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首页> 美国卫生研究院文献>Biophysical Journal >A Transient Diffusion Model Yields Unitary Gap Junctional Permeabilities from Images of Cell-to-Cell Fluorescent Dye Transfer Between Xenopus Oocytes
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A Transient Diffusion Model Yields Unitary Gap Junctional Permeabilities from Images of Cell-to-Cell Fluorescent Dye Transfer Between Xenopus Oocytes

机译：瞬态扩散模型从非洲爪蟾卵母细胞之间的细胞间荧光染料转移的图像中产生单位间隙连接渗透率。

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As ubiquitous conduits for intercellular transport and communication, gap junctional pores have been the subject of numerous investigations aimed at elucidating the molecular mechanisms underlying permeability and selectivity. Dye transfer studies provide a broadly useful means of detecting coupling and assessing these properties. However, given evidence for selective permeability of gap junctions and some anomalous correlations between junctional electrical conductance and dye permeability by passive diffusion, the need exists to give such studies a more quantitative basis. This article develops a detailed diffusion model describing experiments (reported separately) involving transport of fluorescent dye from a “donor” region to an “acceptor” region within a pair of Xenopus oocytes coupled by gap junctions. Analysis of transport within a single oocyte is used to determine the diffusion and binding characteristics of the cellular cytoplasm. Subsequent double-cell calculations then yield the intercellular junction permeability, which is translated into a single-channel permeability using concomitant measurements of intercellular conductance, and known single-channel conductances of gap junctions made up of specific connexins, to count channels. The preceding strategy, combined with use of a graded size series of Alexa dyes, permits a determination of absolute values of gap junctional permeability as a function of dye size and connexin type. Interpretation of the results in terms of pore theory suggests significant levels of dye-pore affinity consistent with the expected order of magnitude of typical (e.g., van der Waals) intermolecular attractions.
机译：作为用于细胞间运输和通讯的无处不在的导管，间隙连接孔已成为旨在阐明通透性和选择性的分子机制的众多研究的主题。染料转移研究提供了检测耦合和评估这些特性的广泛有用的方法。但是，鉴于有证据表明间隙连接具有选择性渗透性，并且通过被动扩散可以使连接电导率和染料渗透性之间存在某些异常相关性，因此有必要为此类研究提供更多的定量依据。本文开发了一个详细的扩散模型，描述了实验（单独报告），其中涉及荧光染料从一对间隙连接的非洲爪蟾卵母细胞中的“供体”区域转移到“受体”区域。分析单个卵母细胞中的转运用于确定细胞质的扩散和结合特征。然后，随后的双细胞计算产生细胞间连接渗透性，使用细胞间电导的伴随测量以及由特定连接蛋白组成的间隙连接的已知单通道电导，将其转换为单通道渗透性以计算通道。前面的策略，结合使用渐变大小的Alexa染料系列，可以确定间隙连接渗透性的绝对值与染料大小和连接蛋白类型的关系。根据孔理论对结果的解释表明，显着水平的染料-孔亲和力与典型的分子间吸引力的预期量级相符。

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