Skip to main content
SpringerLink
Log in

β-Adrenergic receptor stimulation induces endoplasmic reticulum stress in adult cardiac myocytes: role in apoptosis

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Accumulation of misfolded proteins and alterations in calcium homeostasis induces endoplasmic reticulum (ER) stress, leading to apoptosis. In this study, we tested the hypothesis that β-AR stimulation induces ER stress, and induction of ER stress plays a pro-apoptotic role in cardiac myocytes. Using thapsigargin and brefeldin A, we demonstrate that ER stress induces apoptosis in adult rat ventricular myocytes (ARVMs). β-AR-stimulation (isoproterenol; 3h) significantly increased expression of ER stress proteins, such as GRP-78, Gadd-153, and Gadd-34, while activating caspase-12 in ARVMs. In most parts, these effects were mimicked by thapsigargin. β-AR stimulation for 15 min increased PERK and eIF-2α phosphorylation. PERK phosphorylation remained higher, while eIF-2α phosphorylation declined thereafter, reaching to ~50% below basal levels at 3 h after β-AR stimulation. This decline in eIF-2α phosphorylation was prevented by β1-AR, not by β2-AR antagonist. Forskolin, adenylyl cyclase activator, simulated the effects of ISO on eIF-2α phosphorylation. Salubrinal (SAL), an ER stress inhibitor, maintained eIF-2α phosphorylation and inhibited β-AR-stimulated apoptosis. Furthermore, inhibition of caspase-12 using z-ATAD inhibited β-AR-stimulated and thapsigargin-induced apoptosis. In vivo, β-AR stimulation induced ER stress in the mouse heart as evidenced by increased expression of GRP-78 and Gadd-153, activation of caspase-12, and dephosphorylation of eIF-2α. SAL maintained phosphorylation of eIF-2α, inhibited activation of caspase-12, and decreased β-AR-stimulated apoptosis in the heart. Thus, β-AR stimulation induces ER stress in cardiac myocytes and in the heart, and induction of ER stress plays a pro-apoptotic role.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Hasking GJ, Esler MD, Jennings GL, Burton D, Johns JA, Korner PI (1986) Norepinephrine spillover to plasma in patients with congestive heart failure: evidence of increased overall and cardiorenal sympathetic nervous activity. Circulation 73:615–621

    Article  PubMed  CAS  Google Scholar 

  2. Cohn JN, Levine TB, Olivari MT, Garberg V, Lura D, Francis GS, Simon AB, Rector T (1984) Plasma norepinephrine as a guide to prognosis in patients with chronic congestive heart failure. N Engl J Med 311:819–823

    Article  PubMed  CAS  Google Scholar 

  3. Kajstura J, Bolli R, Sonnenblick EH, Anversa P, Leri A (2006) Cause of death: suicide. J Mol Cell Cardiol 40:425–437

    Article  PubMed  CAS  Google Scholar 

  4. Nadal-Ginard B, Kajstura J, Leri A, Anversa P (2003) Myocyte death, growth, and regeneration in cardiac hypertrophy and failure. Circ Res 92:139–150

    Article  PubMed  CAS  Google Scholar 

  5. Krishnamurthy P, Subramanian V, Singh M, Singh K (2007) β1 integrins modulate β-adrenergic receptor-stimulated cardiac myocyte apoptosis and myocardial remodeling. Hypertension 49:865–872

    Article  PubMed  CAS  Google Scholar 

  6. Shizukuda Y, Buttrick PM, Geenen DL, Borczuk AC, Kitsis RN, Sonnenblick EH (1998) Beta-adrenergic stimulation causes cardiocyte apoptosis: influence of tachycardia and hypertrophy. Am J Physiol 275:H961–H968

    PubMed  CAS  Google Scholar 

  7. Communal C, Singh K, Pimentel DR, Colucci WS (1998) Norepinephrine stimulates apoptosis in adult rat ventricular myocytes by activation of the beta-adrenergic pathway. Circulation 98:1329–1334

