Endothelial mitochondria regulate the intracellular Ca2+ response to fluid shear stress

Christopher G. Scheitlin, Justin A. Julian, Santhanam Shanmughapriya, Muniswamy Madesh, Nikolaos M. Tsoukias, B. Rita Alevriadou

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Shear stress is known to stimulate an intracellular free calcium concentration ([Ca2+]i) response in vascular endothelial cells (ECs). [Ca2+]i is a key second messenger for signaling that leads to vasodilation and EC survival. Although it is accepted that the shear-induced [Ca2+]i response is, in part, due to Ca2+ release from the endoplasmic reticulum (ER), the role of mitochondria (second largest Ca2+ store) is unknown. We hypothesized that the mitochondria play a role in regulating [Ca2+]i in sheared ECs. Cultured ECs, loaded with a Ca2+-sensitive fluorophore, were exposed to physiological levels of shear stress. Shear stress elicited [Ca2+]i transients in a percentage of cells with a fraction of them displaying oscillations. Peak magnitudes, percentage of oscillating ECs, and oscillation frequencies depended on the shear level. [Ca2+]i transients/oscillations were present when experiments were conducted in Ca2+-free solution (plus lanthanum) but absent when ECs were treated with a phospholipase C inhibitor, suggesting that the ER inositol 1,4,5-trisphosphate receptor is responsible for the [Ca2+]i response. Either a mitochondrial uncoupler or an electron transport chain inhibitor, but not a mitochondrial ATP synthase inhibitor, prevented the occurrence of transients and especially inhibited the oscillations. Knockdown of the mitochondrial Ca2+ uniporter also inhibited the shear-induced [Ca2+]i transients/oscillations compared with controls. Hence, EC mitochondria, through Ca2+ uptake/release, regulate the temporal profile of shear-induced ER Ca2+ release. [Ca2+]i oscillation frequencies detected were within the range for activation of mechanoresponsive kinases and transcription factors, suggesting that dysfunctional EC mitochondria may contribute to cardiovascular disease by deregulating the shear-induced [Ca2+]i response.

Original languageEnglish (US)
Pages (from-to)C479-C490
JournalAmerican Journal of Physiology - Cell Physiology
Volume310
Issue number6
DOIs
StatePublished - Mar 15 2016

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Mitochondria
Endothelial Cells
Endoplasmic Reticulum
Mitochondrial Proton-Translocating ATPases
Inositol 1,4,5-Trisphosphate Receptors
Lanthanum
Type C Phospholipases
Second Messenger Systems
Electron Transport
Vasodilation
Cultured Cells
Cell Survival
Transcription Factors
Cardiovascular Diseases
Phosphotransferases
Calcium

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cell Biology

Cite this

Scheitlin, Christopher G. ; Julian, Justin A. ; Shanmughapriya, Santhanam ; Madesh, Muniswamy ; Tsoukias, Nikolaos M. ; Alevriadou, B. Rita. / Endothelial mitochondria regulate the intracellular Ca2+ response to fluid shear stress. In: American Journal of Physiology - Cell Physiology. 2016 ; Vol. 310, No. 6. pp. C479-C490.
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Endothelial mitochondria regulate the intracellular Ca2+ response to fluid shear stress. / Scheitlin, Christopher G.; Julian, Justin A.; Shanmughapriya, Santhanam; Madesh, Muniswamy; Tsoukias, Nikolaos M.; Alevriadou, B. Rita.

In: American Journal of Physiology - Cell Physiology, Vol. 310, No. 6, 15.03.2016, p. C479-C490.

Research output: Contribution to journalArticle

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AU - Scheitlin, Christopher G.

AU - Julian, Justin A.

AU - Shanmughapriya, Santhanam

AU - Madesh, Muniswamy

AU - Tsoukias, Nikolaos M.

AU - Alevriadou, B. Rita

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AB - Shear stress is known to stimulate an intracellular free calcium concentration ([Ca2+]i) response in vascular endothelial cells (ECs). [Ca2+]i is a key second messenger for signaling that leads to vasodilation and EC survival. Although it is accepted that the shear-induced [Ca2+]i response is, in part, due to Ca2+ release from the endoplasmic reticulum (ER), the role of mitochondria (second largest Ca2+ store) is unknown. We hypothesized that the mitochondria play a role in regulating [Ca2+]i in sheared ECs. Cultured ECs, loaded with a Ca2+-sensitive fluorophore, were exposed to physiological levels of shear stress. Shear stress elicited [Ca2+]i transients in a percentage of cells with a fraction of them displaying oscillations. Peak magnitudes, percentage of oscillating ECs, and oscillation frequencies depended on the shear level. [Ca2+]i transients/oscillations were present when experiments were conducted in Ca2+-free solution (plus lanthanum) but absent when ECs were treated with a phospholipase C inhibitor, suggesting that the ER inositol 1,4,5-trisphosphate receptor is responsible for the [Ca2+]i response. Either a mitochondrial uncoupler or an electron transport chain inhibitor, but not a mitochondrial ATP synthase inhibitor, prevented the occurrence of transients and especially inhibited the oscillations. Knockdown of the mitochondrial Ca2+ uniporter also inhibited the shear-induced [Ca2+]i transients/oscillations compared with controls. Hence, EC mitochondria, through Ca2+ uptake/release, regulate the temporal profile of shear-induced ER Ca2+ release. [Ca2+]i oscillation frequencies detected were within the range for activation of mechanoresponsive kinases and transcription factors, suggesting that dysfunctional EC mitochondria may contribute to cardiovascular disease by deregulating the shear-induced [Ca2+]i response.

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