Rao R, Brimijoin S
Department of Pharmacology, Mayo Clinic, Rochester, Minnesota 55905, USA.
Neurochem Res. 1995 Feb;20(2):129-35. doi: 10.1007/BF00970536.
In order to examine the molecular basis for regional variation in expression of brain acetylcholinesterase (AChE), an assay using reverse transcription and polymerase chain reaction (RT-PCR) was developed to measure steady state levels of AChE mRNA. The amplification method was designed to be specific for templates derived from AChE mRNA and to avoid potential artifacts induced by the presence of genomic DNA. RT-PCR made it possible to assay AChE mRNA in milligram samples from different regions of the rat brain. Determinations by RT-PCR were faster and more sensitive than Northern blotting. The results, including a surprisingly low level of AChE mRNA in the caudate nucleus, agreed with earlier observations by Northern blot and in-situ hybridization. Quantitative RT-PCR may be useful in future studies on developmental and physiological regulation of AChE expression in the brain.
为了研究脑乙酰胆碱酯酶（AChE）表达区域差异的分子基础，开发了一种使用逆转录和聚合酶链反应（RT-PCR）的检测方法来测量AChE mRNA的稳态水平。该扩增方法设计为对源自AChE mRNA的模板具有特异性，并避免基因组DNA的存在引起的潜在假象。RT-PCR使得能够检测来自大鼠脑不同区域的毫克级样品中的AChE mRNA。通过RT-PCR进行的测定比Northern印迹更快且更灵敏。结果，包括尾状核中AChE mRNA水平出奇的低，与早期Northern印迹和原位杂交的观察结果一致。定量RT-PCR可能在未来关于脑中AChE表达的发育和生理调节的研究中有用。
