Molecular Neurobiology

, Volume 54, Issue 4, pp 2852–2868 | Cite as

The Protective Role of Selenium on Scopolamine-Induced Memory Impairment, Oxidative Stress, and Apoptosis in Aged Rats: The Involvement of TRPM2 and TRPV1 Channels

  • Hasan Balaban
  • Mustafa NazıroğluEmail author
  • Kadir Demirci
  • İshak Suat Övey


Inhibition of Ca2+ entry into the hippocampus and dorsal root ganglion (DRG) through inhibition of N-methyl-d-aspartate (NMDA) receptor antagonist drugs is the current standard of care in neuronal diseases such as Alzheimer’s disease, dementia, and peripheral pain. Oxidative stress activates Ca2+-permeable TRPM2 and TRPV1, and recent studies indicate that selenium (Se) is a potent TRPM2 and TRPV1 channel antagonist in the hippocampus and DRG. In this study, we investigated the neuroprotective properties of Se in primary hippocampal and DRG neuron cultures of aged rats when given alone or in combination with scopolamine (SCOP). Thirty-two aged (18–24 months old) rats were divided into four groups. The first and second groups received a placebo and SCOP (1 mg/kg/day), respectively. The third and fourth groups received intraperitoneal Se (1.5 mg/kg/ over day) and SCOP + Se, respectively. The hippocampal and DRG neurons also were stimulated in vitro with a TRPV1 channel agonist (capsaicin) and a TRPM2 channel agonist (cumene hydroperoxide). We found that Se was fully effective in reversing SCOP-induced TRPM2 and TRPV1 current densities as well as errors in working memory and reference memory. In addition, Se completely reduced SCOP-induced oxidative toxicity by modulating lipid peroxidation, reducing glutathione and glutathione peroxidase. The Se and SCOP + Se treatments also decreased poly (ADP-ribose) polymerase activity, intracellular free Ca2+ concentrations, apoptosis, and caspase 3, caspase 9, and mitochondrial membrane depolarization values in the hippocampus. In conclusion, the current study reports on the cellular level for SCOP and Se on the different endocytotoxic cascades for the first time. Notably, the research indicates that Se can result in remarkable neuroprotective and memory impairment effects in the hippocampal neurons of rats.

Graphical abstract

Possible molecular pathways of involvement of selenium (Se) in scopolamine (SCOP) induced apoptosis, oxidative stress, and calcium accumulation through TRPM2 and TRPV1 channels in the hippocampus neurons of aged rats. The TRPM2 channel is activated by ADP-ribose and oxidative stress, although it is inhibited by ACA. The TRPV1 channel is activated by oxidative stress and capsaicin, and it is blocked by capsazepine (CPZ). The beta-amyloid plaque induces oxidative stress in hippocampus. SCOP can result in augmented ROS release in hippocampal neurons, leading to Ca2+ uptake through TRPM2 and TRPV1 channels. Mitochondria were reported to accumulate Ca2+ provided that intracellular Ca2+ rises, thereby leading to the depolarization of mitochondrial membranes and release of apoptosis-inducing factors such as caspase 3 and caspase 9. Se reduced TRPM2 and TRPV1 channel activation through the modulation of aging oxidative reactions and Se-dependent glutathione peroxidase (GSH-Px) antioxidant pathways.


