Basic Research in Cardiology

, 114:47 | Cite as

Glucocorticoids preserve the t-tubular system in ventricular cardiomyocytes by upregulation of autophagic flux

  • Thomas SeidelEmail author
  • Dominik J. Fiegle
  • Tim J. Baur
  • Anne Ritzer
  • Sandra Nay
  • Christian Heim
  • Michael Weyand
  • Hendrik Milting
  • Robert H. Oakley
  • John A. Cidlowski
  • Tilmann VolkEmail author
Original Contribution


A major contributor to contractile dysfunction in heart failure is remodelling and loss of the cardiomyocyte transverse tubular system (t-system), but underlying mechanisms and signalling pathways remain elusive. It has been shown that dexamethasone promotes t-tubule development in stem cell-derived cardiomyocytes and that cardiomyocyte-specific glucocorticoid receptor (GR) knockout (GRKO) leads to heart failure. Here, we studied if the t-system is altered in GRKO hearts and if GR signalling is required for t-system preservation in adult cardiomyocytes. Confocal and 3D STED microscopy of myocardium from cardiomyocyte-specific GRKO mice revealed decreased t-system density and increased distances between ryanodine receptors (RyR) and L-type Ca2+ channels (LTCC). Because t-system remodelling and heart failure are intertwined, we investigated the underlying mechanisms in vitro. Ventricular cardiomyocytes from failing human and healthy adult rat hearts cultured in the absence of glucocorticoids (CTRL) showed distinctively lower t-system density than cells treated with dexamethasone (EC50 1.1 nM) or corticosterone. The GR antagonist mifepristone abrogated the effect of dexamethasone. Dexamethasone improved RyR–LTCC coupling and synchrony of intracellular Ca2+ release, but did not alter expression levels of t-system-associated proteins junctophilin-2 (JPH2), bridging integrator-1 (BIN1) or caveolin-3 (CAV3). Rather, dexamethasone upregulated LC3B and increased autophagic flux. The broad-spectrum protein kinase inhibitor staurosporine prevented dexamethasone-induced upregulation of autophagy and t-system preservation, and autophagy inhibitors bafilomycin A and chloroquine accelerated t-system loss. Conversely, induction of autophagy by rapamycin or amino acid starvation preserved the t-system. These findings suggest that GR signalling and autophagy are critically involved in t-system preservation and remodelling in the heart.


Autophagy Transverse tubular system Glucocorticoid receptor Excitation–contraction coupling Calcium signalling Remodelling Heart failure 



We would like to thank Celine Grüninger, Lorenz McCargo, Jessica Rinke and Ralf Rinke for excellent technical support, and Philipp Tripal from the Optical Imaging Centre Erlangen (OICE) for excellent assistance with STED microscopy.


This work was supported by grants from the Interdisciplinary Centre for Clinical Research (IZKF) at the University Hospital of the University of Erlangen-Nuremberg (Grant no. J65).

Compliance with ethical standards

Conflict of interest

None declared.

Supplementary material

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Supplementary material 1 (AVI 14392 kb)
395_2019_758_MOESM2_ESM.avi (18.7 mb)
Supplementary material 2 (AVI 19100 kb)
395_2019_758_MOESM3_ESM.pdf (4.4 mb)
Supplementary material 3 (PDF 4470 kb)


