期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 24, 期 17, 页码 -出版社
MDPI
DOI: 10.3390/ijms241713514
关键词
ryanodine receptor; cardiac tissue slices; death staining; sarcoplasmic reticulum; viability assay
The cultivation of beating myocardial slices is an emerging technique in cardiac research for drug screening and studying pathomechanisms. This study focuses on staining viable cardiomyocytes in human and rabbit myocardial slices using immunofluorescence of ryanodine receptors (RyRs). The results suggest that RyR immunofluorescence can indicate viable cardiomyocytes and loss of sarcoplasmic reticulum integrity may be an early event during cardiomyocyte death.
In terms of preserving multicellularity and myocardial function in vitro, the cultivation of beating myocardial slices is an emerging technique in basic and translational cardiac research. It can be used, for example, for drug screening or to study pathomechanisms. Here, we describe staining for viable cardiomyocytes based on the immunofluorescence of ryanodine receptors (RyRs) in human and rabbit myocardial slices. Biomimetic chambers were used for culture and measurements of contractile force. Fixable fluorophore-conjugated dextran, entering cells with a permeable membrane, was used for death staining. RyRs, nuclei and the extracellular matrix, including the t-system, were additionally stained and analyzed by confocal microscopy and image processing. We found the mutual exclusion of the RyR and dextran signals in cultivated slices. T-System density and nucleus size were reduced in RyR-negative/dextran-positive myocytes. The fraction of RyR-positive myocytes and pixels correlated with the contractile force. In RyR-positive/dextran-positive myocytes, we found irregular RyR clusters and SERCA distribution patterns, confirmed by an altered power spectrum. We conclude that RyR immunofluorescence indicates viable cardiomyocytes in vibratome-cut myocardial slices, facilitating the detection and differential structural analysis of living vs. dead or dying myocytes. We suggest the loss of sarcoplasmic reticulum integrity as an early event during cardiomyocyte death.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据