Apoptosis TRPV1 TRPM2 Selenium Oxidative stress Dementia 



Intracellular free calcium ion


N-(p-Amylcinnamoyl)anthranilic acid




Cumene hydroperoxide




Dimethyl sulfoxide


Dorsal root ganglion


Ethylene glycol-bis[2-aminoethyl-ether]-N,N,N,N-tetraacetic acid


Reduced glutathione


Glutathione peroxidase


Hank’s buffered salt solution


Poly (ADP-ribose) polymerase


radial arm-maze


Reference memory error


Reactive oxygen species




Transient receptor potential


Transient receptor potential Mu


Transient receptor potential vanilloid 1


Whole cell


Working memory error



The authors wish to thank researcher Bilal Çiğ and technicians Fatih Şahin and Muhammet Şahin (Neuroscience Research Center, SDU, Isparta, Turkey) for helping with the [Ca2+]i (Fura-2AM), patch-clamp, lipid peroxidation, and antioxidant analyses. The abstract of the study will be partially published in the “8th International Congress on Psychopharmacology & 4th International Symposium on Child and Adolescent Psychopharmacology, April 20–24, 2016, in Antalya, Turkey.”

Financial Disclosure

The study was supported by the Unit of Scientific Research Project (BAP), Süleyman Demirel University, Isparta, Turkey (Project Number BAP: 4257-TU-15). There is no financial disclosure for the current study.

Authorship Contributions

MN and KD formulated the hypothesis and were responsible for writing the report. İSO and HB were also responsible for the animal experiments such as the induction of memory injury through SCOP administration and injection of selenium. İSÖ performed the Ca2+ analyses Western blot, apoptosis, and mitochondrial depolarization analyses.

Compliance with Ethical Standards

The study was approved by the Local Experimental Animal Ethical Committee of Suleyman Demirel University (SDU) (protocol number 10.02.2015-03).

Conflict of Interest

The authors declare that they have no conflicts of interest.

Human and Animal Rights and Informed Consent

All research procedures and animal care complied with the guidelines of the International Association Study Plan for induction of memory injury. The animals were maintained and used according to the Animal Welfare Act and the Guide for the Care and Use of Laboratory.


