微管相关蛋白tau在小鼠卵巢颗粒细胞中的定位及功能
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摘要
目的:探究小鼠卵巢颗粒细胞中微管相关蛋白的表达及功能定位。方法:选取雌性小鼠发生6周成年(成年组,6只)与未成年(未成年组,6只),培养卵巢颗粒细胞,将加入促卵泡刺激素(FSH)培养的颗粒细胞作为FSH组,不加FSH培养的颗粒细胞作为非FSH组。免疫组化染色法观察颗粒细胞中微管相关蛋白tau表达,将tau融合蛋白转染到颗粒细胞上,荧光显微镜下观察tau蛋白在颗粒细胞中的定位。结果:小鼠卵巢颗粒细胞tau蛋白表达阳性数成年组高于未成年组,FSH组高于非FSH组(P<0.05);未成年小鼠中FSH组阳性数高于非FSH组(P<0.05)。成年组小鼠卵巢颗粒细胞中tau蛋白聚集于细胞核,呈放射状。结论:微管相关蛋白tau在成年小鼠卵巢颗粒细胞中高表达,其定位主要集中于细胞核,影响颗粒细胞增殖分化,通过观察tau蛋白表达及定位,可反映微管形成及形态变化。
Objective: To investigate the expression and function localization of microtubule associated protein tau in mouse ovarian granulosa cells. Methods: 12 female mice were selected, which included 6 adult mice and 6 juvenile mice, and their ovarian granulosa cells were collected and cultured. The granulosa cells of these mice were cultured with or without FSH, and immunohistochemical staining was used to observe the expression of microtubule associated protein tau in the granulosa cells. Then Tau fusion protein was transfected into the granulosa cells, and the localization of tau in the granulosa cells was observed by fluorescence microscopy. Results: The positive expression of tau protein in the ovarian granulosa cells of the adult was significant higher than that of juvenile mice. Among the adult mouse, the positive expression of tau protein in the granulosa cells cultured with FSH was significant higher than that of cells cultured without FSH(P<0.05). Among the juvenile mouse, the positive expression of tau protein in ovarian granulosa cells of cultured with FSH was significant higher than that of ovarian granulosa cells cultured without FSH(P<0.05). Tau protein of ovarian granulosa cells of adult mice was aggregated in nucleus and was radiated. Conclusion: The microtubule associated protein Tau can be highly expressed in ovarian granulosa cells of adult mouse, and its localization is mainly concentrated in the nucleus. The expression and localization of tau can reflect the formation and morphological changes of microtubules, which affect the proliferation and differentiation of microtubule of granulosa cells.
Objective: To investigate the expression and function localization of microtubule associated protein tau in mouse ovarian granulosa cells. Methods: 12 female mice were selected, which included 6 adult mice and 6 juvenile mice, and their ovarian granulosa cells were collected and cultured. The granulosa cells of these mice were cultured with or without FSH, and immunohistochemical staining was used to observe the expression of microtubule associated protein tau in the granulosa cells. Then Tau fusion protein was transfected into the granulosa cells, and the localization of tau in the granulosa cells was observed by fluorescence microscopy. Results: The positive expression of tau protein in the ovarian granulosa cells of the adult was significant higher than that of juvenile mice. Among the adult mouse, the positive expression of tau protein in the granulosa cells cultured with FSH was significant higher than that of cells cultured without FSH(P<0.05). Among the juvenile mouse, the positive expression of tau protein in ovarian granulosa cells of cultured with FSH was significant higher than that of ovarian granulosa cells cultured without FSH(P<0.05). Tau protein of ovarian granulosa cells of adult mice was aggregated in nucleus and was radiated. Conclusion: The microtubule associated protein Tau can be highly expressed in ovarian granulosa cells of adult mouse, and its localization is mainly concentrated in the nucleus. The expression and localization of tau can reflect the formation and morphological changes of microtubules, which affect the proliferation and differentiation of microtubule of granulosa cells.
引文
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[13] 洪峰,袁高乐,张辉洁,等.微管不稳定蛋白和CD133在骨肉瘤组织的表达及其临床意义[J].中华实验外科杂志,2017,34(3):516.
[14] 俞雪锋,蒋曦,王国康,等.阿魏酸对脊髓损伤大鼠微管相关蛋白轻链3、Beclin1、Bcl-2和Bax表达的影响[J].中国药理学与毒理学杂志,2016,30(7):714-722.
[15] 赵红,刘振东,王海丰,等.NEP1-40对局灶性脑缺血大鼠生长相关蛋白43和微管相关蛋白2表达的影响[J].中华老年心脑血管病杂志,2016,18(11):1204-1207.
[16] 杨璐,吴海平,李建忠,等.母婴分离对青年期小鼠脑区糖皮质激素受体和微管相关蛋白表达的影响[J].中华行为医学与脑科学杂志,2017,26(2):102-106.
[17] 汤婷婷,刘岿然,李琳,等.NF-YA与LC3在卵巢子宫内膜异位囊肿在位内膜中的表达[J].中国妇幼保健,2017,32(19):4825-4826.
