中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2013年
12期
1100-1104
,共5页
曲露%张晔%衡斌%刘燕强
麯露%張曄%衡斌%劉燕彊
곡로%장엽%형빈%류연강
视觉发育%视皮层%避光%蛋白质组学
視覺髮育%視皮層%避光%蛋白質組學
시각발육%시피층%피광%단백질조학
Visual development%Visual cortex%Dark rearing%Proteomics
背景 以往对视皮层发育关键期的研究仅根据已有的神经信号表达系统,就某一种可能是视皮层发育的关键蛋白质进行实验验证,进行视皮层全蛋白质分析对于了解动物进而研究人类视觉发育的可塑性具有重要意义. 目的 了解避光环境对生长发育期大鼠视觉及视皮层蛋白质组学的影响. 方法 将2只同日分娩12只仔鼠的SD母鼠分别饲养在2个鼠笼中,分娩日将2个母鼠的仔鼠一半进行对换,对换的仔鼠剪耳进行区分,即时起一个鼠笼的大鼠在避光处饲养,一个鼠笼的大鼠在自然光照下饲养作为对照.避光饲养的大鼠在完全避光环境下更换食物、水和垫料,至仔鼠40日龄时测定两组仔鼠对近眼物体瞬目的次数.各组分别处死3只大鼠,分离双侧初级视皮层(枕叶),利用二维电泳和质谱分析的方法,检测仔鼠避光环境与自然光照环境下发育和生长的初级视皮层蛋白质组学的差异,经Swissprot与NCBI数据库比对,鉴定差异蛋白.然后将避光饲养的剩余9只大鼠恢复自然光照l0d至大鼠50日龄,作为避光40 d恢复光照10d组比较与同日龄自然光照组9只大鼠对近眼物体瞬目次数的差异. 结果 大鼠40日龄时,避光饲养组大鼠对近眼物体的瞬目次数为(0.33±0.35)次,明显少于自然光照组大鼠的(6.42±0.68)次,差异有统计学意义(t=24.38,P＜0.01);大鼠50日龄时,自然光照组大鼠对近眼物体的瞬目次数为(6.11±0.59)次,避光饲养组大鼠多于避光40 d后恢复至(5.00±1.22)次,但差异无统计学意义(t=2.09,P＞0.05).二维电泳和质谱分析显示,自然光照组仔鼠与避光40 d组大鼠初级视皮层的差异蛋白共36种,避光40 d组大鼠缺失26种,增加10种,其中经质谱分析和数据库比对3种为已知蛋白,分别是Eps 15同源结构域蛋白3(EHD3)、α-微管蛋白1A链和2’,3’-3’-环核苷酸磷酸二酯酶. 结论 生长发育期避光环境中饲养可造成大鼠可逆的视觉障碍,并使其初级视皮层蛋白质组学发生变化.
揹景 以往對視皮層髮育關鍵期的研究僅根據已有的神經信號錶達繫統,就某一種可能是視皮層髮育的關鍵蛋白質進行實驗驗證,進行視皮層全蛋白質分析對于瞭解動物進而研究人類視覺髮育的可塑性具有重要意義. 目的 瞭解避光環境對生長髮育期大鼠視覺及視皮層蛋白質組學的影響. 方法 將2隻同日分娩12隻仔鼠的SD母鼠分彆飼養在2箇鼠籠中,分娩日將2箇母鼠的仔鼠一半進行對換,對換的仔鼠剪耳進行區分,即時起一箇鼠籠的大鼠在避光處飼養,一箇鼠籠的大鼠在自然光照下飼養作為對照.避光飼養的大鼠在完全避光環境下更換食物、水和墊料,至仔鼠40日齡時測定兩組仔鼠對近眼物體瞬目的次數.各組分彆處死3隻大鼠,分離雙側初級視皮層(枕葉),利用二維電泳和質譜分析的方法,檢測仔鼠避光環境與自然光照環境下髮育和生長的初級視皮層蛋白質組學的差異,經Swissprot與NCBI數據庫比對,鑒定差異蛋白.然後將避光飼養的剩餘9隻大鼠恢複自然光照l0d至大鼠50日齡,作為避光40 d恢複光照10d組比較與同日齡自然光照組9隻大鼠對近眼物體瞬目次數的差異. 結果 大鼠40日齡時,避光飼養組大鼠對近眼物體的瞬目次數為(0.33±0.35)次,明顯少于自然光照組大鼠的(6.42±0.68)次,差異有統計學意義(t=24.38,P＜0.01);大鼠50日齡時,自然光照組大鼠對近眼物體的瞬目次數為(6.11±0.59)次,避光飼養組大鼠多于避光40 d後恢複至(5.00±1.22)次,但差異無統計學意義(t=2.09,P＞0.05).二維電泳和質譜分析顯示,自然光照組仔鼠與避光40 d組大鼠初級視皮層的差異蛋白共36種,避光40 d組大鼠缺失26種,增加10種,其中經質譜分析和數據庫比對3種為已知蛋白,分彆是Eps 15同源結構域蛋白3(EHD3)、α-微管蛋白1A鏈和2’,3’-3’-環覈苷痠燐痠二酯酶. 結論 生長髮育期避光環境中飼養可造成大鼠可逆的視覺障礙,併使其初級視皮層蛋白質組學髮生變化.
