Seniuk-Tatton N A, Henderson J T, Roder J C
Samuel Lunenfeld Research Institute, Mt. Sinai Hospital, Toronto, Canada.
J Neurosci Res. 1995 Aug 1;41(5):663-76. doi: 10.1002/jnr.490410513.
The regional and subcellular localization in the central nervous system (CNS) of postnatal day 5, day 15, and adult rats of ciliary neurotrophic factor (CNTF) mRNA was examined by in situ hybridization with biotinylated riboprobes. Probe specificity was determined by Northern blot analysis of poly(A)+ RNA extracted from adult rat brain using digoxigenin labeled riboprobes and chemiluminescent detection. Both a 4 kb and a 1.2 kb transcript were detected in the cortex and brainstem. In situ hybridization revealed that CNTF mRNA was widely distributed in neurons and glia throughout the CNS at each of the developmental time points. The density of the neuronal hybridization signal was found to be greater in neuronal nuclei than in their cytoplasm. In the nucleus of most neurons, CNTF mRNA distribution was concentrated in a perinucleolar fashion. Alternate sections from the same animals, which were incubated with a specific polyclonal antibody against a CNTF peptide fragment, revealed that both neurons and glia in postnatal day 5, day 15, and adult rat brain were immunoreactive for CNTF.
通过用生物素化的核糖探针进行原位杂交，检测了出生后第5天、第15天和成年大鼠中枢神经系统（CNS）中睫状神经营养因子（CNTF）mRNA的区域和亚细胞定位。使用地高辛标记的核糖探针和化学发光检测，通过对从成年大鼠脑提取的聚腺苷酸（poly(A)+）RNA进行Northern印迹分析来确定探针特异性。在皮质和脑干中检测到了一个4 kb和一个1.2 kb的转录本。原位杂交显示，在每个发育时间点，CNTF mRNA在整个中枢神经系统的神经元和胶质细胞中广泛分布。发现神经元杂交信号在神经元核中的密度高于其细胞质。在大多数神经元的核中，CNTF mRNA分布以核仁周围的方式集中。用针对CNTF肽片段的特异性多克隆抗体孵育同一动物的交替切片，结果显示出生后第5天、第15天和成年大鼠脑中的神经元和胶质细胞对CNTF均具有免疫反应性。
