Organization of intracellular content is affected by multiple simultaneous processes, including diffusion in a viscoelastic and structured environment, intracellular mechanical work and vibrations. The combined effects of these processes on intracellular organization are complex and remain poorly understood. Here, we studied the organization and dynamics of a free Ca++ probe as a small and mobile tracer in live T cells. Ca++, highlighted by Fluo-4, is localized in intracellular organelles. Inhibiting intracellular mechanical work by myosin II through blebbistatin treatment in-creased cellular dis-homogeneity of Ca++-rich features in length scale < 1.1 µm. We detected a similar effect in cells imaged by label-free bright-field (BF) microscopy, in mitochondria-highlighted cells and in ATP-depleted cells. Blebbistatin treatment also reduced the dynamics of the Ca++-rich features and generated prominent negative temporal correlations in their signals. Following Guggenberger et al. and numerical simulations, we suggest that diffusion in the viscoelastic and confined medium of intracellular organelles may promote spatial dis-homogeneity and stability of their content. This may be revealed only after inhibiting intracellular mechanical work and related cell vibrations. Our described mechanisms may allow the cell to control its organization via balancing its viscoelasticity and mechanical activity, with implications to cell physiology in health and disease.
Bibliographical noteFunding Information:
This research was funded by the ISF, grant number 1761/17. We thank Naomi Book (The Silberman Institute at HUJI) for her assistance with confocal microscopy.
organelle medium follows sub-diffusion fBM patterns with dominancy of negative corre-correlations. This is supported by the intra-organelle medium being highly crowded ,
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
- Cell organization
- Fractional Brownian motion (fBM)
- Intracellular work
- Myosin II
- intracellular work
- fractional Brownian motion (fBM)
- cell organization
- myosin II