Cell Biology – Immunology – Pathology
Kidney International (2005) 68, 1071–1077; doi:10.1111/j.1523-1755.2005.00499.x
Scanning ion conductance microscopy reveals how a functional renal epithelial monolayer maintains its integrity
YANJUN ZHANG, JULIA GORELIK, DANIEL SANCHEZ, ANDREW SHEVCHUK, MAX LAB, IGOR VODYANOY, DAVID KLENERMAN, CHRISTOPHER EDWARDS and YURI KORCHEV
Division of Medicine, Imperial College London, London, United Kingdom; The Office of the Vice Chancellor, University of Newcastle, Newcastle upon Tyne, United Kingdom; Department of Chemistry, Cambridge University, Cambridge, United Kingdom; and Office of Naval Research, Arlington, Virginia
Correspondence: Dr Yuri Korchev, Division of Medicine, Imperial College London, MRC Clinical Sciences Center, Hammersmith Campus, Du Cane Road, London W12 0NN, UK. E-mail: y.korchev@ic.ac.uk
Received 28 January 2005; Revised 7 April 2005; Accepted 18 April 2005.
Abstract
Scanning ion conductance microscopy reveals how a functional renal epithelial monolayer maintains its integrity.
Background
To function as a transport barrier a renal tubule epithelial monolayer needs to maintain its integrity when, sudden hypertonic stress causes cell shrinkage, new cells are added, or cells in the monolayer die. However, the mechanism used to achieve this is largely unknown. Scanning ion conductance microscopy (SICM) has been shown to be suitable for imaging the surface of live renal cells with high topographic resolution, and can be used to elucidate how a functional renal epithelial monolayer maintains its integrity.
Methods
SICM was used for high spatial resolution topographic imaging of Xenopus laevis renal epithelial A6 cells cultured on membrane filter inserts.
Results
The SICM images of A6 cells showed that the epithelial monolayer maintains its integrity under hypertonic stress, and during cell division and death. Sequential SICM topographic images revealed detailed structural changes and their time course for these protective processes, which involve highly cooperative cell movement. Some "balloon-like" structures were observed at susceptible tight junction regions, which were proposed to help cell maintaining the monolayer permeability integrity.
Conclusion
SICM is a powerful tool for research on living renal epithelial cells, and has been used to elucidate how a functional epithelial monolayer maintains its integrity. Using this technique we have observed that during hypertonic stress and regeneration, an organized sequence of events protect the loss of integrity of monolayer so that tight junctions and cell-cell contact are maintained and disruption to the function of whole monolayer is prevented.
Keywords:
scanning probe microscopy, renal epithelia monolayer, regeneration, cell division, apoptosis, integrity
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