Calcium signaling in live cells on elastic gels under mechanical vibration at subcellular levels.
A new device was designed to generate a localized mechanical vibration of flexible gels where human umbilical vein endothelial cells (HUVECs) were cultured to mechanically stimulate these cells at subcellular locations. A Fluorescence Resonance Energy Transfer (FRET)-based calcium biosensor (an impr...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3203865?pdf=render |
| _version_ | 1819135182525956096 |
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| author | Wagner Shin Nishitani Taher A Saif Yingxiao Wang |
| author_facet | Wagner Shin Nishitani Taher A Saif Yingxiao Wang |
| author_sort | Wagner Shin Nishitani |
| collection | DOAJ |
| description | A new device was designed to generate a localized mechanical vibration of flexible gels where human umbilical vein endothelial cells (HUVECs) were cultured to mechanically stimulate these cells at subcellular locations. A Fluorescence Resonance Energy Transfer (FRET)-based calcium biosensor (an improved Cameleon) was used to monitor the spatiotemporal distribution of intracellular calcium concentrations in the cells upon this mechanical stimulation. A clear increase in intracellular calcium concentrations over the whole cell body (global) can be observed in the majority of cells under mechanical stimulation. The chelation of extracellular calcium with EGTA or the blockage of stretch-activated calcium channels on the plasma membrane with streptomycin or gadolinium chloride significantly inhibited the calcium responses upon mechanical stimulation. Thapsigargin, an endoplasmic reticulum (ER) calcium pump inhibitor, or U73122, a phospholipase C (PLC) inhibitor, resulted in mainly local calcium responses occurring at regions close to the stimulation site. The disruption of actin filaments with cytochalasin D or inhibition of actomyosin contractility with ML-7 also inhibited the global calcium responses. Therefore, the global calcium response in HUVEC depends on the influx of calcium through membrane stretch-activated channels, followed by the release of inositol trisphosphate (IP3) via PLC activation to trigger the ER calcium release. Our newly developed mechanical stimulation device can also provide a powerful tool for the study of molecular mechanism by which cells perceive the mechanical cues at subcellular levels. |
| first_indexed | 2024-12-22T10:15:01Z |
| format | Article |
| id | doaj.art-50b6dc8a88b94598971825404cbaf3f2 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-22T10:15:01Z |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-50b6dc8a88b94598971825404cbaf3f22022-12-21T18:29:45ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-01610e2618110.1371/journal.pone.0026181Calcium signaling in live cells on elastic gels under mechanical vibration at subcellular levels.Wagner Shin NishitaniTaher A SaifYingxiao WangA new device was designed to generate a localized mechanical vibration of flexible gels where human umbilical vein endothelial cells (HUVECs) were cultured to mechanically stimulate these cells at subcellular locations. A Fluorescence Resonance Energy Transfer (FRET)-based calcium biosensor (an improved Cameleon) was used to monitor the spatiotemporal distribution of intracellular calcium concentrations in the cells upon this mechanical stimulation. A clear increase in intracellular calcium concentrations over the whole cell body (global) can be observed in the majority of cells under mechanical stimulation. The chelation of extracellular calcium with EGTA or the blockage of stretch-activated calcium channels on the plasma membrane with streptomycin or gadolinium chloride significantly inhibited the calcium responses upon mechanical stimulation. Thapsigargin, an endoplasmic reticulum (ER) calcium pump inhibitor, or U73122, a phospholipase C (PLC) inhibitor, resulted in mainly local calcium responses occurring at regions close to the stimulation site. The disruption of actin filaments with cytochalasin D or inhibition of actomyosin contractility with ML-7 also inhibited the global calcium responses. Therefore, the global calcium response in HUVEC depends on the influx of calcium through membrane stretch-activated channels, followed by the release of inositol trisphosphate (IP3) via PLC activation to trigger the ER calcium release. Our newly developed mechanical stimulation device can also provide a powerful tool for the study of molecular mechanism by which cells perceive the mechanical cues at subcellular levels.http://europepmc.org/articles/PMC3203865?pdf=render |
| spellingShingle | Wagner Shin Nishitani Taher A Saif Yingxiao Wang Calcium signaling in live cells on elastic gels under mechanical vibration at subcellular levels. PLoS ONE |
| title | Calcium signaling in live cells on elastic gels under mechanical vibration at subcellular levels. |
| title_full | Calcium signaling in live cells on elastic gels under mechanical vibration at subcellular levels. |
| title_fullStr | Calcium signaling in live cells on elastic gels under mechanical vibration at subcellular levels. |
| title_full_unstemmed | Calcium signaling in live cells on elastic gels under mechanical vibration at subcellular levels. |
| title_short | Calcium signaling in live cells on elastic gels under mechanical vibration at subcellular levels. |
| title_sort | calcium signaling in live cells on elastic gels under mechanical vibration at subcellular levels |
| url | http://europepmc.org/articles/PMC3203865?pdf=render |
| work_keys_str_mv | AT wagnershinnishitani calciumsignalinginlivecellsonelasticgelsundermechanicalvibrationatsubcellularlevels AT taherasaif calciumsignalinginlivecellsonelasticgelsundermechanicalvibrationatsubcellularlevels AT yingxiaowang calciumsignalinginlivecellsonelasticgelsundermechanicalvibrationatsubcellularlevels |