Development of apical hypertrophic cardiomyopathy with age in a transgenic mouse model carrying the cardiac actin E99K mutation

  • Li Wang
  • Fan Bai
  • Qing Zhang
  • Weihua Song
  • Andrew Messer
  • Masataka Kawai
Article
  • 35 Downloads

Abstract

In both humans and mice, the Glu-99-Lys (E99K) mutation in the cardiac actin gene (ACTC) results in little understood apical hypertrophic cardiomyopathy (AHCM). To determine how cross-bridge kinetics change with AHCM development, we applied sinusoidal length perturbations to skinned papillary muscle fibres from 2- and 5-month old E99K transgenic (Tg) and non-transgenic (NTg) mice, and studied tension and its transients. These age groups were chosen because our preliminary studies indicated that AHCM develops with age. Fibres from 5-month old E99K mice showed significant decreases in tension, stiffness, the rate of the medium-speed exponential process and its magnitude compared to non-transgenic control. The nucleotide association constants increased with age, and they were significantly larger in E99K compared to NTg. However, there were no large differences in the rates of the cross-bridge detachment step, the rates of the force generation step, or the phosphate association constant. Our result on force/cross-bridge demonstrates that the decreased active tension of E99K fibres was caused by a decreased amount of force generated per each cross-bridge. The effects were generally less or insignificant at 2 months. A pCa-tension study showed increased Ca2+-sensitivity (pCa50) with age in both the E99K and NTg sample groups, and pCa50 was significantly larger (but only for 0.05–0.06 pCa units) in E99K than in NTg groups. A significant decrease in cooperativity (nH) was observed only in 5-month old E99K mice. We conclude that the AHCM-causing ACTC E99K mutation is associated with progressive alterations in biomechanical parameters, with changes smaller at 2 months but larger at 5 months, correlating with the development of AHCM.

Keyword

Sinusoidal analysis Tension transient Stiffness Rate constant Rigor state Nucleotide association Ca ion sensitivity Pathogenic mechanisms 

Abbreviations

2πb

Apparent rate constant of the delayed tension (exponential process B)

2πc

Apparent rate constant of fast tension recovery (exponential process C)

ACTA1

Skeletal actin gene

ACTC

Cardiac actin gene

AHCM

Apical HCM

B

Magnitude of exponential process B

C

Magnitude of exponential process C

CK

Creatine kinase

D

MgADP or its concentration

E99K

Glu-99-Lys

f

Frequency of length oscillation

HCM

Hypertrophic cardiomyopathy

K0

MgADP association constant

K1

MgATP association constant

k2

Rate constant of the cross-bridge detachment step 2

k− 2

Rate constant of the reversal of step 2

K2

Equilibrium constant of step 2 (= k2/k− 2)

k4

Rate constant of force generation step 4 (isomerization of the AM.ADP.Pi state)

k− 4

Rate constant of the reversal of step 4

K4

Equilibrium constant of step 4 (= k4/k− 4)

K5

Phosphate association constant

kTR

The rate of tension redevelopment

LV

Left ventricle

nH

Cooperativity

NTg

Non-transgenic

P

Pi (phosphate) or its concentration

PCr

Phosphocreatine

pCa50

Ca2+ sensitivity

RV

Right ventricle

S

MgATP or its concentration

SCD

Sudden cardiac death

T5

Tension per cross-bridge supported by the AM*ADP.Pi state

Y(f)

Complex modulus

ΔL

Length change

Notes

Acknowledgements

The authors would like to thank Professor Steven Marston (National Heart and Lung Institute, Imperial College London, London, UK) who developed the Tg mouse model ACTC E99K and made our collaboration possible. The authors also would like to thank Dr. Amy Li in University of Sydney for developing a technique for freezing and thawing muscle samples without causing much damage, and teaching the technique to us. This work was supported by grants from the Natural Science Foundation of Jiangsu Province of China BK20150353 (LW), the National Institutes of Health HL070041 (MK), and The American Heart Association 13GRNT16810043 (MK). The content is solely the responsibility of the authors and does not necessarily reflect the official views of the funding organizations.

Compliance with ethical standards

Conflict of interest

No potential conflict of interest.

