Inhibition of Protein Kinases AKT and ERK1/2 Reduce the Carotid Body Chemoreceptor Response to Hypoxia in Adult Rats

  • Pablo Iturri
  • Vincent Joseph
  • Gloria Rodrigo
  • Aida Bairam
  • Jorge SolizEmail author
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 860)


The carotid body is the main mammalian oxygen-sensing organ regulating ventilation. Despite the carotid body is subjected of extensive anatomical and functional studies, little is yet known about the molecular pathways signaling the neurotransmission and neuromodulation of the chemoreflex activity. As kinases are molecules widely involved in motioning a broad number of neural processes, here we hypothesized that pathways of protein kinase B (AKT) and extracellular signal-regulated kinases ½ (ERK1/2) are implicated in the carotid body response to hypoxia. This hypothesis was tested using the in-vitro carotid body/carotid sinus nerve preparation (“en bloc”) from Sprague Dawley adult rats. Preparations were incubated for 60 min in tyrode perfusion solution (control) or containing 1 μM of LY294002 (AKT inhibitor), or 1 μM of UO-126 (ERK1/2 inhibitor). The carotid sinus nerve chemoreceptor discharge rate was recorded under baseline (perfusion solution bubbled with 5 % CO2 balanced in O2) and hypoxic (perfusion solution bubbled with 5 % CO2 balanced in N2) conditions. Compared to control, both inhibitors significantly decreased the normoxic and hypoxic carotid body chemoreceptor activity. LY294002- reduced carotid sinus nerve discharge rate in hypoxia by about 20 %, while UO-126 reduces the hypoxic response by 45 %. We concluded that both AKT and ERK1/2 pathways are crucial for the carotid body intracellular signaling process in response to hypoxia.


Carotid body Hypoxia Chemoreception ERK AKT 



The authors wish to thank to Melanie Pelletier for her superb assistance and Richard Kinkead for fruitful discussions. J.S. is supported by grants from The Molly Towell Perinatal research Foundation (MTPRF), The Fonds de la Recherche Québec en Santé (FRQ-S: MOP-26974), and The Canadian Institute for Health Research (CIHR: MOP-130258).