  1. 1.
    LaFerla FM (2002) Calcium dyshomeostasis ann intracellular signalling in Alzheimer’s disease. Nat Rev Neurosci 3:862–872CrossRefPubMedGoogle Scholar
  2. 2.
    Pascale A, Etcheberrigaray R (1999) Calcium alterations in Alzheimer’s disease: pathophysiology, models and therapeutic opportunities. Pharmacol Res 39:81–88CrossRefPubMedGoogle Scholar
  3. 3.
    Nishimura I, Takazaki R, Kuwako K, Enokido Y, Yoshikawa K (2003) Upregulation and antiapoptotic role of endogenous Alzheimer amyloid precursor protein in dorsal root ganglion neurons. Exp Cell Res 286:241–251CrossRefPubMedGoogle Scholar
  4. 4.
    Kurz AF (2005) Uncommon neurodegenerative causes of dementia. Int Psychogeriatr 17(Suppl 1):S35–49CrossRefPubMedGoogle Scholar
  5. 5.
    Paula-Lima AC, Adasme T, Hidalgo C (2014) Contribution of Ca2+ release channels to hippocampal synaptic plasticity and spatial memory: potential redox modulation. Antioxid Redox Signal 21:892–914CrossRefPubMedGoogle Scholar
  6. 6.
    Alberdi E, Sánchez-Gómez MV, Cavaliere F, Pérez-Samartín A, Zugaza JL, Trullas R, Domercq M, Matute C (2010) Amyloid beta oligomers induce Ca2+ dysregulation and neuronal death through activation of ionotropic glutamate receptors. Cell Calcium 47:264–272CrossRefPubMedGoogle Scholar
  7. 7.
    Zhou WW, Lu S, Su YJ, Xue D, Yu XL, Wang SW, Zhang H, Xu PX, Xie XX, Liu RT (2014) Decreasing oxidative stress and neuroinflammation with a multifunctional peptide rescues memory deficits in mice with Alzheimer disease. Free Radic Biol Med 74:50–63CrossRefPubMedGoogle Scholar
  8. 8.
    Fonfria E, Marshall IC, Boyfield I, Skaper SD, Hughes JP, Owen DE, Zhang W, Miller BA, Benham CD, McNulty S (2005) Amyloid beta-peptide(1–42) and hydrogen peroxide-induced toxicity are mediated by TRPM2 in rat primary striatal cultures. J Neurochem 95:715–723CrossRefPubMedGoogle Scholar
  9. 9.
    Ostapchenko VG, Chen M, Guzman MS, Xie YF, Lavine N, Fan J, Beraldo FH, Martyn AC, Belrose JC, Mori Y, MacDonald JF, Prado VF, Prado MA, Jackson MF (2015) The Transient receptor potential melastatin 2 (TRPM2) channel contributes to β-amyloid oligomer-related neurotoxicity and memory impairment. J Neurosci 35:15157–15169CrossRefPubMedGoogle Scholar
  10. 10.
    Nazıroğlu M, Dikici DM, Dursun S (2012) Role of oxidative stress and Ca(2)(+) signaling on molecular pathways of neuropathic pain in diabetes: focus on TRP channels. Neurochem Res 37:2065–2075CrossRefPubMedGoogle Scholar
  11. 11.
    Nazıroğlu M (2011) TRPM2 cation channels, oxidative stress and neurological diseases: where are we now? Neurochem Res 36:355–366CrossRefPubMedGoogle Scholar
  12. 12.
    Nazıroglu M (2009) Role of selenium on calcium signaling and oxidative stress-induced molecular pathways in epilepsy. Neurochem Res 34:2181–2191CrossRefPubMedGoogle Scholar
  13. 13.
    Schweizer U, Bräuer AU, Köhrle J, Nitsch R, Savaskan NE (2004) Selenium and brain function: a poorly recognized liaison. Brain Res Brain Res Rev 45:164–178CrossRefPubMedGoogle Scholar
  14. 14.
    Rita Cardoso B, Silva Bandeira V, Jacob-Filho W, Franciscato Cozzolino SM (2014) Selenium status in elderly: relation to cognitive decline. J Trace Elem Med Biol 28:422–6CrossRefPubMedGoogle Scholar
  15. 15.
    Vural H, Demirin H, Kara Y, Eren I, Delibas N (2010) Alterations of plasma magnesium, copper, zinc, iron and selenium concentrations and some related erythrocyte antioxidant enzyme activities in patients with Alzheimer’s disease. J Trace Elem Med Biol 24:169–173CrossRefPubMedGoogle Scholar
  16. 16.
    Lakshmi BV, Sudhakar M, Prakash KS (2015) Protective effect of selenium against aluminum chloride-induced Alzheimer's disease: behavioral and biochemical alterations in rats. Biol Trace Elem Res 165:67–74CrossRefPubMedGoogle Scholar
