A Microfluidics Approach for the Functional Investigation of Signaling Oscillations Governing Somitogenesis
| العنوان: | A Microfluidics Approach for the Functional Investigation of Signaling Oscillations Governing Somitogenesis |
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| المؤلفون: | Wilke H. M. Meijer, Katharina F. Sonnen, Marek J. van Oostrom |
| المساهمون: | Hubrecht Institute for Developmental Biology and Stem Cell Research |
| المصدر: | Journal of Visualized Experiments. MYJoVE Corporation ISSUE=169;ISSN=1940-087X;TITLE=Journal of Visualized Experiments Journal of Visualized Experiments |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Somites/metabolism, General Chemical Engineering, Microfluidics, General Biochemistry, Genetics and Molecular Biology, In vitro model, 03 medical and health sciences, Mice, 0302 clinical medicine, Biological Clocks, Somitogenesis, Paraxial mesoderm, Animals, Microfluidics/methods, 030304 developmental biology, Physics, 0303 health sciences, General Immunology and Microbiology, Oscillation, General Neuroscience, Somites, Biological Clocks/physiology, Signal transduction, Neuroscience, 030217 neurology & neurosurgery, Function (biology), Morphogen |
| الوصف: | Periodic segmentation of the presomitic mesoderm of a developing mouse embryo is controlled by a network of signaling pathways. Signaling oscillations and gradients are thought to control the timing and spacing of segment formation, respectively. While the involved signaling pathways have been studied extensively over the last decades, direct evidence for the function of signaling oscillations in controlling somitogenesis has been lacking. To enable the functional investigation of signaling dynamics, microfluidics is a previously established tool for the subtle modulation of these dynamics. With this microfluidics-based entrainment approach endogenous signaling oscillations are synchronized by pulses of pathway modulators. This enables modulation of, for instance, the oscillation period or the phase-relationship between two oscillating pathways. Furthermore, spatial gradients of pathway modulators can be established along the tissue to study how specific changes in the signaling gradients affect somitogenesis. The present protocol is meant to help establish microfluidic approaches for the first-time users of microfluidics. The basic principles and equipment needed to set up a microfluidic system are described, and a chip design is provided, with which a mold for chip generation can conveniently be prepared using a 3D printer. Finally, how to culture primary mouse tissue on a microfluidic chip and how to entrain signaling oscillations to external pulses of pathway modulators are discussed. This microfluidic system can also be adapted to harbor other in vivo and in vitro model systems such as gastruloids and organoids for functional investigation of signaling dynamics and morphogen gradients in other contexts. |
| اللغة: | English |
| تدمد: | 1940-087X |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::05a1885fd4183dd9a7594b7ec2e9cf1e https://doi.org/10.3791/62318 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....05a1885fd4183dd9a7594b7ec2e9cf1e |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 1940087X |
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