References

  1. AHA (2009) Heart disease and stroke statistics. American Heart Association, DallasGoogle Scholar
  2. Bai F, Caster HM, Dawson JF, Kawai M (2015) The immediate effect of HCM causing actin mutants E99K and A230V on actin-Tm-myosin interaction in thin-filament reconstituted myocardium. J Mol Cell Cardiol 79C:123–132CrossRefGoogle Scholar
  3. Behrmann E, Muller M, Penczek PA, Mannherz HG, Manstein DJ, Raunser S (2012) Structure of the rigor actin-tropomyosin-myosin complex. Cell 150:327–338CrossRefPubMedPubMedCentralGoogle Scholar
  4. Bergen HR, Ajtai K, Burghardt TP, Nepomuceno AI, Muddiman DC (2003) Mass spectral determination of skeletal/cardiac actin isoform ratios in cardiac muscle. Rapid Commun Mass Spectrom 17:1467–1471CrossRefPubMedGoogle Scholar
  5. Bookwalter CS, Trybus KM (2006) Functional consequences of a mutation in an expressed human a-cardiac actin at a site implicated in familial hypertrophic cardiomyopathy. J Biol Chem 281:16777–16784CrossRefPubMedGoogle Scholar
  6. Bremel RD, Weber A (1972) Cooperation within actin filament in vertebrate skeletal muscle. Nat New Biol 238:97–101CrossRefPubMedGoogle Scholar
  7. D’Andrea A, Caso P, Bossone E, Scarafile R, Riegler L, Di Salvo G, Gravino R, Cocchia R, Castaldo F, Salerno G, Golia E, Limongelli G, De Corato G, Cuomo S, Pacileo G, Russo MG, Calabro R (2010) Right ventricular myocardial involvement in either physiological or pathological left ventricular hypertrophy: an ultrasound speckle-tracking two-dimensional strain analysis. Eur J Echocardiogr 11:492–500CrossRefPubMedGoogle Scholar
  8. Dahari M, Dawson JF (2015) Do cardiac actin mutations lead to altered actomyosin interactions? Biochem Cell Biol 93:330–334CrossRefPubMedGoogle Scholar
  9. Dantzig J, Goldman Y, Millar NC, Lacktis J, Homsher E (1992) Reversal of the cross-bridge force-generating transition by the photogeneration of phosphate in rabbit psoas muscle fibers. J Physiol 451:247–278CrossRefPubMedPubMedCentralGoogle Scholar
  10. Durrwang U, Fujita-Becker S, Erent M, Kull FJ, Tsiavaliaris G, Geeves MA, Manstein DJ (2006) Dictyostelium myosin-IE is a fast molecular motor involved in phagocytosis. J Cell Sci 119:550–558CrossRefPubMedGoogle Scholar
  11. Force T, Bonow RO, Houser SR, Solaro RJ, Hershberger RE, Adhikari B, Anderson ME, Boineau R, Byrne BJ, Cappola TP, Kalluri R, LeWinter MM, Maron MS, Molkentin JD, Ommen SR, Regnier M, Tang WH, Tian R, Konstam MA, Maron BJ, Seidman CE (2010) Research priorities in hypertrophic cardiomyopathy: report of a Working Group of the National Heart, Lung, and Blood Institute. Circulation 122:1130–1133CrossRefPubMedPubMedCentralGoogle Scholar
  12. Fortune NS, Geeves MA, Ranatunga KW (1991) Tension responses to rapid pressure release in glycerinated rabbit muscle fibers. Proc Natl Acad Sci (USA) 88:7323–7327CrossRefGoogle Scholar
  13. Furch M, Geeves MA, Manstein DJ (1998) Modulation of actin affinity and actomyosin adenosine triphosphatase by charge changes in the myosin motor domain. Biochemistry 37:6317–6326CrossRefPubMedGoogle Scholar
  14. Geeves MA, Holmes KC (1999) Structural mechanism of muscle contraction. Ann Rev Biochem 68:687–728CrossRefPubMedGoogle Scholar
  15. Holmes KC, Schroder RR, Sweeney HL, Houdusse A (2004) The structure of the rigor complex and its implications for the power stroke. Philos Trans R Soc Lond B Biol Sci 359:1819–1828CrossRefPubMedPubMedCentralGoogle Scholar
  16. Huxley HE, Stewart A, Sosa H, Irving T (1994) X-ray diffraction measurements of the extensibility of actin and myosin filaments in contracting muscle. Biophys J 67:2411–2421CrossRefPubMedPubMedCentralGoogle Scholar
