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Articles in PresS, published online ahead of print January 28, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00356.2001
Submitted on December 6, 2001
Accepted on January 16, 2002
1 Nephrology Research and Training Center, Department of Medicine, Division of Nephrology, University of Alabama at Birmingham, Birmingham, AL, USA; CREST, Japan Science and Technology Corporation, Okazaki, Japan
2 Department of Cell Physiology, National Institute for Physiological Sciences, Okazaki, Japan; CREST, Japan Science and Technology Corporation, Okazaki, Japan
Recently, multi-photon excitation fluorescence microscopy has been developed that offers important advantages over confocal imaging, particularly for in vivo visualization of thick tissue samples. We used this state-of-the-art technique to capture high quality images and study the function of otherwise inaccessible cell types and complex cell structures of the juxtaglomerular apparatus (JGA) in living preparations of the kidney. This structure has multiple cell types that exhibit a complex array of functions that regulate the process of filtrate formation and renal hemodynamics. We now report, for the first time, high-resolution 3D morphology and Z-sectioning through isolated, perfused kidney glomeruli, tubules, and JGA. Time-series images show how alterations in tubular fluid composition causes striking changes in single cell volume of the unique macula densa tubular epithelium in situ and how it also affects glomerular filtration through alterations in associated structures within the JGA. In addition, calcium imaging of the glomerulus and JGA demonstrates the utility of this system in capturing the complexity of events and effects that are exerted by the specific hypertensive autacoid angiotensin II. This imaging approach, to study isolated perfused live tissue with multi-photon microscopy may be applied in other biological systems where multiple cell types form a functionally integrated syncytium.
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