Biology subjects

Piolot, T.

Publications and source records attributed to Piolot, T..

4 recordsLinked to original sources

bioRxiv2025

Adaptation of endothelial cells to microenvironment topographical cues through lysyl oxidase like-2-mediated basement membrane scaffolding

Basement membrane (BM) provides structural support and signaling platform for blood vessels. While its major structural components are required for vascular morphogenesis, integrating BM regulators, like the lysyl oxidase LOXL2, and BM assembly in cell response to microenvironement cues remain poorly understood. Here we study the early deposition and supramolecular assembly of BM components using correlative atomic force and fluorescence microscopy. The fibrillar deposition of fibronectin is gradually remodeled and associates with the collagen IV meshwork as it organizes into BM. We demonstrate that LOXL2 is deposited with both proteins and participates in their remodeling. Alteration of BM scaffolding by LOXL2-depletion affects focal adhesion maturation and cytoskeleton remodeling. This altered BM organization maintains stress fibers, affects the distribution and activation of mechanosensors and alters cell barrier properties. Furthermore, using 3D micro-printed substrates, we demonstrate that BM assembly regulates endothelial cell response to topographical constraint. We therefore propose a mechanism directly linking the scaffolding of BM components and adaptation to the topographical signals from the microenvironment.

cell biology↗

bioRxiv2022

The filipodia-like protrusions of adjacent somatic cells shape the developmental potential of mouse oocytes

The oocyte must grow and mature before fertilization, thanks to a close dialogue with the somatic cells which surround it. Part of this communication is through filopodialike protrusions, called transzonal projections (TZPs), sent by the somatic cells to the oocyte membrane. To investigate the contribution of TZPs to oocyte quality, we impaired their structure by generating a full knockout mouse of the TZP structural component Myosin-X (MYO10). Using spinning disk and super-resolution microscopy combined with a machine learning approach to phenotype oocyte morphology, we show that the lack of Myo10 decreases TZP density during oocyte growth. Reduction in TZPs does not prevent oocyte growth but impairs oocyte-matrix integrity. Importantly, we reveal by transcriptomic analysis that gene expression is altered in TZP-deprived oocytes, and that oocyte maturation and subsequent early embryonic development are partially affected, effectively reducing mouse fertility. We propose that TZPs play a role in the structural integrity of the germline-somatic complex, which is essential for regulating gene expression in the oocyte and thus its developmental potential.

cell biology↗

bioRxiv2022

Topology of connective tissues: a key parameter in cellular heterogeneity, beyond composition and stiffness

Cellular plasticity is essential in physiological contexts, including pathological ones. It is the basis of morphogenesis and organogenesis, as well as tumorigenesis and metastasis. The extracellular matrix (ECM) is a key player in the generation of cellular heterogeneity. Advances in our understanding of cell plasticity rely on our ability to provide relevant in vitro models. This requires to catch the characteristics of the tissues that are essential for controlling cell fate. To do this, we must consider the diversity of tissues, the diversity of physiological contexts, and the constant remodeling of ECM along these processes. To this aim, we have fabricated a library of ECM models for reproducing the scaffold of connective tissues and basement membrane with different biofabrication routes based on the electrospining and drop casting of biopolymers. Using a combination of multiphoton imaging and nanoindentation, we show that we can vary independently protein composition, topology of connective tissues and stiffness of ECM models. Reproducing the features of a tissue and physiological context in turns allows to generate the complexity of the phenotypic landscape associated with the epithelial-to-mesenchymal transition (EMT) in human ovarian cancer. We show that EMT shift cannot be directly correlated with a unique ECM feature, which reflects the multidimensionality of living environments. Very importantly, our combinatorial approach allows us to provide in vitro models, where the impact of the topological cues on cellular phenotypes can be revealed, beyond protein composition and stiffness of the ECM matrix. On this line, this work is a further step towards the development of ECM models recapitulating the constantly remodeled scaffolding environment that cells face and provides new insights for the development of cell-free matrices.

bioengineering↗

bioRxiv2018

Active fluctuations modulate gene expression in mouse oocytes

In mammals, the nucleus is central in oocytes, not defining the future embryo axis. Nucleus centring depends on an F-actin mediated pressure gradient. In Fmn2-/- oocytes, lacking the F-actin nucleator Formin 2, the nucleus is off-centre and can be centred by re-expressing Formin 2. Here, we addressed the biological significance of nucleus positioning in mammalian oocytes. Using a dedicated computational 3D imaging approach, we observed nuclear architecture alterations in mouse Fmn2-/- oocytes. RNA sequencing of control versus Fmn2-/- oocytes detected 2285 mis-regulated genes. Rescue experiments showed that the process of nuclear positioning impacts nuclear architecture and gene expression. Using signal processing methods coupled to biophysical modelling allowing the extraction of in vivo mechanical properties of the nuclear envelope, we showed that F-actin-mediated activity promotes nuclear envelope shape fluctuations and chromatin motion. We thus propose a mechano-transduction model whereby nucleus positioning via microfilaments modulates oocyte transcriptome, essential for further embryo development.

cell biology↗