    PubMed  CAS  Google Scholar 

  8. Iwai-Kanai E, Hasegawa K, Araki M, Kakita T, Morimoto T, Sasayama S (1999) Alpha- and beta-adrenergic pathways differentially regulate cell type-specific apoptosis in rat cardiac myocytes. Circulation 100:305–311

    PubMed  CAS  Google Scholar 

  9. Remondino A, Kwon SH, Communal C, Pimentel DR, Sawyer DB, Singh K, Colucci WS (2003) Beta-adrenergic receptor-stimulated apoptosis in cardiac myocytes is mediated by reactive oxygen species/c-Jun NH2-terminal kinase-dependent activation of the mitochondrial pathway. Circ Res 92:136–138

    Article  PubMed  CAS  Google Scholar 

  10. Menon B, Johnson JN, Ross RS, Singh M, Singh K (2007) Glycogen synthase kinase-3beta plays a pro-apoptotic role in beta-adrenergic receptor-stimulated apoptosis in adult rat ventricular myocytes: role of beta1 integrins. J Mol Cell Cardiol 42:653–661

    Article  PubMed  CAS  Google Scholar 

  11. Rao RV, Ellerby HM, Bredesen DE (2004) Coupling endoplasmic reticulum stress to the cell death program. Cell Death Differ 11:372–380

    Article  PubMed  CAS  Google Scholar 

  12. Szegezdi E, FitzGerald U, Samali A (2003) Caspase-12 and ER-stress-mediated apoptosis: the story so far. Ann N Y Acad Sci 1010:186–194

    Article  PubMed  CAS  Google Scholar 

  13. Rutkowski DT, Kaufman RJ (2004) A trip to the ER: coping with stress. Trends Cell Biol 14:20–28

    Article  PubMed  CAS  Google Scholar 

  14. Okada K, Minamino T, Tsukamoto Y, Liao Y, Tsukamoto O, Takashima S, Hirata A, Fujita M, Nagamachi Y, Nakatani T, Yutani C, Ozawa K, Ogawa S, Tomoike H, Hori M, Kitakaze M (2004) Prolonged endoplasmic reticulum stress in hypertrophic and failing heart after aortic constriction: possible contribution of endoplasmic reticulum stress to cardiac myocyte apoptosis. Circulation 110:705–712

    Article  PubMed  Google Scholar 

  15. Kostin S, Pool L, Elsasser A, Hein S, Drexler HC, Arnon E, Hayakawa Y, Zimmermann R, Bauer E, Klovekorn WP, Schaper J (2003) Myocytes die by multiple mechanisms in failing human hearts. Circ Res 92:715–724

    Article  PubMed  CAS  Google Scholar 

  16. Xu J, Wang G, Wang Y, Liu Q, Xu W, Tan Y, Cai L (2009) Diabetes- and angiotensin II-induced cardiac endoplasmic reticulum stress and cell death: metallothionein protection. J Cell Mol Med 13:1499–1512

    Article  PubMed  CAS  Google Scholar 

  17. Hamada H, Suzuki M, Yuasa S, Mimura N, Shinozuka N, Takada Y, Suzuki M, Nishino T, Nakaya H, Koseki H, Aoe T (2004) Dilated cardiomyopathy caused by aberrant endoplasmic reticulum quality control in mutant KDEL receptor transgenic mice. Mol Cell Biol 24:8007–8017

    Article  PubMed  CAS  Google Scholar 

  18. Liang CS, Mao W, Liu J (2008) Pro-apoptotic effects of anti-beta1-adrenergic receptor antibodies in cultured rat cardiomyocytes: actions on endoplasmic reticulum and the prosurvival PI3K-Akt pathway. Autoimmunity 41:434–441

    Article  PubMed  CAS  Google Scholar 

  19. Menon B, Singh M, Ross RS, Johnson JN, Singh K (2006) Beta-adrenergic receptor-stimulated apoptosis in adult cardiac myocytes involves MMP-2-mediated disruption of beta1 integrin signaling and mitochondrial pathway. Am J Physiol Cell Physiol 290:C254–C261

