摘要
为了检测单侧注射ＭＰＴＰ制备的帕金森病恒河猴模型纹状体内多巴胺及其代谢产物的含量变化，本研究采用脑内微透析技术和高效液相色谱－ 电化学方法检测了双侧尾状核头部多巴胺及其代谢产物３，４－二羟基苯乙酸和高香草酸的含量。结果证明，ＭＰＴＰ注射侧与注射对侧相比，多巴胺、３，４－二羟基笨乙酸和高香草酸的含量分别降低８５．７％ ， ９５．１％ 和６７．８％ 。这与动物呈现单侧帕金森病症状，核磁共振检查显示单侧黑质区域密度减低、面积缩小以及经抗酪氨酸羟化酶免疫组化方法显示脑切片单侧多巴胺能神经元明显减少等现象一致。以上结果表明，脑内微透析技术是活体检测脑内神经递质含量变化的有效方法，可以反映模型的病程变化，可用于评价帕金森病的治疗效果。
Molecular biological techniques were used to produce a monoclonal antibody against the central type of ChAT (cChAT). Rat cChAT fragment (containing exons 6, 7, 8, 9 and 10) in a length of 715 bp was generated from rat striatum cDNA and inserted into pGEM T vector. After sequencing analysis, the fragment was subcloned into pMAL c2 expression vector. The cChAT cDNA was expressed in E.coli as a fusion protein with maltose binding protein (MBP). The fusion protein gave a band of 69 kDa on SDS polyacrylamide gel. The size of this fusion protein was almost identical to the sum of those deduced from the amino acid sequences of MBP and cChAT. A clone, Mab676, was successfully obtained by immunizing Balb/c mice with purified cChAT fusion protein. Western blot analysis indicated that Mab676 recognized a single band of about 62 kDa in rat brain tissue extracts. When Mab676 was applied to rat brain for immunohistochemical staining, many neuronal structures in various brain regions were stained, e.g., striatum, globus pallidus, vertical and horizontal limbs of the diagonal band of Broca, substantia innominata, medial septal nucleus as well as the cranial motor nuclei of the brainstem. The above mentioned results confirmed the specificity of this monoclonal antibody against rat ChAT.
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
1999年第4期315-318,共4页
Chinese Journal of Neuroanatomy
基金
国家自然科学基金!(No. C03030307)
北京市自然科学基金!(No. 7971001)
北京市科技项目基金!(No. 952
关键词
微透析
尾状核
猴
帕金森病
疾病模型
多巴胺
choline acetyltransferase, fusion protein, western blot, immunohistochemistry, monoclonal antibody, rat
