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Metabolic compartmentation in rainbow trout cardiomyocytes: coupling of hexokinase but not creatine kinase to mitochondrial respiration

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Abstract

Rainbow trout (Oncorhynchus mykiss) cardiomyocytes have a simple morphology with fewer membrane structures such as sarcoplasmic reticulum and t-tubules penetrating the cytosol. Despite this, intracellular ADP diffusion is restricted. Intriguingly, although diffusion is restricted, trout cardiomyocytes seem to lack the coupling between mitochondrial creatine kinase (CK) and respiration. Our aim was to study the distribution of diffusion restrictions in permeabilized trout cardiomyocytes and verify the role of CK. We found a high activity of hexokinase (HK), which led us to reassess the situation in trout cardiomyocytes. We show that diffusion restrictions are more prominent than previously thought. In the presence of a competitive ADP-trapping system, ADP produced by HK, but not CK, was channeled to the mitochondria. In agreement with this, we found no positively charged mitochondrial CK in trout heart homogenate. The results were best fit by a simple mathematical model suggesting that trout cardiomyocytes lack a functional coupling between ATPases and pyruvate kinase. The model simulations show that diffusion is restricted to almost the same extent in the cytosol and by the outer mitochondrial membrane. Furthermore, they confirm that HK, but not CK, is functionally coupled to respiration. In perspective, our results suggest that across a range of species, cardiomyocyte morphology and metabolism go hand in hand with cardiac performance, which is adapted to the circumstances. Mitochondrial CK is coupled to respiration in adult mammalian hearts, which are specialized to high, sustained performance. HK associates with mitochondria in hearts of trout and neonatal mammals, which are more hypoxia-tolerant.

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References

  • Aho E, Vornanen M (1999) Contractile properties of atrial and ventricular myocardium of the heart of rainbow trout Oncorhynchus mykiss: effects of thermal acclimation. J Exp Biol 202(Pt 19):2663–2677

    PubMed  Google Scholar 

  • Anmann T, Eimre M, Kuznetsov AV et al (2005) Calcium-induced contraction of sarcomeres changes the regulation of mitochondrial respiration in permeabilized cardiac cells. FEBS J 272:3145–3161

    Article  CAS  PubMed  Google Scholar 

  • Appaix F, Kuznetsov A, Usson Y et al (2003) Possible role of cytoskeleton in intracellular arrangement and regulation of mitochondria. Exp Physiol 88:175–190

    Article  CAS  PubMed  Google Scholar 

  • Balaban RS (2009) The role of Ca2+ signaling in the coordination of mitochondrial ATP production with cardiac work. Biochim Biophys Acta BBA Bioenerg 1787:1334–1341. doi:10.1016/j.bbabio.2009.05.011

    Article  CAS  Google Scholar 

  • Birkedal R, Gesser H (2006) Intracellular compartmentation of cardiac fibres from rainbow trout and Atlantic cod—a general design of heart cells. BBA Bioenerg 1757:764–772

    Article  CAS  Google Scholar 

  • Birkedal R, Shiels HA (2007) High [Na+]i in cardiomyocytes from rainbow trout. Am J Physiol Regul Integr Comp Physiol 293:R861–R866

    Article  CAS  PubMed  Google Scholar 

  • Birkedal R, Shiels HA, Vendelin M (2006) Three-dimensional mitochondrial arrangement in ventricular myocytes: from chaos to order. Am J Physiol Cell Physiol 291:C1148

    Article  CAS  PubMed  Google Scholar 

  • Birkedal R, Laasmaa M, Vendelin M (2014) The location of energetic compartments affects energetic communication in cardiomyocytes. Front Physiol 5:376. doi:10.3389/fphys.2014.00376

    Article  PubMed  PubMed Central  Google Scholar 

  • Bose S, French S, Evans FJ et al (2003) Metabolic network control of oxidative phosphorylation. J Biol Chem 278:39155–39165. doi:10.1074/jbc.M306409200

    Article  CAS  PubMed  Google Scholar 

  • Boudina S, Laclau MN, Tariosse L et al (2002) Alteration of mitochondrial function in a model of chronic ischemia in vivo in rat heart. Am J Physiol Heart Circ Physiol 282:H821–H831. doi:10.1152/ajpheart.00471.2001

    Article  CAS  PubMed  Google Scholar 

  • Branovets J, Sepp M, Kotlyarova S et al (2013) Unchanged mitochondrial organization and compartmentation of high-energy phosphates in creatine-deficient GAMT−/− mouse hearts. Am J Physiol Heart Circ Physiol 305:H506–H520. doi:10.1152/ajpheart.00919.2012

