We investigated the expression of prion protein gene both on mRNA and protein levels in bovine and ovine female reproductive organs during gestation and various tissues of their fetuses. The fetal tissues of both species included brain, cotyledon, heart, intestine, kidney, liver, lung, and muscle. In cattle, prion protein gene (PRNP) transcripts were detected by semiquantitative RT-PCR in reproductive tissues such as ovary, oviduct, endometrium, myometrium, follicles, and granulosa cells. In various tissues of 2-month-old fetuses, higher expression levels were found in brain and cotyledon compared to the other tissues. To detect the expression of the gene transcript in in vivo preimplantation embryos and 1-month-old fetuses, real-time PCR was performed showing that the level of gene expression in zygote stage was significantly higher (p < or = 0.05) than that of the other stages. Sheep were categorized as resistant (RI) or high susceptible (R5) to scrapie according to their PRNP genotype. In both genotype groups, the PRNP mRNA was detectable in all tissues studied including ovary, oviduct, endometrium, myometrium, and caruncle of ewes and all tissues of 2-month-old fetuses of both groups. Comparison between reproductive organs demonstrates the highest expression level in caruncle tissue of R1 ewes, whereas the level was high in brain and low in liver of both R1 and R5 fetuses. In addition, real-time RT-PCR was performed in immature oocytes, mature oocytes, in vivo embryos at morula stage, and 1-month-old fetuses. The results showed that the relative expression levels of the ovine PRNP mRNA in mature oocytes and morula stage embryos were significantly lower than those in immature oocytes and 1-month-old fetuses (p < or = 0.05). Western blot analyses revealed the immunoreactive bands corresponding to the cellular prion protein (PrPc) in all maternal and fetal tissues examined of both cattle and sheep. Moreover, immunohistochemical staining implicated localization of the PrPc in ovarian cortex and ovarian medulla of both species. However, PrPc was not detected in oocyte, granulosa cells, theca cells, and corpus luteum in this study.

译文

我们研究了妊娠期间牛和绵羊雌性生殖器官及其胎儿各个组织中prion蛋白基因在mRNA和蛋白质水平上的表达。这两个物种的胎儿组织包括大脑,子叶,心脏,肠,肾脏,肝脏,肺和肌肉。在牛中,通过半定量rt-pcr在生殖组织 (例如卵巢,输卵管,子宫内膜,子宫肌层,卵泡和颗粒细胞) 中检测到prion蛋白基因 (PRNP) 转录本。在2个月大的胎儿的各种组织中,与其他组织相比,大脑和子叶中的表达水平更高。为了检测基因转录本在体内植入前胚胎和1个月大的胎儿中的表达,进行了实时PCR,显示受精卵阶段的基因表达水平显着高于其他阶段 (p <或 = 0.05)。根据绵羊的PRNP基因型,绵羊被分为对瘙痒病的抗性 (RI) 或高敏感性 (R5)。在两个基因型组中,在所有研究的组织中都可以检测到PRNP mRNA,包括卵巢,输卵管,子宫内膜,子宫肌层和母羊以及两组2个月大胎儿的所有组织。生殖器官之间的比较表明,R1母羊的肉鸡组织中表达水平最高,而R1和R5胎儿的大脑中表达水平较高,肝脏中表达水平较低。此外,在未成熟卵母细胞,成熟卵母细胞,桑胚期的体内胚胎和1个月大的胎儿中进行了实时rt-pcr。结果表明,绵羊PRNP mRNA在成熟卵母细胞和桑胚期胚胎中的相对表达水平明显低于未成熟卵母细胞和1月龄胎儿 (p <或 = 0.05)。Western印迹分析显示,在所有检查的牛和羊的母体和胎儿组织中,与细胞病毒蛋白 (PrPc) 相对应的免疫反应带。此外,免疫组织化学染色暗示PrPc在两种物种的卵巢皮质和卵巢髓质中的定位。然而,在本研究中未在卵母细胞,颗粒细胞,卵泡膜细胞和黄体中检测到PrPc。

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