鸡YAP1基因在不同生长发育时期卵泡中的时空表达分析
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摘要
为探讨YAP1基因在鸡卵泡生长发育中的调控作用,以150 d海兰褐蛋鸡为供试材料,采用相对定量RT-PCR方法对YAP1基因在鸡卵巢组织不同发育时期卵泡中的mRNA转录水平进行检测分析,并用免疫组织化学方法对YAP1蛋白质分子在鸡前等级卵泡中的时空表达模式进行蛋白定位分析。结果表明:鸡YAP1基因mRNA在卵巢间质、前等级卵泡和不同等级化的卵泡中均有表达,但在不同等级的卵泡中表达差异不显著(P>0.05),呈现出组成型表达的特点。鸡YAP1蛋白质分子主要在卵巢间质组织、前等级卵泡卵母细胞和颗粒细胞中有表达,但在膜层细胞中未检测到其表达信号。此结果表明,YAP1分子可能主要以自分泌或旁分泌的方式在前等级卵泡的生长发育过程中发挥着重要调节作用。
To investigate the regulatory role of chicken YAP1 gene in ovarian follicle growth and development, the expression abundances of YAP1 gene mRNA in different developmental phase follicles sampled from 150 d Hylan brown layers were determined by using relative quantitative RT-PCR method, and molecule location of YAP1 protein in the ovarian prehierarchical follicles was examined by immunohistochemistry. The present results revealed that chicken YAP1 gene mRNA was expressed in ovarian stroma, prehierarchical follicles and different hierarchical follicles, but there was no significant difference among the follicles of different growth and development phases(P>0.05). The YAP1 protein was dominantly expressed in ovarian stroma tissue, oocytes and granulosa cells, but the positive signal of YAP1 protein expression was not detected in theca cells of the follicles sampled. The current data indicated that chicken YAP1 molecule might exert a critical biological function in regulating follicular growth and development in an autocrine and/or paracrine manner.
To investigate the regulatory role of chicken YAP1 gene in ovarian follicle growth and development, the expression abundances of YAP1 gene mRNA in different developmental phase follicles sampled from 150 d Hylan brown layers were determined by using relative quantitative RT-PCR method, and molecule location of YAP1 protein in the ovarian prehierarchical follicles was examined by immunohistochemistry. The present results revealed that chicken YAP1 gene mRNA was expressed in ovarian stroma, prehierarchical follicles and different hierarchical follicles, but there was no significant difference among the follicles of different growth and development phases(P>0.05). The YAP1 protein was dominantly expressed in ovarian stroma tissue, oocytes and granulosa cells, but the positive signal of YAP1 protein expression was not detected in theca cells of the follicles sampled. The current data indicated that chicken YAP1 molecule might exert a critical biological function in regulating follicular growth and development in an autocrine and/or paracrine manner.
引文
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[12] 牛小天. 鸡LATS2基因调控卵巢等级前卵泡生长发育的作用及其分子机制[D].长春:吉林农业大学, 2017:80-82.
[13] 韩香玉, 吴炎, 李璐,等. 梅花鹿VEGF基因的克隆及其在鹿茸顶端组织的表达[J].吉林农业大学学报, 2017,39(4):454-459.
[14] Wu W, Xu R F, Xu H, et al. Expression of the β-catenin gene in the skin of embryonic geese during feather bud development[J]. Poult Sci, 2008,87(1): 204-211.
[15] 王丽君,孙永峰,隋玉健,等. 不同杂交组合鹅胸肌和腿肌GHR、MSTN基因mRNA表达量与屠宰性状间的相关分析[J]. 吉林农业大学学报, 2016,38(4):460-466.
[16] 刘德会,秦宁,牛小天,等.鸡PAPPA2基因在卵泡中的时空表达特性分析[J].吉林农业大学学报, 2018,40(1):110-114.
[17] 柳俭强, 隋玉健, 武惠岩,等. PDGF及其受体mRNA在鹅不同等级前卵泡中的表达差异[J]. 吉林农业大学学报, 2017, 39 (1): 89-93.
[18] 刘强,徐日福,张英英,等.鸡MYCN 基因不同组织mRNA表达及其遗传多态性与免疫性状的关联分析[J].吉林农业大学学报,2014,36(4):469-474.
[19] Qin N, Fan X C, Zhang Y Y, et al. New insights into implication of the SLIT/ROBO pathway in the prehierarchical follicle development of hen ovary[J]. Poult Sci,2015, 94 (9): 2235-2246.
[20] Johnson A L. Ovarian follicle selection and granulosa cell differentiation[J]. Poult Sci, 2015, 94(4):781-785.
[21] Onagbesan O, Bruggeman V, Decuypere E. Intra-ovarian growth factors regulating ovarian function in avian species: a review[J]. Anim Reprod Sci, 2009, 111(2-4): 121-140.
[22] Abe W, Nasu K, Nakada C,et al. miR-196b targets c-myc and Bcl-2 expression, inhibits proliferation and induces apoptosis in endometriotic stromal cells[J]. Hum Reprod, 2013, 28(3):750-761.
[23] Gao F, Zhang J, Wang X,et al. Wt1 functions in ovarian follicle development by regulating granulosa cell differentiation[J]. Hum Mol Genet, 2014, 23(2): 333-341.
