Neuropeptide Y suppresses epileptiform discharges by regulating AMPA receptor GluR2 subunit in rat hippocampal neurons

Bu,Wei; Zhao,Wen‑Qing; Li,Wen‑Ling; Dong,Chang‑Zheng; Zhang,Zhe; Li,Qi‑Jun

doi:10.3892/mmr.2017.6567

Neuropeptide Y suppresses epileptiform discharges by regulating AMPA receptor GluR2 subunit in rat hippocampal neurons

Authors:
- Wei Bu
- Wen‑Qing Zhao
- Wen‑Ling Li
- Chang‑Zheng Dong
- Zhe Zhang
- Qi‑Jun Li
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Affiliations: Department of Neurosurgery, The Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China, Department of Neurosurgery, Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China, Department of Functional Neurosurgery, Hebei General Hospital, Shijiazhuang, Hebei 050051, P.R. China, Department of Orthopaedics, Shijiazhuang Third Hospital, Shijiazhuang, Hebei 050017, P.R. China
Published online on: May 10, 2017 https://doi.org/10.3892/mmr.2017.6567
Pages: 387-395

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Abstract

The present study aimed to investigate the effects of neuropeptide Y (NPY) on the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor glutamate receptor 2 (GluR2) subunit in epileptiform discharge hippocampal neurons. Hippocampal neurons were harvested from neonatal Sprague‑Dawley rats aged <24 h and primarily cultured in vitro. At day 12 following culture, hippocampal neurons were divided into the following groups: Control, Mg2+‑free, NPY+Mg2+‑free and BIBP3226+NPY+Mg2+‑free. The action potential of neurons was measured using the whole cell patch clamp technique in the control, Mg2+‑free and NPY+Mg2+‑free groups. AMPA current (IAMPA) was detected and peak current density was calculated in each group. Alterations in total protein and phosphorylation of the GluR2 subunit were detected by western blot analysis, and GluR2 mRNA expression levels were detected by reverse transcription‑quantitative polymerase chain reaction, in each group. The whole cell patch clamp technique demonstrated an abnormal action potential in the Mg2+‑free group. The frequency and amplitude of the action potential were significantly greater in the Mg2+‑free group compared with the control group, and significantly reduced in the NPY+Mg2+‑free group compared with the Mg2+‑free group (P<0.05). In the Mg2+‑free group, compared with the control group, peak current density was significantly reduced (P<0.05), GluR2 subunit protein content was slightly reduced (P>0.05), phosphorylation levels of GluR2 subunit were significantly greater (P<0.05) and GluR2 mRNA was significantly reduced (P<0.05). In the NPY+Mg2+‑free group, compared with the Mg2+‑free group, peak current density was significantly greater (P<0.05), phosphorylation levels of GluR2 subunit were significantly reduced (P<0.05) and GluR2 mRNA expression was significantly greater (P<0.05). In the BIBP3226+NPY+Mg2+‑free group, compared with the NPY+Mg2+‑free group, peak current density was significantly reduced (P<0.05), phosphorylation levels of GluR2 subunit were significantly greater (P<0.05) and GluR2 mRNA expression was significantly reduced (P<0.05). After 3 h of treatment with Mg2+‑free extracellular fluid, epileptiform discharge was detected in the cells. NPY inhibited the discharge and its underlying mechanism may be that epileptiform discharge suppressed the function of the AMPA receptor GluR2 subunit. NPY relieved the inhibition of the GluR2 subunit via the Y1 receptor. This may provide a novel direction for future studies on the pathogenesis and treatment of epilepsy.

