目的:利用神经细胞原代培养体系研究内质网钙通道蛋白三磷酸肌醇受体2(IP3R2)的拮抗剂对小鼠少突胶质细胞增殖分化的作用。方法:分离并纯化新生小鼠大脑皮质来源的少突胶质前体细胞(OPC)。利用real time RT-PCR检测不同分化阶段少突胶...目的:利用神经细胞原代培养体系研究内质网钙通道蛋白三磷酸肌醇受体2(IP3R2)的拮抗剂对小鼠少突胶质细胞增殖分化的作用。方法:分离并纯化新生小鼠大脑皮质来源的少突胶质前体细胞(OPC)。利用real time RT-PCR检测不同分化阶段少突胶质细胞中IP3R2 mRNA的表达水平;利用细胞钙成像观察给予IP3R2拮抗剂2-氨基乙氧基二苯基硼酸盐(2-APB)对OPC的Ca^(2+)活动影响;利用免疫荧光染色法检测2-APB对OPC中Ki67、2′,3′-环核苷酸3′-磷酸二酯酶(CNPase)和髓鞘碱性蛋白(MBP)的表达情况以及对细胞周期的影响。结果:IP3R2 mRNA水平在未成熟少突胶质细胞阶段最高;2-APB处理OPC可显著降低细胞内Ca^(2+)浓度;Ki67标记的增殖OPC比例在2-APB处理组显著降低;且2-APB的处理可以延长OPC的细胞周期进程。在诱导分化的少突胶质细胞培养物中,2-APB处理可以使CNPase^(+)与MBP^(+)细胞数量显著增加。结论:IP3R2介导的细胞内钙库释放对于OPC的增殖与分化具有重要的调控作用,为治疗其异常相关疾病提供了重要的思路。展开更多
Food deprivation can rescue obesity and overweight-induced mood disorders,and promote mood performance in normal subjects.Animal studies and clinical research have revealed the antidepressant-like effect of calorie re...Food deprivation can rescue obesity and overweight-induced mood disorders,and promote mood performance in normal subjects.Animal studies and clinical research have revealed the antidepressant-like effect of calorie restriction,but little is known about the mechanism of calorie restriction-induced mood modification.Previous studies have found that astrocytes modulate depressive-like behaviors.Inositol 1,4,5-trisphosphate receptor type 2(IP3R2)is the predominant isoform in mediating astrocyte Ca2+signals and its genetic knockout mice are widely used to study astrocyte function in vivo.In this study,we showed that deletion of IP3R2 blocked the antidepressant-like effect induced by calorie restriction.In vivo microdialysis experiments demonstrated that calorie restriction induced an increase in ATP level in the medial prefrontal cortex(mPFC)in naïve mice but this effect disappeared in IP3R2-knockout mice,suggesting a role of astrocytic ATP in the calorie restriction-induced antidepressant effect.Further experiments showed that systemic administration and local infusion of ATP into the mPFC induced an antidepressant effect,whereas decreasing ATP by Apyrase in the mPFC blocked calorie restriction-induced antidepressant regulation.Together,these findings support a role for astrocytic ATP in the antidepressant–like effect caused by calorie restriction.展开更多
文摘目的:利用神经细胞原代培养体系研究内质网钙通道蛋白三磷酸肌醇受体2(IP3R2)的拮抗剂对小鼠少突胶质细胞增殖分化的作用。方法:分离并纯化新生小鼠大脑皮质来源的少突胶质前体细胞(OPC)。利用real time RT-PCR检测不同分化阶段少突胶质细胞中IP3R2 mRNA的表达水平;利用细胞钙成像观察给予IP3R2拮抗剂2-氨基乙氧基二苯基硼酸盐(2-APB)对OPC的Ca^(2+)活动影响;利用免疫荧光染色法检测2-APB对OPC中Ki67、2′,3′-环核苷酸3′-磷酸二酯酶(CNPase)和髓鞘碱性蛋白(MBP)的表达情况以及对细胞周期的影响。结果:IP3R2 mRNA水平在未成熟少突胶质细胞阶段最高;2-APB处理OPC可显著降低细胞内Ca^(2+)浓度;Ki67标记的增殖OPC比例在2-APB处理组显著降低;且2-APB的处理可以延长OPC的细胞周期进程。在诱导分化的少突胶质细胞培养物中,2-APB处理可以使CNPase^(+)与MBP^(+)细胞数量显著增加。结论:IP3R2介导的细胞内钙库释放对于OPC的增殖与分化具有重要的调控作用,为治疗其异常相关疾病提供了重要的思路。
基金This work was supported by grants from the National Natural Science Foundation of China(31830033,82090032,and 81971080)NSFC-Guangdong Joint Fund-U20A6005+1 种基金the Program for Changjiang Scholars and Innovative Research Team in University(IRT_16R37)the Key-Area Research and Development Program of Guangdong Province(2018B030334001).
文摘Food deprivation can rescue obesity and overweight-induced mood disorders,and promote mood performance in normal subjects.Animal studies and clinical research have revealed the antidepressant-like effect of calorie restriction,but little is known about the mechanism of calorie restriction-induced mood modification.Previous studies have found that astrocytes modulate depressive-like behaviors.Inositol 1,4,5-trisphosphate receptor type 2(IP3R2)is the predominant isoform in mediating astrocyte Ca2+signals and its genetic knockout mice are widely used to study astrocyte function in vivo.In this study,we showed that deletion of IP3R2 blocked the antidepressant-like effect induced by calorie restriction.In vivo microdialysis experiments demonstrated that calorie restriction induced an increase in ATP level in the medial prefrontal cortex(mPFC)in naïve mice but this effect disappeared in IP3R2-knockout mice,suggesting a role of astrocytic ATP in the calorie restriction-induced antidepressant effect.Further experiments showed that systemic administration and local infusion of ATP into the mPFC induced an antidepressant effect,whereas decreasing ATP by Apyrase in the mPFC blocked calorie restriction-induced antidepressant regulation.Together,these findings support a role for astrocytic ATP in the antidepressant–like effect caused by calorie restriction.
