温度对不同年龄大鼠海马器官型脑片长期培养中细胞活性和tau蛋白表达的影响  

Effect of Temperature on Viability and tau Expression in Hippocampal Slice Culture of Rat with Different Ages

作　　者：廖晓梅[1] 张迎春[1] 王建枝[1] 

机构地区：[1]华中科技大学同济医学院病理生理系,神经科学研究所,武汉430070

出　　处：《中国生物化学与分子生物学报》2005年第5期637-641,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金重点项目(No.30430270);国家自然科学基金(No.30170221)资助~~

摘　　要：探讨在海马器官型脑片的长期培养过程中,温度对不同年龄大鼠的海马脑片细胞活性和tau蛋白表达的影响,并以此为依据建立一种研究tau相关疾病的模型.选用出生后1周、2周、4周和8周的Wistar大鼠制备海马器官型脑片,培养温度分别为34℃和37℃,培养时间为21d,在培养过程中,检测培养基中的乳酸脱氢酶的含量以判断脑片的活性,采用免疫印迹技术检测细胞骨架蛋白tau的含量的变化.结果如下:(1)温度对海马脑片的细胞活性影响:34℃较37℃能在较长的时间内保持细胞活性,而在同一培养温度时,不同年龄鼠的脑片的细胞活性变化趋势一致;(2)温度对海马脑片的tau蛋白表达的影响:成年鼠(4周和8周)的海马脑片tau蛋白在34℃时能维持较长时间的稳定表达,而在37℃时tau的表达量随培养时间的延长而显著下降,且随鼠龄的增加,这种影响越明显.温度对1周和2周龄乳鼠的海马脑片tau蛋白的表达无影响.结论为:34℃培养条件下,4周和8周龄大鼠制备的海马器官型脑片能更长时间维持脑片的活性和tau蛋白的稳定表达,从而可望成为研究与tau蛋白相关疾病(如老年性痴呆)的理想模型.To investigate microtubule associated protein tau-related diseases, many studies have employed various cultured cell lines to study the aberrant tau modification. However, the cultured cells lack the natural extracellular matrix, neuronal connectivity and the normal neuronal expression of proteins. The use of cultured brain slices has become an accepted technique for the ex vivo tool, yet the viability of this preparation is not routinely measured. The effect of temperature on the cell viability and the expression of tan was investigated in hippocampi slice culture of rat brain of different ages. Hippocampal slices of male Wister rats of 1, 2, 4 and 8 week-old were cultured at 34 ℃ and 37 ℃ for 3 weeks, respectively. The lactate dehydrogenase （LDH） activity and Western blotting were measured to evaluate the cell viability and expression of tan protein. Results were followed as （ 1 ） Effect of temperature on the viability of brain slice ： Data from LDH assay showed that the slices could maintain activity for longer time at 34℃ than at 37℃, and at the same culture temperature, no obvious difference was observed in groups with different age; （2） Effect of temperature on expression of tau： The expression of tau in 4 and 8 week-old rat brain slices was more stable at 34℃ than at 37℃. Tau expression was significantly decreased at 37℃ and this decrease was becoming more obvious with increase of the age of rats. No effect of temperature was observed in 1 and 2 week-old rats. The results indicate that temperature at 34℃ is more suitable than at 37℃ for the survival and stable expression of tau in adult brain slice culture. On other hand, use of culture temperature and animal ages as signifcant functional parameter of regional slice viability also provides an inexpensive and expeditious means to enhances the feasibility of this preparation for function studies.

关 键 词：海马器官型脑片 温度 乳酸脱氢酶 TAU蛋白 

分 类 号：R745.7[医药卫生—神经病学与精神病学] Q513[医药卫生—临床医学]