CAJ | 학술논문

目的观察reelin阳性Cajal-Retzius细胞(CR细胞)在正常小鼠及APPswe转基因小鼠海马发育中的变化,探讨CR细胞在阿尔茨海默病(AD)发生发展过程中所起的作用,为研究AD发病机制和临床治疗提供新的思路和方法 .方法 80只实验小鼠分为APPswe转基因模型组和对照组,每一组内分E16、P0、P7 、P15 、P30、P90、P180和P360 8个年龄段,每一年龄段小鼠各取5只.另取12月龄模型组和对照组小鼠各3只,用硫黄素S染色技术检测APPswe转基因小鼠脑内沉积的老年斑;免疫荧光技术标记正常及模型组小鼠齿状回分子层内reelin阳性CR细胞,同时采用谷氨酸、γ-氨基丁酸(GABA)、活化型Caspase-3分别与reelin双重标记以研究CR细胞的组织化学特点及凋亡情况;最后利用免疫印迹方法对海马组织内reelin的活化片段进行半定量分析.结果随着海马发育CR细胞逐渐减少,不同时期的reelin阳性CR细胞可以分别被谷氨酸、GABA、Caspase-3标记;APPswe转基因小鼠海马内CR细胞的数量明显少于正常对照组,免疫印迹法结果　与免疫细胞化学统计结果吻合.结论出生后CR细胞的丢失是由于凋亡所致,在海马发育的不同时期CR细胞分泌兴奋性或抑制性神经递质,以此来调节突触间的信息传递与突触可塑性.APPswe转基因小鼠海马内CR细胞低于正常对照组,提示CR细胞丢失可能与AD相关的神经元退行性病变有关.
목적 관찰reelin양성Cajal-Retzius세포(CR세포)재정상소서급APPswe전기인소서해마발육중적변화,탐토CR세포재아이자해묵병(AD)발생발전과정중소기적작용,위연구AD발병궤제화림상치료제공신적사로화방법 .방법 80지실험소서분위APPswe전기인모형조화대조조,매일조내분E16、P0、P7 、P15 、P30、P90、P180화P360 8개년령단,매일년령단소서각취5지.령취12월령모형조화대조조소서각3지,용류황소S염색기술검측APPswe전기인소서뇌내침적적노년반;면역형광기술표기정상급모형조소서치상회분자층내reelin양성CR세포,동시채용곡안산、γ-안기정산(GABA)、활화형Caspase-3분별여reelin쌍중표기이연구CR세포적조직화학특점급조망정황;최후이용면역인적방법 대해마조직내reelin적활화편단진행반정량분석.결과 수착해마발육CR세포축점감소,불동시기적reelin양성CR세포가이분별피곡안산、GABA、Caspase-3표기;APPswe전기인소서해마내CR세포적수량명현소우정상대조조,면역인적법결과　여면역세포화학통계결과문합.결론 출생후CR세포적주실시유우조망소치,재해마발육적불동시기CR세포분비흥강성혹억제성신경체질,이차래조절돌촉간적신식전체여돌촉가소성.APPswe전기인소서해마내CR세포저우정상대조조,제시CR세포주실가능여AD상관적신경원퇴행성병변유관.
Objective In order to compare the alteration of reelin-immunoreactive Cajal-Retzius cells (CR cells) in molecular layer of dentate gyrus of APPswe transgenic mice with wild type, the histochemical and developmental characteristics of CR cells were studied, therefore, the roles of CR cells in Alzheimer's disease would be revealed further.Methods The Thioflavine S staining, reelin immunofluorescence with or without reelin/glutamate and reelin/GABA immuno-double staining were carried out in the study. In the meantime, Western blotting was used to study the expression of reelin in hippocampi of the both wild type and transgenic mice. Results Reelin positive CR cells could be double-labeled with either glutamate or GABA immunostaining. Caspase-3 immunofluorescence demonstrated that some CR cells went through apoptosis during their development. Compared with wild type, CR cells in APPswe transgenic mice had significantly decreased in the molecular layer of the dentate gyrus. The result was supported with Western blotting analysis of reelin expression in hippocampus. Conclusion Reelin could be co-expressed with either glutamate or GABA, suggesting CR cells would be glutamatergic exciting neurons and GABAergic interneurons. The loss of CR cells during development probably was caused by the neuroapoptosis. Significant decrease of CR cells in hippocampus of APPswe transgenic mice indicated reelin may play an important role in AD pathological alterations.