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Brdiczka DG, Zorov DB, Sheu S-S (2006) Mitochondrial contact sites: their role in energy metabolism and apoptosis. Biochim Biophys Acta 1762:148–163. doi:10.1016/j.bbadis.2005.09.007

    Article  CAS  PubMed  Google Scholar 

  • Christensen M, Hartmund T, Gesser H (1994) Creatine kinase, energy-rich phosphates and energy metabolism in heart muscle of different vertebrates. J Comp Physiol B 164:118–123

    Article  CAS  PubMed  Google Scholar 

  • England PJ, Randle PJ (1967) Effectors of rat-heart hexokinases and the control of rates of glucose phosphorylation in the perfused rat heart. Biochem J 105:907–920

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Franzini-Armstrong C (2007) ER-mitochondria communication. How privileged? Physiol Bethesda 22:261–268. doi:10.1152/physiol.00017.2007

    Article  CAS  Google Scholar 

  • Franzini-Armstrong C, Protasi F, Tijskens P (2005) The assembly of calcium release units in cardiac muscle. Ann N Y Acad Sci 1047:76–85

    Article  CAS  PubMed  Google Scholar 

  • Guerrero K, Monge C, Brückner A et al (2009) Study of possible interactions of tubulin, microtubular network, and STOP protein with mitochondria in muscle cells. Mol Cell Biochem 337:239–249. doi:10.1007/s11010-009-0304-1

    Article  PubMed  Google Scholar 

  • Haverinen J, Vornanen M (2009) Comparison of sarcoplasmic reticulum calcium content in atrial and ventricular myocytes of three fish species. Am J Physiol Regul Integr Comp Physiol 297:R1180–R1187. doi:10.1152/ajpregu.00022.2009

    Article  CAS  PubMed  Google Scholar 

  • Hoerter JA, Kuznetsov A, Ventura-Clapier R (1991) Functional development of the creatine kinase system in perinatal rabbit heart. Circ Res 69:665–676

    Article  CAS  PubMed  Google Scholar 

  • Hoerter JA, Ventura-Clapier R, Kuznetsov A (1994) Compartmentation of creatine kinases during perinatal development of mammalian heart. Mol Cell Biochem 133–134:277–286

    Article  PubMed  Google Scholar 

  • Hove-Madsen L (1992) The influence of temperature on ryanodine sensitivity and the force-frequency relationship in the myocardium of rainbow trout. J Exp Biol 167:47–60

    CAS  PubMed  Google Scholar 

  • Huang J, Hove-Madsen L, Tibbits GF (2008) Ontogeny of Ca2+ -induced Ca2+ release in rabbit ventricular myocytes. Am J Physiol Cell Physiol 294:C516–C525

    Article  CAS  PubMed  Google Scholar 

  • Illaste A, Laasmaa M, Peterson P, Vendelin M (2012) Analysis of molecular movement reveals latticelike obstructions to diffusion in heart muscle cells. Biophys J 102:739–748. doi:10.1016/j.bpj.2012.01.012

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jayasinghe ID, Cannell MB, Soeller C (2009) Organization of ryanodine receptors, transverse tubules, and sodium-calcium exchanger in rat myocytes. Biophys J 97:2664–2673. doi:10.1016/j.bpj.2009.08.036

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kaasik A, Veksler V, Boehm E et al (2001) Energetic crosstalk between organelles: architectural integration of energy production and utilization. Circ Res 89:153–159. doi:10.1161/hh1401.093440

    Article  CAS  PubMed  Google Scholar 

  • Khuchua ZA, Qin W, Boero J et al (1998) Octamer formation and coupling of cardiac sarcomeric mitochondrial creatine kinase are mediated by charged N-terminal residues. J Biol Chem 273:22990–22996. doi:10.1074/jbc.273.36.22990

    Article  CAS  PubMed  Google Scholar 

  • Kinsey ST, Moerland TS (2002) Metabolite diffusion in giant muscle fibers of the spiny lobster Panulirus argus. J Exp Biol 205:3377–3386

    CAS  PubMed  Google Scholar 

  • Laasmaa M, Vendelin M, Peterson P (2011) Application of regularized Richardson-Lucy algorithm for deconvolution of confocal microscopy images. J Microsc 243:124–140. doi:10.1111/j.1365-2818.2011.03486.x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Laclau MN, Boudina S, Thambo JB et al (2001) Cardioprotection by ischemic preconditioning preserves mitochondrial function and functional coupling between adenine nucleotide translocase and creatine kinase. J Mol Cell Cardiol 33:947–956