  1. 1.
    Al-Qusairi L, Laporte J (2011) T-tubule biogenesis and triad formation in skeletal muscle and implication in human diseases. Skelet Muscle 1:26. CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Banyasz T, Lozinskiy I, Payne CE, Edelmann S, Norton B, Chen B, Chen-Izu Y, Izu LT, Balke CW (2008) Transformation of adult rat cardiac myocytes in primary culture. Exp Physiol 93:370–382. CrossRefPubMedGoogle Scholar
  3. 3.
    Beavers DL, Landstrom AP, Chiang DY, Wehrens XH (2014) Emerging roles of junctophilin-2 in the heart and implications for cardiac diseases. Cardiovasc Res 103:198–205. CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Bishu K, Ogut O, Kushwaha S, Mohammed SF, Ohtani T, Xu X, Brozovich FV, Redfield MM (2013) Anti-remodeling effects of rapamycin in experimental heart failure: dose response and interaction with angiotensin receptor blockade. PLoS One 8:e81325. CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Bryant SM, Kong CHT, Watson JJ, Gadeberg HC, Roth DM, Patel HH, Cannell MB, James AF, Orchard CH (2018) Caveolin-3 KO disrupts t-tubule structure and decreases t-tubular ICa density in mouse ventricular myocytes. Am J Physiol Heart Circ Physiol 315:H1101–H1111. CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Caldwell JL, Smith CE, Taylor RF, Kitmitto A, Eisner DA, Dibb KM, Trafford AW (2014) Dependence of cardiac transverse tubules on the BAR domain protein amphiphysin II (BIN-1). Circ Res 115:986–996. CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Checchia PA, Backer CL, Bronicki RA, Baden HP, Crawford SE, Green TP, Mavroudis C (2003) Dexamethasone reduces postoperative troponin levels in children undergoing cardiopulmonary bypass. Crit Care Med 31:1742–1745. CrossRefPubMedGoogle Scholar
  8. 8.
    Crossman DJ, Shen X, Jullig M, Munro M, Hou Y, Middleditch M, Shrestha D, Li A, Lal S, Dos Remedios CG, Baddeley D, Ruygrok PN, Soeller C (2017) Increased collagen within the transverse tubules in human heart failure. Cardiovasc Res 113:879–891. CrossRefPubMedGoogle Scholar
  9. 9.
    Crossman DJ, Young AA, Ruygrok PN, Nason GP, Baddelely D, Soeller C, Cannell MB (2015) T-tubule disease: relationship between t-tubule organization and regional contractile performance in human dilated cardiomyopathy. J Mol Cell Cardiol 84:170–178. CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Cruz-Topete D, Oakley RH, Carroll NG, He B, Myers PH, Xu X, Watts MN, Trosclair K, Glasscock E, Dominic P, Cidlowski JA (2019) Deletion of the cardiomyocyte glucocorticoid receptor leads to sexually dimorphic changes in cardiac gene expression and progression to heart failure. J Am Heart Assoc 8:e011012. CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    De La Mata A, Tajada S, O’Dwyer S, Matsumoto C, Dixon RE, Hariharan N, Moreno CM, Santana LF (2019) BIN1 induces the formation of T-tubules and adult-like Ca(2+) release units in developing cardiomyocytes. Stem Cells 37:54–64. CrossRefGoogle Scholar
  12. 12.
    Frisk M, Ruud M, Espe EK, Aronsen JM, Roe AT, Zhang L, Norseng PA, Sejersted OM, Christensen GA, Sjaastad I, Louch WE (2016) Elevated ventricular wall stress disrupts cardiomyocyte t-tubule structure and calcium homeostasis. Cardiovasc Res 112:443–451. CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Guo X, Huang C, Lian K, Wang S, Zhao H, Yan F, Zhang X, Zhang J, Xie H, An R, Tao L (2016) BCKA down-regulates mTORC2-Akt signal and enhances apoptosis susceptibility in cardiomyocytes. Biochem Biophys Res Commun 480:106–113. CrossRefPubMedGoogle Scholar
  14. 14.
    Heinzel FR, Bito V, Biesmans L, Wu M, Detre E, von Wegner F, Claus P, Dymarkowski S, Maes F, Bogaert J, Rademakers F, D’Hooge J, Sipido K (2008) Remodeling of T-tubules and reduced synchrony of Ca2+ release in myocytes from chronically ischemic myocardium. Circ Res 102:338–346. CrossRefPubMedGoogle Scholar
  15. 15.
