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活细胞和体外环境中哺乳动物细胞核的通透性特性

Permeability properties of mammalian cell nuclei in living cells and in vitro.

作者信息

Reynolds C R, Tedeschi H

出版信息

J Cell Sci. 1984 Aug;70:197-207. doi: 10.1242/jcs.70.1.197.

DOI:10.1242/jcs.70.1.197
PMID:6501436
Abstract

The present study evaluates the role of the nuclear envelope in mammalian cells by applying two different approaches using either intact cells of mouse liver or isolated nuclei. In one approach the nuclei were studied with microelectrodes. The transmembrane voltage drop produced by passing current through an impaling microelectrode was measured with a second impaling microelectrode. In the second approach, the permeability of the nuclear envelope was studied by injection of a series of fluorescent probes. Lucifer Yellow CH and a variety of exogenous proteins labelled by conjugation with Lucifer Yellow VS were delivered into either the cytoplasm or the nucleus in situ. The fluorescence of the probe was followed either with a video camera or photographically. The results agree with the idea that the mammalian nuclear envelope is permeable to rather large molecules. Molecules with estimated radii below 2.4 nm seem to exchange rapidly, whereas molecules with estimated radii of 2.8 nm or above are excluded. The low electrical resistance of the envelope yields an estimate of pore radius, in the range of 3.4-6.5 nm.

摘要

本研究通过使用两种不同方法来评估核膜在哺乳动物细胞中的作用,这两种方法分别使用小鼠肝脏的完整细胞或分离的细胞核。在一种方法中,用微电极研究细胞核。通过刺入微电极通电流产生的跨膜电压降,用第二个刺入微电极进行测量。在第二种方法中,通过注射一系列荧光探针来研究核膜的通透性。将荧光黄CH以及通过与荧光黄VS偶联标记的多种外源蛋白原位递送至细胞质或细胞核中。用摄像机或通过摄影跟踪探针的荧光。结果支持这样一种观点,即哺乳动物的核膜对相当大的分子具有通透性。估计半径小于2.4nm的分子似乎能快速交换,而估计半径为2.8nm或更大的分子则被排除在外。核膜的低电阻得出的孔径估计值在3.4 - 6.5nm范围内。

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