| Title: |
Three‐dimensional spatial quantitative analysis of cardiac lymphatics in the mouse heart |
| Authors: |
Phillips, Evan H.; Bindokas, Vytautas P.; Jung, Dahee; Teamer, Jay; Kitajewski, Jan K.; Solaro, R. John; Wolska, Beata M.; Lee, Steve Seung‐Young |
| Contributors: |
National Heart, Lung, and Blood Institute; National Institute of Biomedical Imaging and Bioengineering; National Institute of General Medical Sciences |
| Source: |
Microcirculation ; volume 30, issue 7 ; ISSN 1073-9688 1549-8719 |
| Publisher Information: |
Wiley |
| Publication Year: |
2023 |
| Collection: |
Wiley Online Library (Open Access Articles via Crossref) |
| Description: |
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. |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| DOI: |
10.1111/micc.12826 |
| Availability: |
https://doi.org/10.1111/micc.12826; https://onlinelibrary.wiley.com/doi/pdf/10.1111/micc.12826 |
| Rights: |
http://creativecommons.org/licenses/by/4.0/ |
| Accession Number: |
edsbas.BB20CC09 |
| Database: |
BASE |