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培养的海马神经元中rRNA和多聚腺苷酸RNA的亚细胞分布

Subcellular distribution of rRNA and poly(A) RNA in hippocampal neurons in culture.

作者信息

Kleiman R, Banker G, Steward O

机构信息

Department of Neuroscience, University of Virginia, Charlottesville 22908.

出版信息

Brain Res Mol Brain Res. 1993 Dec;20(4):305-12. doi: 10.1016/0169-328x(93)90057-v.

Abstract

In situ hybridization was used to assess the subcellular distribution of rRNA and poly(A) RNA in hippocampal neurons maintained in culture. Labeling produced with 35S-labeled probes to either rRNA or poly(A) was heaviest over the cell body with lighter, patchy labeling of proximal dendrites. In contrast, 3H-labeled probes labeled dendrites throughout their length, and the ratio of dendritic to cell body labeling was higher with 3H-labeled probes. There was no detectable labeling of axons of mature neurons with either probe. The pattern of hybridization produced by 35S-labeled oligonucleotide probes to rRNA varied depending on the concentration of the oligonucleotide. These studies provide the first detailed study of the subcellular distribution of rRNA and poly(A) RNA in neurons, and highlight technical issues to consider when evaluating results of hybridizations carried out with 35S- and 3H-labeled probes on cells in culture.

摘要

原位杂交用于评估培养的海马神经元中rRNA和多聚腺苷酸RNA(poly(A) RNA)的亚细胞分布。用35S标记的rRNA或poly(A) 探针产生的标记在细胞体上最重,近端树突有较轻的斑块状标记。相比之下,3H标记的探针标记了整个树突长度,并且3H标记的探针的树突与细胞体标记的比率更高。两种探针均未检测到成熟神经元轴突的标记。35S标记的寡核苷酸探针与rRNA产生的杂交模式因寡核苷酸浓度而异。这些研究首次详细研究了神经元中rRNA和poly(A) RNA的亚细胞分布,并突出了在评估用35S和3H标记的探针在培养细胞上进行杂交的结果时需要考虑的技术问题。

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