  17. Kaski JP, Syrris P, Esteban MT, Jenkins S, Pantazis A, Deanfield JE, McKenna WJ, Elliott PM (2009) Prevalence of sarcomere protein gene mutations in preadolescent children with hypertrophic cardiomyopathy. Circ Cardiovasc Genet 2:436–441CrossRefPubMedGoogle Scholar
  18. Kawai M (2003) What do we learn by studying the temperature effect on isometric tension and tension transients in mammalian striated muscle fibres? J Muscle Res Cell Motil 24:127–138CrossRefPubMedGoogle Scholar
  19. Kawai M, Brandt PW (1980) Sinusoidal analysis: a high resolution method for correlating biochemical reactions with physiological processes in activated skeletal muscles of rabbit, frog and crayfish. J Muscle Res Cell Mot 1:279–303CrossRefGoogle Scholar
  20. Kawai M, Halvorson H (1989) Role of MgATP and MgADP in the crossbridge kinetics in chemically skinned rabbit psoas fibers. Study of a fast exponential process C. Biophys J 55:595–603CrossRefPubMedPubMedCentralGoogle Scholar
  21. Kawai M, Halvorson HR (1991) Two step mechanism of phosphate release and the mechanism of force generation in chemically skinned fibers of rabbit psoas. Biophys J 59:329–342CrossRefPubMedPubMedCentralGoogle Scholar
  22. Kawai M, Zhao Y (1993) Cross-bridge scheme and force per cross-bridge state in skinned rabbit psoas muscle fibers. Biophys J 65:638–651CrossRefPubMedPubMedCentralGoogle Scholar
  23. Kawai M, Saeki Y, Zhao Y (1993) Cross-bridge scheme and the kinetic constants of elementary steps deduced from chemically skinned papillary and trabecular muscles of the ferret. Circ Res 73:35–50CrossRefPubMedGoogle Scholar
  24. Lorenz M, Holmes KC (2010) The actin-myosin interface. Proc Natl Acad Sci USA 107:12529–12534CrossRefPubMedPubMedCentralGoogle Scholar
  25. Lu X, Bryant MK, Bryan KE, Rubenstein PA, Kawai M (2005) Role of the N-terminal negative charges of actin in force generation and cross-bridge kinetics in reconstituted bovine cardiac muscle fibres. J Physiol 564:65–82CrossRefPubMedPubMedCentralGoogle Scholar
  26. Marston SB (2011) How do mutations in contractile proteins cause the primary familial cardiomyopathies? J Cardiovasc Transl Res 4:245–255CrossRefPubMedGoogle Scholar
  27. Miller CJ, Wong WW, Bobkova E, Rubenstein PA, Reisler E (1996) Mutational analysis of the role of the N terminus of actin in actomyosin interactions. Comparison with other mutant actins and implications for the cross-bridge cycle. Biochemistry 35:16557–16565CrossRefPubMedGoogle Scholar
  28. Mogensen J, Klausen IC, Pedersen AK, Egeblad H, Bross P, Kruse TA, Gregersen N, Hansen PS, Baandrup U, Borglum AD (1999) Alpha-cardiac actin is a novel disease gene in familial hypertrophic cardiomyopathy. J Clin Investig 103:R39–R43CrossRefPubMedPubMedCentralGoogle Scholar
  29. Mogensen J, Perrot A, Andersen PS, Havndrup O, Klausen IC, Christiansen M, Bross P, Egeblad H, Bundgaard H, Osterziel KJ, Haltern G, Lapp H, Reinecke P, Gregersen N, Borglum AD (2004) Clinical and genetic characteristics of alpha cardiac actin gene mutations in hypertrophic cardiomyopathy. J Med Genet 41:e10CrossRefPubMedPubMedCentralGoogle Scholar
  30. Mozaffarian D, Caldwell JH (2001) Right ventricular involvement in hypertrophic cardiomyopathy: a case report and literature review. Clin Cardiol 24:2–8CrossRefPubMedGoogle Scholar
  31. Mundia MM, Demers RW, Chow ML, Perieteanu AA, Dawson JF (2012) Subdomain location of mutations in cardiac actin correlate with type of functional change. PLoS One 7:e36821CrossRefPubMedPubMedCentralGoogle Scholar
  32. Murphy KP, Zhao Y, Kawai M (1996) Molecular forces involved in force generation during skeletal muscle contraction. J Exp Biol 199:2565–2571PubMedGoogle Scholar