배경 이왕대시피층발육관건기적연구부근거이유적신경신호표체계통,취모일충가능시시피층발육적관건단백질진행실험험증,진행시피층전단백질분석대우료해동물진이연구인류시각발육적가소성구유중요의의. 목적 료해피광배경대생장발육기대서시각급시피층단백질조학적영향. 방법 장2지동일분면12지자서적SD모서분별사양재2개서롱중,분면일장2개모서적자서일반진행대환,대환적자서전이진행구분,즉시기일개서롱적대서재피광처사양,일개서롱적대서재자연광조하사양작위대조.피광사양적대서재완전피광배경하경환식물、수화점료,지자서40일령시측정량조자서대근안물체순목적차수.각조분별처사3지대서,분리쌍측초급시피층(침협),이용이유전영화질보분석적방법,검측자서피광배경여자연광조배경하발육화생장적초급시피층단백질조학적차이,경Swissprot여NCBI수거고비대,감정차이단백.연후장피광사양적잉여9지대서회복자연광조l0d지대서50일령,작위피광40 d회복광조10d조비교여동일령자연광조조9지대서대근안물체순목차수적차이. 결과 대서40일령시,피광사양조대서대근안물체적순목차수위(0.33±0.35)차,명현소우자연광조조대서적(6.42±0.68)차,차이유통계학의의(t=24.38,P＜0.01);대서50일령시,자연광조조대서대근안물체적순목차수위(6.11±0.59)차,피광사양조대서다우피광40 d후회복지(5.00±1.22)차,단차이무통계학의의(t=2.09,P＞0.05).이유전영화질보분석현시,자연광조조자서여피광40 d조대서초급시피층적차이단백공36충,피광40 d조대서결실26충,증가10충,기중경질보분석화수거고비대3충위이지단백,분별시Eps 15동원결구역단백3(EHD3)、α-미관단백1A련화2’,3’-3’-배핵감산린산이지매. 결론 생장발육기피광배경중사양가조성대서가역적시각장애,병사기초급시피층단백질조학발생변화.
Background Previous study on critical period plasticity in local cortical circuits primarily was only to test the role of some proteins in visual cortex on visual development based on the existed neural signals expression system,but whole containing proteins analysis in cortical circuits is lack.To perform a whole containing proteins analysis has an important significance for critical period plasticity study.Objective This study was to investigate the influence of dark rearing on visual sense and proteomics in visual cortex for growing rat.Methods Two SD female rats were fed in two cages together with 12 newborn rats on the same day respectively,and half number of newborn rats were exchanged each other from first day after delivering and marked by eartipping.The newborn rats in a cage were bred in the dark environment for 40 days,and newborn rats in other cage were bred in the nature environment as controls.The blink response of rats to nearby object was examined and compared between the two groups of rats.Then three rats from two cages were sacrificed respectively and bilateral primary visual cortex tissue was isolated.Proteomics in rat primary visual cortex was detected by two-dimensional electrophoresis and mass spectrometry and the result was checked from database.Then the survival rats in the dark environment returned to the nature environment for 10 days,and the blink response of rats to nearby object were compared with that of agematched rats in the natural environment.The use and care of experimental rats followed the instruction of Ethic Committee of Nankai University.Results The blink response of rats was (0.33 ± 0.35) times in the dark environment for 40-day group,and that in the natural environment for 40-day group was (6.42±0.68) times,with a significant difference between them (t =24.38,P＜0.01).After returned to natural environment for 10 days,the blink response times of rats were less than those of the natural environment group ([5.00±1.22] times vs.[6.11±0.59]times),but this change was not statistically significant (t =2.09,P＞0.05).Two-dimensional electrophoresis and mass spectrometry assay revealed that 36 different proteins in visual cortex were found in the dark-feed rats compared with the natural environment rats,including 26 loss proteins and 10 extra proteins.Among the different proteins,Eps 15 homology domain-containing protein-3 (EHD3),tubulin alpha-1A chain and 2 ',3 '-cyclic-nucleotide 3 ' phosphordiesterase were the known proteins.Conclusions Dark rearing cause reversible visual loss in critical period plasticity newborn rat,and the change of proteomics in visual cortex is probably an affecting factor.