  1. Bairam A, Carroll JL (2005) Neurotransmitters in carotid body development. Respir Physiol Neurobiol 149(1–3):217–232PubMedGoogle Scholar
  2. Balasubramanian S, Teissere JA, Raju DV et al (2004) Hetero-oligomerization between GABAA and GABAB receptors regulates GABAB receptor trafficking. J Biol Chem 279(18):18840–18850PubMedGoogle Scholar
  3. Beaulieu JM, Sotnikova TD, Gainetdinov RR et al (2006) Paradoxical striatal cellular signaling responses to psychostimulants in hyperactive mice. J Biol Chem 281(43):32072–32080PubMedGoogle Scholar
  4. Beitner-Johnson D, Rust RT, Hsieh TC, Millhorn DE (2001) Hypoxia activates Akt and induces phosphorylation of GSK-3 in PC12 cells. Cell Signal 13(1):23–27PubMedGoogle Scholar
  5. Borowiec AS, Hague F, Harir N, Guenin S, Guerineau F, Gouilleux F, Roudbaraki M, Lassoued K, Ouadid-Ahidouch H (2007) IGF-1 activates hEAG K(+) channels through an Akt-dependent signaling pathway in breast cancer cells: role in cell proliferation. J Cell Physiol 212(3):690–701PubMedGoogle Scholar
  6. Buckler KJ, Vaughan-Jones RD (1994) Effects of hypoxia on membrane potential and intracellular calcium in rat neonatal carotid body type I cells. J Physiol 476(3):423–428PubMedPubMedCentralGoogle Scholar
  7. Chambard JC, Lefloch R, Pouyssegur J, Lenormand P (2007) ERK implication in cell cycle regulation. Biochim Biophys Acta 1773(8):1299–1310PubMedGoogle Scholar
  8. Chan WS, Sideris A, Sutachan JJ et al (2013) Differential regulation of proliferation and neuronal differentiation in adult rat spinal cord neural stem/progenitors by ERK1/2, Akt, and PLCgamma. Front Mol Neurosci 6:23PubMedPubMedCentralGoogle Scholar
  9. Cowen DS (2007) Serotonin and neuronal growth factors – a convergence of signaling pathways. J Neurochem 101(5):1161–1171PubMedGoogle Scholar
  10. Dey N, Howell BW, De PK et al (2005) CSK negatively regulates nerve growth factor induced neural differentiation and augments AKT kinase activity. Exp Cell Res 307(1):1–14PubMedGoogle Scholar
  11. Engelman JA, Luo J, Cantley LC (2006) The evolution of phosphatidylinositol 3-kinases as regulators of growth and metabolism. Nat Rev Genet 7(8):606–619PubMedGoogle Scholar
  12. Fang W, Gao G, Zhao H et al (2014) Role of the Akt/GSK-3beta/CRMP-2 pathway in axon degeneration of dopaminergic neurons resulting from MPP+ toxicity. Brain Res 1602:9–19PubMedGoogle Scholar
  13. Ganfornina MD, Lopez-Barneo J (1991) Single K+ channels in membrane patches of arterial chemoreceptor cells are modulated by O2 tension. Proc Natl Acad Sci U S A 88(7):2927–2930PubMedPubMedCentralGoogle Scholar
  14. Hirai K, Hayashi T, Chan PH, Zeng J, Yang GY, Basus VJ, James TL, Litt L (2004) P13K inhibition in neonatal rat brain slices during and after hypoxia reduces phospho-Akt and increases cytosolic cytochrome c and apoptosis. Brain Res Mol Brain Res 124(1):51–61PubMedGoogle Scholar
  15. Joseph V, Niane LM, Bairam A (2012) Antagonism of progesterone receptor suppresses carotid body responses to hypoxia and nicotine in rat pups. Neuroscience 207:103–109PubMedPubMedCentralGoogle Scholar
  16. Kelly A, Lynch MA (2000) Long-term potentiation in dentate gyrus of the rat is inhibited by the phosphoinositide 3-kinase inhibitor, wortmannin. Neuropharmacology 39(4):643–651PubMedGoogle Scholar
  17. Lee SM, Lee CT, Kim YW, Han SK, Shim YS, Yoo CG (2006) Hypoxia confers protection against apoptosis via PI3K/Akt and ERK pathways in lung cancer cells. Cancer Lett 242(2):231–238PubMedGoogle Scholar
  18. Lopez-Barneo J, Lopez-Lopez JR, Urena J et al (1988) Chemotransduction in the carotid body: K+ current modulated by PO2 in type I chemoreceptor cells. Science 241(4865):580–582PubMedGoogle Scholar
  19. Lopez-Barneo J, Pardal R, Ortega-Saenz P (2001) Cellular mechanism of oxygen sensing. Annu Rev Physiol 63:259–287PubMedGoogle Scholar
  20. Man HY, Wang Q, Lu WY, Ju W, Ahmadian G, Liu L, D’Souza S, Wong TP, Taghibiglou C, Lu J, Becker LE, Pei L, Liu F, Wymann MP, MacDonald JF, Wang YT (2003) Activation of PI3-kinase is required for AMPA receptor insertion during LTP of mEPSCs in cultured hippocampal neurons. Neuron 38(4):611–624PubMedGoogle Scholar
