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Inhibition of the NMDA Currents by Probenecid in Amygdaloid Kindling Epilepsy Model

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Abstract

Epilepsy is characterized by a sustained depolarization and repeated discharge of neurons, attributed to overstimulation of N-methyl-D-aspartate receptors (NMDAr). Herein, we propose that probenecid (PROB), an inhibitor of the activity of some ATP binding-cassette transporters (ABC-transporters) can modify NMDAr activity and expression in amygdaloid kindled model. Some studies have suggested that NMDAr expression could be regulated by inhibiting the activity of P-glycoprotein (MDR1) and drug resistance protein-1 (MRP1). Besides, PROB was found to interact with other proteins with proven activity in the kindling model, such as TRPV2 channels, OAT1, and Panx1. Administering PROB at two doses (100 and 300 mg/kg/d) for 5 d decreased after-discharge duration and Racine behavioral scores. It also reduced the expression of NR2B and the activity of total NOS and the expression of nNOS with respect to the kindling group. In a second protocol, voltage-clamp measurements of NMDA-evoked currents were performed in CA1 hippocampal cells dissociated from control and kindled rats. PROB produced a dose-dependent reduction in NMDA-evoked currents. In neurons from kindled rats, a residual NMDA-evoked current was registered with respect to control animals, while a reduction in NMDA-evoked currents was observed in the presence of 20 mM PROB. Finally, we evaluated the expression of MRP1 and MDR1 in order to establish a relationship between the reduction of kindling parameters, the inhibition of NMDA-type currents, and the expression of these transporters. Based on our results, we conclude that at the concentrations used, PROB inhibits currents evoked by NMDA in dissociated neurons of control and kindled rats. In the kindling model, at the tested doses, PROB decreases the after-discharge duration and Racine behavioral score in the kindling model. We propose a mechanism that could be dependent on the expression of ABC-type transporters.

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Data Availability

The datasets used and/or analyses during the current study are available from the corresponding author upon reasonable request.

References

  1. MacDermott AB, Mayer ML, Westbrook GL, Smith SJ, Barker JL (1986) NMDA-receptor activation increases cytoplasmic calcium concentration in cultured spinal cord neurones. Nature 321(6069):519–522

    Article  CAS  PubMed  Google Scholar 

  2. Johnson JW, Ascher P (1987) Glycine potentiates the NMDA response in cultured mouse brain neurons. Nature 325(6104):529–531

    Article  CAS  PubMed  Google Scholar 

  3. Nowak L, Bregestovski P, Ascher P, Herbet A, Prochiantz A (1984) Magnesium gates glutamate-activated channels in mouse central neurones. Nature 307(5950):462–465

    Article  CAS  PubMed  Google Scholar 

  4. Mayer ML, Westbrook GL, Guthrie PB (1984) Voltage-dependent block by Mg2+ of NMDA responses in spinal cord neurones. Nature 309(5965):261–263

    Article  CAS  PubMed  Google Scholar 

  5. Collingridge GL, Bliss TV (1995) Memories of NMDA receptors and LTP. Trends Neurosci 18(2):54–56

    Article  CAS  PubMed  Google Scholar 

  6. Bliss TV, Collingridge GL (2013) Expression of NMDA receptor-dependent LTP in the hippocampus: bridging the divide. Mol Brain 6:5

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Mody I, Heinemann U (1987) NMDA receptors of dentate gyrus granule cells participate in synaptic transmission following kindling. Nature 326(6114):701–4

    Article  CAS  PubMed  Google Scholar 

  8. Sprengel R, Suchanek B, Amico C et al (1998) Importance of the intracellular domain of NR2 subunits for NMDA receptor function in vivo. Cell 92(2):279–289

    Article  CAS  PubMed  Google Scholar 

  9. Bankstahl JP, Hoffmann K, Bethmann K, Loscher W (2008) Glutamate is critically involved in seizure-induced overexpression of P-glycoprotein in the brain. Neuropharmacology 54(6):1006–1016

    Article  CAS  PubMed  Google Scholar 

  10. Avemary J, Salvamoser JD, Peraud A et al (2013) Dynamic regulation of P-glycoprotein in human brain capillaries. Mol Pharm 10(9):3333–3341

    Article  CAS  PubMed  Google Scholar 

  11. Tishler DM, Weinberg KI, Hinton DR, Barbaro N, Annett GM, Raffel C (1995) MDR1 gene expression in brain of patients with medically intractable epilepsy. Epilepsia 36(1):1–6

