Stopa E G, Uhl G R, O'Hara B F, Chorsky R L, King J C, Bird E D, Wolfe H
Department of Pathology (Neuropathology), SUNY, HSC, Syracuse 13210.
Pept Res. 1992 Jul-Aug;5(4):201-5.
We have examined the utility of in situ hybridization for detecting pre-prosomatostatin mRNA in postmortem human brain. In preliminary studies, Northern blot analysis using a rat model, which simulates the normal pattern of human post-mortem brain cooling, revealed retention of significant amounts of hybridizable somatostatin mRNA relative to control levels between 12 and 24 hours after death. mRNA extracted from postmortem fetal human brain specimens showed hybridization to cRNA probes directed against pre-prosomatostatin mRNA. We thus undertook in situ hybridization studies. Antisense RNA probes were hybridized to neurons that expressed pre-prosomatostatin in 10-microns sections of adult and fetal human brain. The distribution of pre-prosomatostatin mRNA-containing neurons was similar to that observed for somatostatin-like immunoreactivity; however, the in situ hybridization technique was a more sensitive marker of neuronal perikarya. Our results indicate that hybridization to pre-prosomatostatin mRNA is a useful method for localizing these peptidergic neurons in postmortem human brain tissue.
我们已经研究了原位杂交技术在检测死后人类大脑中前促生长抑素原mRNA方面的实用性。在初步研究中，使用大鼠模型进行的Northern印迹分析模拟了人类死后大脑冷却的正常模式，结果显示，在死亡后12至24小时之间，相对于对照水平，仍保留了大量可杂交的生长抑素mRNA。从死后人类胎儿脑标本中提取的mRNA与针对前促生长抑素原mRNA的cRNA探针发生杂交。因此，我们进行了原位杂交研究。将反义RNA探针与成人和胎儿人类大脑10微米切片中表达前促生长抑素原的神经元进行杂交。含前促生长抑素原mRNA的神经元分布与生长抑素样免疫反应性观察到的分布相似；然而，原位杂交技术是神经元胞体更敏感的标记物。我们的结果表明，与前促生长抑素原mRNA杂交是在死后人类脑组织中定位这些肽能神经元的一种有用方法。
