 27(20):11-13.
[31]	金慧慧, 刘秋月, 狄冉, 胡文萍, 王翔宇, 储明星. Kiss1基因调控哺乳动物繁殖性能的研究进展[J]. 家畜生态学报, 2015, 36(8):1-6.doi: 10.3969/j.issn.1673-1182.2015.08.001. doi: 10.3969/j.issn.1673-1182.2015.08.001
	Jin H H, Liu Q Y, Di R, Hu W P, Wang X Y, Chu M X. Research progress on reproduction regulation of Kiss1 gene in mammals[J]. Journal of Domestic Animal Ecology, 2015, 36(8):1-6.
[32]	郭若婷. 绵羊kisspeptin介导的GnRH基因表达调控机制的研究[D]. 石河子: 石河子大学, 2015,doi: 10.7666/d.D718147. doi: 10.7666/d.D718147
	Guo R T. Genetic mechanisms mediating kisspeptin regulation of GnRH gene expression in sheep[D]. Shihezi: Shihezi University, 2015,
[33]	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.doi: 10.3969/j.issn.1674-7968.2017.07.006. doi: 10.3969/j.issn.1674-7968.2017.07.006
	Yang Y, Chen L, Lu L Z. Review of regulation on the pineal gland of animal seasonal reproduction[J]. Journal of Agricultural Biotechnology, 2017, 25(7):1086-1101.
[34]	欧科鹏, 李尤简, 郭若婷, 王颖, 顾真真, 刘乙, 刘小军. 绵羊甲状腺自主合成并分泌褪黑素的研究[J]. 畜牧兽医学报, 2015, 46(4):576-582.doi: 10.11843/j.issn.0366-6964.2015.04.010. doi: 10.11843/j.issn.0366-6964.2015.04.010
	Ou K P, Li Y J, Guo R T, Wang Y, Gu Z Z, Liu Y, Liu X J. Investigation into synthesis and secretion of melatonin in thyroid gland in sheep[J]. Chinese Journal of Animal and Veterinary Sciences, 2015, 46(4):576-582.

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甘加藏羊发情周期血浆Kisspeptin动态变化和PHPT轴中Kiss1/GPR54 的表达与分布

Dynamic Changes of Plasma Kisspeptin and Expression and Distribution of Kiss1/GPR54 in PHPT Axis of Ganjia Tibetan Sheep During Estrus Cycle

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甘加藏羊发情周期血浆Kisspeptin动态变化和PHPT轴中Kiss1/GPR54 的表达与分布

Dynamic Changes of Plasma Kisspeptin and Expression and Distribution of Kiss1/GPR54 in PHPT Axis of Ganjia Tibetan Sheep During Estrus Cycle

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