학술논문
Three-dimensional spatial quantitative analysis of cardiac lymphatics in the mouse heart.
Document Type
Academic Journal
Author
Phillips EH; Department of Pharmaceutical Sciences, University of Illinois Chicago, Chicago, Illinois, USA.; Department of Physiology and Biophysics, University of Illinois Chicago, Chicago, Illinois, USA.; Bindokas VP; Integrated Light Microscopy Facility, The University of Chicago, Chicago, Illinois, USA.; Jung D; Department of Pharmaceutical Sciences, University of Illinois Chicago, Chicago, Illinois, USA.; Teamer J; Department of Pharmaceutical Sciences, University of Illinois Chicago, Chicago, Illinois, USA.; Kitajewski JK; Department of Physiology and Biophysics, University of Illinois Chicago, Chicago, Illinois, USA.; Solaro RJ; Department of Physiology and Biophysics, University of Illinois Chicago, Chicago, Illinois, USA.; Wolska BM; Department of Physiology and Biophysics, University of Illinois Chicago, Chicago, Illinois, USA.; Department of Medicine, Division of Cardiology, Center for Cardiovascular Research, University of Illinois Chicago, Chicago, Illinois, USA.; Lee SS; Department of Pharmaceutical Sciences, University of Illinois Chicago, Chicago, Illinois, USA.
Source
Publisher: Wiley-Blackwell Country of Publication: United States NLM ID: 9434935 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1549-8719 (Electronic) Linking ISSN: 10739688 NLM ISO Abbreviation: Microcirculation Subsets: MEDLINE
Subject
Language
English
Abstract
Objective: Three-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).
Methods: We 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.
Results: Chamber-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.
Conclusions: This 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.
(© 2023 The Authors. Microcirculation published by John Wiley & Sons Ltd.)
Methods: We 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.
Results: Chamber-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
Conclusions: This 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.
(© 2023 The Authors. Microcirculation published by John Wiley & Sons Ltd.)