An analysis of the time-dependent changes in intracellular calcium concentration in endothelial cells in culture induced by mechanical stimulation.
When bovine pulmonary artery endothelial cells in culture are subjected to mechanical strain, their physiology is altered. Experimentally, this mechanical strain is generated by increased tension in the substrate to which the cells are attached and results in altered levels of fibronectin. Studies of the structural response of the endothelial cell suggest that this stimulus is transmitted to the cell membrane, organelles, and cytoskeleton by natural cell attachments in a quantifiable and predictable manner. This report examines altered intracellular calcium homeostasis as a possible messenger for the observed strain-induced physiologic response. In particular, using the intracellular trapped calcium indicator dyes, Quin2 and Fura2, we observed changes in cytosolic free calcium ion concentration in response to biaxial strain of bovine pulmonary artery endothelial cells in culture. The magnitude and time course of this calcium transient resemble that produced by treatment with the calcium ionophore, Ionomycin, indicating that mechanical stimulation may alter cell membrane permeability to calcium. Additional experiments in the presence of EDTA indicated that calcium was also released from intracellular stores in response to strain. In order to explain the stretch-induced calcium transients, a first-order species conservation model is presented that takes into account both the cell's structural response and the calcium homeostatic mechanisms of the cell. It is hypothesized that the cell's calcium sequestering and pumping capabilities balanced with its mechanically induced changes in calcium ion permeability will determine the level and time course of calcium accumulation in the cytosol.
Department of Bioengineering, University of Pennsylvania, Philadelphia 19104.
F K
FK
Winston
L E
LE
Thibault
E J
EJ
Macarak
D000818:Animals; D002118:Calcium / Q000032:analysis* / Q000493:pharmacokinetics; D015220:Calcium Channels / Q000502:physiology*; D000252:Calcium-Transporting ATPases / Q000502:physiology*; D002417:Cattle; D002463:Cell Membrane Permeability / Q000502:physiology*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000737:chemistry*; D005069:Evaluation Studies as Topic; D005353:Fibronectins / Q000032:analysis; D008958:Models, Molecular; D010812:Physical Stimulation; D011651:Pulmonary Artery / Q000166:cytology; D013314:Stress, Mechanical; D013997:Time Factors
D015220:Calcium Channels; D005353:Fibronectins; D000252:Calcium-Transporting ATPases; D002118:Calcium