摘要
背景哺乳动物视觉适应机制与其视皮质发育密切相关，在不同发育阶段的视皮质中，有不同生物学功能的多种基因参与视觉发育的调控。研究大鼠视皮质发育不同阶段视皮质中差异基因的表达对视觉发育的研究可提供理论依据。目的应用基因芯片技术研究SD（Sprague-Dawley）大鼠出生后不同时间点视皮质中差异基因表达谱的变化。方法60只清洁级雌性sD大鼠分为刚出生组（0d，20只）、睁眼前组（生后10d，15只）和视皮质发育关键期前组（生后20d，15只）、视皮质发育完成后组（生后45d，10只）。所有大鼠在相对应的时间点处死并取出新鲜大脑视皮质，用基因芯片技术检测差异基因的表达，用实时定量PCR（RT-PCR）法验证基因芯片技术检测出的差异表达比率≥2．0的基因的mRNA表达水平。对相邻鼠龄12个基因进行了36组时间点配对，即每个基因3组配对，分别为P45／P0、P20／P0、P10／P0，用基因芯片技术和RT-PCR法分析大鼠视皮质差异基因表达与鼠龄的关联性。结果在上述不同时间点，基因芯片技术检出的差异表达比率≥2．0倍的基因包括Akap7、Asam、Casp3、Cxcr4、Egr1、Ennp2、Fabp7、印r88、Inpp5d、Rpsa、Stk32c和Vampl共12个基因，real—timePCR验证结果表明，26个探针的24个基因（其中有2对探针代表相同的基因）具有相同的变化趋势，其中20个探针表现为正向调控，6个探针为负向调控。Akap7、Asam、Casp3、Cxcr4、Egrl、Ennp2、Fabp7、lnpp5d、Rpsa和Vampl基因与其mRNA表达趋势一致，Gpr88和Stk32c基因在P45／P0中显示为相反的变化。在基因芯片中为正向调控，而在RT—PCR中为负向调控。RT-PCR检测结果表明，12个基因的36组不同时间点配对中34个配对表达出与基因芯片检测结果同向的调控，一致率为94.44％。不同鼠龄大鼠视皮质表达的不同基因的生物功能主要包括神经系统发育、（�
Background Visual adaptive mechanism of mammalian is close responsible for the development of visual cortex. The various genes with different biological functions in different developing stages of visual cortex participate in regulation of visual development. To investigate the differential expression profiles of various genes in different ages of rat cortex can offer basis and evidence for the study of visual development. Objective Present study aimed to investigate the genes that changed continuously in the postnatal developmental process of rat visual cortex by microarray analysis of visual cortex RNA. Methods Sixty clean SD rats were grouped numbered and randomized into the postnatal day 0 group （ P0, n = 20 ） , before eye opening group （ postnatal day 10, P10, n = 15 ） , before the critical period of visual cortex growth group （ postnatal day 20, P20,n = 15 ） and the end of development of visual cortex group（ postnatal day 45, P45 ,n = 15 ）. The rats were sacrificed at corresponding time point respectively, and the fresh visual cortex were obtained for the extraction of total RNA and microarray analysis. Genes exhibiting changes in expression by≥2.0 folds were further confirmed using real-time PCR（RT-PCR）. In order to evaluate theassociation of differential gene expression with growth,the postnatal stages were paired as 36 groups with the 3 pairs for each target gene （P45/P0, P20/P0, P10/P0）. Results Microarray analysis showed that the gene with differential rati≥2.0 folds in rat visual cortex included Akap7, Asam, Casp3, Cxcr4, Egrl, Ennp2, Fabp7, Gpr88, Inpp5d,Rpsa, Stk32c and Vamp1. Real-time PCR verified that 24 genes form 26 probe sets had the same-phase regulating tendency, including 20 up-regulating probe sets and 6 down-regulating probe sets. The homodromous expressing tendency was seen in Akap7 , Asam, Casp3 , Cxcr4 , Egrl , Ennp2 , Fabp7 , Inpp5d, Rpsa and Vamp1 genes between microarray analysis and RT-PCR. However, reverse expressions were found in the P45/P0 of Gpr88 a
出处
《中华实验眼科杂志》
CAS
CSCD
北大核心
2014年第8期682-687,共6页
Chinese Journal Of Experimental Ophthalmology
基金
北京市卫生系统高层次卫生技术人才培养计划项目（2013-03-051）
关键词
基因表达谱
大鼠
视皮质
生长和发育
寡核苷酸阵列序列分析
神经元可塑性
聚合酶链反应
Gene expression profiling
Rat, Sprague-Dawley
Visual cortex/growth and development
Oligonucleotide array sequence analysis
Neuronal plasticity
Polymerase chain reaction