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1	Rocha AK, de Lima E, Amaral FG, Peres R, Cipolla-Neto J and Amado D: Pilocarpine-induced epilepsy alters the expression and daily variation of the nuclear receptor RORα in the hippocampus of rats. Epilepsy Behav. 55:38–46. 2015. View Article : Google Scholar : PubMed/NCBI
2	Muldoon SF, Villette V, Tressard T, Malvache A, Reichinnek S, Bartolomei F and Cossart R: GABAergic inhibition shapes interictal dynamics in awake epileptic mice. Brain. 138:2875–2890. 2015. View Article : Google Scholar : PubMed/NCBI
3	Tsamis KI, Mytilinaios DG, Njau SN and Baloyannis SJ: Glutamate receptors in human caudate nucleus in normal aging and Alzheimer's disease. Curr Alzheimer Res. 10:469–475. 2013. View Article : Google Scholar : PubMed/NCBI
4	Ettinger AB, LoPresti A, Yang H, Williams B, Zhou S, Fain R and Laurenza A: Psychiatric and behavioral adverse events in randomized clinical studies of the noncompetitive AMPA receptor antagonist perampanel. Epilepsia. 56:1252–1263. 2015. View Article : Google Scholar : PubMed/NCBI
5	Harlow DE, Saul KE, Komuro H and Macklin WB: Myelin proteolipid protein complexes with αv integrin and AMPA receptors in vivo and regulates AMPA-dependent oligodendrocyte progenitor cell migration through the modulation of cell-surface GluR2 expression. J Neurosci. 35:12018–12032. 2015. View Article : Google Scholar : PubMed/NCBI
6	Friedman LK, Velísková J, Kaur J, Magrys BW and Liu H: GluR2(B) knockdown accelerates CA3 injury after kainate seizures. J Neuropathol Exp Neurol. 62:733–750. 2003. View Article : Google Scholar : PubMed/NCBI
7	Tatemoto K, Carlquist M and Mutt V: Neuropeptide Y-a novel brain peptide with structural similarities to peptide YY and pancreatic polypeptide. Nature. 296:659–660. 1982. View Article : Google Scholar : PubMed/NCBI
8	Woldbye DP and Kokaia M: Neuropeptide Y and seizures: Effects of exogenously applied ligands. Neuropeptides. 38:253–260. 2004. View Article : Google Scholar : PubMed/NCBI
9	Malva JO, Xapelli S, Baptista S, Valero J, Agasse F, Ferreira R and Silva AP: Multifaces of neuropeptide Y in the brain-neuroprotection, neurogenesis and neuroinflammation. Neuropeptides. 46:299–308. 2012. View Article : Google Scholar : PubMed/NCBI
10	Vezzani A and Sperk G: Overexpression of NPY and Y2 receptors in epileptic brain tissue: An endogenous neuroprotective mechanism in temporal lobe epilepsy? Neuropeptides. 38:245–252. 2004. View Article : Google Scholar : PubMed/NCBI
11	Yang S, Zhou W, Zhang Y, Yan C and Zhao Y: Effects of Liuwei Dihuang decoction on ion channels and synaptic transmission in cultured hippocampal neuron of rat. J Ethnopharmacol. 106:166–172. 2006. View Article : Google Scholar : PubMed/NCBI
12	Atalay B, Caner H, Can A and Cekinmez M: Attenuation of microtubule associated protein-2 degradation after mild head injury by mexiletine and calpain-2 inhibitor. Br J Neurosurg. 21:281–287. 2007. View Article : Google Scholar : PubMed/NCBI
13	DeLorenzo RJ, Sombati S and Coulter DA: Effects of topiramate on sustained repetitive firing and spontaneous recurrent seizure discharges in cultured hippocampal neurons. Epilepsia. 41:(Suppl 1). S40–S44. 2000. View Article : Google Scholar : PubMed/NCBI
14	Rozov A, Sprengel R and Seeburg PH: GluA2-lacking AMPA receptors in hippocampal CA1 cell synapses: Evidence from gene-targeted mice. Front Mol Neurosci. 5:222012. View Article : Google Scholar : PubMed/NCBI
15	Rozov A, Zivkovic AR and Schwarz MK: Homer1 gene products orchestrate Ca⁽²⁺⁾-permeable AMPA receptor distribution and LTP expression. Front Synaptic Neurosci. 4:42012. View Article : Google Scholar : PubMed/NCBI
16	Curtis KM, Gomez LA, Rios C, Garbayo E, Raval AP, Perez-Pinzon MA and Schiller PC: EF1alpha and RPL13a represent normalization genes suitable for RT-qPCR analysis of bone marrow derived mesenchymal stem cells. BMC Mol Biol. 11:612010. View Article : Google Scholar : PubMed/NCBI