    Hong T, Yang H, Zhang SS, Cho HC, Kalashnikova M, Sun B, Zhang H, Bhargava A, Grabe M, Olgin J, Gorelik J, Marban E, Jan LY, Shaw RM (2014) Cardiac BIN1 folds T-tubule membrane, controlling ion flux and limiting arrhythmia. Nat Med 20:624–632. CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Humeres C, Frangogiannis NG (2019) Fibroblasts in the infarcted, remodeling, and failing heart. JACC Basic Transl Sci 4:449–467. CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Jayasinghe ID, Baddeley D, Kong CH, Wehrens XH, Cannell MB, Soeller C (2012) Nanoscale organization of junctophilin-2 and ryanodine receptors within peripheral couplings of rat ventricular cardiomyocytes. Biophys J 102:L19–L21. CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Jones PP, MacQuaide N, Louch WE (2018) Dyadic plasticity in cardiomyocytes. Front Physiol 9:1773. CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Kawai M, Hussain M, Orchard CH (1999) Excitation–contraction coupling in rat ventricular myocytes after formamide-induced detubulation. Am J Physiol Heart Circ Physiol 277:H603–H609. CrossRefGoogle Scholar
  20. 20.
    Kim J, Kundu M, Viollet B, Guan KL (2011) AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol 13:132–141. CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Kong CHT, Bryant SM, Watson JJ, Roth DM, Patel HH, Cannell MB, James AF, Orchard CH (2019) Cardiac-specific overexpression of caveolin-3 preserves t-tubular ICa during heart failure in mice. Exp Physiol. CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Kostin S, Scholz D, Shimada T, Maeno Y, Mollnau H, Hein S, Schaper J (1998) The internal and external protein scaffold of the T-tubular system in cardiomyocytes. Cell Tissue Res 294:449–460CrossRefGoogle Scholar
  23. 23.
    Lawless M, Caldwell JL, Radcliffe EJ, Smith CER, Madders GWP, Hutchings DC, Woods LS, Church SJ, Unwin RD, Kirkwood GJ, Becker LK, Pearman CM, Taylor RF, Eisner DA, Dibb KM, Trafford AW (2019) Phosphodiesterase 5 inhibition improves contractile function and restores transverse tubule loss and catecholamine responsiveness in heart failure. Sci Rep 9:6801. CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Li H, Lichter JG, Seidel T, Tomaselli GF, Bridge JH, Sachse FB (2015) Cardiac resynchronization therapy reduces subcellular heterogeneity of ryanodine receptors, T-tubules, and Ca2+ sparks produced by dyssynchronous heart failure. Circ Heart Fail 8:1105–1114. CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Lipsett DB, Frisk M, Aronsen JM, Norden ES, Buonarati OR, Cataliotti A, Hell JW, Sjaastad I, Christensen G, Louch WE (2019) Cardiomyocyte substructure reverts to an immature phenotype during heart failure. J Physiol. CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Lother A, Deng L, Huck M, Furst D, Kowalski J, Esser JS, Moser M, Bode C, Hein L (2018) Endothelial cell mineralocorticoid receptors oppose VEGF-induced gene expression and angiogenesis. J Endocrinol. CrossRefPubMedGoogle Scholar
  27. 27.
    Louch WE, Bito V, Heinzel FR, Macianskiene R, Vanhaecke J, Flameng W, Mubagwa K, Sipido KR (2004) Reduced synchrony of Ca2+ release with loss of T-tubules-a comparison to Ca2+ release in human failing cardiomyocytes. Cardiovasc Res 62:63–73. CrossRefPubMedGoogle Scholar
  28. 28.
    Manfra O, Frisk M, Louch WE (2017) Regulation of cardiomyocyte T-tubular structure: opportunities for therapy. Curr Heart Fail Rep 14:167–178. CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    McMullen JR, Sherwood MC, Tarnavski O, Zhang L, Dorfman AL, Shioi T, Izumo S (2004) Inhibition of mTOR signaling with rapamycin regresses established cardiac hypertrophy induced by pressure overload. Circulation 109:3050–3055. CrossRefPubMedGoogle Scholar
  30. 30.
    Mizushima N, Yoshimori T, Levine B (2010) Methods in mammalian autophagy research. Cell 140:313–326. CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Nakai A, Yamaguchi O, Takeda T, Higuchi Y, Hikoso S, Taniike M, Omiya S, Mizote I, Matsumura Y, Asahi M, Nishida K, Hori M, Mizushima N, Otsu K (2007) The role of autophagy in cardiomyocytes in the basal state and in response to hemodynamic stress. Nat Med 13:619–624. CrossRefPubMedGoogle Scholar
  32. 32.
    Nishida K, Otsu K (2016) Autophagy during cardiac remodeling. J Mol Cell Cardiol 95:11–18. CrossRefPubMedGoogle Scholar
  33. 33.