    Article  CAS  PubMed  Google Scholar 

  12. Smolarz B, Makowska M, Romanowicz H (2021) Pharmacogenetics of Drug-Resistant Epilepsy (Review of Literature). Int J Mol Sci 22(21):11696

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Lazarowski A, Sevlever G, Taratuto A, Massaro M, Rabinowicz A (1999) Tuberous sclerosis associated with MDR1 gene expression and drug-resistant epilepsy. Pediatr Neurol 21(4):731–734

    Article  CAS  PubMed  Google Scholar 

  14. Sisodiya SM, Heffernan J, Squier MV (1999) Over-expression of P-glycoprotein in malformations of cortical development. NeuroReport 10(16):3437–3441

    Article  CAS  PubMed  Google Scholar 

  15. Dombrowski SM, Desai SY, Marroni M et al (2001) Overexpression of multiple drug resistance genes in endothelial cells from patients with refractory epilepsy. Epilepsia 42(12):1501–1506

    Article  CAS  PubMed  Google Scholar 

  16. Lazarowski A, Lubieniecki F, Camarero S et al (2004) Multidrug resistance proteins in tuberous sclerosis and refractory epilepsy. Pediatr Neurol 30(2):102–106

    Article  PubMed  Google Scholar 

  17. Lazarowski A, Massaro M, Schteinschnaider A, Intruvini S, Sevlever G, Rabinowicz A (2004) Neuronal MDR-1 gene expression and persistent low levels of anticonvulsants in a child with refractory epilepsy. Ther Drug Monit 26(1):44–46

    Article  PubMed  Google Scholar 

  18. Zhu HJ, Liu GQ (2004) Glutamate up-regulates P-glycoprotein expression in rat brain microvessel endothelial cells by an NMDA receptor-mediated mechanism. Life Sci 75(11):1313–1322

    Article  CAS  PubMed  Google Scholar 

  19. Yang T, Kong B, Kuang Y et al (2015) Emodin plays an interventional role in epileptic rats via multidrug resistance gene 1 (MDR1). Int J Clin Exp Pathol 8(3):3418–3425

    PubMed  PubMed Central  Google Scholar 

  20. Cheng MH, Kim SJ (2020) Inhibitory Effect of Probenecid on Osteoclast Formation via JNK, ROS and COX-2. Biomol Ther (Seoul) 28(1):104–109

    Article  CAS  PubMed  Google Scholar 

  21. Du L, Empey PE, Ji J et al (2016) Probenecid and N-Acetylcysteine Prevent Loss of Intracellular Glutathione and Inhibit Neuronal Death after Mechanical Stretch Injury In Vitro. J Neurotrauma 33(20):1913–1917

    Article  PubMed  PubMed Central  Google Scholar 

  22. Jiang B, Zhao Y, Cao J et al (2021) Synthesis and preliminary biological evaluation of naproxen-probenecid conjugate for central nervous system (CNS) delivery. Pak J Pharm Sci 34(6):2197–2203

    CAS  PubMed  Google Scholar 

  23. Vamos E, Voros K, Zadori D, Vecsei L, Klivenyi P (2009) Neuroprotective effects of probenecid in a transgenic animal model of Huntington’s disease. J Neural Transm (Vienna) 116(9):1079–1086

    Article  CAS  PubMed  Google Scholar 

  24. Bang S, Kim KY, Yoo S, Lee SH, Hwang SW (2007) Transient receptor potential V2 expressed in sensory neurons is activated by probenecid. Neurosci Lett 425:120–125

    Article  CAS  PubMed  Google Scholar 

  25. SilvermanW LS, Dahl G (2008) Probenecid, a gout remedy, inhibits pannexin 1 channels. Am J Physiol Cell Physiol 295:C761–C767

    Article  Google Scholar 

  26. Taylor DL, Urenjak J, Zilkha E, Obrenovitch TP (1997) Effects of probenecid on the elicitation of spreading depression in the rat striatum. Brain Res 764(1–2):117–125

    Article  CAS  PubMed  Google Scholar 

  27. Urenjak J, Obrenovitch TP, Zilkha E (1997) Effect of probenecid on depolarizations evoked by N-methyl-D-aspartate (NMDA) in the rat striatum. Naunyn Schmiedebergs Arch Pharmacol 355(1):36–42

