title: Mitochondrial Ca²⁺ handling is crucial for generation of rhythmical Ca²⁺ waves in vascular interstitial cells from rabbit portal vein.
abstract: Vasomotion is the rhythmical changes in vascular tone of various blood vessels. It was proposed that in rabbit portal vein (RPV) the spontaneous contractile activity is driven by vascular interstitial cells (VICs), since RPV VICs generate rhythmical changes in intracellular Ca(2+) concentration ([Ca(2+)]i) associated with membrane depolarisation in these cells. In this work, using confocal imaging in Fluo-3 loaded RPV VICs we studied if generation of rhythmical [Ca(2+)]i changes is affected when Ca(2+) handling by mitochondria is compromised. We also visualised mitochondria in VICs using Mito Tracker Green fluorescent dye. Our results showed that freshly dispersed RPV VICs generated rhythmical [Ca(2+)]i oscillations with a frequency of 0.2-0.01 Hz. Imaging of VICs stained with Mito Tracker Green revealed abundant mitochondria in these cells with a higher density of the organelles in sub-plasmalemmar region compared to the central region of the cell. Oligomycin, an ATP synthase inhibitor, did not affect the amplitude and frequency of rhythmical [Ca(2+)]i oscillations. In contrast, two uncoupling agents, carbonylcyanide-3-chlorophenylhydrazone (CCCP) and carbonylcyanide-4-trifluoromethoxyphenylhydrazone (FCCP) effectively abolished rhythmical [Ca(2+)]i changes with simultaneous increase in basal [Ca(2+)]i in RPV VICs. These data suggest that in RPV VICs mitochondrial Ca(2+) handling is important for the generation of rhythmical [Ca(2+)]i changes which underlie the spontaneous rhythmical contractile activity in this vessel.
authors.affiliation: Division of Biomedical Sciences, St. George's, University of London, London, United Kingdom; Laboratory of Molecular Pharmacology and Biophysics of Cell Signalling, Bogomoletz Institute of Physiology, Kyiv, Ukraine. Electronic address: m.harhun@gmail.com.
authors.forename: Maksym I
authors.initials: MI
authors.lastname: Harhun
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology*; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D003239:Connective Tissue Cells / Q000378:metabolism / Q000648:ultrastructure; D008928:Mitochondria / Q000378:metabolism*; D009119:Muscle Contraction; D009840:Oligomycins / Q000494:pharmacology; D010507:Periodicity; D011169:Portal Vein / Q000378:metabolism / Q000648:ultrastructure; D011817:Rabbits; D014475:Uncoupling Agents / Q000494:pharmacology
chemical_list: D009840:Oligomycins; D014475:Uncoupling Agents; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D002118:Calcium
keywords: Mitochondria; Rhythmical calcium waves; Vascular interstitial cell; Vasomotion