Bending the MDCK cell primary cilium increases intracellular calcium.
We tested the hypothesis that the primary cilium of renal epithelia is mechanically sensitive and serves as a flow sensor in MDCK cells using differential interference contrast and fluorescence microscopy. Bending the cilium, either by suction with a micropipette or by increasing the flow rate of perfusate, causes intracellular calcium to substantially increase as indicated by the fluorescent indicator, Fluo-4. This calcium signal is initiated by Ca2+-influx through mechanically sensitive channels that probably reside in the cilium or its base. The influx is followed by calcium release from IP3-sensitive stores. The calcium signal then spreads as a wave from the perturbed cell to its neighbors by diffusion of a second messenger through gap junctions. This spreading of the calcium wave points to flow sensing as a coordinated event within the tissue, rather than an isolated phenomenon in a single cell. Measurement of the membrane potential difference by microelectrode during perfusate flow reveals a profound hyperpolarization during the period of elevated intracellular calcium. We conclude that the primary cilium in MDCK cells is mechanically sensitive and responds to flow by greatly increasing intracellular calcium.
Praetorius
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NIH, NHLBI, LKEM, 10 Center Drive, Bldg. 10, Bethesda, MD 20892-1603, USA. praetorh@nhlbi.nih.gov
Spring
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D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020013:Calcium Signaling / Q000502:physiology*; D002460:Cell Line; D016764:Cell Polarity; D002923:Cilia / Q000378:metabolism*; D004285:Dogs; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D019850:Heptanol / Q000494:pharmacology; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000378:metabolism; D008564:Membrane Potentials / Q000502:physiology; D018613:Microscopy, Confocal; D009568:Nitrendipine / Q000494:pharmacology; D013314:Stress, Mechanical; D014700:Verapamil / Q000494:pharmacology
D002121:Calcium Channel Blockers; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D019850:Heptanol; D009568:Nitrendipine; D014700:Verapamil; D002118:Calcium