    Article  CAS  PubMed  Google Scholar 

  28. Paxinos G, Watson CH (2013) The Rat Brain in Stereotaxic Coordinates, 7th edn. Academic Press, New York

    Google Scholar 

  29. Hernandez-Ceron M, Martinez-Lazcano JC, Rubio C et al (2017) Participation of the dentate-rubral pathway in the kindling model of epilepsy. J Neurosci Res 95(7):1495–1502

    Article  CAS  PubMed  Google Scholar 

  30. Goddard GV, McIntyre DC, Leech CK (1969) A permanent change in brain function resulting from daily electrical stimulation. Exp Neurol 25(3):295–330

    Article  CAS  PubMed  Google Scholar 

  31. Racine RJ (1972) Modification of seizure activity by electrical stimulation. II. Motor seizure. Electroencephalogr Clin Neurophysiol 32(3):281–94

    Article  CAS  PubMed  Google Scholar 

  32. Martinez-Lazcano JC, Gonzalez-Guevara E, Custodio V et al (2018) Activity of nitric oxide synthase isoforms in acute brain oxidative damage induced by ozone exposure. Nitric Oxide 75:42–52

    Article  CAS  PubMed  Google Scholar 

  33. Martinez-Lazcano JC, Perez-Severiano F, Escalante B et al (2007) Selective protection against oxidative damage in brain of mice with a targeted disruption of the neuronal nitric oxide synthase gene. J Neurosci Res 85(7):1391–1402

    Article  CAS  PubMed  Google Scholar 

  34. Bargas J, Howe A, Eberwine J, Cao Y, Surmeier DJ (1994) Cellular and molecular characterization of Ca2+ currents in acutely isolated, adult rat neostriatal neurons. J Neurosci 14(11 Pt 1):6667–6686

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Flores-Hernandez J, Galarraga E, Pineda JC, Bargas J (1994) Patterns of excitatory and inhibitory synaptic transmission in the rat neostriatum as revealed by 4-AP. J Neurophysiol 72(5):2246–2256

    Article  CAS  PubMed  Google Scholar 

  36. Rendon-Ochoa EA, Hernandez-Flores T, Aviles-Rosas VH et al (2018) Calcium currents in striatal fast-spiking interneurons: dopaminergic modulation of CaV1 channels. BMC Neurosci 19(1):42

    Article  PubMed  PubMed Central  Google Scholar 

  37. Flores-Hernandez J, Cepeda C, Hernandez-Echeagaray E et al (2002) Dopamine enhancement of NMDA currents in dissociated medium-sized striatal neurons: role of D1 receptors and DARPP-32. J Neurophysiol 88(6):3010–3020

    Article  CAS  PubMed  Google Scholar 

  38. Campos-Arroyo D, Maldonado V, Bahena I et al (2016) Probenecid Sensitizes Neuroblastoma Cancer Stem Cells to Cisplatin. Cancer Invest 34(3):155–66

    Article  CAS  PubMed  Google Scholar 

  39. Leino E, MacDonald E, Airaksinen MM, Riekkinen PJ (1980) Homovanillic acid and 5-hydroxyindoleacetic acid levels in cerebrospinal fluid of patients with progressive myoclonus epilepsy. Acta Neurol Scand 62(1):41–54

    Article  CAS  PubMed  Google Scholar 

  40. Shaywitz BA, Cohen DJ, Leckman JF, Young JG, Bowers MB (1980) Ontogeny of dopamine and serotonin metabolites in the cerebrospinal fluid of children with neurological disorders. Dev Med Child Neurol 22(6):748–754

    Article  CAS  PubMed  Google Scholar 

  41. Fukuyama Y, Ochiai Y (1982) Therapeutic trial by taurine for intractable childhood epilepsies. Brain Dev 4(1):63–69

    Article  CAS  PubMed  Google Scholar 

  42. Tang CM, Dichter M, Morad M (1990) Modulation of the N-methyl-D-aspartate channel by extracellular H+. Proc Natl Acad Sci U S A 87(16):6445–6449

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Lipton SA, Rosenberg PA (1994) Excitatory amino acids as a final common pathway for neurologic disorders. N Engl J Med 330(9):613–622

    Article  CAS  PubMed  Google Scholar 

  44. West AE, Chen WG et al (2001) Calcium regulation of neuronal gene expression. Proc Natl Acad Sci U S A 98(20):11024–11031

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Banach M, Piskorska B, Czuczwar SJ, Borowicz KK (2011) Nitric oxide, epileptic seizures, and action of antiepileptic drugs. CNS Neurol Disord Drug Targets 10(7):808–819