[24] Fu D, Lv X, Hua G,et al. YAP regulates cell proliferation, migration, and steroidogenesis in adult granulosa cell tumors[J]. Endocr Relat Cancer,2014, 21(2):297-310.
[2] Sudol M, Recinos C C, Abraczinskas J, et al. WW or WoW: the WWdomains in a union of bliss[J].IUBMB Life, 2005(57):773-778.
[3] Xie Q, Chen J, Feng H, et al. YAP/TEAD-mediated transcription controls cellular senescence[J].Cancer Res, 2013 (73):3615-3624.
[4] Zhang X, George J, Deb S, et al. The Hippo pathway transcriptional co-activator, YAP, is an ovarian cancer oncogene[J]. Oncogene,2011, 30(25): 2810-2822.
[5] Liu Z, Castrillon D H, Zhou W, et al. FOXO1/3 depletion in granulosa cells alters follicle growth, death and regulation of pituitary FSH[J]. Mol Endocrinol, 2013, 27(2):238-252.
[6] Pisarska M D, Kuo F T, Bentsi-Barnes I K,et al. LATS1 phosphorylates forkhead L2 and regulates its transcriptional activity[J]. Am J Physiol Endocrinol Metab, 2010, 299(1): E101-9. doi: 10.1152/ajpendo.00534.
[7] Kim M, Kim T, Johnson R L, et al. Transcriptional co-repressor function of the hippo pathway transducers YAP and TAZ[J]. Cell Rep, 2015,11(2): 270-282.
[8] Santucci M, Vignudelli T, Ferrari S,et al. The hippo pathway and YAP/TAZ-TEAD protein-protein interaction as targets for regenerative medicine and cancer treatment[J]. J Med Chem, 2015, 58(12):4857-4873.
[9] 周芳月,李佳,潘泽政,等.Hippo-YAP通路在多种干细胞尤其是卵巢生殖干细胞自我更新中作用的研究[J].中国细胞生物学学报,2015,37(7):1053-1058.
[10] Lyu Z, Qin N, Tyasi T L, et al. The Hippo/MST pathway member SAV1 plays a suppressive role in development of the prehierarchical follicles in hen ovary[J]. PLoS one, 2016, 11(8): e0160896.
[11] 吕智超. Hippo/MST信号通路核心因子在鸡前等级卵泡发育中的作用及分子机制[D]. 长春:吉林农业大学, 2016:59-65.
[12] 牛小天. 鸡LATS2基因调控卵巢等级前卵泡生长发育的作用及其分子机制[D].长春:吉林农业大学, 2017:80-82.
[13] 韩香玉, 吴炎, 李璐,等. 梅花鹿VEGF基因的克隆及其在鹿茸顶端组织的表达[J].吉林农业大学学报, 2017,39(4):454-459.
[14] Wu W, Xu R F, Xu H, et al. Expression of the β-catenin gene in the skin of embryonic geese during feather bud development[J]. Poult Sci, 2008,87(1): 204-211.
[15] 王丽君,孙永峰,隋玉健,等. 不同杂交组合鹅胸肌和腿肌GHR、MSTN基因mRNA表达量与屠宰性状间的相关分析[J]. 吉林农业大学学报, 2016,38(4):460-466.
[16] 刘德会,秦宁,牛小天,等.鸡PAPPA2基因在卵泡中的时空表达特性分析[J].吉林农业大学学报, 2018,40(1):110-114.
[17] 柳俭强, 隋玉健, 武惠岩,等. PDGF及其受体mRNA在鹅不同等级前卵泡中的表达差异[J]. 吉林农业大学学报, 2017, 39 (1): 89-93.
[18] 刘强,徐日福,张英英,等.鸡MYCN 基因不同组织mRNA表达及其遗传多态性与免疫性状的关联分析[J].吉林农业大学学报,2014,36(4):469-474.
[19] Qin N, Fan X C, Zhang Y Y, et al. New insights into implication of the SLIT/ROBO pathway in the prehierarchical follicle development of hen ovary[J]. Poult Sci,2015, 94 (9): 2235-2246.
[20] Johnson A L. Ovarian follicle selection and granulosa cell differentiation[J]. Poult Sci, 2015, 94(4):781-785.
[21] Onagbesan O, Bruggeman V, Decuypere E. Intra-ovarian growth factors regulating ovarian function in avian species: a review[J]. Anim Reprod Sci, 2009, 111(2-4): 121-140.
[22] Abe W, Nasu K, Nakada C,et al. miR-196b targets c-myc and Bcl-2 expression, inhibits proliferation and induces apoptosis in endometriotic stromal cells[J]. Hum Reprod, 2013, 28(3):750-761.
[23] Gao F, Zhang J, Wang X,et al. Wt1 functions in ovarian follicle development by regulating granulosa cell differentiation[J]. Hum Mol Genet, 2014, 23(2): 333-341.
[24] Fu D, Lv X, Hua G,et al. YAP regulates cell proliferation, migration, and steroidogenesis in adult granulosa cell tumors[J]. Endocr Relat Cancer,2014, 21(2):297-310.