Three‐dimensional spatial quantitative analysis of cardiac lymphatics in the mouse heart
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Published:2023-08-22
Issue:7
Volume:30
Page:
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ISSN:1073-9688
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Container-title:Microcirculation
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language:en
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Short-container-title:Microcirculation
Author:
Phillips Evan H.12ORCID,
Bindokas Vytautas P.3,
Jung Dahee1,
Teamer Jay1,
Kitajewski Jan K.2,
Solaro R. John2,
Wolska Beata M.24,
Lee Steve Seung‐Young1ORCID
Affiliation:
1. Department of Pharmaceutical Sciences University of Illinois Chicago Chicago Illinois USA
2. Department of Physiology and Biophysics University of Illinois Chicago Chicago Illinois USA
3. Integrated Light Microscopy Facility The University of Chicago Chicago Illinois USA
4. Department of Medicine, Division of Cardiology, Center for Cardiovascular Research University of Illinois Chicago Chicago Illinois USA
Abstract
AbstractObjectiveThree‐dimensional (3D) microscopy and image data analysis are necessary for studying the morphology of cardiac lymphatic vessels (LyVs) and their association with other cell types. We aimed to develop a methodology for 3D multiplexed lightsheet microscopy and highly sensitive and quantitative image analysis to identify pathological remodeling in the 3D morphology of LyVs in young adult mouse hearts with familial hypertrophic cardiomyopathy (HCM).MethodsWe developed a 3D lightsheet microscopy workflow providing a quick turn‐around (as few as 5–6 days), multiplex fluorescence detection, and preservation of LyV structure and epitope markers. Hearts from non‐transgenic and transgenic (TG) HCM mice were arrested in diastole, retrograde perfused, immunolabeled, optically cleared, and imaged. We built an image‐processing pipeline to quantify LyV morphological parameters at the chamber and branch levels.ResultsChamber‐specific pathological alterations of LyVs were identified, and significant changes were seen in the right atrium (RA). TG hearts had a higher volume percent of ER‐TR7+ fibroblasts and reticular fibers. In the RA, we found associations between ER‐TR7+ volume percent and both LyV segment density and median diameter.ConclusionsThis workflow and study enabled multi‐scale analysis of pathological changes in cardiac LyVs of young adult mice, inviting ideas for research on LyVs in cardiac disease.
Funder
National Heart, Lung, and Blood Institute
National Institute of Biomedical Imaging and Bioengineering
National Institute of General Medical Sciences
Subject
Physiology (medical),Cardiology and Cardiovascular Medicine,Molecular Biology,Physiology
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