Fluctuating Shear Stress Effects on Stress Fiber Architecture and Energy Metabolism of Cultured Renal Cells

The project investigates the relationship between the external shear force and the actin cytoskeleton along with the metabolic changes occurring inside the cells due to this force. Anchorage‐dependent Madin Darby canine kidney (MDCK) cells were placed in spinner flasks with paddle‐type stirrers agit...

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Bibliographic Details
Published in:Biotechnology progress 1995-09, Vol.11 (5), p.596-600
Main Authors: Bhat, Vinayak D., Windridge, Paula A., Cherry, Robert S., Mandel, Lazaro J.
Format: Article
Language:English
Subjects:
Actins - metabolism
Adenosine Triphosphate - metabolism
Animal cells
Animals
Biological and medical sciences
Biotechnology
Biotechnology - methods
Cell Count
Cell Survival - physiology
Cells, Cultured
Cytoskeleton - metabolism
Cytoskeleton - physiology
Dogs
Energy Metabolism
Epithelial Cells
Eukaryotic cell cultures
Fundamental and applied biological sciences. Psychology
Kidney - cytology
Kidney - metabolism
Kidney Tubules, Proximal - cytology
Kidney Tubules, Proximal - physiology
Methods. Procedures. Technologies
Miscellaneous
Proteins - metabolism
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Summary:The project investigates the relationship between the external shear force and the actin cytoskeleton along with the metabolic changes occurring inside the cells due to this force. Anchorage‐dependent Madin Darby canine kidney (MDCK) cells were placed in spinner flasks with paddle‐type stirrers agitated at 20 rpm, where they experienced shear stress fluctuations from 0.02 to 0.27 dyn/cm2 in magnitude. Following fixation, permeabilization, and staining with rhodamine—phalloidin, the relative amounts and distribution of F‐actin stress fibers in the 1 μm basal layer of the cells were visualized by confocal microscopy. These structures disappeared after 12–15 h of exposure to shear stress. Previous results showed that the stress fibers disappear, leading to loss of epithelial attachment, after only 1 h of starvation‐induced energy depletion. Therefore, in this study, the energy metabolism of the cells was established by measuring adenosine triphosphate (ATP) levels at different time intervals. No statistical difference in ATP content was found between the shear‐stressed cells and the controls, showing that shear stresses cause cytoskeletal reorganization by a mechanism other than ATP depletion.
ISSN:8756-7938
1520-6033
DOI:10.1021/bp00035a016