    Article  PubMed  CAS  Google Scholar 

  20. Communal C, Singh K, Sawyer DB, Colucci WS (1999) Opposing effects of beta(1)- and beta(2)-adrenergic receptors on cardiac myocyte apoptosis : role of a pertussis toxin-sensitive G protein. Circulation 100:2210–2212

    PubMed  CAS  Google Scholar 

  21. Boyce M, Bryant KF, Jousse C, Long K, Harding HP, Scheuner D, Kaufman RJ, Ma D, Coen DM, Ron D, Yuan J (2005) A selective inhibitor of eIF2alpha dephosphorylation protects cells from ER stress. Science 307:935–939

    Article  PubMed  CAS  Google Scholar 

  22. Luthra S, Dong J, Gramajo AL, Chwa M, Kim DW, Neekhra A, Kuppermann BD, Kenney MC (2008) 7-Ketocholesterol activates caspases-3/7, -8, and -12 in human microvascular endothelial cells in vitro. Microvasc Res 75:343–350

    Article  PubMed  CAS  Google Scholar 

  23. Germain M, Mathai JP, McBride HM, Shore GC (2005) Endoplasmic reticulum BIK initiates DRP1-regulated remodelling of mitochondrial cristae during apoptosis. EMBO J 24:1546–1556

    Article  PubMed  CAS  Google Scholar 

  24. Zhu Y, Fenik P, Zhan G, Sanfillipo-Cohn B, Naidoo N, Veasey SC (2008) Eif-2a protects brainstem motoneurons in a murine model of sleep apnea. J Neurosci 28:2168–2178

    Article  PubMed  CAS  Google Scholar 

  25. Wiseman RL, Balch WE (2005) A new pharmacology–drugging stressed folding pathways. Trends Mol Med 11:347–350

    Article  PubMed  CAS  Google Scholar 

  26. Nakagawa T, Zhu H, Morishima N, Li E, Xu J, Yankner BA, Yuan J (2000) Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-beta. Nature 403:98–103

    Article  PubMed  CAS  Google Scholar 

  27. Hetz C, Russelakis-Carneiro M, Maundrell K, Castilla J, Soto C (2003) Caspase-12 and endoplasmic reticulum stress mediate neurotoxicity of pathological prion protein. EMBO J 22:5435–5445

    Article  PubMed  CAS  Google Scholar 

  28. Shizukuda Y, Buttrick PM (2002) Subtype specific roles of beta-adrenergic receptors in apoptosis of adult rat ventricular myocytes. J Mol Cell Cardiol 34:823–831

    Article  PubMed  CAS  Google Scholar 

  29. Kessel D (2006) Protection of Bcl-2 by salubrinal. Biochem Biophys Res Commun 346:1320–1323

    Article  PubMed  CAS  Google Scholar 

  30. Fu HY, Minamino T, Tsukamoto O, Sawada T, Asai M, Kato H, Asano Y, Fujita M, Takashima S, Hori M, Kitakaze M (2008) Overexpression of endoplasmic reticulum-resident chaperone attenuates cardiomyocyte death induced by proteasome inhibition. Cardiovasc Res 79:600–610

    Article  PubMed  CAS  Google Scholar 

  31. Singh K, Xiao L, Remondino A, Sawyer DB, Colucci WS (2001) Adrenergic regulation of cardiac myocyte apoptosis. J Cell Physiol 189:257–265

    Article  PubMed  CAS  Google Scholar 

  32. Zaugg M, Xu W, Lucchinetti E, Shafiq SA, Jamali NZ, Siddiqui MA (2000) Beta-adrenergic receptor subtypes differentially affect apoptosis in adult rat ventricular myocytes. Circulation 102:344–350

    PubMed  CAS  Google Scholar 

  33. Colucci WS, Sawyer DB, Singh K, Communal C (2000) Adrenergic overload and apoptosis in heart failure: implications for therapy. J Card Fail 6:1–7

