Visualization Of Protein Dynamics Research Articles

A β-catenin chromobody-based probe highlights endothelial maturation during vascular morphogenesis in vivo.

Visualization of protein dynamics is a crucial step in understanding cellular processes. Chromobodies, fluorescently labelled single-domain antibodies, have emerged as versatile probes for live cell imaging of endogenous proteins. However, how these chromobodies behave in vivo and how accurately they monitor tissue changes remain poorly explored. Here, we generated an endothelial-specific β-catenin chromobody-derived probe and analyzed its expression pattern during cardiovascular development in zebrafish. Using high-resolution confocal imaging, we show that the chromobody signal correlates with the localization of β-catenin in the nucleus and at cell-cell junctions, and thereby can be used to assess endothelial maturation. Loss of Cadherin 5 strongly affects the localization of the chromobody at the cell membrane, confirming the cadherin-based adherens junction role of β-catenin. Furthermore, using a genetic model to block blood flow, we observed that cell junctions are compromised in most endothelial cells but not in the endocardium, highlighting the heterogeneous response of the endothelium to the lack of blood flow. Overall, our data further expand the use of chromobodies for in vivo applications and illustrate their potential to monitor tissue morphogenesis at high resolution.

Dynamics of gene expression regulatory proteins in the living cell nucleus

The intricate physical interaction of transcription factors with specific target genes in the genome has long been regarded as a fundamental mechanism of cell-type specific gene expression. However, due to insufficient spatiotemporal resolution of microscopy techniques, direct visualization of protein dynamics within the nuclei of living cells had not been achieved for decades. Resolving existing limitations, recent advances in imaging techniques enabled the direct observation of protein dynamics, even at a single-molecule level. In addition, the new imaging techniques accomplished capturing higher-order chromatin structures that influence gene expression regulation along with transcription factor dynamics. This review discusses the recent applications of microscopy techniques to investigate the dynamics of nuclear proteins in living cells and their achievements.

Visualization of Protein Dynamics using High-Speed Atomic Force Microscopy and Image Analysis

Visualization of Protein Dynamics using High-Speed Atomic Force Microscopy and Image Analysis

フォトクロミック蛍光タンパク質を用いた細胞内物質運動の可視化

フォトクロミック蛍光タンパク質を用いた細胞内物質運動の可視化