黄体期和卵泡期小尾寒羊DIO2与DIO3组织表达
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摘要
为分析DIO2和DIO3基因在小尾寒羊不同繁殖时期(黄体期和卵泡期)下丘脑-垂体-性腺轴各组织中的表达差异,阐明这2个基因在绵羊发情转换中的表达模式,实验采用实时荧光定量PCR(Real-time PCR)技术对比分析DIO2和DIO3基因在黄体期和卵泡期小尾寒羊下丘脑-垂体-性腺轴各组织中的表达差异。结果表明:DIO2和DIO3基因在下丘脑、垂体、松果体、大脑、小脑、卵巢、子宫、输卵管、肾脏、肾上腺10种组织中均表达;DIO2基因在黄体期和卵泡期的垂体组织中表达量显著高于其他组织(P<0.05),其中黄体期垂体、子宫、下丘脑、松果体、输卵管和卵巢DIO2的表达量显著高于卵泡期(P<0.05);DIO3在卵泡期松果体、下丘脑、子宫、垂体和输卵管的表达量显著高于黄体期(P<0.05)。综上,DIO2能抑制绵羊发情,而DIO3促进发情。
The aim of this study was to analyze the expression of DIO2 and DIO3 genes in the tissues of the hypothalamuspituitary-gonadal axis in Small Tail Han sheep during different reproductive periods(luteal and follicular periods), and to elucidate the expression pattern of these two genes in estrus conversion of sheep. Real-time PCR method was used to detect the expression of DIO2 and DIO3 genes in tissues of hypothalamus-pituitary-gonadal axis during luteal and follicular periods in Small Tail Han sheep. The results showed that DIO2 and DIO3 genes were all expressed in ten tissues such as brain, cerebellum, hypothalamus, pituitary, pineal body, uterus, ovary and oviduct. The expression of DIO2 in pituitary was significantly higher(P<0.05) than that in other tissues at both luteal and follicular periods. The expression of DIO2 in pituitary, uterus, hypothalamus, pineal body, fallopian tubes and ovaries was significantly higher(P<0.05) in luteal period than that in follicular period. The expression of DIO3 in pineal gland, hypothalamus, uterus, pituitary and fallopian tubes was significantly higher(P<0.05) in follicular period than that in luteal period. In conclusion, DIO2 could inhibit sheep estrus but DIO3 promote sheep estrus.
The aim of this study was to analyze the expression of DIO2 and DIO3 genes in the tissues of the hypothalamuspituitary-gonadal axis in Small Tail Han sheep during different reproductive periods(luteal and follicular periods), and to elucidate the expression pattern of these two genes in estrus conversion of sheep. Real-time PCR method was used to detect the expression of DIO2 and DIO3 genes in tissues of hypothalamus-pituitary-gonadal axis during luteal and follicular periods in Small Tail Han sheep. The results showed that DIO2 and DIO3 genes were all expressed in ten tissues such as brain, cerebellum, hypothalamus, pituitary, pineal body, uterus, ovary and oviduct. The expression of DIO2 in pituitary was significantly higher(P<0.05) than that in other tissues at both luteal and follicular periods. The expression of DIO2 in pituitary, uterus, hypothalamus, pineal body, fallopian tubes and ovaries was significantly higher(P<0.05) in luteal period than that in follicular period. The expression of DIO3 in pineal gland, hypothalamus, uterus, pituitary and fallopian tubes was significantly higher(P<0.05) in follicular period than that in luteal period. In conclusion, DIO2 could inhibit sheep estrus but DIO3 promote sheep estrus.
引文
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[16]王喜英.基于家系的DIO2基因多态性与秦巴山区人群精神发育迟滞的关联研究[D].西安:西北大学,2010.
[17]Torriani M,Srinivasa S,Fitch K V,et al.Dysfunctional subcutaneous fat with reduced Dicer and brown adipose tissue gene expression in HIV-infected patients[J].J Clin Endocr Metab,2016,101(3):1225.
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[22]Miera C S D,Hanon E A,Dardente H,et al.Circannual variation in thyroid hormone deiodinases in a short-day breeder[J].J Neuroendocrinol,2013,25(4):412.
[23]丁吉顺.Dio2/Dio3系统对黑像仓鼠繁殖的调控机制[D].曲阜:曲阜师范大学,2015.
