大鼠视网膜中不同亚型无长突细胞对N-甲基-D-天冬氨酸损伤敏感性的差异  

Different susceptibility of amacrine cell subtypes in rat retina to N-methyl-D-aspartate injury

作　　者：乔侨 肖虹蕾[1] 李艳[1] 周国民[1] Qiao Qiao;Xiao Honglei;Li Yan;Zhou Guomin(Department of Anatomy,Histology and Embryology,Shanghai Medical College,Fudan University,Shanghai 200032,China)

机构地区：[1]复旦大学上海医学院解剖与组织胚胎学系,上海200032

出　　处：《解剖学杂志》2024年第2期101-106,170,F0002,共8页Chinese Journal of Anatomy

基　　金：国家自然科学基金(81700866、31571238)。

摘　　要：目的:探究大鼠视网膜中对N-甲基-D天冬氨酸(NMDA)敏感的无长突细胞(AC)亚型。方法:H-E染色观察NMDA损伤后视网膜组织病理变化,免疫荧光染色观察大鼠视网膜中神经节细胞标志物(Brn3a)、AC标志物(syntaxin)阳性细胞数量变化;在NMDA致AC损伤的剂量-效应变化关系和时间-效应变化关系的研究中用免疫荧光染色检测大鼠视网膜中GABA能AC标志物(GAD65/67)、甘氨酸能AC标志物(GlyT1)、胆碱能AC标志物(ChAT)、多巴胺能AC标志物(TH)、AⅡAC标志物(PV)阳性细胞的形态和数目变化;利用碘化丙啶(PI)和cleaved casapse-3染色确定死亡的AC亚型。结果:NMDA可损伤视网膜节细胞层(GCL)中的神经节细胞和移位的AC以及内核层(INL)中的AC。在NMDA致AC损伤的剂量-效应关系变化中,所有AC均随着NMDA剂量的增加而显著减少,且在100 nmol NMDA中未见胞体位于INL中的胆碱能AC和多巴胺能AC(DAC)的胞体;在NMDA致AC损伤的时间-效应关系变化中,INL中的GAD65/67^(+)、GlyT1^(+)、ChAT^(+)、TH^(+)细胞的数量均在损伤后6 h开始减少,GCL中的GAD65/67^(+)细胞、视网膜中GAD65/67^(+)细胞总数以及PV+细胞均在损伤后12 h开始减少,且均随着NMDA损伤时间的推移而减少。结论:视网膜GABA能AC和甘氨酸能AC均在NMDA作用下出现损伤,其中以INL中的胆碱能AC和DAC对NMDA损伤敏感。Objective:To explore the susceptibility of amacrine cell(AC)types to N-methyl-D-aspartate(NMDA)excitotoxicity in the rat retina.Methods:H-E staining was used to observe the pathological changes in retinal tissue after NMDA injury,while immunofluorescence staining was used to observe the changes in the quantity of Brn3a(the marker of retinal ganglion cell)and syntaxin(the marker of AC).In the study of the dose-effect relationship and time-effect relationship of NMDA-induced AC injury,immunofluorescence staining was used to detect the morphological and numerical changes of GAD65/67^(+)(the marker of GABAergic AC),GlyT1^(+)(the marker of glycinergic AC),ChAT^(+)(the marker of cholinergic AC),TH^(+)(the marker of dopaminergic AC),and PV^(+)cells(the marker of AⅡAC)in the rat retina.Propidium iodide(PI)and cleaved caspase-3 staining were employed to determine the subtypes of dead AC.Results:NMDA can damage both retinal ganglion cells and displaced ACs in the retinal ganglion cell layer(GCL),as well as ACs in the inner nuclear layer(INL).In the dose-effect relationship of AC damage induced by NMDA,all ACs significantly decreased with the increase of NMDA dose,and the cholinergic ACs in the INL and dopaminergic ACs(DAC)did not show their cell somas in 100 nmol NMDA.In the time-effect relationship of AC damage induced by NMDA,the number of GAD65/67^(+)in INL,GlyT1^(+),ChAT^(+)and TH^(+)cells began to decrease at 6 h after injury,while the number of GAD65/67^(+)cells in GCL,the total number of GAD65/67^(+)cells in the retina,and PV^(+)cells all began to decrease at 12 h after injury and continued to decrease with the progression of NMDA-induced damage.Conclusion:The GABAergic ACs and glycinergic ACs in the retina both show damage under the influence of NMDA,with cholinergic ACs in the INL and DACs being particularly sensitive to NMDA-induced damage.

关 键 词：视网膜 无长突细胞 兴奋性毒性 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]