    Article  PubMed  CAS  Google Scholar 

  34. Wang X, Robbins J (2006) Heart failure and protein quality control. Circ Res 99:1315–1328

    Article  PubMed  CAS  Google Scholar 

  35. Nickson P, Toth A, Erhardt P (2007) PUMA is critical for neonatal cardiomyocyte apoptosis induced by endoplasmic reticulum stress. Cardiovasc Res 73:48–56

    Article  PubMed  CAS  Google Scholar 

  36. Kajstura J, Mansukhani M, Cheng W, Reiss K, Krajewski S, Reed JC, Quaini F, Sonnenblick EH, Anversa P (1995) Programmed cell death and expression of the protooncogene bcl-2 in myocytes during postnatal maturation of the heart. Exp Cell Res 219:110–121

    Article  PubMed  CAS  Google Scholar 

  37. Simpson P (1985) Stimulation of hypertrophy of cultured neonatal rat heart cells through an alpha 1-adrenergic receptor and induction of beating through an alpha 1- and beta 1-adrenergic receptor interaction. Evidence for independent regulation of growth and beating. Circ Res 56:884–894

    PubMed  CAS  Google Scholar 

  38. Clark WA, Rudnick SJ, LaPres JJ, Andersen LC, LaPointe MC (1993) Regulation of hypertrophy and atrophy in cultured adult heart cells. Circ Res 73:1163–1176

    PubMed  CAS  Google Scholar 

  39. Armstrong JL, Flockhart R, Veal GJ, Lovat PE, Redfern CP (2010) Regulation of endoplasmic reticulum stress-induced cell death by ATF4 in neuroectodermal tumor cells. J Biol Chem 285:6091–6100

    Article  PubMed  CAS  Google Scholar 

  40. Suragani RN, Ghosh S, Ehtesham NZ, Ramaiah KV (2006) Expression and purification of the subunits of human translational initiation factor 2 (eIF2): phosphorylation of eIF2 alpha and beta. Protein Expr Purif 47:225–233

    Article  PubMed  CAS  Google Scholar 

  41. Sokka AL, Putkonen N, Mudo G, Pryazhnikov E, Reijonen S, Khiroug L, Belluardo N, Lindholm D, Korhonen L (2007) Endoplasmic reticulum stress inhibition protects against excitotoxic neuronal injury in the rat brain. J Neurosci 27:901–908

    Article  PubMed  CAS  Google Scholar 

  42. Sperelakis N, Xiong Z, Haddad G, Masuda H (1994) Regulation of slow calcium channels of myocardial cells and vascular smooth muscle cells by cyclic nucleotides and phosphorylation. Mol Cell Biochem 140:103–117

    Article  PubMed  CAS  Google Scholar 

  43. Hoppe V, Hoppe J (2004) Mutations dislocate caspase-12 from the endoplasmatic reticulum to the cytosol. FEBS Lett 576:277–283

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

Technical help received from Barbara A. Connelly is appreciated. This study is supported by the National Institutes of Health (Grant numbers HL-091405 and HL-092459) and a Merit Review Grant (award number IO1BX000640) from the Biomedical Laboratory Research & Development Service of the VA office of Research and Development.

Conflicts of interest

None.

Author information

Authors and Affiliations

  1. Department of Physiology, James H Quillen College of Medicine, James H Quillen Veterans Affairs Medical Center, East Tennessee State University, PO Box 70576, Johnson City, TN, 37614, USA

    Suman Dalal, Cerrone R. Foster, Mahipal Singh & Krishna Singh

  2. Dr. B.R. Ambedkar Research Centre for Biomedical Research, University of Delhi (North Campus), Delhi, 110 007, India

    Bhudev C. Das

Authors
  1. Suman Dalal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Cerrone R. Foster
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bhudev C. Das
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mahipal Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Krishna Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Krishna Singh.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dalal, S., Foster, C.R., Das, B.C. et al. β-Adrenergic receptor stimulation induces endoplasmic reticulum stress in adult cardiac myocytes: role in apoptosis. Mol Cell Biochem 364, 59–70 (2012). https://doi.org/10.1007/s11010-011-1205-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-011-1205-7

Keywords

Search

Navigation