Diffusion Behavior of Gap Junction Hemichannels in Living Cells

M.      Gerken; E.      Thews; C.      Tietz; J.      Wrachtrup; R.      Eckert

Abstract

Due to its non-invasive character, fluorescence correlation spectroscopy (FCS) is particularly suited for the investigation of diffusion behavior of proteins in living cells. In this study we have investigated the diffusion properties of CFP-labeled gap junction hemichannels in the plasma membrane of living HeLa cells. Gap junction hemichannels or connexons are the precursors for the cell-cell- or gap junction channels that form large plaques at the contact areas between two adjacent cells. It has been proposed that new channels are recruited into a gap junction structure from a pool of hemichannels that can freely diffuse over the entire plasma membrane. The statistical approach shows that the geometry of the membrane within the focus is the most important property for the form of the autocorrelation curve and in turn for the determination of the diffusion coefficient. On the other hand binding-unbinding events which lead to anomalous diffusion have only a minor effect to the position and shape of the correlation curve compared to the geometry of the membrane.

Keywords: diffusion, fluorescence correlation spectroscopy (fcs), cytoplasm, cell-cell channels, connexins, gfp-labeling, transient transfection, brownian diffusion models