To investigate how physical forces and changes in mechanical properties of cells contribute to development and disease, we designed the fluorescent reporter GenEPi for visualizing dynamics and mechanical stimuli of Piezo1, an essential mechanosensitive ion channel found in plants and animals(10). Throughout an organism’s lifetime, cell mechanosensation (i.e., the ability to perceive and respond to mechanical stimuli in the form of shear stress, tension, or compression) is essential in a myriad of developmental, physiological, and pathophysiological processes including embryogenesis, homeostasis, metastasis, and wound healing. Using engineered optimized primed convertible FPs (pr-FPs), we (and others) have applied primed conversion also in non-toxic single molecule dynamic analysis using super-resolution imaging(9). PhoCl spontaneously dissociates into two fragments after light-induced cleavage of a specific bond in the protein backbone, opening the path to transcriptional manipulation of cells in vivo at single cells resolution(8). Primed conversion has also been successfully extended to manipulate the pcFP-based optogenetic effector, photocleavable protein (PhoCl). The combination of primed conversion and a spatial drift correction algorithm, primed Track, allowed us to accomplish high-fidelity volumetric lineage tracing in mouse pre-implantation embryos(6,7). Using confined primed conversion, we revealed the complex anatomy of individual neurons packed between neighboring cells in zebrafish(2,5). As two-photon-based photoconversion is extremely inefficient, primed conversion is the only way to precisely photoconvert in 3D pcFPs for real-time in vivo studies aiming to unravel complex structural and dynamic information. To get more insight into the elaborate cell and protein dynamics that underlie development and disease(1), the lab introduced a unique optical mechanism, primed conversion, where dual-wavelength illumination results in pronounced photoconversion of photoconvertible fluorescent proteins (pcFPs)(2-4). 1) to establish an effective acquisition and interpretation workflow i) for the mechanistic analysis of biological systems in animal models such as mouse and zebrafish and ii) for the use in novel diagnostic and therapeutic strategies. Recent major research contributions Since establishing the lab, the aim of Dr Pantazis' research activity was to develop advanced imaging technologies (probes, imaging modality, and quantitative analysis see Fig.
The introduction of advanced imaging tools and automated instrumentation is the main focus of my laboratory, which will enable us to apply imaging for both hypothesis-driven research and high-throughput analysis. Live imaging offers the unique advantage of observing biological processes with high spatiotemporal resolution in whole organisms, offering a path to more refined, quantitative dynamic models. However, rough static models are relatively coarse, error prone, and provide limited information of biological dynamics during development and disease. These approaches give significant insight into the components and interactions that comprise biological networks on an unprecedented scale in several organisms. Using genome-wide, high-throughput -omics analyses, structures of biological circuits are increasingly being uncovered. The fields of systems biology and recently precision medicine were made possible by genomics as well as complementary high-throughput approaches such as microarrays and proteomics.
The PANTAZIS LAB captures the chemical and mechanical dynamics of development and disease by using cutting-edge approaches to live imaging.
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