Terenghi G, Polak J M, Hamid Q, O'Brien E, Denny P, Legon S, Dixon J, Minth C D, Palay S L, Yasargil G
Department of Histochemistry, Royal Postgraduate Medical School, London, United Kingdom.
Proc Natl Acad Sci U S A. 1987 Oct;84(20):7315-8. doi: 10.1073/pnas.84.20.7315.
The distribution of mRNA encoding neuropeptide Y (NPY) in neurons of the normal human cerebral cortex in surgical biopsy specimens and postmortem brain was studied by in situ hybridization techniques. A 32P-labeled complementary RNA (cRNA) probe was used on cryostat sections of 13 formaldehyde-fixed cortical biopsy specimens. Hybridization to NPY mRNA was found in all samples: after autoradiography, discrete deposits of silver granules were observed on neuronal cell bodies abundantly distributed in the deep layers of the cortex, particularly laminae IV and VI, and on smaller cell bodies in the white matter. The localization of the neurons hybridized for NPY mRNA was comparable to that of NPY-immunoreactive cells as shown in sections from the same tissue blocks immunostained by using NPY antibodies. The specificity of the in situ hybridization technique was confirmed by blot hybridization analysis of electrophoretically fractionated RNA. This study clearly demonstrated the consistent localization of NPY gene transcription and expression in normal mature human cortical neurons.
采用原位杂交技术研究了手术活检标本和死后大脑中正常人类大脑皮质神经元中编码神经肽Y(NPY)的mRNA的分布情况。使用32P标记的互补RNA(cRNA)探针检测13份甲醛固定的皮质活检标本的低温恒温器切片。在所有样本中均发现与NPY mRNA的杂交信号:放射自显影后,在大量分布于皮质深层尤其是IV层和VI层的神经元细胞体以及白质中的较小细胞体上观察到离散的银颗粒沉积。如使用NPY抗体对同一组织块切片进行免疫染色所示,杂交到NPY mRNA的神经元的定位与NPY免疫反应性细胞的定位相当。通过对电泳分离的RNA进行印迹杂交分析证实了原位杂交技术的特异性。本研究清楚地证明了NPY基因转录和表达在正常成熟人类皮质神经元中的一致定位。