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Updated: Nov 4, 2025
Simultaneous Visualization of the Dynamics of Crosslinked and Single Microtubules In Vitro by TIRF Microscopy
Published on: February 18, 2022
Live-Cell Fluorescence Imaging of Microtubules by Using a Tau-Derived Peptide
Hiroshi Inaba1,2, Kazunori Matsuura3,4
1Department of Chemistry and Biotechnology, Graduate School of Engineering, Tottori University, Tottori, Japan. hinaba@tottori-u.ac.jp.
Abstract:
Microtubules (MTs) are important targets for imaging in living cells because of their vital roles in cellular processes. The dynamics (polymerization/depolymerization) of MTs has been imaged in living cells by utilizing MT-targeted drugs as scaffolds. We previously developed a unique MT-binding motif derived from a MT-associated protein, Tau. The Tau-derived peptide (TP) binds to the inner surface of MTs without inhibiting the dynamics of MTs. We introduce a new protocol for live-cell imaging of MTs by using fluorescently labeled TP. We exemplify that tetramethylrhodamine (TMR)-labeled TP (TP-TMR) is spontaneously internalized into HepG2 cells and binds to intracellular MTs, enabling visualization of MTs in living cells. TP-TMR shows no apparent effects on polymerization/depolymerization of MTs and no cytotoxicity. Thus, the peptide-based approach is useful for long-term imaging of MTs.
Insights
Researchers developed a novel peptide probe for live-cell imaging of microtubules (MTs). This Tau-derived peptide (TP) allows long-term visualization of MT dynamics without affecting cell health.
Area of Science:
- Cell Biology
- Biophysics
- Molecular Imaging
Background:
- Microtubules (MTs) are crucial for cellular functions and are key targets for live-cell imaging.
- Existing imaging methods often rely on MT-targeting drugs, which can interfere with MT dynamics.
Purpose of the Study:
- To develop a new, non-disruptive method for live-cell imaging of microtubules.
- To utilize a novel Tau-derived peptide (TP) for visualizing MTs in living cells.
Main Methods:
- Developed a Tau-derived peptide (TP) that binds to the inner surface of microtubules.
- Fluorescently labeled TP with tetramethylrhodamine (TP-TMR) for visualization.
- Assessed spontaneous internalization, MT binding, MT dynamics, and cytotoxicity in HepG2 cells.
Main Results:
- TP-TMR was spontaneously internalized into HepG2 cells.
- TP-TMR successfully bound to intracellular MTs, enabling their visualization.
- TP-TMR did not inhibit MT polymerization/depolymerization and exhibited no cytotoxicity.
Conclusions:
- The peptide-based approach using fluorescently labeled TP is effective for live-cell imaging of MTs.
- This method allows for long-term visualization of MT dynamics without adverse effects.
- TP-TMR offers a promising tool for studying microtubule functions in living cells.
