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1.
EMBO J ; 42(17): e113280, 2023 09 04.
Artigo em Inglês | MEDLINE | ID: mdl-37522872

RESUMO

Embryo implantation into the uterus marks a key transition in mammalian development. In mice, implantation is mediated by the trophoblast and is accompanied by a morphological transition from the blastocyst to the egg cylinder. However, the roles of trophoblast-uterine interactions in embryo morphogenesis during implantation are poorly understood due to inaccessibility in utero and the remaining challenges to recapitulate it ex vivo from the blastocyst. Here, we engineer a uterus-like microenvironment to recapitulate peri-implantation development of the whole mouse embryo ex vivo and reveal essential roles of the physical embryo-uterine interaction. We demonstrate that adhesion between the trophoblast and the uterine matrix is required for in utero-like transition of the blastocyst to the egg cylinder. Modeling the implanting embryo as a wetting droplet links embryo shape dynamics to the underlying changes in trophoblast adhesion and suggests that the adhesion-mediated tension release facilitates egg cylinder formation. Light-sheet live imaging and the experimental control of the engineered uterine geometry and trophoblast velocity uncovers the coordination between trophoblast motility and embryo growth, where the trophoblast delineates space for embryo morphogenesis.


Assuntos
Blastocisto , Implantação do Embrião , Feminino , Camundongos , Animais , Trofoblastos , Útero , Desenvolvimento Embrionário , Mamíferos
2.
Sci Rep ; 10(1): 1942, 2020 02 06.
Artigo em Inglês | MEDLINE | ID: mdl-32029815

RESUMO

Three-dimensional live imaging has become an indispensable technique in the fields of cell, developmental and neural biology. Precise spatio-temporal manipulation of biological entities is often required for a deeper functional understanding of the underlying biological process. Here we present a home-built integrated framework and optical design that combines three-dimensional light-sheet imaging over time with precise spatio-temporal optical manipulations induced by short infrared laser pulses. We demonstrate their potential for sub-cellular ablation of neurons and nuclei, tissue cauterization and optogenetics by using the Drosophila melanogaster and zebrafish model systems.


Assuntos
Microscopia , Animais , Drosophila melanogaster/fisiologia , Imageamento Tridimensional/métodos , Raios Infravermelhos , Lasers , Peixe-Zebra/fisiologia
3.
Elife ; 72018 10 30.
Artigo em Inglês | MEDLINE | ID: mdl-30375972

RESUMO

Extraembryonic tissues contribute to animal development, which often entails spreading over embryo or yolk. Apart from changes in cell shape, the requirements for this tissue spreading are not well understood. Here, we analyze spreading of the extraembryonic serosa in the scuttle fly Megaselia abdita. The serosa forms from a columnar blastoderm anlage, becomes a squamous epithelium, and eventually spreads over the embryo proper. We describe the dynamics of this process in long-term, whole-embryo time-lapse recordings, demonstrating that free serosa spreading is preceded by a prolonged pause in tissue expansion. Closer examination of this pause reveals mechanical coupling to the underlying yolk sac, which is later released. We find mechanical coupling prolonged and serosa spreading impaired after knockdown of M. abdita Matrix metalloprotease 1. We conclude that tissue-tissue interactions provide a critical functional element to constrain spreading epithelia.


Assuntos
Dípteros/embriologia , Embrião não Mamífero/metabolismo , Membranas Extraembrionárias/metabolismo , Saco Vitelino/embriologia , Âmnio/citologia , Âmnio/embriologia , Animais , Blastoderma/citologia , Forma Celular , Dípteros/genética , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Membrana Serosa/citologia , Membrana Serosa/embriologia , Imagem com Lapso de Tempo
4.
J Biophotonics ; 6(8): 645-55, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23420601

RESUMO

Understanding the cellular response to DNA strand breaks is crucial to decipher the mechanisms maintaining the integrity of our genome. We present a novel method to visualize how the mobility of nuclear proteins changes in response to localized DNA damage. DNA strand breaks are induced via nonlinear excitation with femtosecond laser pulses at λ = 1050 nm in a 3D-confined subnuclear volume. After a time delay of choice, protein mobility within this volume is analysed by two-photon photoactivation of PA-GFP fusion proteins at λ = 775 nm. By changing the position of the photoactivation spot with respect to the zone of lesion the influence of chromatin structure and of the distance from damage are investigated. As first applications we demonstrate a locally confined, time-dependent mobility increase of histone H1.2, and a progressive retardation of the DNA repair factor XRCC1 at damaged sites. This assay can be used to map the response of nuclear proteins to DNA damage in time and space.


Assuntos
Dano ao DNA , Raios Infravermelhos , Lasers , Imagem Molecular , Dinâmica não Linear , Proteínas Nucleares/metabolismo , Cromatina/metabolismo , Cromatina/efeitos da radiação , Proteínas de Ligação a DNA/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Células HeLa , Histonas/metabolismo , Humanos , Microscopia de Fluorescência por Excitação Multifotônica , Fótons , Transporte Proteico/efeitos da radiação , Proteína 1 Complementadora Cruzada de Reparo de Raio-X
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