不同发育阶段大鼠脊髓中Olig1的表达及表达规律

发布时间：2018-01-04 16:30

本文关键词：不同发育阶段大鼠脊髓中Olig1的表达及表达规律　出处：《安徽医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：(1)建立SD大鼠从胚胎到成年不同发育阶段脊髓的组织标本库。 (2)观察Olig1在SD大鼠不同发育阶段脊髓中的表达及细胞定位。 (3)探讨Olig1在SD大鼠不同发育阶段脊髓中的表达规律。方法：建立SD大鼠从胚胎到成年不同发育阶段的脊髓组织标本库,选取E14.5d(胚胎期14.5天),E18.5d(胚胎期18.5天),POd(出生0天),P3d(出生后3天),P7d(出生后7天),P2w(出生后2周),P4w(出生后4周),Adult(成年)几个不同发育时期的SD大鼠的脊髓。(1)使用Trizol试剂抽提出RNA,逆转录为cDNA检测Olig1表达情况；(2)使用荧光定量PCR,免疫印迹检测中枢神经系统不同发育阶段Olig1在信使核糖核酸(messenger RNA, mRNA)和蛋白水平的表达变化规律；(3)使用免疫荧光组织化学技术,通过Oligl单染检测出其在大鼠脊髓发育上述几个时期的表达情况；同时使用Oligl/O4, Oligl/MBP, Oligl/GFAP, Oligl/β-Tublin双标记染色法检测大鼠脊髓发育的P7d(出生后第7天),P2w(出生后2周)时Oligl在不同类型神经细胞上的表达和定位；最后总结出Oligl在大鼠脊髓发育过程中的表达规律。结果：(1)RT-PCR,免疫印迹法,免疫荧光组织化学单染显示,Oligl在E14.5d,E18.5d, POd, P3d, P7d, P2w, P4w, Adult几个不同发育时期的SD大鼠脊髓中均有表达；(2)免疫荧光双标记染色检测显示,在P7d和P2w的SD大鼠的脊髓中,Oligl在04阳性细胞和β-Tublin阳性细胞中表达,在MBP阳性细胞和GFAP阳性细胞中不表达；(3)荧光定量PCR、免疫印迹法检测进一步显示,Oligl在大鼠脊髓从E14.5d时期即有表达并且一直持续到成年,但是其mRNA和蛋白的表达水平在E14.5d, E18.5d, POd, P3d, P7d, P2w, P4w, Adult的8个不同时期不是一直保持在同—水平,也不是随着发育逐渐增加或减少,而是在胚胎早期表达水平较高以后逐渐减低,至P7d时低表达,而后又渐增高。结论：(1)Olig1在E14.5d到成年不同发育时期的SD大鼠脊髓中均表达；(2)Oligl在SD大鼠脊髓中的表达水平在胚胎早期较高,以后逐渐减弱,至P7d时呈最低水平,以后又逐渐升高；(3)Olig1主要是在少突胶质前体细胞和神经元中表达,在成熟少突胶质细胞中不表达或表达很低,在星形胶质细胞中也不表达。
[Abstract]:Objective to establish a tissue library of spinal cord of SD rats at different stages of development from embryo to adulthood. (2) to observe the expression and cellular localization of Olig1 in the spinal cord of SD rats at different developmental stages. To investigate the expression of Olig1 in the spinal cord of SD rats at different developmental stages. Methods: the spinal cord tissue library of SD rats from embryonic to adult stages was established. E14.5 days (14.5 days of embryo) and 18.5 days of embryo (18.5 days of embryo) were selected. POD (0 days after birth) P3d (3 days after birth) P7d (7 days after birth) (2 weeks after birth P4w (4 weeks after birth). The spinal cord of several adult SD rats at different developmental stages) Trizol reagent was used to extract RNAs and reverse transcription was used to detect the expression of Olig1 in the spinal cord of SD rats. (2) fluorescent quantitative PCR and Western blot were used to detect Olig1 in messenger RNA messenger RNA at different stages of development of central nervous system (CNS). Expression of mRNAs and proteins; (3) Immunofluorescence histochemical technique was used to detect the expression of Oligl in the above stages of spinal cord development in rats. And use Olig / O _ 4, Olig / MBP, Oligl/GFAP. Oligl- 尾 -Tublin double labeling staining was used to detect P7 days of spinal cord development in rats (7 days after birth). Expression and localization of Oligl in different types of nerve cells at P2w (2 weeks after birth); Finally, the expression of Oligl in the development of rat spinal cord was summarized. Results the expression of Oligl in E14.5dE 18.5d, POD, P3d, P7d was detected by Western blot and Western blotting. P2w, P4wand Adult were all expressed in spinal cord of SD rats at different developmental stages. (2) Immunofluorescence double labeling staining showed that Oligl was expressed in 04 positive cells and 尾 -Tublin positive cells in the spinal cord of P7d and P2w SD rats. It was not expressed in MBP positive cells and GFAP positive cells. Fluorescence quantitative PCR and Western blotting further showed that Oligl was expressed in the spinal cord of rats from E14.5d and lasted until adulthood. But the expression levels of mRNA and protein were at 14.5 days, 18.5 days, POD, P3d, P7d, P2wand P4w. The eight different stages of Adult were not maintained at the same level, nor increased or decreased with the development, but decreased gradually after the early embryonic stage was higher, and the expression was low at P7 days. And then it went up again. Conclusion the expression of Olig1 in the spinal cord of Sprague-Dawley rats from 14. 5 d to 14. 5 d was found in adult SD rats. The expression of Oligl in spinal cord of Sprague-Dawley rats was higher at the early stage of embryo, then gradually decreased, and then decreased to the lowest level at P7 day, and then increased gradually in the Sprague-Dawley rat spinal cord. Olig1 was mainly expressed in oligodendrocytes and neurons, but not in mature oligodendrocytes or astrocytes.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

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