17	Dudek FE and Rogawski MA: The epileptic neuron redux. Epilepsy Curr. 2:151–152. 2002. View Article : Google Scholar : PubMed/NCBI
18	Bernard C, Anderson A, Becker A, Poolos NP, Beck H and Johnston D: Acquired dendritic channelopathy in temporal lobe epilepsy. Science. 305:532–535. 2004. View Article : Google Scholar : PubMed/NCBI
19	Kato K, Sekino Y, Takahashi H, Yasuda H and Shirao T: Increase in AMPA receptor-mediated miniature EPSC amplitude after chronic NMDA receptor blockade in cultured hippocampal neurons. Neurosci Lett. 418:4–8. 2007. View Article : Google Scholar : PubMed/NCBI
20	Solger J, Heinemann U and Behr J: Electrical and chemical long-term depression do not attenuate low-Mg2+-induced epileptiform activity in the entorhinal cortex. Epilepsia. 46:509–516. 2005. View Article : Google Scholar : PubMed/NCBI
21	Hu Y, Jiang L, Chen H and Zhang X: Expression of AMPA receptor subunits in hippocampus after status convulsion. Childs Nerv Syst. 28:911–918. 2012. View Article : Google Scholar : PubMed/NCBI
22	Ma YX, Yin JB, Fan XT, Xu HW, An N, Wan ZB, Li ZF, Liu GL, Zhang YH and Yang Hui: Establishment of kainic acid induced temporal lobe epilepsy in rat and study of its neurogenesis. Acta Academiae Medicinae Militaris Tertiae. 29:872–875. 2007.
23	Sombati S and Delorenzo RJ: Recurrent spontaneous seizure activity in hippocampal neuronal networks in culture. J Neurophysiol. 73:1706–1711. 1995.PubMed/NCBI
24	Furtinger S, Pirker S, Czech T, Baumgartner C, Ransmayr G and Sperk G: Plasticity of Y1 and Y2 receptors and neuropeptide Y fibers in patients with temporal lobe epilepsy. J Neurosci. 21:5804–5812. 2001.PubMed/NCBI
25	Sørensen G and Woldbye DP: Mice lacking neuropeptide Y show increased sensitivity to cocaine. Synapse. 66:840–843. 2012. View Article : Google Scholar : PubMed/NCBI
26	Yamamoto BK and Raudensky J: The role of oxidative stress, metabolic compromise, and inflammation in neuronal injury produced by amphetamine-related drugs of abuse. J Neuroimmune Pharmacol. 3:203–217. 2008. View Article : Google Scholar : PubMed/NCBI
27	Cadet JL and Krasnova IN: Molecular bases of methamphetamine-induced neurodegeneration. Int Rev Neurobiol. 88:101–119. 2009. View Article : Google Scholar : PubMed/NCBI
28	Nunes AF, Montero M, Franquinho F, Santos SD, Malva J, Zimmer J and Sousa MM: Transthyretin knockout mice display decreased susceptibility to AMPA-induced neurodegeneration. Neurochem Int. 55:454–457. 2009. View Article : Google Scholar : PubMed/NCBI
29	Hoy KC, Huie JR and Grau JW: AMPA receptor mediated behavioral plasticity in the isolated rat spinal cord. Behav Brain Res. 236:319–326. 2013. View Article : Google Scholar : PubMed/NCBI
30	Blair RE, Sombati S, Churn SB and Delorenzo RJ: Epileptogenesis causes an N-methyl-d-aspartate receptor/Ca2+-dependent decrease in Ca²⁺/calmodulin-dependent protein kinase II activity in a hippocampal neuronal culture model of spontaneous recurrent epileptiform discharges. Eur J Pharmacol. 588:64–71. 2008. View Article : Google Scholar : PubMed/NCBI
31	Blair RE, Deshpande LS, Sombati S, Elphick MR, Martin BR and DeLorenzo RJ: Prolonged exposure to WIN55,212-2 causes downregulation of the CB1 receptor and the development of tolerance to its anticonvulsant effects in the hippocampal neuronal culture model of acquired epilepsy. Neuropharmacology. 57:208–218. 2009. View Article : Google Scholar : PubMed/NCBI
32	Gardner SM, Takamiya K, Xia J, Suh JG, Johnson R, Yu S and Huganir RL: Calcium-permeable AMPA receptor plasticity is mediated by subunit-specific interactions with PICK1 and NSF. Neuron. 45:903–915. 2005. View Article : Google Scholar : PubMed/NCBI