  33. Nowak KJ, Ravenscroft G, Jackaman C, Filipovska A, Davies SM, Lim EM, Squire SE, Potter AC, Baker E, Clément S, Sewry CA, Fabian V, Crawford K, Lessard JL, Griffiths LM, Papadimitriou JM, Shen Y, Morahan G, Bakker AJ, Davies KE, Laing NG (2009) Rescue of skeletal muscle α-actin–null mice by cardiac (fetal) α-actin. J Cell Biol 185:903–915CrossRefPubMedPubMedCentralGoogle Scholar
  34. Olson TM, Michels VV, Thibodeau SN, Tai YS, Keating MT (1998) Actin mutations in dilated cardiomyopathy, a heritable form of heart failure. Science 280:750–752CrossRefPubMedGoogle Scholar
  35. Olson TM, Doan TP, Kishimoto NY, Whitby FG, Ackerman MJ, Fananapazir L (2000) Inherited and de novo mutations in the cardiac actin gene cause familial hypertrophic cardiomyopathy. J Mol Cell Cardiol 32:1687–1694CrossRefPubMedGoogle Scholar
  36. Opie LH, Mansford KR, Owen P (1971) Effects of increased heart work on glycolysis and adenine nucleotides in the perfused heart of normal and diabetic rats. Biochem J 124:475–490CrossRefPubMedPubMedCentralGoogle Scholar
  37. Perhonen MA, Franco F, Lane LD, Buckey JC, Blomqvist CG, Zerwekh JE, Peshock RM, Weatherall PT, Levine BD (2001) Cardiac atrophy after bed rest and spaceflight. J Appl Physiol 91:645–653CrossRefPubMedGoogle Scholar
  38. Rall JA, Woledge RC (1990) Influence of temperature on mechanics and energetics of muscle contraction. Am J Physiol 259:R197-203PubMedGoogle Scholar
  39. Rayment I, Holden HM, Whittaker M, Yohn CB, Lorenz M, Holmes KC, Milligan RA (1993) Structure of the actin-myosin complex and its implications for muscle contraction. Science 261:58–65CrossRefPubMedGoogle Scholar
  40. Roth K, Hubesch B, Meyerhoff DJ, Naruse S, Gober JR, Lawry TJ, Boska MD, Matson GB, Weiner MW (1989) Noninvasive quantitation of phosphorus metabolites in human tissue by NMR spectroscopy. J Magn Reson 81:299–311Google Scholar
  41. Rowlands CT, Owen T, Lawal S, Cao S, Pandey SP, Yang H-Y, Song W, Wilkinson R, Alvarez-Laviada A, Gehmlich K, Marston SB, MacLeod KT (2017) Age- and strain-related aberrant Ca2+ release is associated with sudden cardiac death in the ACTC E99K mouse model of hypertrophic cardiomyopathy. Am J Physiol Heart Circ Physiol 313:H1213–H1226CrossRefPubMedGoogle Scholar
  42. Schober KE, Savino SI, Yildiz V (2016) Right ventricular involvement in feline hypertrophic cardiomyopathy. J Vet Cardiol 18:297–309CrossRefPubMedGoogle Scholar
  43. Seidman CE, Seidman JG (2011) Identifying sarcomere gene mutations in hypertrophic cardiomyopathy: a personal history. Circ Res 108:743–750CrossRefPubMedGoogle Scholar
  44. Semsarian C, Ingles J, Maron MS, Maron BJ (2015) New perspectives on the prevalence of hypertrophic cardiomyopathy. J Am Coll Cardiol 65:1249–1254CrossRefPubMedGoogle Scholar
  45. Song W, Dyer E, Stuckey DJ, Copeland O, Leung MC, Bayliss C, Messer A, Wilkinson R, Tremoleda JL, Schneider MD, Harding SE, Redwood CS, Clarke K, Nowak K, Monserrat L, Wells D, Marston SB (2011) Molecular mechanism of the E99K mutation in cardiac actin (ACTC Gene) that causes apical hypertrophy in man and mouse. J Biol Chem 286:27582–27593CrossRefPubMedPubMedCentralGoogle Scholar
  46. Song W, Vikhorev PG, Kashyap MN, Rowlands C, Ferenczi MA, Woledge RC, MacLeod K, Marston S, Curtin NA (2013) Mechanical and energetic properties of papillary muscle from ACTC E99K transgenic mouse models of hypertrophic cardiomyopathy. Am J Physiol Heart Circ Physiol 304:H1513–H1524CrossRefPubMedPubMedCentralGoogle Scholar
  47. Sutoh K (1983) Mapping of actin-binding sites on the heavy chain of myosin subfragment 1. Biochemistry 22:1579–1585CrossRefPubMedGoogle Scholar
