Publications
6 results found
Yaganoglu S, Kalyviotis K, Vagena-Pantoula C, et al., 2023, Author Correction: Highly specific and non-invasive imaging of Piezo1-dependent activity across scales using GenEPi, Nature Communications, Vol: 14, ISSN: 2041-1723
Pantazis P, Pantazis P, Yaganoglu S, et al., 2023, Highly specific and non-invasive imaging of Piezo1-dependent activity across scales using GenEPi, Nature Communications, Vol: 14, Pages: 1-16, ISSN: 2041-1723
Mechanosensing is a ubiquitous process to translate external mechanical stimuliinto biological responses. Piezo1 ion channels are directly gated by mechanical forces and playan essential role in cellular mechanotransduction. However, readouts of Piezo1 activity aremainly examined by invasive or indirect techniques, such as electrophysiological analyses andcytosolic calcium imaging. Here, we introduce GenEPi, a genetically-encoded fluorescentreporter for non-invasive optical monitoring of Piezo1-dependent activity. We demonstrate thatGenEPi has high spatiotemporal resolution for Piezo1-dependent stimuli from the single-celllevel to that of the entire organism. GenEPi reveals transient, local mechanical stimuli in theplasma membrane of single cells, resolves repetitive contraction-triggered stimulation ofbeating cardiomyocytes within microtissues, and allows for robust and reliable monitoring of Piezo1-dependent activity in vivo. GenEPi will enable non-invasive optical monitoring ofPiezo1 activity in mechanochemical feedback loops during development, homeostaticregulation, and disease.
Steib E, Vagena-Pantoula C, Vermot J, 2023, TissUExM protocol for ultrastructure expansion microscopy of zebrafish larvae and mouse embryos, STAR Protocols, Vol: 4, ISSN: 2666-1667
Expansion microscopy of millimeter-large mechanically heterogeneous tissues, such as whole vertebrate embryos, has been limited, particularly when combined with post-expansion immunofluorescence. Here, we present a protocol to perform ultrastructure expansion microscopy of whole vertebrate embryos, optimized to perform post-expansion labeling. We describe steps for embedding and denaturing zebrafish larvae or mouse embryos. We then detail procedures for hydrogel handling and mounting. This protocol is particularly well suited for super-resolution imaging of macromolecular protein complexes in situ but does not preserve lipids. For complete details on the use and execution of this protocol, please refer to Steib et al.1.
Vignes H, Vagena-Pantoula C, Vermot J, 2022, Mechanical control of tissue shape: Cell-extrinsic and -intrinsic mechanisms join forces to regulate morphogenesis, SEMINARS IN CELL & DEVELOPMENTAL BIOLOGY, Vol: 130, Pages: 45-55, ISSN: 1084-9521
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- Citations: 4
Vignes H, Vagena-Pantoula C, Prakash M, et al., 2022, Extracellular mechanical forces drive endocardial cell volume decrease during zebrafish cardiac valve morphogenesis, DEVELOPMENTAL CELL, Vol: 57, Pages: 598-+, ISSN: 1534-5807
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- Citations: 12
Garefalaki V, Kontomina E, Ioannidis C, et al., 2019, The actinobacterium Tsukamurella paurometabola has a functionally divergent arylamine N-acetyltransferase (NAT) homolog, World Journal of Microbiology and Biotechnology, Vol: 35, Pages: 1-18, ISSN: 0959-3993
Actinobacteria in the Tsukamurella genus are aerobic, high-GC, Gram-positive mycolata, considered as opportunistic pathogens and isolated from various environmental sources, including sites contaminated with oil, urban or industrial waste and pesticides. Although studies look into xenobiotic biotransformation by Tsukamurella isolates, the relevant enzymes remain uncharacterized. We investigated the arylamine N-acetyltransferase (NAT) enzyme family, known for its role in the xenobiotic metabolism of prokaryotes and eukaryotes. Xenobiotic sensitivity of Tsukamurella paurometabola type strain DSM 20162T was assessed, followed by cloning, recombinant expression and functional characterization of its single NAT homolog (TSUPD)NAT1. The bacterium appeared quite robust against chloroanilines, but more sensitive to 4-anisidine and 2-aminophenol. However, metabolic activity was not evident towards those compounds, presumably due to mechanisms protecting cells from xenobiotic entry. Of the pharmaceutical arylhydrazines tested, hydralazine was toxic, but the bacterium was less sensitive to isoniazid, a drug targeting mycolic acid biosynthesis in mycobacteria. Although (TSUPD)NAT1 protein has an atypical Cys-His-Glu (instead of the expected Cys-His-Asp) catalytic triad, it is enzymatically active, suggesting that this deviation is likely due to evolutionary adaptation potentially serving a different function. The protein was indeed found to use malonyl-CoA, instead of the archetypal acetyl-CoA, as its preferred donor substrate. Malonyl-CoA is important for microbial biosynthesis of fatty acids (including mycolic acids) and polyketide chains, and the corresponding enzymatic systems have common evolutionary histories, also linked to xenobiotic metabolism. This study adds to accummulating evidence suggesting broad phylogenetic and functional divergence of microbial NAT enzymes that goes beyond xenobiotic metabolism and merits investigation.
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