[24]Kampflassin A,Prendergast B J.Acute downregulation of Type II and Type III iodothyronine deiodinases by photoperiod in peripubertal male and female Siberian hamsters[J].Gen Comp Endocr,2013,193(4):72.
[25]Janssen R,Zuidwijk M J,Muller A,et al.MicroRNA 214 is a potential regulator of thyroid hormone levels in the mouse heart following myocardial infarction,by targeting the thyroid-hormoneinactivating enzyme deiodinase type III[J].Front Endocrinol,2016,7:22.
[26]吴茜红.牛Dio3、Dio3os和Wif1基因印记及Wif1基因DNA甲基化状态分析[D].保定:河北农业大学,2015.
[27]Koopdonk-Kool J M,de Vijlder J J,Veenboer G J,et al.Type II and type III deiodinase activity in human placenta as a function of gestational age[J].J Clin Endocr Metab,1996,81(6):2154-2158.
[28]Deng W B,Liang X H,Liu J L,et al.Regulation and function of deiodinases during decidualization in female mice[J].Endocrinology,2014,155(7):2704-2717.
[29]杨宗林.猪DLK1-DIO3印记域4个候选印记基因的分离及其单等位基因表达分析[D].重庆:西南大学,2009.
[30]Kester M H,Martinez D M R,Obregon M J,et al.Iodoth-yronine levels in the human developing brain:major regulatory roles of iodothyronine deiodinases in different areas[J].J Clin Endocr Metab,2004,89(7):3117-3128.
[31]Ono H,Hoshino Y,Yasuo S,et al.Involvement of thyrotropin in photoperiodic signal transduction in mice[J].P Natl Acad Sci USA,2008,105(47):18238-18242.
[32]Nakao N,Ono H,Yamamura T,et al.Thyrotrophin in the pars tuberalis triggers photoperiodic response[J].Nature,2008,452(7185):317.
[33]Hanon E A,Routledge K,Dardente H,et al.Effect of photoperiod on the thyroid-stimulating hormone neuroendocrine system in the European hamster(Cricetus cricetus)[J].J Neuroendocrinol,2010,22(1):51-55.
[34]Ebling F J.Photoperiodic regulation of puberty in seasonal species[J].Mol Cell Endocrinol,2010,324(1):95-101.
[2]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.
[3]Barrett P,Ebling F J,Schuhler S,et al.Hypothalamic thyroid hormone catabolism acts as a gatekeeper for the seasonal control of body weight and reproduction[J].Endocrinology,2007,148(8):3608-3617.
[4]Revel F G,Saboureau M,Pévet P,et al.Melatonin regulates type 2 deiodinase gene expression in the Syrian hamster[J].Endocrinology,2006,147(10):4680-4687.
[5]Yasuo S,Watanabe M,Nakao N,et al.The reciprocal switching of two thyroid hormone-activating and-inactivating enzyme genes is involved in the photoperiodic gonadal response of Japanese quail[J].Endocrinology,2005,146(6):2551-2554.
[6]Majumdar G,Trivedi A K,Gupta N J,et al.Circadian synchronization deter mines critical day length for seasonal responses[J].Physiol Behav,2015,147:282-290.
[7]Bianco A C,Salvatore D,Gereben B,et al.Biochemistry,cellular and molecular biology,and physiological roles of the iodothyronine selenodeiodinases[J].Endocr Rev,2002,23(23):38-89.
[8]Yoshimura T,Yasuo S,Watanabe M,et al.Light-induced hormone conversion of T4 to T3 regulates photoperiodic response of gonads in birds[J].Nature,2003,426(6963):178-181.
[9]Yasuo S,Nakao N,Ohkura S,et al.Long-day suppressed expression of type 2 deiodinase gene in the mediobasal hypothalamus of the Saanen goat,a shor t-day breeder:implication for seasonal window of thyroid hormone action on reproductive neuroendocrine axis[J].Endocrinology,2006,147(1):432-440.
[10]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.
[11]Schmittgen T D,Livak K J.Analyzing real-time PCR data by the comparative CT method[J].Nat Protoc,2008,3(6):1101-1108.
[12]Xu M,Sulkowski Z L,Parekh P,et al.Effects of perinatal lipopolysaccharide(LPS)exposure on the developing rat brain;modeling the effect of mater nal infection on the developing human CNS[J].Cerebellum,2013,12(4):572-586.
[13]Wei W,Wang Y,Dong J,et al.Hypothyroxinemia induced by mild iodine deficiency deregulats thyroid proteins during gestation and lactation in dams[J].Int J Env Res Pub He,2013,10(8):3233-3245.