  48. Taylor EW (1979) Mechanism of actomyosin ATPase and the problem of muscle contraction. CRC Crit Rev Biochem 6:103–164CrossRefPubMedGoogle Scholar
  49. Thirlwell H, Corrie JE, Reid GP, Trentham DR, Ferenczi MA (1994) Kinetics of relaxation from rigor of permeabilized fast-twitch skeletal fibers from the rabbit using a novel caged ATP and apyrase. Biophys J 67:2436–2447CrossRefPubMedPubMedCentralGoogle Scholar
  50. Thirlwell H, Sleep JA, Ferenczi MA (1995) Inhibition of unloaded shortening velocity in permeabilized muscle fibres by caged ATP compounds. J Muscle Res Cell Motil 16:131–137CrossRefPubMedGoogle Scholar
  51. Van Driest SL, Ellsworth EG, Ommen SR, Tajik AJ, Gersh BJ, Ackerman MJ (2003) Prevalence and spectrum of thin filament mutations in an outpatient referral population with hypertrophic cardiomyopathy. Circulation 108:445–451CrossRefPubMedGoogle Scholar
  52. Vandekerckhove J, Bugaisky G, Buckingham M (1986) Simultaneous expression of skeletal muscle and heart actin proteins in various striated muscle tissues and cells. A quantitative determination of the two actin isoforms. J Biol Chem 261:1838–1843PubMedGoogle Scholar
  53. Wakabayashi K, Sugimoto Y, Tanaka H, Ueno Y, Takezawa Y, Amemiya Y (1994) X-ray diffraction evidence for the extensibility of actin and myosin filaments during muscle contraction. Biophys J 67:2422–2435CrossRefPubMedPubMedCentralGoogle Scholar
  54. Wang L, Kawai M (2013) A re-interpretation of the rate of tension redevelopment (kTR) in active muscle. J Muscle Res Cell Motil 34:407–415CrossRefPubMedPubMedCentralGoogle Scholar
  55. Wang G, Ding W, Kawai M (1999) Does thin filament compliance diminish the cross-bridge kinetics? A study in rabbit psoas fibers. Biophys J 76:978–984CrossRefPubMedPubMedCentralGoogle Scholar
  56. Wang L, Muthu P, Szczesna-Cordary D, Kawai M (2013) Diversity and similarity of motor function and cross-bridge kinetics in papillary muscles of transgenic mice carrying myosin regulatory light chain mutations D166Vand R58Q. J Mol Cell Cardiol 62:153–163CrossRefPubMedGoogle Scholar
  57. Wang L, Sadayappan S, Kawai M (2014) Cardiac myosin binding protein C phosphorylation affects cross-bridge cycle’s elementary steps in a site-specific manner. PLoS One 9:1–21Google Scholar
  58. Wang L, Bahadir A, Kawai M (2015) High ionic strength depresses muscle contractility by decreasing both force per cross-bridge and the number of strongly attached cross-bridges. J Muscle Res Cell Motil 36:227–241CrossRefPubMedPubMedCentralGoogle Scholar
  59. Wannenburg T, Heijne GH, Geerdink JH, Van-Den-Dool HW, Janssen PM, DeTombe PP (2000) Cross-bridge kinetics in rat myocardium: effect of sarcomere length and calcium activation. Am J Physiol 279:H779-H790Google Scholar
  60. Zhao Y, Kawai M (1994) Kinetic and thermodynamic studies of the cross-bridge cycle in rabbit psoas muscle fibers. Biophys J 67:1655–1668CrossRefPubMedPubMedCentralGoogle Scholar
  61. Zhao Y, Swamy PMG, Humphries KA, Kawai M (1996) The effect of partial extraction of troponin C on the elementary steps of the cross-bridge cycle in rabbit psoas fibers. Biophys J 71:2759–2773CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  • Li Wang
    • 1
    • 2
  • Fan Bai
    • 2
  • Qing Zhang
    • 1
  • Weihua Song
    • 3
  • Andrew Messer
    • 3
  • Masataka Kawai
    • 2
  1. 1.School of Nursing, Medical CollegeSoochow UniversitySuzhouChina
  2. 2.Department of Anatomy and Cell BiologyUniversity of IowaIowa CityUSA
  3. 3.National Heart and Lung InstituteImperial College LondonLondonUK

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