Marine Laporte, a previous research and mentor fellow in the UNIGE group and first author of the study, used expansion microscopy to examine the place of 24 proteins in the 6 domains in over a thousand centrioles at various phases of growth.Reorganizing Images To Set Them in Motion” This really laborious work was followed by a pseudo-temporal kinematic reconstruction. In other words, we were able to put these thousands of images taken at random throughout centriole biogenesis back into chronological order, to rebuild the various stages in the formation of centriole substructures, using a computer system analysis we developed,” discusses Virginie Hamel, co-leader of the study.This distinct technique, which integrates the really high resolution of expansion microscopy and kinematic reconstruction, has allowed us to design the very first 4D assembly of the human centriole. “Our work will not just deepen our understanding of centriole development, however likewise open up extraordinary potential customers in cellular and molecular biology, considering that this technique can be applied to cellular structures and other macromolecules to study their assembly in area and time,” concludes Paul Guichard.Reference: “Time-series restoration of the molecular architecture of human centriole assembly” by Marine H. Laporte, Davide Gambarotto, Éloïse Bertiaux, Lorène Bournonville, Vincent Louvel, José M. Nunes, Susanne Borgers, Virginie Hamel and Paul Guichard, 10 April 2024, Cell.DOI: 10.1016/ j.cell.2024.03.025.
Model of a human centriole cut along its longitudinal axis and viewed from above. Credit: © CentrioleLabUNIGE scientists have rebuilded for the very first time a movie of the assembly of the human centriole, one of the important structures that constitute our cells.Cells consist of numerous specialized structures– such as the nucleus, mitochondria, or peroxisomes– called “organelles.” Tracing their genesis and identifying their structure is fundamental to understanding cell function and the pathologies linked to their dysfunction. Researchers at the University of Geneva (UNIGE) have actually combined high resolution microscopy and kinematic restoration strategies to picture, in movement, the genesis of the human centriole. This organelle, vital to the organization of the cell skeleton, is associated– in case of dysfunction– with specific cancers, brain disorders or retinal diseases. This work, published in the journal Cell, elucidates the intricacies of centriole assembly. It also opens up many new avenues for the research study of other cell organelles.The Genesis of Our Cellular Skeleton, Image by ImageOrganelle genesis continues according to an exact series of succeeding protein recruitment occasions. Picturing this assembly in real-time offers a better understanding of the role of these proteins in organelle structure or function. However, acquiring a video series with adequate resolution to distinguish such complex microscopic elements faces a number of technical limitations.Inflating Cells for Better ObservationThis is particularly real of the centriole. This organelle, measuring less than 500 nanometers (half a thousandth of a millimeter), is constituted of around 100 different proteins arranged into six substructural domains. Up until a few years back, it was difficult to imagine the structure of the centriole in detail. The laboratory of Paul Guichard and Virginie Hamel, co-directors of research in the Department of Cellular and molecular Biology at the UNIGE Faculty of Science, has altered this situation by utilizing the strategy of expansion microscopy. This advanced strategy allows cells and their constituents to be progressively inflated without being warped, so that they can then be observed– using standard microscopic lens– with really high resolution.Obtaining images of the centriole with such high resolution allows the precise location of proteins at a given time but gives no information on the order of appearance of substructural domains or of specific proteins. Marine Laporte, a previous research and teaching fellow in the UNIGE group and very first author of the research study, utilized expansion microscopy to analyze the place of 24 proteins in the 6 domains in over a thousand centrioles at different phases of growth.Reorganizing Images To Set Them in Motion” This really laborious work was followed by a pseudo-temporal kinematic reconstruction. Simply put, we were able to put these thousands of images taken at random during centriole biogenesis back into sequential order, to reconstruct the numerous stages in the development of centriole bases, using a computer analysis we established,” describes Virginie Hamel, co-leader of the study.This distinct approach, which integrates the very high resolution of growth microscopy and kinematic reconstruction, has enabled us to model the first 4D assembly of the human centriole. “Our work will not just deepen our understanding of centriole development, however also open incredible prospects in molecular and cellular biology, given that this technique can be applied to other macromolecules and cellular structures to study their assembly in space and time,” concludes Paul Guichard.Reference: “Time-series reconstruction of the molecular architecture of human centriole assembly” by Marine H. Laporte, Davide Gambarotto, Éloïse Bertiaux, Lorène Bournonville, Vincent Louvel, José M. Nunes, Susanne Borgers, Virginie Hamel and Paul Guichard, 10 April 2024, Cell.DOI: 10.1016/ j.cell.2024.03.025.