[14]Romano R M,Gomes S N,Cardoso N C,et al.New insights for male infertility revealed by alterations in spermatic function and differential testicular expression of thyroid-related genes[J].Endocrine,2016,55(2):607-617.
[15]Liu L,Chen Y,Wang D,et al.Cloning and expression characterization in hypothalamic Dio2/3 under a natural photoperiod in the domesticated Brandt's vole(Lasiopodomys brandtii)[J].Gen Comp Endocr,2018,259:45-53.
[16]王喜英.基于家系的DIO2基因多态性与秦巴山区人群精神发育迟滞的关联研究[D].西安:西北大学,2010.
[17]Torriani M,Srinivasa S,Fitch K V,et al.Dysfunctional subcutaneous fat with reduced Dicer and brown adipose tissue gene expression in HIV-infected patients[J].J Clin Endocr Metab,2016,101(3):1225.
[18]Detti L,Uhlmann R A,Fletcher N M,et al.Endometrial signaling pathways during ovarian stimulation for assisted reproduction technology[J].Int J Fertil Steril,2013,100(3):889-894.
[19]刘静.中华大蟾蜍胚胎发育中脱碘酶基因DIO2、DIO3表达特性的研究[D].西安:陕西师范大学,2013.
[20]黄冬维,狄冉,胡文萍,等.山羊繁殖季节性相关基因DIO2与DIO3的表达分析[J].农业生物技术学报,2016,24(10):1536-1543.
[21]Tavolaro F M,Thomson L M,Ross A W,et al.Photoperiodic effects on seasonal physiology,reproductive status and hypothalamic gene expression in young male F344 rats[J].J Neuroendocrinol,2015,27(2):79-87.
[22]Miera C S D,Hanon E A,Dardente H,et al.Circannual variation in thyroid hormone deiodinases in a short-day breeder[J].J Neuroendocrinol,2013,25(4):412.
[23]丁吉顺.Dio2/Dio3系统对黑像仓鼠繁殖的调控机制[D].曲阜:曲阜师范大学,2015.
[24]Kampflassin A,Prendergast B J.Acute downregulation of Type II and Type III iodothyronine deiodinases by photoperiod in peripubertal male and female Siberian hamsters[J].Gen Comp Endocr,2013,193(4):72.
[25]Janssen R,Zuidwijk M J,Muller A,et al.MicroRNA 214 is a potential regulator of thyroid hormone levels in the mouse heart following myocardial infarction,by targeting the thyroid-hormoneinactivating enzyme deiodinase type III[J].Front Endocrinol,2016,7:22.
[26]吴茜红.牛Dio3、Dio3os和Wif1基因印记及Wif1基因DNA甲基化状态分析[D].保定:河北农业大学,2015.
[27]Koopdonk-Kool J M,de Vijlder J J,Veenboer G J,et al.Type II and type III deiodinase activity in human placenta as a function of gestational age[J].J Clin Endocr Metab,1996,81(6):2154-2158.
[28]Deng W B,Liang X H,Liu J L,et al.Regulation and function of deiodinases during decidualization in female mice[J].Endocrinology,2014,155(7):2704-2717.
[29]杨宗林.猪DLK1-DIO3印记域4个候选印记基因的分离及其单等位基因表达分析[D].重庆:西南大学,2009.
[30]Kester M H,Martinez D M R,Obregon M J,et al.Iodoth-yronine levels in the human developing brain:major regulatory roles of iodothyronine deiodinases in different areas[J].J Clin Endocr Metab,2004,89(7):3117-3128.
[31]Ono H,Hoshino Y,Yasuo S,et al.Involvement of thyrotropin in photoperiodic signal transduction in mice[J].P Natl Acad Sci USA,2008,105(47):18238-18242.
[32]Nakao N,Ono H,Yamamura T,et al.Thyrotrophin in the pars tuberalis triggers photoperiodic response[J].Nature,2008,452(7185):317.
[33]Hanon E A,Routledge K,Dardente H,et al.Effect of photoperiod on the thyroid-stimulating hormone neuroendocrine system in the European hamster(Cricetus cricetus)[J].J Neuroendocrinol,2010,22(1):51-55.
[34]Ebling F J.Photoperiodic regulation of puberty in seasonal species[J].Mol Cell Endocrinol,2010,324(1):95-101.