    Article  CAS  PubMed  Google Scholar 

  • Llach A, Molina CE, Alvarez-Lacalle E et al (2011) Detection, properties, and frequency of local calcium release from the sarcoplasmic reticulum in teleost cardiomyocytes. PLoS One 6:e23708. doi:10.1371/journal.pone.0023708

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lygate CA, Aksentijevic D, Dawson D et al (2013) Living without creatine unchanged exercise capacity and response to chronic myocardial infarction in creatine-deficient mice. Circ Res 112:945–955. doi:10.1161/CIRCRESAHA.112.300725

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mergenthaler P, Kahl A, Kamitz A et al (2012) Mitochondrial hexokinase II (HKII) and phosphoprotein enriched in astrocytes (PEA15) form a molecular switch governing cellular fate depending on the metabolic state. Proc Natl Acad Sci USA 109:1518–1523. doi:10.1073/pnas.1108225109

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • O’Brien KM, Mueller IA, Orczewska JI et al (2014) Hearts of some Antarctic fishes lack mitochondrial creatine kinase. Comp Biochem Physiol A Mol Integr Physiol 178:30–36. doi:10.1016/j.cbpa.2014.08.003

    Article  PubMed  Google Scholar 

  • Ostadal B, Ostadalova I, Dhalla NS (1999) Development of cardiac sensitivity to oxygen deficiency: comparative and ontogenetic aspects. Physiol Rev 79:635–659

    CAS  PubMed  Google Scholar 

  • Postic C, Leturque A, Printz RL et al (1994) Development and regulation of glucose transporter and hexokinase expression in rat. Am J Physiol Endocrinol Metab 266:E548–E559

    CAS  Google Scholar 

  • Ramay HR, Vendelin M (2009) Diffusion restrictions surrounding mitochondria: a mathematical model of heart muscle fibers. Biophys J 97:443–452. doi:10.1016/j.bpj.2009.04.062

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rostovtseva TK, Sheldon KL, Hassanzadeh E et al (2008) Tubulin binding blocks mitochondrial voltage-dependent anion channel and regulates respiration. Proc Natl Acad Sci USA 105:18746–18751. doi:10.1073/pnas.0806303105

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Saks VA, Vasil’eva E, Belikova YO et al (1993) Retarded diffusion of ADP in cardiomyocytes: possible role of mitochondrial outer membrane and creatine kinase in cellular regulation of oxidative phosphorylation. Biochim Biophys Acta 1144:134–148

    Article  CAS  PubMed  Google Scholar 

  • Saks VA, Kuznetsov AV, Khuchua ZA et al (1995) Control of cellular respiration in vivo by mitochondrial outer membrane and by creatine kinase. A new speculative hypothesis: possible involvement of mitochondrial-cytoskeleton interactions. J Mol Cell Cardiol 27:625–645

    Article  CAS  PubMed  Google Scholar 

  • Saks VA, Kongas O, Vendelin M, Kay L (2000) Role of the creatine/phosphocreatine system in the regulation of mitochondrial respiration. Acta Physiol Scand 168:635–641

    Article  CAS  PubMed  Google Scholar 

  • Saks VA, Kaambre T, Sikk P et al (2001) Intracellular energetic units in red muscle cells. Biochem J 356:643–657

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Santer RM (1985) Morphology and innervation of the fish heart. Adv Anat Embryol Cell Biol 89:1–102

    Article  CAS  PubMed  Google Scholar 

  • Schlattner U, Wallimann T (2000) Octamers of mitochondrial creatine kinase isoenzymes differ in stability and membrane binding. J Biol Chem 275:17314–17320. doi:10.1074/jbc.M001919200

    Article  CAS  PubMed  Google Scholar 

  • Schlattner U, Tokarska-Schlattner M, Wallimann T (2006) Mitochondrial creatine kinase in human health and disease. Biochim Biophys Acta 1762:164–180. doi:10.1016/j.bbadis.2005.09.004

    Article  CAS  PubMed  Google Scholar 

  • Schurmann H, Steffensen JF, Lomholt JP (1991) The influence of hypoxia on the preferred temperature of Rainbow Trout, Oncorhyncus mykiss. JExpBiol 157:75–86

    Google Scholar 

  • Sepp M, Vendelin M, Vija H, Birkedal R (2010) ADP compartmentation analysis reveals coupling between pyruvate kinase and ATPases in heart muscle. Biophys J 98:2785–2793. doi:10.1016/j.bpj.2010.03.025

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Seppet EK, Kaambre T, Sikk P et al (2001) Functional complexes of mitochondria with Ca, MgATPases of myofibrils and sarcoplasmic reticulum in muscle cells. Biochim Biophys Acta 1504:379–395