33	Essin K, Nistri A and Magazanik L: Evaluation of GluR2 subunit involvement in AMPA receptor function of neonatal rat hypoglossal motoneurons. Eur J Neurosci. 15:1899–1906. 2002. View Article : Google Scholar : PubMed/NCBI
34	Palmer CL, Cotton L and Henley JM: The molecular pharmacology and cell biology of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors. Pharmacol Rev. 57:253–277. 2005. View Article : Google Scholar : PubMed/NCBI
35	Montero M, Nielsen M, Rønn LC, Møller A, Noraberg J and Zimmer J: Neuroprotective effects of the AMPA antagonist PNQX in oxygen-glucose deprivation in mouse hippocampal slice cultures and global cerebral ischemia in gerbils. Brain Res. 1177:124–135. 2007. View Article : Google Scholar : PubMed/NCBI
36	Sager C, Tapken D, Kott S and Hollmann M: Functional modulation of AMPA receptors by transmembrane AMPA receptor regulatory proteins. Neuroscience. 158:45–54. 2009. View Article : Google Scholar : PubMed/NCBI
37	Colbourne F, Grooms SY, Zukin RS, Buchan AM and Bennett MV: Hypothermia rescues hippocampal CA1 neurons and attenuates down-regulation of the AMPA receptor GluR2 subunit after forebrain ischemia. Proc Natl Acad Sci USA. 100:2906–2910. 2003. View Article : Google Scholar : PubMed/NCBI
38	Calderone A, Jover T, Mashiko T, Noh KM, Tanaka H, Bennett MV and Zukin RS: Late calcium EDTA rescues hippocampal CA1 neurons from global ischemia-induced death. J Neurosci. 24:9903–9913. 2004. View Article : Google Scholar : PubMed/NCBI
39	Grooms SY, Opitz T, Bennett MV and Zukin RS: Status epilepticus decreases glutamate receptor 2 mRNA and protein expression in hippocampal pyramidal cells before neuronal death. Proc Natl Acad Sci USA. 97:3631–3636. 2000. View Article : Google Scholar : PubMed/NCBI
40	Sanchez RM, Koh S, Rio C, Wang C, Lamperti ED, Sharma D, Corfas G and Jensen FE: Decreased glutamate receptor 2 expression and enhanced epileptogenesis in immature rat hippocampus after perinatal hypoxia-induced seizures. J Neurosci. 21:8154–8163. 2001.PubMed/NCBI
41	Hanley JG and Henley JM: PICK1 is a calcium-sensor for NMDA-induced AMPA receptor trafficking. EMBO J. 24:3266–3278. 2005. View Article : Google Scholar : PubMed/NCBI
42	Zhang Y, Venkitaramani DV, Gladding CM, Zhang Y, Kurup P, Molnar E, Collingridge GL and Lombroso PJ: The tyrosine phosphatase STEP mediates AMPA receptor endocytosis after metabotropic glutamate receptor stimulation. J Neurosci. 28:10561–10566. 2008. View Article : Google Scholar : PubMed/NCBI
43	Dev KK, Nakanishi S and Henley JM: The PDZ domain of PICK1 differentially accepts protein kinase C-alpha and GluR2 as interacting ligands. J Biol Chem. 279:41393–41397. 2004. View Article : Google Scholar : PubMed/NCBI
44	Alvaro AR, Martins J, Costa AC, Fernandes E, Carvalho F, Ambrósio AF and Cavadas C: Neuropeptide Y protects retinal neural cells against cell death induced by ecstasy. Neuroscience. 152:97–105. 2008. View Article : Google Scholar : PubMed/NCBI
45	Baptista S, Bento AR, Gonçalves J, Bernardino L, Summavielle T, Lobo A, Fontes-Ribeiro C, Malva JO, Agasse F and Silva AP: Neuropeptide Y promotes neurogenesis and protection against methamphetamine-induced toxicity in mouse dentate gyrus-derived neurosphere cultures. Neuropharmacology. 62:2413–2423. 2012. View Article : Google Scholar : PubMed/NCBI
46	Jinde S, Masui A, Morinobu S, Noda A and Kato N: Differential changes in messenger RNA expressions and binding sites of neuropeptide Y Y1, Y2 and Y5 receptors in the hippocampus of an epileptic mutant rat: Noda epileptic rat. Neuroscience. 115:1035–1045. 2002. View Article : Google Scholar : PubMed/NCBI
47	Woldbye DP, Nanobashvili A, Sørensen AT, Husum H, Bolwig TG, Sørensen G, Ernfors P and Kokaia M: Differential suppression of seizures via Y2 and Y5 neuropeptide Y receptors. Neurobiol Dis. 20:760–772. 2005. View Article : Google Scholar : PubMed/NCBI