Biology subjects

Michels, L.

Publications and source records attributed to Michels, L..

5 recordsLinked to original sources

bioRxiv2024

CarboTag: a modular approach for live and functional imaging of plant cell walls

Plant cells are contained inside a rigid network of cell walls. Cell walls are highly dynamic structures that act both as a structural material and as a hub for a wide range of signaling processes. Despite its crucial role in all aspects of the plant life cycle, live dynamical imaging of the cell wall and its functional properties has remained challenging. Here, we introduce CarboTag, a modular toolbox for live functional imaging of plant walls. CarboTag relies on a small molecular motif, a pyridine boronic acid, that targets its cargo to the cell wall, is non-toxic and ensures rapid tissue permeation. We designed a suite of cell wall imaging probes based on CarboTag in any desired color for multiplexing. Moreover, we created new functional reporters for live quantitative imaging of key cell wall features: network porosity, cell wall pH and the presence of reactive oxygen species. CarboTag opens the way to dynamical and quantitative mapping of cell wall responses at subcellular resolution.

plant biology↗

bioRxiv2023

Regional assessment of male murine bone exposes spatial heterogeneity in osteocyte lacunar volume associated with intracortical canals and regulation by VEGF

The porous bone cortex comprises an interconnected network of intracortical vascular canals and osteocyte lacunae, embedded within the bone mineral. Increases in cortical porosity reduce bone strength and increase fracture risk. To date, our understanding of mechanisms coupling the arrangements of the vascular: lacunar network in the bone cortex is poorly understood yet it could be key in establishing regulation of cortical porosity evident with age. Using synchrotron radiation-based computed tomography we develop automated tools to characterise the 3D spatial organisation and morphology of osteocyte lacunae, and the bone vasculature at the tibiofibular junction (TFJ), defining posterior, medial, lateral, and anterior regions in male C57BL/6 mice (n = 3). We also investigate the role of osteoblast-derived VEGF in regulating the 3D spatial arrangement by conditional disruption of VEGF in osteocalcin-expressing cells (OcnVEGFKO versus WT, n = 3). Regional lacunar phenotypes were assessed by 3D distance mapping of lacunar organisation surrounding the vascular compartments, including endosteal and periosteal surfaces, or intracortical canals. Surface-associated lacunae were indistinct in size across posterior, medial, lateral and anterior regions. However, lacunae associated with intracortical canals were significantly larger exclusively within the posterior region. In the absence of VEGF, the increased lacunar volume associated with posterior intracortical canals was lost. Our results suggest that the influence of intracortical canals on lacunar volumes is spatially regulated and sensitive to locally produced growth factors such as osteoblast-derived VEGF.

bioengineering↗

bioRxiv2022

Direct measurement of appressorium turgor using a molecular mechanosensor in the rice blast fungus Magnaporthe oryzae

Many plant pathogenic fungi forcibly enter their hosts to cause disease. The rice blast fungus Magnaporthe oryzae, for example, infects plants using a specialised infection cell called an appressorium, which generates enormous turgor to drive a rigid penetration peg through the rice leaf cuticle. While these vast internal pressures are a critical weapon in fungal host penetration, they have remained very challenging to probe directly during host invasion, leaving our understanding of these extreme cellular mechanics incomplete. Here, we combine Fluorescence Lifetime Imaging (FLIM) with a membrane-targeting molecular mechanoprobe to quantify changes in membrane tension as a direct proxy for appressorial turgor in M. oryzae. We report that mature melanin-pigmented M. oryzae appressoria display a heterogeneous low fluorescence lifetime and high membrane tension, consistent with enormous turgor. These extreme pressures lead to large-scale spatial heterogeneities in membrane mechanics, much greater than observed in any other cell type previously, highlighting the extreme mechanics of turgor-driven appressorium-mediated plant infection. By contrast, appressoria of non-pathogenic melanin-deficient mutants, alb1 and buf1, or immature non-melanised appressoria, exhibit high fluorescence lifetime, consistent with low membrane tension and turgor, that remain spatially homogeneous. To evaluate the method, we investigated turgor dynamics in a range of mutants impaired in appressorium function. We show that the turgor sensor kinase mutant{Delta} sln1, recently proposed to generate excess appressorium turgor, displayed a significantly higher membrane tension compared to an isogenic wild type M. oryzae strain. This non-invasive, live cell imaging technique allows direct quantification and visualization of the enormous turgor pressures deployed during pathogen infection.

cell biology↗

bioRxiv2021

Long-term effects of network-based fMRI neurofeedback training for sustained attention

Neurofeedback allows for learning voluntary control over ones own brain activity, aiming to enhance cognition and clinical symptoms. A recent study improved sustained attention temporarily by training healthy participants to up-regulate the differential activity of the sustained attention network (SAN) minus the default mode network (DMN). However, long-term learning effects of functional magnetic resonance imaging (fMRI) neurofeedback training remain under-explored. Here, we evaluate the effects of network-based fMRI neurofeedback training for sustained attention by assessing behavioral and brain measures before, one day after, and two months after training. The behavioral measures include task as well as questionnaire scores, and the brain measures include activity and connectivity during self-regulation runs without feedback (i.e., transfer runs) and during resting-state runs. Neurally, we found that participants maintained their ability to control the differential activity during follow-up sessions. Further, exploratory analyses showed that the training-induced increase in FC between the DMN and occipital gyrus was maintained during follow-up transfer runs, but not during follow-up resting-state runs. Behaviorally, we found that enhanced sustained attention right after training returned to baseline level during follow-up. The discrepancy between lasting regulation-related brain changes but transient behavioral and resting-state effects raises the question of how neural changes induced by neurofeedback training translate to potential behavioral improvements. Since neurofeedback directly targets brain measures to indirectly improve behavior long-term, a better understanding of the brain-behavior associations during and after neurofeedback training is needed to develop its full potential as a promising scientific and clinical tool. Key pointsO_LIParticipants were still able to self-regulate the differential activity between large-scale networks two months after the end of neurofeedback training and this during transfer runs without feedback. C_LIO_LILasting brain changes were also observed in the functional connectivity of trained regions in runs during which participants engaged in active self-regulation as well as during resting-state runs without concomitant self-regulation. C_LIO_LIThe increased sustained attention we observed right after the end of neurofeedback training did not persist two months later. C_LI

neuroscience↗

bioRxiv2018

Shared neural mechanisms between imagined and perceived egocentric motion

Many cognitive and social processes involve mental simulations of a change in perspective. Behavioral studies suggest that such egocentric mental rotations rely on brain areas that are also involved in processing actual self-motion, thus depending on vestibular input. In a combined galvanic vestibular stimulation (GVS) and functional Magnetic Resonance Imaging (fMRI) study, we investigated the brain areas that underlie both simulated changes in self-location and the processing of vestibular stimulation within the same individuals. Participants performed an egocentric mental rotation task, an object-based mental rotation task, or a pure lateralization task during GVS or sham stimulation. At the neural level, we expected an overlap between brain areas activated during vestibular processing and egocentric mental rotation (against object-based mental rotation) within area OP2 and the Posterior Insular Cortex (PIC), two core brain regions involved in vestibular processing. The fMRI data showed a small overlap within area OP2 and a larger overlap within the PIC for both egocentric mental rotation against object-based mental rotation and vestibular processing. GVS did not influence the ability to perform egocentric mental rotation.\n\nOur results provide evidence for shared neural mechanisms underlying perceived and simulated self-motion. We conclude that mental rotation of ones body involves neural activity in the PIC and area OP2, but the behavioral results also suggest that those mental simulations of ones body might be robust to modulatory input from vestibular stimulation.

neuroscience↗