    Article  CAS  PubMed  Google Scholar 

  • Shiels HA, Vornanen M, Farrell AP (2002) Temperature dependence of cardiac sarcoplasmic reticulum function in rainbow trout myocytes. J Exp Biol 205:3631–3639

    CAS  PubMed  Google Scholar 

  • Simson P, Jepihhina N, Laasmaa M et al (2016) Restricted ADP movement in cardiomyocytes: cytosolic diffusion obstacles are complemented with a small number of open mitochondrial voltage-dependent anion channels. J Mol Cell Cardiol 97:197–203. doi:10.1016/j.yjmcc.2016.04.012

    Article  CAS  PubMed  Google Scholar 

  • Sokolova N, Vendelin M, Birkedal R (2009) Intracellular diffusion restrictions in isolated cardiomyocytes from rainbow trout. BMC Cell Biol 10:90. doi:10.1186/1471-2121-10-90

    Article  PubMed  PubMed Central  Google Scholar 

  • Southworth R, Davey KAB, Warley A, Garlick PB (2007) A reevaluation of the roles of hexokinase I and II in the heart. Am J Physiol Heart Circ Physiol 292:H378–H386. doi:10.1152/ajpheart.00664.2006

    Article  CAS  PubMed  Google Scholar 

  • Vendelin M, Kongas O, Saks V (2000) Regulation of mitochondrial respiration in heart cells analyzed by reaction-diffusion model of energy transfer. Am J Physiol Cell Physiol 278:C747–C764

    CAS  PubMed  Google Scholar 

  • Vendelin M, Eimre M, Seppet E et al (2004a) Intracellular diffusion of adenosine phosphates is locally restricted in cardiac muscle. Mol Cell Biochem 256(257):229–241. doi:10.1023/B:MCBI.0000009871.04141.64

    Article  PubMed  Google Scholar 

  • Vendelin M, Lemba M, Saks VA (2004b) Analysis of functional coupling: mitochondrial creatine kinase and adenine nucleotide translocase. Biophys J 87:696–713. doi:10.1529/biophysj.103.036210

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ventura-Clapier R, Kuznetsov A, Veksler V et al (1998) Functional coupling of creatine kinases in muscles: species and tissue specificity. Mol Cell Biochem 184:231–247

    Article  CAS  PubMed  Google Scholar 

  • Ventura-Clapier R, Garnier A, Veksler V (2004) Energy metabolism in heart failure. J Physiol 555:1–13. doi:10.1113/jphysiol.2003.055095

    Article  CAS  PubMed  Google Scholar 

  • Vornanen M (2006) Temperature and Ca2+ dependence of [3H]ryanodine binding in the burbot (Lota lota L.) heart. Am J Physiol Regul Integr Comp Physiol 290:R345–R351

    Article  CAS  PubMed  Google Scholar 

  • Wallimann T, Wyss M, Brdiczka D et al (1992) Intracellular compartmentation, structure and function of creatine kinase isoenzymes in tissues with high and fluctuating energy demands: the “phosphocreatine circuit” for cellular energy homeostasis. Biochem J 281:21–40

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wu F, Zhang EY, Zhang J et al (2008) Phosphate metabolite concentrations and ATP hydrolysis potential in normal and ischaemic hearts. J Physiol 586:4193–4208. doi:10.1113/jphysiol.2008.154732

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

We wish to acknowledge Ms. Merle Mandel for her assistance with the spectrophotometric recordings. We also thank Dr. Erkki Truve and Dr. Cecilia Sarmiento for letting us use their gel electrophoresis system. This study was funded by the Estonian Science Foundation (Grant No. ETF8041), the European Union through the European Regional Development Fund (CENS Estonian Center of Excellence in Research) and the Estonian Research Council (IUT 33-7).

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  1. Institute of Cybernetics, Tallinn University of Technology, Akadeemia 21, 12618, Tallinn, Estonia

    Niina Karro, Mervi Sepp, Svetlana Jugai, Martin Laasmaa, Marko Vendelin & Rikke Birkedal

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  1. Niina Karro
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  2. Mervi Sepp
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  3. Svetlana Jugai
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Correspondence to Rikke Birkedal.

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Communicated by G. Heldmaier.

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Karro, N., Sepp, M., Jugai, S. et al. Metabolic compartmentation in rainbow trout cardiomyocytes: coupling of hexokinase but not creatine kinase to mitochondrial respiration. J Comp Physiol B 187, 103–116 (2017). https://doi.org/10.1007/s00360-016-1025-x

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