    Oakley RH, Cruz-Topete D, He B, Foley JF, Myers PH, Xu X, Gomez-Sanchez CE, Chambon P, Willis MS, Cidlowski JA (2019) Cardiomyocyte glucocorticoid and mineralocorticoid receptors directly and antagonistically regulate heart disease in mice. Sci Signal. CrossRefPubMedGoogle Scholar
  34. 34.
    Oakley RH, Ren R, Cruz-Topete D, Bird GS, Myers PH, Boyle MC, Schneider MD, Willis MS, Cidlowski JA (2013) Essential role of stress hormone signaling in cardiomyocytes for the prevention of heart disease. Proc Natl Acad Sci USA 110:17035–17040. CrossRefPubMedGoogle Scholar
  35. 35.
    Oakley RH, Revollo J, Cidlowski JA (2012) Glucocorticoids regulate arrestin gene expression and redirect the signaling profile of G protein-coupled receptors. Proc Natl Acad Sci USA 109:17591–17596. CrossRefPubMedGoogle Scholar
  36. 36.
    Parikh SS, Blackwell DJ, Gomez-Hurtado N, Frisk M, Wang L, Kim K, Dahl CP, Fiane A, Tonnessen T, Kryshtal DO, Louch WE, Knollmann BC (2017) Thyroid and glucocorticoid hormones promote functional T-tubule development in human-induced pluripotent stem cell-derived cardiomyocytes. Circ Res 121:1323–1330. CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Pinali C, Malik N, Davenport JB, Allan LJ, Murfitt L, Iqbal MM, Boyett MR, Wright EJ, Walker R, Zhang Y, Dobryznski H, Holt CM, Kitmitto A (2017) Post-myocardial infarction T-tubules form enlarged branched structures with dysregulation of junctophilin-2 and bridging integrator 1 (BIN-1). J Am Heart Assoc. CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Poindexter BJ, Smith JR, Buja LM, Bick RJ (2001) Calcium signaling mechanisms in dedifferentiated cardiac myocytes: comparison with neonatal and adult cardiomyocytes. Cell Calcium 30:373–382. CrossRefPubMedGoogle Scholar
  39. 39.
    Quinn TA, Camelliti P, Rog-Zielinska EA, Siedlecka U, Poggioli T, O’Toole ET, Knopfel T, Kohl P (2016) Electrotonic coupling of excitable and nonexcitable cells in the heart revealed by optogenetics. Proc Natl Acad Sci USA 113:14852–14857. CrossRefPubMedGoogle Scholar
  40. 40.
    Richardson RV, Batchen EJ, Thomson AJ, Darroch R, Pan X, Rog-Zielinska EA, Wyrzykowska W, Scullion K, Al-Dujaili EA, Diaz ME, Moran CM, Kenyon CJ, Gray GA, Chapman KE (2017) Glucocorticoid receptor alters isovolumetric contraction and restrains cardiac fibrosis. J Endocrinol 232:437–450. CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Riquelme JA, Chavez MN, Mondaca-Ruff D, Bustamante M, Vicencio JM, Quest AF, Lavandero S (2016) Therapeutic targeting of autophagy in myocardial infarction and heart failure. Expert Rev Cardiovasc Ther 14:1007–1019. CrossRefPubMedGoogle Scholar
  42. 42.
    Rog-Zielinska EA, Thomson A, Kenyon CJ, Brownstein DG, Moran CM, Szumska D, Michailidou Z, Richardson J, Owen E, Watt A, Morrison H, Forrester LM, Bhattacharya S, Holmes MC, Chapman KE (2013) Glucocorticoid receptor is required for foetal heart maturation. Hum Mol Genet 22:3269–3282. CrossRefPubMedGoogle Scholar
  43. 43.
    Rohdewald P, Mollmann H, Barth J, Rehder J, Derendorf H (1987) Pharmacokinetics of dexamethasone and its phosphate ester. Biopharm Drug Dispos 8:205–212CrossRefGoogle Scholar
  44. 44.
    Sainte-Marie Y, Nguyen Dinh Cat A, Perrier R, Mangin L, Soukaseum C, Peuchmaur M, Tronche F, Farman N, Escoubet B, Benitah JP, Jaisser F (2007) Conditional glucocorticoid receptor expression in the heart induces atrio-ventricular block. FASEB J 21:3133–3141. CrossRefPubMedGoogle Scholar
  45. 45.
    Sarbassov DD, Ali SM, Sengupta S, Sheen JH, Hsu PP, Bagley AF, Markhard AL, Sabatini DM (2006) Prolonged rapamycin treatment inhibits mTORC2 assembly and Akt/PKB. Mol Cell 22:159–168. CrossRefPubMedGoogle Scholar
  46. 46.
    Seidel T, Navankasattusas S, Ahmad A, Diakos NA, Xu WD, Tristani-Firouzi M, Bonios MJ, Taleb I, Li DY, Selzman CH, Drakos SG, Sachse FB (2017) Sheet-like remodeling of the transverse tubular system in human heart failure impairs excitation–contraction coupling and functional recovery by mechanical unloading. Circulation 135:1632–1645. CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Seidel T, Sankarankutty AC, Sachse FB (2017) Remodeling of the transverse tubular system after myocardial infarction in rabbit correlates with local fibrosis: a potential role of biomechanics. Prog Biophys Mol Biol 130:302–314. CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Shah SJ, Aistrup GL, Gupta DK, O’Toole MJ, Nahhas AF, Schuster D, Chirayil N, Bassi N, Ramakrishna S, Beussink L, Misener S, Kane B, Wang D, Randolph B, Ito A, Wu M, Akintilo L, Mongkolrattanothai T, Reddy M, Kumar M, Arora R, Ng J, Wasserstrom JA (2014) Ultrastructural and cellular basis for the development of abnormal myocardial mechanics during the transition from hypertension to heart failure. Am J Physiol Heart Circ Physiol 306:H88–H100. CrossRefPubMedGoogle Scholar
