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. 2022 Jan 19;12:978. doi: 10.1038/s41598-022-04929-1

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Comparison of cristae density and dimensions in each of the mitochondrial subpopulations using TEM images of thin sections of the WT and GENA348 myocardium. (A) There is no difference between the cristae density within the mitochondrial subpopulations. Each data point represents the mean of the cristae density for a mitochondrial subpopulation in each TEM image analysed. (B) Left: Example of a mitochondrion with ordered cristae (WT) used to analyse cristae dimensions. Right: The same mitochondrion after applying the (ImageJ) threshold function. Scale bar = 100 nm. The red line illustrates the position of the ‘line plot’ which spans 6 cristae. (C) Density analysis of the line spanning the cristae highlighted in (B) using ‘plot line’ function (ImageJ), each cristae is labelled 1–6. Each crista has a characteristic doublet representing the outer and inner membrane surface. Parameters measured were W_O, the distance between the outer membrane leaflet; W_I, the distance between the cristae inner membrane (lumen) and S, the distance between adjacent crista. (D) There is no difference between the WT and GENA348 cristae widths (W_O/W_I) or spacing (S); (see Table 2 for dimensions). (E) Examples of mitochondria in WT and GENA348 micrographs assigned according to ‘quality’; 1 = mitochondria has ordered cristae; 3 = mitochondria with areas of partial cristae organization; 5 = no visible cristae. N nucleus. (F) WT SSM and IFM mitochondrial subpopulations have a greater percentage of type 1 mitochondria with ordered cristae. (G) GENA348 SSM and PNM have more mitochondria with only partially ordered cristae and mitochondria without defined cristae. (H) A comparison of the ‘quality’ scoring between WT and GENA348 subpopulations shows that there are less ‘type 1’ mitochondria with more ‘type 3 and 5’ in the GENA348 SSM and IFM compared to WT. All data is presented as mean ± SEM. Data were analysed using a one-way ANOVA (with Tukey’s multiple comparison test) n = 3 animals per group.