- 作者：Niels Erik Olesen1 ; Johannes P. Hofgaard1 ; Niels-Henrik Holstein-Rathlou ; Morten Schak Nielsen ; Jens Christian Brings Jacobsen ; jcbrings@sund.ku.dk
- 关键词：Gap junction ; Connexin ; Dye diffusion ; Cellular coupling ; Mathematical model
- 刊名：European Journal of Pharmaceutical Sciences
- 出版年：2012
- 期刊代码：15_09280987
- 类别：pha
- 出版时间：16 July, 2012
- 卷：46
- 期：4
- 页码：222-232
- 文件大小：757 K
Objective
A recently developed dye-based assay to study gap junction permeability is analysed. The assay is based on electroporation of dye into a large number of connexin 43 expressing cells, grown to confluency on electrically conductive slides. The subsequent intercellular spread of dye to non-electroporated parts of the monolayer enables estimation of the intercellular coupling. So far, the extent of dye spread has been analyzed in qualitative terms only and not in a manner based directly on the physics of the underlying diffusion process.Methods
We apply a continuum approximation assuming that the observed dye spread can be described by Fick鈥檚 law of diffusion. Deduced from Fick鈥檚 law, new measures are presented which directly relate the observed spread of dye to the diffusion coefficient.
Results
The theoretical framework enables the estimation of an effective diffusion coefficient from Fick鈥檚 law independently of the specific indicator substance used in the assay. For Lucifer Yellow, diffusion stops within few minutes after the electroporation. Therefore only an order-of-magnitude estimate of the diffusion coefficient can be given for this dye.
Conclusion
In terms of the underlying diffusion coefficient, the hitherto used measures give a relatively poor degree of quantification. In contrast, the present methods may yield direct information on the effective intercellular diffusion coefficient and hence provide additional and more precise information as to the permeability modulating effect of various substances.