卵泡抑素、激活素与BMP-4在大鼠脑发育过程中表达的初步研究

发布时间：2018-03-26 09:44

本文选题：卵泡抑素　切入点：激活素　出处：《辽宁医学院》2012年硕士论文

【摘要】：目的本实验观察了Follistatin、Activin A与BMP-4在大鼠大脑皮质、纹状体、海马和嗅球发育过程中的表达变化规律,籍以探讨Follistatin、Activin A与BMP-4在神经系统发育过程中的表达特点和调控机制。方法随机取同期合笼后查有阴道栓的SD入鼠30只,随机分成6组,每组5只。分别为E8.5天组、E13天组、E18天组、P3d(出生后3天)组、P7d组、P30d组。取E8.5组、E13组、E18组孕鼠(各5只)。每只孕鼠取5只胎鼠,取出胎脑固定。其他出生后P3d组、P7d组、P30d组每组随机取5只大鼠灌注固定,切取包含皮质、纹状体、海马、嗅球组织的脑片,每个部位切8张切片,用于免疫组化FS、Act、BMP-4染色。分别取E8.5、E13、E18、生后P3、1周(记作P3d、PIW)和生后1月(记作P1M)共6组,每组取大鼠5只,分别在冰台上分离提取大脑皮层、纹状体、海马和嗅球总RNA。应用免疫组织化学ABC法和RT-PCR法(半定量)分析Follistatin、Activin A与BMP-4在出生前后大鼠大脑皮质、纹状体、海马、嗅球表达水平的变化。结果进行免疫组织化学染色及光镜观察Follistatin与Activin A在大鼠脑内广泛分布,二者在胚胎期E8.5表达最高,并以胚胎期E13表达强度开始降低,在出生后P1M降至最低,同一发育阶段各脑区表达无明显差异。在大鼠不同发育阶段的相应脑区BMP-4亦广泛表达,但从胚胎期至新生期P1W持续低表达,尤以海马表达极弱,在生后P1M在不同脑区呈高表达,各发育阶段以大脑皮质和纹状体表达略强。RT-PCR结果显示：Follistatin mRNA与Activin A mRNA二者在胚胎期E8.5表达最强,胚胎期E13表达水平开始降低,在E18至生后P1W呈下降趋势,P1M表达降至最低。并在大鼠同一发育年龄各脑区表达无明显差异。BMP-4mRNA在胚胎期至新生期P3呈较弱表达,在生后P1W表达开始升高,P1M达到峰值,在大鼠各发育阶段以大脑皮质和纹状体表达略强。结论 Follistatin、Activin A与BMP-4在大鼠不同发育年龄各脑区具有波动性表达,表达水平与发育年龄密切相关。Follistatin、Activin A、BMP-4与大脑皮质、纹状体、海马、嗅球的发育密切相关。
[Abstract]:Purpose. The expression of Follistatin A and BMP-4 during the development of cerebral cortex, striatum, hippocampus and olfactory bulb in rats was observed in order to investigate the expression characteristics and regulation mechanism of FollistatinActivin A and BMP-4 during the development of nervous system. Method. Thirty SD rats with vaginal suppository were randomly divided into 6 groups. Five rats in each group were divided into E8.5-day group, E13-day group, E18-day group, P3-day group (3 days after birth), P7-day group, P30d group, E8.5 group, E13 group, E18 group (5 rats each). Other postnatal P3 d group, P7 d group, P30 d group, 5 rats in each group were randomly selected for perfusion fixation, and the brain slices containing cortex, striatum, hippocampus and olfactory bulb were cut into 8 sections. Six groups of E8.5E13E18 were used for immunohistochemical staining. The rats were divided into 6 groups: one week after birth (recorded as P3dPIW) and one month after birth (recorded as P1M). Five rats in each group were isolated and extracted from the cerebral cortex and striatum on the ice platform, respectively. The expression of Follistatin A and BMP-4 in cerebral cortex, striatum, hippocampus and olfactory bulb of rats before and after birth were analyzed by immunohistochemical ABC method and RT-PCR method (semi-quantitative). Results. Immunohistochemical staining and light microscopy were used to observe the wide distribution of Follistatin and Activin A in the brain of rats. The expression of E8. 5 was the highest in the embryonic phase, and the expression intensity of E13 began to decrease in the embryonic stage, and the P1m decreased to the lowest after birth. There was no significant difference in the expression of P1W in different brain regions of rats at the same developmental stage, but the expression of P1W continued to be low from embryonic stage to neonatal stage, especially in hippocampus, and the expression of P1M was high in different brain regions after birth. The results of RT-PCR showed that the expression of mRNA and Activin A mRNA were the strongest in embryonic E8.5, and the expression level of E13 in embryonic phase began to decrease. From E18 to postnatal, the expression of P1M decreased to the lowest. There was no significant difference in the expression of BMP-4 mRNA in different brain regions of the rats at the same developmental age. The expression of BMP-4 mRNA was weakly expressed from embryonic stage to newborn stage, and the expression of P1W began to increase after birth, and the expression of P1M reached its peak. The expression of cerebral cortex and striatum was slightly stronger in all developmental stages of rats. Conclusion. The expression level of FollistatinA and BMP-4 in different brain regions of rats at different developmental ages was closely related to the development of cerebral cortex, striatum, hippocampus and olfactory bulb, and the expression level was closely related to the development of the brain cortex, striatum, hippocampus and olfactory bulb, and the level of expression was closely related to the development of the cerebral cortex, striatum, hippocampus and olfactory bulb.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R338

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