  17. 17.
    Uğuz AC, Nazıroğlu M (2012) Effects of selenium on calcium signaling and apoptosis in rat dorsal root ganglion neurons induced by oxidative stress. Neurochem Res 37:1631–1638CrossRefPubMedGoogle Scholar
  18. 18.
    Kahya MC, Nazıroğlu M, Çiğ B. Modulation of diabetes-induced oxidative stress, apoptosis, and Ca2+ entry through TRPM2 and TRPV1 channels in dorsal root ganglion and hippocampus of diabetic rats by melatonim and selenium. Mol Neurobiol. 2016 [Epub ahead of print]. DOI  10.1007/s12035-016-9727-3.
  19. 19.
    Caterina MJ, Schumacher MA, Tominaga M, Rosen TA, Levine JD, Julius D (1997) The capsaicin receptor: a heat-activated ion channel in the pain pathway. Nature 389:816–824CrossRefPubMedGoogle Scholar
  20. 20.
    Perraud AL, Fleig A, Dunn CA, Bagley LA, Launay P, Schmitz C, Stokes AJ, Zhu Q, Bessman MJ, Penner R, Kinet JP, Scharenberg AM (2001) ADP-ribose gating of the calcium-permeable LTRPC2 channel revealed by Nudix motif homology. Nature 411:595–599CrossRefPubMedGoogle Scholar
  21. 21.
    Nazıroğlu M, Lückhoff A (2008) A calcium influx pathway regulated separately by oxidative stress and ADP-Ribose in TRPM2 channels: single channel events. Neurochem Res 33:1256–1262CrossRefPubMedGoogle Scholar
  22. 22.
    Shimizu S, Takahashi N, Mori Y (2014) TRPs as chemosensors (ROS, RNS, RCS, gasotransmitters). Handb Exp Pharmacol 223:767–794CrossRefPubMedGoogle Scholar
  23. 23.
    Cristino L, de Petrocellis L, Pryce G, Baker D, Guglielmotti V, Di Marzo V (2006) Immunohistochemical localization of cannabinoid type 1 and vanilloid transient receptor potential vanilloid type 1 receptors in the mouse brain. Neuroscience 139:1405–1415CrossRefPubMedGoogle Scholar
  24. 24.
    Bai JZ, Lipski J (2010) Differential expression of TRPM2 and TRPV4 channels and their potential role in oxidative stress-induced cell death in organotypic hippocampal culture. Neurotoxicology 31:204–214CrossRefPubMedGoogle Scholar
  25. 25.
    Gupta S, Sharma B (2014) Pharmacological benefits of agomelatine and vanillin in experimental model of Huntington's disease. Pharmacol Biochem Behav 122:122–35CrossRefPubMedGoogle Scholar
  26. 26.
    Gupta S, Sharma B, Singh P, Sharma BM (2014) Modulation of transient receptor potential vanilloid subtype 1 (TRPV1) and norepinephrine transporters (NET) protect against oxidative stress, cellular injury, and vascular dementia. Curr Neurovasc Res 11:94–106CrossRefPubMedGoogle Scholar
  27. 27.
    Park L, Wang G, Moore J, Girouard H, Zhou P, Anrather J, Iadecola C (2014) The key role of transient receptor potential melastatin-2 channels in amyloid-β-induced neurovascular dysfunction. Nat Commun 5:5318CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Jiang X, Jia LW, Li XH, Cheng XS, Xie JZ, Ma ZW, Xu WJ, Liu Y, Yao Y, Du LL, Zhou XW (2013) Capsaicin ameliorates stress-induced Alzheimer’s disease-like pathological and cognitive impairments in rats. J Alzheimers Dis 35:91–105PubMedGoogle Scholar
  29. 29.
    Nazıroğlu M, Özgül C, Küçükayaz M, Çiğ B, Hebeisen S, Bal R (2013) Selenium modulates oxidative stress-induced TRPM2 cation channel currents in transfected Chinese hamster ovary cells. Basic Clin Pharmacol Toxicol 112:96–102CrossRefPubMedGoogle Scholar
  30. 30.
    Köse SA, Nazıroğlu M (2014) Selenium reduces oxidative stress and calcium entry through TRPV1 channels in the neutrophils of patients with polycystic ovary syndrome. Biol Trace Elem Res 158:136–142CrossRefPubMedGoogle Scholar
  31. 31.
    Chen C, Li XH, Zhang S, Tu Y, Wang YM, Sun HT (2014) 7,8-dihydroxyflavone ameliorates scopolamine-induced Alzheimer-like pathologic dysfunction. Rejuvenation Res 17:249–254CrossRefPubMedGoogle Scholar
