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Biology subjects

Brillard, C.

Publications and source records attributed to Brillard, C..

3 recordsLinked to original sources

Embryo-eggshell interaction counteracts chiral bias in early Drosophila morphogenesis

Morphogenetic processes during animal development are remarkably invariant (Duboule, 1994; Hall, 1997; Kalinka et al., 2010; Raff, 1996). This stability is established by the interaction between genetic determination of developmental progression and the constraints imposed by the surrounding embryonic environment (Busby and Steventon, 2021; Gilmour et al., 2017; Gorfinkiel and Martinez Arias, 2021). We discovered that the germ band extension process in Drosophila is rather variable: instead of extending straight towards the head, the germ band tends to twist to the side. Through a combination of experiments and theory, we demonstrated that Scab integrin-mediated attachment to the vitelline envelope stabilizes the germ band and supports its straight extension. Our quantification of germ band extension dynamics also revealed a consistent handedness to the twist of the germ band. We showed that this left-right asymmetry can be altered by manipulating the expression of Myo1D, the molecular determinant of chirality in Drosophila (Lebreton et al., 2018). Our data thus suggest that Myo1D expression causes the early gastrulating blastoderm epithelium to already exhibit inherent chirality and that the resulting destabilization of germ band extension is suppressed by Scab-mediated friction between the blastoderm and the vitelline envelope.

developmental biology↗

Time-resolved scanning ion conductance microscopy for three-dimensional tracking of nanoscale cell surface dynamics

Nanocharacterization plays a vital role in understanding the complex nanoscale organization of cells and organelles. Understanding cellular function requires high-resolution information about how the cellular structures evolve over time. A number of techniques exist to resolve static nanoscale structure of cells in great detail (super-resolution optical microscopy1, EM2, AFM3). However, time-resolved imaging techniques tend to either have lower resolution, are limited to small areas, or cause damage to the cells thereby preventing long-term time-lapse studies. Scanning probe microscopy methods such as atomic force microscopy (AFM) combine high-resolution imaging with the ability to image living cells in physiological conditions. The mechanical contact between the tip and the sample, however, deforms the cell surface, disturbs the native state, and prohibits long-term time-lapse imaging. Here, we develop a scanning ion conductance microscope (SICM) for high-speed and long-term nanoscale imaging. By utilizing recent advances in nanopositioning4, nanopore fabrication5, microelectronics6, and controls engineering7 we developed a microscopy method that can resolve spatiotemporally diverse three-dimenional processes on the cell membrane at sub-5nm axial resolution. We tracked dynamic changes in live cell morphology with nanometer details and temporal ranges of sub-second to days, imagining diverse processes ranging from endocytosis, micropinocytosis, and mitosis, to bacterial infection and cell differentiation in cancer cells. This technique enables a detailed look at membrane events and may offer insights into cell-cell interactions for infection, immunology, and cancer research.

biophysics↗

Surface-catalyzed SAS-6 self-assembly directs centriole formation through kinetic and structural mechanisms

Discovering the physical principles directing organelle assembly is a fundamental pursuit in biology. Centrioles are evolutionarily conserved organelles with a 9-fold rotational symmetry of chiral microtubules imparted onto the cilia they template1. Centriole assemble from likewise symmetrical ring polymers of SAS-6 proteins, orthogonal to a toroidal surface surrounding the resident centriole2-4. How surface properties ensure ring assembly with proper symmetry and orthogonal arrangement is not known. Here, we deployed photothermally-actuated off-resonance tapping high-speed atomic force microscopy (PORT-HS-AFM) to decipher physical principles of surface-guided SAS-6 self-assembly. Using machine learning to quantify the polymerization reaction and developing a coagulation-fragmentation model, we discovered that the surface shifts the reaction equilibrium by [~]104 compared to the solution situation, explaining orthogonal organelle emergence. Moreover, molecular dynamics and PORT-HS-AFM revealed that the surface converts helical SAS-6 polymers into 9-fold ring polymers with residual asymmetry, which may impart chiral features to centrioles and cilia. Overall, we discovered two fundamental physical principles directing robust centriole organelle assembly.

biophysics↗