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. 2018 Oct 30;7:e39427. doi: 10.7554/eLife.39427

Figure 2. RyRs are dispersed in failing cardiomyocytes.

Alterations in nanoscale RyR organization were examined in cardiomyocytes from rats with post-infarction heart failure (HF). Representative images show that macroscale organization of RyRs was similar in HF and Sham-operated controls (A), upper panels). However, nanoscale examination revealed that RyR clusters were broken apart in HF. For the magnified regions in (A), conversion from raw dSTORM to binary images is shown in the middle and lower panels (saturation levels indicated by high-low look-up table). Mean measurements showed fewer RyRs per cluster in failing cells, with an increased fraction of small clusters (B). Dispersion of RyR clusters into smaller fragments resulted in an increased overall number of clusters (D), reduced inter-cluster distances (E) and inclusion of more clusters in each CRU (F). Overall CRU composition became less solid in failing cells ((G), assessed by convex-hull analysis), as the average CRU contained fewer RyRs (C). See Figure 2—source data 1 for analysis of 100 nm vs 150 nm CRU inclusion criterion (ncells = 46, 50 in Sham, HF; *=P < 0.05 vs Sham).

Figure 2—source data 1. Quantification of RyR organization using 150 vs 100 nm CRU inclusion criteria.
RyR cluster and CRU data were compared in Sham and HF myocytes, with data cumulated across cells or animals. Two CRU definitions were also compared, with maximum edge-to-edge distances of 150 nm or 100 nm. Significant differences within data cumulated across cells were determined by t-test, while data cumulated across animals were tested with linear mixed effects models (Lindstrom and Bates, 1988).
elife-39427-fig2-data1.pdf (357.3KB, pdf)
DOI: 10.7554/eLife.39427.006

Figure 2.

Figure 2—figure supplement 1. HF in post-infarction rats is not associated with altered expression of RyR, BIN1, or Junctophilin-2.

Figure 2—figure supplement 1.

Western blotting was performed on homogenates of left ventricles from post-MI HF rats and Sham-operated controls. Representative immunoblots are shown at left, with mean values at right, normalized to Sham. (JPH2 = junctophilin 2; nhearts = 5, 5 in Sham, HF; P = NS).