Department of Cell and Molecular Physiology, and Cardiovascular Research Institute, Stritch School of Medicine, Loyola University Chicago, 2160 S. First Ave, Maywood, IL, USA.
Department of Cell and Molecular Physiology, and Cardiovascular Research Institute, Stritch School of Medicine, Loyola University Chicago, 2160 S. First Ave, Maywood, IL, USA.
Cell Calcium. 2024 May;119:102873. doi: 10.1016/j.ceca.2024.102873. Epub 2024 Mar 20.
Calcium signaling is a critical process required for cellular mechanisms such as cardiomyocyte contraction. The inability of the cell to properly activate or regulate calcium signaling can lead to contractile dysfunction. In isolated cardiomyocytes, calcium signaling has been primarily studied using calcium fluorescent dyes, however these dyes have limited applicability to whole organs. Here, we crossed the Salsa6f mouse which expresses a genetically encoded ratiometric cytosolic calcium indicator with a cardiomyocyte specific inducible cre to temporally-induce expression and studied cytosolic calcium transients in isolated cardiomyocytes and modified Langendorff heart preparations. Isolated cardiomyocytes expressing Salsa6f or Fluo-4AM loaded were compared. We also crossed the Salsa6f mouse with a floxed Polycystin 2 (PC2) mouse to test the feasibility of using the Salsa6f mouse to measure calcium transients in PC2 heterozygous or homozygous knock out mice. Although there are caveats in the applicability of the Salsa6f mouse, there are clear advantages to using the Salsa6f mouse to measure whole heart calcium signals.
钙信号是细胞机制（如心肌细胞收缩）所必需的关键过程。细胞不能正确激活或调节钙信号会导致收缩功能障碍。在分离的心肌细胞中，钙信号主要使用钙荧光染料进行研究，然而这些染料在整个器官中的应用有限。在这里，我们将表达基因编码的比率型细胞质钙指示剂的 Salsa6f 小鼠与心肌细胞特异性诱导型 cre 进行杂交，以在时间上诱导表达，并研究分离的心肌细胞和改良的 Langendorff 心脏制剂中的细胞质钙瞬变。比较了表达 Salsa6f 或 Fluo-4AM 负载的分离的心肌细胞。我们还将 Salsa6f 小鼠与 floxed Polycystin 2 (PC2) 小鼠杂交，以测试使用 Salsa6f 小鼠测量 PC2 杂合或纯合敲除小鼠钙瞬变的可行性。尽管 Salsa6f 小鼠的适用性存在一些限制，但使用 Salsa6f 小鼠测量整个心脏钙信号具有明显的优势。