  32. 32.
    Neha X, Sodhi RK, Jaggi AS, Singh N (2014) Animal models of dementia and cognitive dysfunction. Life Sci 109:73–86CrossRefPubMedGoogle Scholar
  33. 33.
    Hritcu L, Stefan M, Brandsch R, Mihasan M (2015) Enhanced behavioral response by decreasing brain oxidative stress to 6-hydroxy-l-nicotine in Alzheimer's disease rat model. Neurosci Lett 591:41–7CrossRefPubMedGoogle Scholar
  34. 34.
    Uğuz AC, Naziroğlu M, Espino J, Bejarano I, González D, Rodríguez AB, Pariente JA (2009) Selenium modulates oxidative stress-induced cell apoptosis in human myeloid HL-60 cells through regulation of calcium release and caspase-3 and −9 activities. J Membr Biol 232:15–23CrossRefPubMedGoogle Scholar
  35. 35.
    Grynkiewicz C, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450PubMedGoogle Scholar
  36. 36.
    Espino J, Bejarano I, Redondo PC, Rosado JA, Barriga C, Reiter RJ, Pariente JA, Rodríguez AB (2010) Melatonin reduces apoptosis induced by calcium signaling in human leukocytes: evidence for the involvement of mitochondria and Bax activation. J Membr Biol 233:105–118CrossRefPubMedGoogle Scholar
  37. 37.
    Espino J, Bejarano I, Paredes SD, Barriga C, Rodríguez AB, Pariente JA (2011) Protective effect of melatonin against human leukocyte apoptosis induced by intracellular calcium overload: relation with its antioxidant actions. J Pineal Res 51:195–206CrossRefPubMedGoogle Scholar
  38. 38.
    Bejarano I, Redondo PC, Espino J, Rosado JA, Paredes SD, Barriga C, Reiter RJ, Pariente JA, Rodríguez AB (2009) Melatonin induces mitochondrial-mediated apoptosis in human myeloid HL-60 cells. J Pineal Res 46:392–400CrossRefPubMedGoogle Scholar
  39. 39.
    Placer ZA, Cushman L, Johnson BC (1966) Estimation of products of lipid peroxidation (malonyl dialdehyde) in biological fluids. Analytical Biochem 16:359–364CrossRefGoogle Scholar
  40. 40.
    Sedlak J, Lindsay RHC (1968) Estimation of total, protein bound and non-protein sulfhydryl groups in tissue with Ellmann’ s reagent. Analytical Biochem 25:192–205CrossRefGoogle Scholar
  41. 41.
    Lawrence RA, Burk RF (1976) Glutathione peroxidase activity in selenium-deficient rat liver. Biochem Biophys Res Com 71:952–958CrossRefPubMedGoogle Scholar
  42. 42.
    Bihaqi SW, Singh AP, Tiwari M (2012) Supplementation of Convolvulus pluricaulis attenuates scopolamine-induced increased tau and amyloid precursor protein (AβPP) expression in rat brain. Indian J Pharmacol 44:593–598CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Barbero-Camps E, Fernández A, Martínez L, Fernández-Checa JC, Colell A (2013) APP/PS1 mice overexpressing SREBP-2 exhibit combined Aβ accumulation and tau pathology underlying Alzheimer’s disease. Hum Mol Genet 22:3460–3476CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Kumar VS, Gopalakrishnan A, Naziroğlu M, Rajanikant GK (2014) Calcium ion—the key player in cerebral ischemia. Curr Med Chem 21:2065–2075CrossRefPubMedGoogle Scholar
  45. 45.
    Hajnóczky G, Csordás G, Das S, Garcia-Perez C, Saotome M, Sinha Roy S, Yi M (2006) Mitochondrial calcium signalling and cell death: approaches for assessing the role of mitochondrial Ca2+ uptake in apoptosis. Cell Calcium 40:553–560CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Hasan Balaban
    • 1
  • Mustafa Nazıroğlu
    • 2
    • 3
    Email author
  • Kadir Demirci
    • 1
  • İshak Suat Övey
    • 2
  1. 1.Department of Psychiatry, Faculty of MedicineSuleyman Demirel UniversityIspartaTurkey
  2. 2.Department of Neuroscience, Institute of Health ScienceSuleyman Demirel UniversityIspartaTurkey
  3. 3.Neuroscience Research CenterUniversity of Suleyman DemirelIspartaTurkey

Personalised recommendations