Real time mitochondrial dimension measurements

Main Article Content

Joseph M. Leichner
Evgene Konyukhov
David Kamoun
Yael Yaniv

Keywords

energetics, mitochondrial deformation, mitochondrial volume, real time, sarcomere length

Abstract

Mitochondrial volume is correlated with cell function and internal cell processes. Changes in mitochondrial volume were associated with advanced states of cardiac disease. Thus, measurements of mitochondrial dimension deformations are important to the understanding of cell function and its deterioration. Existing methods either allow measurements of the volume of isolated mitochondria, which are an inferior model to that of isolated cells, or they allow short time measurements that are toxic to the cells. Recent studies have discovered that mitochondrial deformation along a given cell axis can be measured by using the Fourier transformation on the variation in transmitted light intensity induced by the periodic lattice of myofilaments alternating with mitochondrial rows. However, this method was used only offline and in a line scan mode, making it impossible to measure both axes. We designed an open source program in LabVIEW to take advantage of the transmitted light diffraction technique and quantify mitochondrial two dimension (2D) deformation in cardiomyocytes, in situ in real time for long periods (more than several seconds). We validated the program on synthetic and on experimental images from rabbit and rat ventricular myocytes. The program can analyze offline and real time simultaneous 2D mitochondrial deformation dynamics as well as also sarcomere length dynamics. Moreover, the program can accurately analyze images acquired from different cameras. Quantification of mitochondrial 2D deformations is a powerful tool for exploring cell biophysics and bioenergetics mechanisms and will lay the foundation for a future clinical tool for quantifying mitochondrial volume changes associated with different cardiac diseases.

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References

[1] Lim KHH, Javadov SA, Das M, Clarke SJ, Suleiman M-S, Halestrap AP. The effects of ischaemic preconditioning, diazoxide and 5-hydroxydecanoate on rat heart mitochondrial volume and respiration. J Physiol 2002;545:961–74.
[2] Juhaszova M, Zorov DB, Kim SH, Pepe S, Fu Q, Fishbein KW, et al. Glycogen synthase kinase-3beta mediates convergence of protection signaling to inhibit the mitochondrial permeability transition pore. J Clin Invest 2004;113:1535–49. doi:10.1172/JCI19906.
[3] Gogvadze V, Robertson JD, Enoksson M, Zhivotovsky B, Orrenius S. Mitochondrial cytochrome c release may occur by volume-dependent mechanisms not involving permeability transition. Biochem J 2004;378:213–7.
[4] Kaasik A, Joubert F, Ventura-Clapier R, Veksler V. A novel mechanism of regulation of cardiac contractility by mitochondrial functional state. FASEB J 2004;18:1219–27. doi:10.1096/fj.04-1508com.
[5] Tedeschi H, Harris DL. Some observations on the photometric estimation of mitochondrial volume. Biochim Biophys Acta 1958;28:392–402.
[6] Yaniv Y, Juhaszova M, Nuss HB, Wang S, Zorov DB, Lakatta EG, et al. Matching ATP supply and demand in mammalian heart In vivo, in vitro, and in silico perspectives. Anal Card Dev From Embryo to Old Age 2010;1188:133–42. doi:DOI 10.1111/j.1749-6632.2009.05093.x.
[7] Safiulina D, Veksler V, Zharkovsky A, Kaasik A. Loss of mitochondrial membrane potential is associated with increase in mitochondrial volume: Physiological role in neurones. J Cell Physiol 2006;206:347–53. doi:10.1002/jcp.20476.
[8] Hayashi T, Martone ME, Yu Z, Thor A, Doi M, Holst MJ, et al. Three-dimensional electron microscopy reveals new details of membrane systems for Ca2+ signaling in the heart. J Cell Sci 2009;122.
[9] Yaniv Y, Juhaszova M, Wang S, Fishbein KW, Zorov DB, Sollott SJ. Analysis of mitochondrial 3D-deformation in cardiomyocytes during active contraction reveals passive structural anisotropy of orthogonal short axes. PLoS One 2011;6:e21985. doi:10.1371/journal.pone.0021985PONE-D-11-08642 [pii].
[10] Davoodi M, Segal S, Kirschner Peretz N, Kamoun D, Yaniv Y. Semi-automated program for analysis of local Ca2+ spark release with application for classification of heart cell type. Cell Calcium 2017;64:83–90. doi:10.1016/j.ceca.2017.02.003.
[11] de Tombe PP, ter Keurs HEDJ. Cardiac muscle mechanics: Sarcomere length matters. J Mol Cell Cardiol 2016;91:148–50. doi:10.1016/j.yjmcc.2015.12.006.
[12] Nguyen JS, Lakkis NM, Bobek J, Goswami R, Dokainish H. Systolic and Diastolic Myocardial Mechanics in Patients with Cardiac Disease and Preserved Ejection Fraction: Impact of Left Ventricular Filling Pressure. J Am Soc Echocardiogr 2010;23:1273–80. doi:10.1016/j.echo.2010.09.008.
[13] Gao B, Qu Y, Sutherland W, Chui RW, Hoagland K, Vargas HM. Decreased contractility and altered responses to inotropic agents in myocytes from tachypacing-induced heart failure canines. J Pharmacol Toxicol Methods 2018. doi:10.1016/j.vascn.2018.06.001.
[14] Liu T, Chen L, Kim E, Tran D, Phinney BS, Knowlton AA. Mitochondrial proteome remodeling in ischemic heart failure. Life Sci 2014;101:27–36. doi:10.1016/j.lfs.2014.02.004.