  21. Manning BD, Cantley LC (2007) AKT/PKB signaling: navigating downstream. Cell 129(7):1261–1274PubMedPubMedCentralGoogle Scholar
  22. Montoro RJ, Urena J, Fernandez-Chacon R et al (1996) Oxygen sensing by ion channels and chemotransduction in single glomus cells. J Gen Physiol 107(1):133–143PubMedGoogle Scholar
  23. Nishio H, Matsui K, Tsuji H, Tamura A, Suzuki K (2001) Immunolocalization of the mitogen-activated protein kinase signaling pathway in Hassall’s corpuscles of the human thymus. Acta Histochem 103(1):89–98PubMedGoogle Scholar
  24. Opazo P, Watabe AM, Grant SG, O’Dell TJ (2003) Phosphatidylinositol 3-kinase regulates the induction of long-term potentiation through extracellular signal-related kinase-independent mechanisms. J Neurosci 23(9):3679–3688PubMedGoogle Scholar
  25. Porzionato A, Macchi V, Parenti A, De Caro R (2010) Extracellular signal-regulated kinase and phosphatidylinositol-3-kinase/AKT signalling pathways in the human carotid body and peripheral ganglia. Acta Histochem 112(4):305–316PubMedGoogle Scholar
  26. Prabhakar NR, Overholt JL (2000) Cellular mechanisms of oxygen sensing at the carotid body: heme proteins and ion channels. Respir Physiol 122(2–3):209–221PubMedGoogle Scholar
  27. Rosenblum K, Futter M, Jones M et al (2000) ERKI/II regulation by the muscarinic acetylcholine receptors in neurons. J Neurosci 20(3):977–985PubMedGoogle Scholar
  28. Sanna PP, Cammalleri M, Berton F, Simpson C, Lutjens R, Bloom FE, Francesconi W (2002) Phosphatidylinositol 3-kinase is required for the expression but not for the induction or the maintenance of long-term potentiation in the hippocampal CA1 region. J Neurosci 22(9):3359–3365PubMedGoogle Scholar
  29. Su B, Karin M (1996) Mitogen-activated protein kinase cascades and regulation of gene expression. Curr Opin Immunol 8(3):402–411PubMedGoogle Scholar
  30. Taraviras S, Olli-Lahdesmaki T, Lymperopoulos A et al (2002) Subtype-specific neuronal differentiation of PC12 cells transfected with alpha2-adrenergic receptors. Eur J Cell Biol 81(6):363–374PubMedGoogle Scholar
  31. Teng L, Kou C, Lu C et al (2014) Involvement of the ERK pathway in the protective effects of glycyrrhizic acid against the MPP+-induced apoptosis of dopaminergic neuronal cells. Int J Mol Med 34(3):742–748PubMedPubMedCentralGoogle Scholar
  32. Urena J, Fernandez-Chacon R, Benot AR et al (1994) Hypoxia induces voltage-dependent Ca2+ entry and quantal dopamine secretion in carotid body glomus cells. Proc Natl Acad Sci U S A 91(21):10208–10211PubMedPubMedCentralGoogle Scholar
  33. Waskiewicz AJ, Cooper JA (1995) Mitogen and stress response pathways: MAP kinase cascades and phosphatase regulation in mammals and yeast. Curr Opin Cell Biol 7(6):798–805PubMedGoogle Scholar
  34. Weir EK, Lopez-Barneo J, Buckler KJ et al (2005) Acute oxygen-sensing mechanisms. N Engl J Med 353(19):2042–2055PubMedPubMedCentralGoogle Scholar
  35. Wiese S, Jablonka S, Holtmann B et al (2007) Adenosine receptor A2A-R contributes to motoneuron survival by transactivating the tyrosine kinase receptor TrkB. Proc Natl Acad Sci U S A 104(43):17210–17215PubMedPubMedCentralGoogle Scholar
  36. Yoshii A, Constantine-Paton M (2014) Postsynaptic localization of PSD-95 is regulated by all three pathways downstream of TrkB signaling. Front Synaptic Neurosci 6:6PubMedPubMedCentralGoogle Scholar
  37. Zhang X, Zhang Q, Tu J et al (2014) Prosurvival NMDA 2A receptor signaling mediates postconditioning neuroprotection in the hippocampus. Hippocampus 25(3):286–296PubMedGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Pablo Iturri
    • 1
    • 2
  • Vincent Joseph
    • 1
  • Gloria Rodrigo
    • 2
  • Aida Bairam
    • 3
    • 4
  • Jorge Soliz
    • 1
    Email author
  1. 1.Department of Pediatrics, Centre de Recherche de l’Hôpital St-François d’Assise (CR-SFA), Faculty of MedicineCentre Hospitalier Universitaire de Québec (CHUQ), Laval UniversityQuébecCanada
  2. 2.Molecular Biology and Biotechnology InstituteUniversidad Mayor de San AndresLa PazBolivia
  3. 3.Pediatric Department, Laval University, Research Center of Centre HospitalierUniversitaire de QuebecQuebec CityCanada
  4. 4.Centre de recherche du CHUQHSFAQuébecCanada

Personalised recommendations