Vibration of osteoblastic cells using a novel motion‐control platform does not acutely alter cytosolic calcium, but desensitizes subsequent responses to extracellular ATP
| Autor: | Hristo N. Nikolov, S. Jeffrey Dixon, Daniel Lorusso, David W. Holdsworth |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Cell type Physiology Clinical Biochemistry Stimulation Cell morphology Mechanotransduction Cellular Vibration Mice 03 medical and health sciences Adenosine Triphosphate Cytosol 0302 clinical medicine Calcium imaging Cell Movement Accelerometry Fluorescence microscope medicine Extracellular Animals Mechanotransduction Osteoblasts Receptors Purinergic P2 Chemistry Osteoblast Cell Biology 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis Biophysics Calcium |
| Zdroj: | Journal of Cellular Physiology. 235:5096-5110 |
| ISSN: | 1097-4652 0021-9541 |
| DOI: | 10.1002/jcp.29378 |
| Popis: | Low-magnitude high-frequency mechanical vibration induces biological responses in many tissues. Like many cell types, osteoblasts respond rapidly to certain forms of mechanostimulation, such as fluid shear, with transient elevation in the concentration of cytosolic free calcium ([Ca2+ ]i ). However, it is not known whether vibration of osteoblastic cells also induces acute elevation in [Ca2+ ]i . To address this question, we built a platform for vibrating live cells that is compatible with microscopy and microspectrofluorometry, enabling us to observe immediate responses of cells to low-magnitude high-frequency vibrations. The horizontal vibration system was mounted on an inverted microscope, and its mechanical performance was evaluated using optical tracking and accelerometry. The platform was driven by a sinusoidal signal at 20-500 Hz, producing peak accelerations from 0.1 to 1 g. Accelerometer-derived displacements matched those observed optically within 10%. We then used this system to investigate the effect of acceleration on [Ca2+ ]i in rodent osteoblastic cells. Cells were loaded with fura-2, and [Ca2+ ]i was monitored using microspectrofluorometry and fluorescence ratio imaging. No acute changes in [Ca2+ ]i or cell morphology were detected in response to vibration over the range of frequencies and accelerations studied. However, vibration did attenuate Ca2+ transients generated subsequently by extracellular ATP, which activates P2 purinoceptors and has been implicated in mechanical signaling in bone. In summary, we developed and validated a motion-control system capable of precisely delivering vibrations to live cells during real-time microscopy. Vibration did not elicit acute elevation of [Ca2+ ]i , but did desensitize responses to later stimulation with ATP. |
| Databáze: | OpenAIRE |
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