Ryanodine Receptor Staining Identifies Viable Cardiomyocytes in Human and Rabbit Cardiac Tissue Slices.
Ryanodine Receptor Staining Identifies Viable Cardiomyocytes in Human and Rabbit Cardiac Tissue Slices.
复制标题
DOI：
10.3390/ijms241713514
复制
发表时间：
2023-08-31
影响因子：
5.6
通讯作者：
Seidel, Thomas
中科院分区：
文献类型：
--
作者：
Pfeuffer, Ann-Katrin M.;Kuepfer, Linda K.;Shankar, Thirupura S.;Drakos, Stavros G.;Volk, Tilmann;Seidel, Thomas
在体外保存多细胞和心肌功能方面，搏动心肌切片的培养是基础和转化心脏研究中的新兴技术。例如，它可以用于药物筛选或研究病理机制。在这里，我们描述了染色的基础上，在人类和兔心肌切片的兰尼碱受体（RyRs）的免疫荧光存活心肌细胞。仿生室用于培养和测量收缩力。可固定的荧光团缀合的葡聚糖，进入细胞的渗透膜，用于死亡染色。RyR，细胞核和细胞外基质，包括t-系统，另外染色，并通过共聚焦显微镜和图像处理分析。我们发现在培养切片中RyR和葡聚糖信号的相互排斥。RyR阴性/葡聚糖阳性肌细胞的T-系统密度和细胞核大小减少。RyR阳性心肌细胞的比例和像素与收缩力相关。在RyR阳性/葡聚糖阳性的肌细胞，我们发现不规则的RyR集群和SERCA分布模式，证实了改变功率谱。我们的结论是，RyR免疫荧光表明在振动切片机切割的心肌切片中有存活的心肌细胞，有利于活的与死的或垂死的心肌细胞的检测和差异结构分析。我们认为肌浆网完整性的丧失是心肌细胞死亡的早期事件。
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.
登录
查看更多内容
DOI：
10.1016/j.pbiomolbio.2017.07.006
发表时间：
2017-11
影响因子：
3.8
作者：
Seidel T;Sankarankutty AC;Sachse FB
通讯作者：
Sachse FB
影响因子：
16.6
作者：
Esfandyari D;Idrissou BMG;Hennis K;Avramopoulos P;Dueck A;El-Battrawy I;Grüter L;Meier MA;Näger AC;Ramanujam D;Dorn T;Meitinger T;Hagl C;Milting H;Borggrefe M;Fenske S;Biel M;Dendorfer A;Sassi Y;Moretti A;Engelhardt S
通讯作者：
Engelhardt S
影响因子：
3.7
作者：
Crossman DJ;Ruygrok PN;Soeller C;Cannell MB
通讯作者：
Cannell MB
影响因子：
15.9
作者：
GOTTLIEB, RA;BURLESON, KO;ENGLER, RL
通讯作者：
ENGLER, RL
影响因子：
5.5
作者：
Lukyanenko, V;Györke, S
通讯作者：
Györke, S