基于RNA-seq技术的单眼形觉剥夺模型大鼠视皮层差异基因筛选、鉴定及功能分析  

Screening,identification and functional analysis of differential expression genes in rat visual cortex after monocular deprivation by RNA-Seq technique

作　　者：孙娅玲 严兴科 刘安国 Sun Yaling;Yan Xingke;Liu Anguo(School of The First Clinical Medical,Gansu University of Chinese Medicine,Lanzhou 730000,China;School of Acupuncture-Moxibustion and Tuina,Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学第一临床医学院,兰州730000 [2]甘肃中医药大学针灸推拿学院,兰州730000

出　　处：《中华实验眼科杂志》2022年第3期210-215,共6页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目(81660816);甘肃省高等学校创新能力项目(2019A-073);甘肃中医药大学研究生创新基金项目(2021CX51)。

摘　　要：目的借助全转录组测序(RNA-seq)技术对右眼形觉剥夺模型大鼠视皮层差异表达基因进行筛选和生物学功能分析。方法将18只14日龄SD幼鼠按照随机数字表法随机分为空白对照组和单眼形觉剥夺组,每组9只。其中单眼形觉剥夺组幼鼠采用右眼眼睑缝合方法制作单眼形觉剥夺模型,连续14 d。分别于造模前和造模后14 d记录各组大鼠右眼图形视觉诱发电位P 100波的潜伏期和振幅。分别提取各组大鼠双侧视皮质组织,通过RNA-seq技术,筛选出弱视相关发病基因,利用基因本体论(GO)富集分析对上述基因进行生物学功能描述,并进行京都基因与基因组百科全书(KEGG)通路分析。结果与空白对照组相比,单眼形觉剥夺组P 100波潜伏期明显延长,振幅明显下降(均P<0.05),表明造模成功。共筛选出左侧视皮质差异表达基因40个,右侧视皮质差异表达基因63个,其中9个基因重叠。GO富集分析表明,差异表达基因主要参与转录DNA模板化、谷氨酸分泌、RNA聚合酶Ⅱ启动子的转录调控、蛋白磷酸化等生物学过程,参与DNA结合、ATP结合、蛋白丝氨酸/苏氨酸激酶活性、钙离子结合、锌离子结合、磷脂酶A 2活性、核酸绑定等分子功能,参与细胞内、内质网的膜等细胞组分。其中,Grm2、Pla2g2a基因的异常表达可能与视功能损伤改变过程密切相关,Grm2基因主要参与谷氨酸突触、长时程增强(LTP)、长时程抑制(LTD)等视觉信号通路过程,Pla2g2a基因主要参与α-亚麻酸代谢通路和花生四烯酸代谢通路。结论单眼形觉剥夺大鼠视觉发育敏感期内存在双侧视皮质基因的异常表达,导致视觉信号传导功能异常。基于特定响应基因调控的代谢通路改变可能是弱视发病的重要的分子生物学机制之一。Objective To screen differentially expressed genes(DEGs)in rat visual cortex after monocular deprivation by RNA sequencing technology,and to analyze the function of the DEGs.Methods Eighteen 14-day-old SD rats were randomly divided into blank control group and monocular deprivation model group according to random number table method,with 9 rats in each group.The monocular deprivation model was established through lid suture of the right eye for 14 days.Patten visual evoked potential(PVEP)in the right eyes of the rats was recorded before and 14 days after modeling,respectively.Bilateral visual cortex tissues of the rats were dissected from the two groups,and specific genes associated with the pathogenesis of amblyopia were screened out for RNA-seq analysis.The biological functions of differentially expressed genes were evaluated by Gene Ontology(GO)enrichment analysis,and metabolic pathways involved were analyzed by Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis.The use and care of the animals complied with ARVO statement.This study protocol was approved by an Ethics Committee of Gansu University of Chinese Medicine(No.2016-58).Results Compared with blank control group,the latency of P 100 wave was significantly prolonged,and the amplitude was reduced in the eyes of monocular deprivation model group(both at P<0.05).Forty DEGs in the left visual cortex and 63 DEGs in the right visual cortex were determined,among which 9 genes were overlapped.GO analysis indicated that the DEGs were mainly involved in biological processes,such as DNA-templated transcription,glutamate secretion,transcriptional regulation of RNA polymeraseⅡpromoter,protein phosphorylation etc.,as well as molecular functions,such as DNA binding,ATP binding,protein serine/threonine kinase activity,calcium ion binding,zinc ion binding,phospholipase A 2 activity,nucleic acid binding and cell components involved in the formation of intracellular and membrane of endoplasmic reticulum.The abnormal expressions of Grm 2 and Pla2g2a genes might

关 键 词：全转录组测序 大鼠模型 图形视觉诱发电位 形觉剥夺 生物信息学 

分 类 号：R777.44[医药卫生—眼科]