  49. 49.
    Shimizu N, Yoshikawa N, Ito N, Maruyama T, Suzuki Y, Takeda S, Nakae J, Tagata Y, Nishitani S, Takehana K, Sano M, Fukuda K, Suematsu M, Morimoto C, Tanaka H (2011) Crosstalk between glucocorticoid receptor and nutritional sensor mTOR in skeletal muscle. Cell Metab 13:170–182. CrossRefPubMedGoogle Scholar
  50. 50.
    Smith CER, Trafford AW, Caldwell JL, Dibb KM (2018) Physiology and patho-physiology of the cardiac transverse tubular system. Curr Opin Physiol 1:153–160. CrossRefGoogle Scholar
  51. 51.
    Spath JA, Lefer AM (1975) Effects of dexamethasone on myocardial cells in the early phase of acute myocardial infarction. Am Heart J 90:50–55. CrossRefPubMedGoogle Scholar
  52. 52.
    Tato I, Bartrons R, Ventura F, Rosa JL (2011) Amino acids activate mammalian target of rapamycin complex 2 (mTORC2) via PI3K/Akt signaling. J Biol Chem 286:6128–6142. CrossRefPubMedGoogle Scholar
  53. 53.
    Tian Q, Pahlavan S, Oleinikow K, Jung J, Ruppenthal S, Scholz A, Schumann C, Kraegeloh A, Oberhofer M, Lipp P, Kaestner L (2012) Functional and morphological preservation of adult ventricular myocytes in culture by sub-micromolar cytochalasin D supplement. J Mol Cell Cardiol 52:113–124. CrossRefPubMedGoogle Scholar
  54. 54.
    Troncoso R, Paredes F, Parra V, Gatica D, Vasquez-Trincado C, Quiroga C, Bravo-Sagua R, Lopez-Crisosto C, Rodriguez AE, Oyarzun AP, Kroemer G, Lavandero S (2014) Dexamethasone-induced autophagy mediates muscle atrophy through mitochondrial clearance. Cell Cycle 13:2281–2295. CrossRefPubMedPubMedCentralGoogle Scholar
  55. 55.
    Vlahakis A, Graef M, Nunnari J, Powers T (2014) TOR complex 2-Ypk1 signaling is an essential positive regulator of the general amino acid control response and autophagy. Proc Natl Acad Sci USA 111:10586–10591. CrossRefPubMedGoogle Scholar
  56. 56.
    Wei S, Guo A, Chen B, Kutschke W, Xie YP, Zimmerman K, Weiss RM, Anderson ME, Cheng H, Song LS (2010) T-tubule remodeling during transition from hypertrophy to heart failure. Circ Res 107:520–531. CrossRefPubMedPubMedCentralGoogle Scholar
  57. 57.
    Xu B, Strom J, Chen QM (2011) Dexamethasone induces transcriptional activation of Bcl-xL gene and inhibits cardiac injury by myocardial ischemia. Eur J Pharmacol 668:194–200. CrossRefPubMedPubMedCentralGoogle Scholar
  58. 58.
    Yoshii SR, Mizushima N (2017) Monitoring and measuring autophagy. Int J Mol Sci. CrossRefPubMedPubMedCentralGoogle Scholar
  59. 59.
    Zhang C, Chen B, Guo A, Zhu Y, Miller JD, Gao S, Yuan C, Kutschke W, Zimmerman K, Weiss RM, Wehrens XH, Hong J, Johnson FL, Santana LF, Anderson ME, Song LS (2014) Microtubule-mediated defects in junctophilin-2 trafficking contribute to myocyte transverse-tubule remodeling and Ca2+ handling dysfunction in heart failure. Circulation 129:1742–1750. CrossRefPubMedPubMedCentralGoogle Scholar

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© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Cellular and Molecular PhysiologyFriedrich-Alexander-Universität Erlangen-NürnbergErlangenGermany
  2. 2.Muscle Research Center Erlangen (MURCE), Friedrich-Alexander-Universität Erlangen-NürnbergErlangenGermany
  3. 3.Department of Cardiac SurgeryFriedrich-Alexander-Universität Erlangen-NürnbergErlangenGermany
  4. 4.Erich and Hanna Klessmann Institute, Clinic for Thoracic and Cardiovascular Surgery, Heart and Diabetes Centre NRWRuhr-University BochumBad OeynhausenGermany
  5. 5.Signal Transduction Laboratory, Department of Health and Human ServicesNational Institute of Environmental Health Sciences, National Institutes of HealthResearch Triangle ParkUSA

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