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Published on: April 6, 2011
Microtubule-dependent PKC-alpha localization in A7r5 smooth muscle cells
A C Dykes1, M E Fultz, M L Norton
1Joan Edwards School of Medicine, Marshall University, Huntington, WV 25704, USA.
American Journal of Physiology. Cell Physiology
|March 15, 2003
Summary
Stimulus causes protein kinase C-alpha (PKC-alpha) to associate with perinuclear microtubules, forming specialized structures. These structures may increase microtubule mass and contribute to dynamic activity or act as scaffolds.
Area of Science:
- Cell Biology
- Molecular Biology
- Biophysics
Background:
- Protein kinase C-alpha (PKC-alpha) plays a role in smooth muscle cell signaling.
- Microtubules are dynamic cytoskeletal components involved in various cellular processes.
- The perinuclear region is a critical area for cellular organization and signaling.
Purpose of the Study:
- To investigate the association of PKC-alpha with microtubules in A7r5 smooth muscle cells.
- To understand the role of stimulus-induced relocalization of PKC-alpha and its interaction with microtubules.
- To characterize the structural and dynamic properties of perinuclear microtubules.
Main Methods:
- Laser scanning confocal microscopy
- Fluorescence resonance energy transfer (FRET) microscopy
- Atomic force microscopy (AFM)
- Immunostaining techniques
- Colchicine treatment to disrupt microtubules
Main Results:
- PKC-alpha colocalizes with beta-tubulin in perinuclear microtubules, both in unstimulated and stimulated cells.
- FRET confirmed heterogeneous association of PKC-alpha with perinuclear microtubules, persisting even after microtubule disruption.
- AFM revealed a fivefold increase in the volume of the perinuclear microtubule structure after stimulation, indicating increased mass.
- Perinuclear microtubules showed higher sensitivity to colchicine-induced dissolution compared to other cellular regions.
Conclusions:
- Perinuclear microtubules form a specialized, dynamically robust structure in stimulated smooth muscle cells.
- PKC-alpha may contribute to the dynamic activity of these perinuclear microtubule structures.
- These structures might serve as scaffolds for regulatory molecule interactions at the cell center.
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