共存递质囊泡循环动力学研究细胞模型的建立

摘要

Objective To prove directly that PC12 cells can synthesize and secrete histamine (HA) and to provide experimental evidence for researching vesicle recycle kinetics of sympathetic cotransmitter. Methods PC12 cells were cultured in the conventional way. RT-PCR and Western blot techniques were used to detect histidine decarboxylase (HDC), the key enzyme, in HA synthesis process. Immunofluorescence technique was used to detect HA, HDC and HA vesicles in the differentiated PC12 cells. Results HDC expression could be detected in PC12 cells at both the mRNA and protein levels. Furthermore, immunofluorescece data showed that detectable levels of HA, HDC and HA vesicles were found in PC12 cells. Conclusion Since PC12 cells have HA expression and the characteristic similar to that of the sympathetic neuron, they can be used as an ideal cell model for researching vesicle recycling kinetics of sympathetic cotransmitter.%目的 直接证明PC12细胞能够合成和分泌组胺(histamine,HA),为将其应用于交感神经共存递质囊泡循环动力学研究提供实验依据.方法 常规培养PC12细胞,用嵌套RT PCR(nRT PCR)和Western blot技术检测其合成HA的关键酶组氨酸脱羧酶(histidine decarboxylase,HDC)的mRNA和蛋白表达,利用免疫荧光技术观测HA、HDC以及HA囊泡在细胞中的定位.结果 从基因和蛋白水平均证实了该细胞中表达HDC,并利用形态学方法在细胞中检测到HA、HDC以及HA囊泡的存在.结论 PC12细胞确实含有HA,结合分化型PC12细胞与交感神经元相似的特征,可将其作为交感神经共存递质囊泡循环动力学研究的理想细胞模型.