Biology subjects

Quesson, B.

Publications and source records attributed to Quesson, B..

3 recordsLinked to original sources

bioRxiv2026

Structural Characterization of Cardiac Purkinje Fibers Using Inhomogeneous Magnetization Transfer (ihMT): A proof of Concept MRI-Histology Approach

BACKGROUNDThe cardiac Purkinje network plays a critical role in maintaining synchronized activation of ventricles but remains challenging to image due to its fine and unique structure. Conventional MRI techniques lack sufficient contrast to distinguish the underlying structural composition of Purkinje Fibers (PF). PURPOSEThis study investigates the potential of inhomogeneous Magnetization Transfer (ihMT) as a novel contrast mechanism for visualizing and differentiating subregions of the PF. METHODSFive fixed ex-vivo sheep hearts (n = 5) containing free running PF were scanned with a 9.4T MRI using a 2D ihMT RARE sequence. ASSESSMENTihMTR maps were analyzed using manually defined regions-of-interest (ROIs) corresponding to free-running PF, insertion points, and myocardium. Histological analysis (light and polarized microscopy) was performed on matched sections to quantify collagen types I and III, adipocytes, Purkinje cells, and cardiomyocytes. RESULTSThree ihMT protocols, which produced high ihMTR values in free-running PF (9.25-10.83%) and strong absolute contrast relative to the myocardium (2.00-2.17%) and insertion points (2.99-3.40%) in one sample were selected and applied to all samples. Across all samples, mean ihMTR in free-running was consistently higher than in insertion points (11.5 {+/-} 1.5% vs. 9.0 {+/-} 2.9%). Histological analysis revealed a significantly greater collagen content in free-running regions compared with insertion points (72.4 {+/-} 15.9% vs. 31.1 {+/-} 13.1%; p = 0.001), along with higher adipocyte content at insertion points vs. free-running regions (12.3 {+/-} 6.1% vs. 3.8 {+/-} 2.7%, non-significant). Collagen type III was more prominent at insertion points but remained a minor component overall. CONCLUSIONihMT imaging can distinguish PF subregions based on microstructural differences, particularly collagen and adipocyte distribution. This study lays the groundwork for developing biophysical models to interpret ihMT signals in terms of tissue composition and microstructure, providing a foundation for future studies. SponsorThis study received financial support from the French Government by the National Research Agency (ANR; SYNATRA ANR-21-CE19-0014-01) and Region Nouvelle Aquitaine (convention N{degrees}AAPR2022-2021-16609210).

biophysics↗

bioRxiv2023

Increased spontaneous Ca2+ activity in Cardiac Purkinje cells after myocardial infarction; A consequence of a dramatic shift of SERCA isoforms as potential adaptation to acute ischemia?

BackgroundStudies of Purkinje cells (Pcells) from canine hearts have suggested an increase of Ca2+-release by the sarcoplasmic reticulum (SR) but also reported a potential augmentation of SR-Ca2+-uptake after MI. Abnormal increase of SR-Ca2+-uptake in heart cells is novel and contrasts with the reduction of this function in cells of failing heart. Our study examined the origin of this increased SR-Ca2+-uptake by considering a change in SR-Ca2+ pump (SERCA2) expression in Purkinje fibers (PFs) post MI. MethodsPcells were isolated from canine hearts 48Hrs post MI. Intracellular Ca2+-activity was captured by confocal microscopy. Purkinje-typical Ca2+ events were analyzed to probe the regional Ca2+-dynamics within Pcells. A Purkinje-specific numerical model assisted in the interpretation of Ca2+-anomalies detected in Pcells Ca2+-transients. SR-Ca2+-uptake system was studied by immunofluorescence in Pcells from canine, ovine and human hearts post MI. SERCA protein and gene expressions in PFs and myocardium were measured by Western Blots and RT-qPCR in a classical porcine model of MI. Results48Hrs after MI, Pcells showed 60% increase in spark-rate and 37% acceleration of Ca2+ wave decay. In the model of normal wave, 35% increase of Ca2+-uptake rate reproduced the actual post-MI wave alterations. In apparent contrast with increased Ca2+-uptake rate, SERCA2 protein expression was reduced in canine, sheep, and human Pcells after MI. In pig MI model, the protein level of cardiac-specific SERCA2-splicing variant SERCA2a was reduced by 52% in the whole infarcted ventricle whereas the "non-cardiac" SERCA2b level was increased by 120%. In the infarcted regions, PFs showed 30% downregulation of SERCA2a gene expression and 630% upregulation of SERCA2b. ConclusionOur results confirm that elevated spontaneous Ca2+-activity in post-MI PFs is due to increased SR-Ca2+-uptake within Pcells. Data suggest that a replacement of "cardiac" SERCA2a by the "non-cardiac" SERCA2b sub-isoform in cardiac cells in response to ischemia is implicated in this alteration.

cell biology↗

bioRxiv2021

Optimizing large organ scale micro computed tomography imaging in pig and human hearts using a novel air-drying technique

Underlying electrical propagation in the heart and potentially fatal arrhythmia is the cardiac microstructure. Despite the critical role of muscle architecture, a non-destructive approach to examine not only myocyte orientation, but cellular arrangement in to laminar organization is lacking in hearts from translational animal models and humans. X-ray micro computed tomography using contrast enhancing agents achieves three-dimensional images at near-histological resolutions. However, imaging large mammalian hearts presents challenges including X-ray over-attenuation and loss of image contrast. The goal of this study was to rethink tissue pre-treatment to optimize, and benefit from micro computed tomography imaging resolution in large tissues. Whole pig and human hearts were dehydrated and perfused with a tissue reinforcing agent, hexamethyldisilazane, and slowly air-dried. Heart morphology was conserved and temporally stable. This enabled direct air-mounting for micro computed tomography imaging. Moreover, the desiccated tissue density was significantly reduced compared to the initial hydrated state (P=0.04). Three-dimensional image reconstructions of air-dried hearts segmented using a single intensity threshold revealed detailed microstructural architecture of myolaminae. Conversely, one-step segmentation of hearts loaded with contrast agents poorly estimated the gross anatomical morphology of the heart and lacked identification of tissue microarchitecture. Air-drying large mammalian hearts optimizes X-ray imaging of cardiac microstructure.

physiology↗