Hammond P I, Jelacic T, Padilla S, Brimijoin S
Department of Pharmacology, Mayo Clinic, Rochester, Minnesota 55905, USA.
Anal Biochem. 1996 Oct 1;241(1):82-92. doi: 10.1006/abio.1996.0381.
Acetylcholinesterase histochemistry was coupled to inexpensive and widely available apparatus for video microscopy and densitometry to study enzyme activity and inhibition in different parts of the rat brain. Quantitative histochemistry, under properly defined conditions, yielded an output that increased linearly with incubation time and section thickness and was a smooth hyperbolic function of substrate concentration. The time-course of staining after in vivo exposure to eserine revealed no sign that carbamate-induced cholinesterase inhibition was readily reversed in vitro. Brains from rats treated either with a carbamate or an organophosphate anticholinesterase pesticide showed significant regional variation in cholinesterase inhibition. The histochemical data corresponded well with data from biochemical assays of acetylcholinesterase activity (overall correlation coefficient of absolute values, r = 0.95). Also, a comparison of assay types by two-way analysis of variance showed no significant main effect. These results support the conclusion that video-based histochemistry is suitable for detailed studies of developmental and toxicological influences on cholinesterases in multiple microscopic regions of the rat brain.
乙酰胆碱酯酶组织化学与用于视频显微镜和光密度测定的廉价且广泛可用的仪器相结合,以研究大鼠大脑不同部位的酶活性和抑制作用。在适当定义的条件下,定量组织化学产生的输出随孵育时间和切片厚度呈线性增加,并且是底物浓度的平滑双曲线函数。体内暴露于毒扁豆碱后的染色时间进程显示,没有迹象表明氨基甲酸酯诱导的胆碱酯酶抑制在体外很容易逆转。用氨基甲酸酯或有机磷酸酯抗胆碱酯酶农药处理的大鼠大脑显示胆碱酯酶抑制存在显著的区域差异。组织化学数据与乙酰胆碱酯酶活性生化测定的数据非常吻合(绝对值的总体相关系数,r = 0.95)。此外,通过双向方差分析对测定类型进行比较,未显示出显著的主效应。这些结果支持以下结论:基于视频的组织化学适用于详细研究发育和毒理学对大鼠大脑多个微观区域胆碱酯酶的影响。