9 research outputs found
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The role of hydrodynamic forces in synchronisation and alignment of mammalian motile cilia
Fluid flow generated by a ciliated epithelium is a fascinating evidence of collective behaviour in nature. In many organs and eukaryotic organisms, thousands of microscale whip-like structures called `motile cilia' beat aligned at the same frequency and in a coordinated fashion. This dynamics, known as `metachronal wave', has fundamental physiological roles in microorganisms and many organs of vertebrates. In the airways, the coordinated beatings of motile cilia generate a fluid flow that pushes mucus to the pharynx, and so protects the lungs from inhaled contaminants. The failure of this collective dynamics can precipitate or exacerbate severe infections and chronic inflammatory conditions such as cystic fibrosis (CF), primary ciliary dyskinesia (PCD) or asthma. In the brain, the multiciliated ependymal cells cover all the ventricles. Their cilia beat in a coordinated fashion to ensure the cerebrospinal fluid circulation necessary for brain homoeostasis, toxin washout and orientation of the migration of newborn neurons. Despite the fundamental role in nature, the mechanism underpinning such collective behaviour is still unknown.
A recent hypothesis, supported by simulations, experiments with microorganisms and with cilia models, proposed that hydrodynamic interactions between cilia could provide a physical mechanism for their coordination. In contrast, others have proposed a role of the cytoskeletal elastic coupling between cilia. While previous works mainly focused on algae and protists, investigating the conditions that are required for the emergence of the metachronal wave in mammalian tissues can provide important progress in the diagnosis and treatment of human medical diseases. Specifically, I tackled this broad topic by studying the hydrodynamic forces necessary for the synchronisation and alignment of motile cilia from brain and airways. This question was addressed experimentally by measuring cilia motility during treatment with oscillatory and constant external fluid flows. We found that synchronisation and alignment of mammalian cilia in the brain is achieved with flows of similar magnitude of the ones generated by cilia themselves. Our results suggest that hydrodynamic forces between cilia are sufficient for the emergence of their collective behaviour.
The first chapter provides basic knowledge on motile cilia structure and functions in microorganisms and humans. Additionally, I introduce the reader to the open questions related to the coordination of a pair and a carpet of cilia, with specific attention on previous works on mammals. This first chapter is followed by a description of a novel microfluidic device that I developed to grow airway and brain cells and apply controlled viscous forces.
In Chapter 3, I describe how we have investigated cilia synchronisation of mammalian cilia. Applying external oscillatory flow on brain cells, we studied the susceptibility of cilia motility to hydrodynamic forces similar to the ones generated by cilia themselves. We found that cells with few cilia (up to five) can be entrained at flows comparable to the cilia-driven flows reported in vivo. We suggest that hydrodynamic forces between mammalian cilia are sufficiently strong to be the mechanism underpinning frequency synchronisation.
In the second part of my thesis, I looked into the hydrodynamic shear forces needed to align permanently the cilia direction of beating. We tackled this problem by using cultures of mouse brain and human airway cells grown in custom flow channels.
We found that cilia from mouse brain do not lock their beating direction after \emph{ciliogenesis}, but can respond and align to physiological shear stress found \emph{in vivo} at any time, in contrast with was previously believed. Moreover, we suggest that cilia alignment depends on the density of cilia, in agreement with a hydrodynamic screening effect of the external flow by the nearby cilia that we aim to investigate in the future. These results are described in Chapter 4. Successively in Chapter 5, I report our approach to study whether physiological shear stress can induce cilia alignment in airway cell cultures. The current hypothesis is that these cilia may also be able to align with external hydrodynamic forces - however, experimental evidence is still needed. There is a lack of experiments on this topic mainly because airway cells are cultured in an air-liquid interface, and so shear stress has to be applied with airflows. We developed novel setups for applying long term shear stress with air and fluid flow on this system, leaving further experiments for the future.EU Horizon 2020 research and innovation program under Marie Sklodowska-Curie 641639 ITN BioPol and ERC CoG HydroSyn
Generalized energy equipartition in harmonic oscillators driven by active baths
We study experimentally and numerically the dynamics of colloidal beads
confined by a harmonic potential in a bath of swimming E. coli bacteria. The
resulting dynamics is well approximated by a Langevin equation for an
overdamped oscillator driven by the combination of a white thermal noise and an
exponentially correlated active noise. This scenario leads to a simple
generalization of the equipartition theorem resulting in the coexistence of two
different effective temperatures that govern dynamics along the flat and the
curved directions in the potential landscape.Comment: 4 pages, 3 figure
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Entrainment of mammalian motile cilia in the brain with hydrodynamic forces.
Motile cilia are widespread across the animal and plant kingdoms, displaying complex collective dynamics central to their physiology. Their coordination mechanism is not generally understood, with previous work mainly focusing on algae and protists. We study here the entrainment of cilia beat in multiciliated cells from brain ventricles. The response to controlled oscillatory external flows shows that flows at a similar frequency to the actively beating cilia can entrain cilia oscillations. We find that the hydrodynamic forces required for this entrainment strongly depend on the number of cilia per cell. Cells with few cilia (up to five) can be entrained at flows comparable to cilia-driven flows, in contrast with what was recently observed in Chlamydomonas Experimental trends are quantitatively described by a model that accounts for hydrodynamic screening of packed cilia and the chemomechanical energy efficiency of the flagellar beat. Simulations of a minimal model of cilia interacting hydrodynamically show the same trends observed in cilia
Light Controlled Biohybrid Microbots
Biohybrid microbots integrate biological actuators and sensors into synthetic chassis with the aim of providing the building blocks of next-generation micro-robotics. One of the main challenges is the development of self-assembled systems with consistent behavior and such that they can be controlled independently to perform complex tasks. Herein, it is shown that, using light-driven bacteria as propellers, 3D printed microbots can be steered by unbalancing light intensity over different microbot parts. An optimal feedback loop is designed in which a central computer projects onto each microbot a tailor-made light pattern, calculated from its position and orientation. In this way, multiple microbots can be independently guided through a series of spatially distributed checkpoints. By exploiting a natural light-driven proton pump, these bio-hybrid microbots are able to extract mechanical energy from light with such high efficiency that, in principle, hundreds of these systems can be controlled simultaneously with a total optical power of just a few milliwatts. © 2023 The Authors. Advanced Functional Materials published by Wiley-VCH GmbH
Cilia density and flow velocity affect alignment of motile cilia from brain cells
In many organs, thousands of microscopic ‘motile cilia’ beat in a coordinated fashion generating fluid flow. Physiologically, these flows are important in both development and homeostasis of ciliated tissues. Combining experiments and simulations, we studied how cilia from brain tissue align their beating direction. We subjected cilia to a broad range of shear stresses, similar to the fluid flow that cilia themselves generate, in a microfluidic setup. In contrast to previous studies, we found that cilia from mouse ependyma respond and align to these physiological shear stress at all maturation stages. Cilia align more easily earlier in maturation, and we correlated this property with the increase in multiciliated cell density during maturation. Our numerical simulations show that cilia in densely packed clusters are hydrodynamically screened from the external flow, in agreement with our experimental observation. Cilia carpets create a hydrodynamic screening that reduces the susceptibility of individual cilia to external flows