[18] 宋甜甜,肖雨,白春梅,等.微管相关蛋白2在胰腺神经内分泌肿瘤组织中的表达及预后意义[J].中国肿瘤临床,2017,44(11):532-538.
[19] 刘富兵,马欢欢,徐珊,等.新生期重复吸入七氟烷的成年大鼠海马组织CA3区Aβ、Tau蛋白表达变化[J].山东医药,2016,56(27):30-32.
[20] Feinstein HE,Benbow SJ,Lapointe NE.Oligomerization of the microtubule-associated protein tau is mediated by its N-terminal sequences implications for normal and pathological tau action[J].Journal of Neurochemistry,2016,137(6):939-954.
[21] 王亚男,余意,冯成强.tau蛋白与阿尔茨海默病[J].中国老年学杂志,2016,36(3):740-741.
[2] Pouwer AW,Farquhar C,Kremer JA,et al.Long-acting follicle-stimulating hormone versus daily follicle-stimulating hormone for women undergoing assisted reproduction[J].Fertility & Sterility,2016,105(6):1454-1456.
[3] Follicle-Stimulating Hormone (FSH)-dependent Regulation of Extracellular Regulated Kinase (ERK) Phosphorylation by the Mitogen-activ.
[4] 肖英,程爱兰.细胞骨架在肿瘤侵袭转移中的研究进展[J].中国肿瘤临床,2016,43(22):1007-1011.
[5] 王军,程萍萍,袁俊英,等.缺氧缺血性脑损伤新生大鼠脑组织神经细胞凋亡情况及Tau、p-Tau蛋白表达变化[J].山东医药,2016,56(15):36-38.
[6] 高雅楠,袁婷,张伟伟,等.二仙汤水提物、醇提物对小鼠脾脏和卵巢细胞作用的比较研究[J].中华中医药学刊,2017,35(04):958-961.
[7] Stoenescu VE,Niculescu M,Novac L,et al.Immunohistochemical reaction of the glandular epithelium in endometrial hyperplasia compared to endometrial carcinoma[J].Rom J Morphol Embryol,2017,58(3):791-800.
[8] 杨贞,郭新宇,邓伟民,等.益气血法对超排卵大鼠卵巢颗粒细胞凋亡及Bim表达的影响[J].实用医学杂志,2017,33(10):1593-1596.
[9] 袁丽娟,任君旭,姬宏宇,等.转化生长因子-β和Smad4在绝经过渡期大鼠卵巢颗粒细胞中的表达[J].解剖学报,2018,49(1):108-112.
[10] Bridges AA,Jentzsch MS,Oakes PW,et al.Micron-scale plasma membrane curvature is recognized by the septin cytoskeleton[J].Journal of Cell Biology,2016,213(1):23-32.
[11] Sohn PD,Tracy TE,Son HI,et al.Acetylated tau destabilizes the cytoskeleton in the axon initial segment and is mislocalized to the somatodendritic compartment[J].Molecular Neurodegeneration,2016,11(1):47.
[12] 修成奎,雷燕,王强,等.自然衰老大鼠血管老化中细胞骨架的改变及人参三七川芎提取物的干预作用[J].中国中西医结合杂志,2017,37(8):968-972.
[13] 洪峰,袁高乐,张辉洁,等.微管不稳定蛋白和CD133在骨肉瘤组织的表达及其临床意义[J].中华实验外科杂志,2017,34(3):516.
[14] 俞雪锋,蒋曦,王国康,等.阿魏酸对脊髓损伤大鼠微管相关蛋白轻链3、Beclin1、Bcl-2和Bax表达的影响[J].中国药理学与毒理学杂志,2016,30(7):714-722.
[15] 赵红,刘振东,王海丰,等.NEP1-40对局灶性脑缺血大鼠生长相关蛋白43和微管相关蛋白2表达的影响[J].中华老年心脑血管病杂志,2016,18(11):1204-1207.
[16] 杨璐,吴海平,李建忠,等.母婴分离对青年期小鼠脑区糖皮质激素受体和微管相关蛋白表达的影响[J].中华行为医学与脑科学杂志,2017,26(2):102-106.
[17] 汤婷婷,刘岿然,李琳,等.NF-YA与LC3在卵巢子宫内膜异位囊肿在位内膜中的表达[J].中国妇幼保健,2017,32(19):4825-4826.
[18] 宋甜甜,肖雨,白春梅,等.微管相关蛋白2在胰腺神经内分泌肿瘤组织中的表达及预后意义[J].中国肿瘤临床,2017,44(11):532-538.
[19] 刘富兵,马欢欢,徐珊,等.新生期重复吸入七氟烷的成年大鼠海马组织CA3区Aβ、Tau蛋白表达变化[J].山东医药,2016,56(27):30-32.
[20] Feinstein HE,Benbow SJ,Lapointe NE.Oligomerization of the microtubule-associated protein tau is mediated by its N-terminal sequences implications for normal and pathological tau action[J].Journal of Neurochemistry,2016,137(6):939-954.
[21] 王亚男,余意,冯成强.tau蛋白与阿尔茨海默病[J].中国老年学杂志,2016,36(3):740-741.
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