    Article  CAS  PubMed  Google Scholar 

  46. Bankstahl JP, Kuntner C, Abrahim A et al (2008) Langer, Tariquidar-induced P-glycoprotein inhibition at the rat blood-brain barrier studied with (R)-11C-verapamil and PET. J Nucl Med 49(8):1328–1335

    Article  CAS  PubMed  Google Scholar 

  47. Sutula TP (2004) Mechanisms of epilepsy progression: current theories and perspectives from neuroplasticity in adulthood and development. Epilepsy Res 60(2–3):161–171

    Article  PubMed  Google Scholar 

  48. Sutula TP, Dudek FE (2007) Unmasking recurrent excitation generated by mossy fiber sprouting in the epileptic dentate gyrus: an emergent property of a complex system. Prog Brain Res 163:541–563

    Article  CAS  PubMed  Google Scholar 

  49. McNamara JO (1995) Analyses of the molecular basis of kindling development. Psychiatry Clin Neurosci 49(3):S175–S178

    Article  CAS  PubMed  Google Scholar 

  50. Kohr G, De Koninck Y, Mody I (1993) Properties of NMDA receptor channels in neurons acutely isolated from epileptic (kindled) rats. J Neurosci 13(8):3612–3627

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Behr J, Heinemann U, Mody I (2000) Glutamate receptor activation in the kindled dentate gyrus. Epilepsia 41(Suppl 6):S100–S103

    PubMed  Google Scholar 

  52. Kamphuis W, Monyer H, De Rijk TC, Lopes da Silva FH (1992) Hippocampal kindling increases the expression of glutamate receptor-A Flip and -B Flip mRNA in dentate granule cells. Neurosci Lett 148(1–2):51–54

    Article  CAS  PubMed  Google Scholar 

  53. Chen YH, Wang CC, Xiao X, Wei L, Xu G (2013) Multidrug resistance-associated protein 1 decreases the concentrations of antiepileptic drugs in cortical extracellular fluid in amygdale kindling rats. Acta Pharmacol Sin 34(4):473–479

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Bauer B, Hartz AM, Pekcec A, Toellner K, Miller DS, Potschka H (2008) Seizure-induced up-regulation of P-glycoprotein at the blood-brain barrier through glutamate and cyclooxygenase-2 signaling. Mol Pharmacol 73(5):1444–1453

    Article  CAS  PubMed  Google Scholar 

  55. García-Rodríguez C, Mujica P, Illanes-González J, López A, Vargas C et al (2023) Probenecid, an Old Drug with Potential New Uses for Central Nervous System Disorders and Neuroinflammation. Biomedicines 11(6):1516

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The authors would like to thank Dr. Carlos Paz Tres for facilitating the infrastructure of his laboratory, Dr. Denise Campos-Arroyo for her supervision in the design and development of the semi-quantitative RT-PCR technique, Francisco Gutiérrez-Baeza by his technical support, and Juan Francisco Rodriguez for copyediting the manuscript.

Funding

The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.

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Authors and Affiliations

  1. Departamento de Neurofisiología, Instituto Nacional de Neurología y Neurocirugía MVS, Mexico City, 14629, México

    Edith González-Guevara, Esther Lara-González, Javier Franco-Pérez, Miguel Hernández-Cerón, Cesar Martínez-de los Santos, Verónica Custodio & Juan Carlos Martínez-Lazcano

  2. Laboratorio de Neurofarmacología Molecular y Nanotecnología, Instituto Nacional de Neurología y Neurocirugía MVS, Insurgentes Sur 3877, La Fama, Mexico City, 14629, México

    Edith González-Guevara, Francisca Pérez-Severiano & Juan Carlos Martínez-Lazcano

  3. División Neurociencias, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Mexico City, 04510, México

    Esther Lara-González, Ernesto Rendon-Ochoa, Antonio Laville & José Bargas

  4. Department of Neuroscience, Feinberg School of Medicine, Northwestern University, Chicago, IL, 60611, USA

    Esther Lara-González

  5. Laboratorio de Psicofarmacología, Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México, Tlalnepantla, 54090, México

    Ernesto Rendon-Ochoa

  6. Laboratorio de Neuropatología Vascular, Instituto Nacional de Neurología y Neurocirugía MVS, Mexico City, 14629, México

    Javier Franco-Pérez

  7. Laboratorio de Neuroquímica, Instituto Nacional de Neurología y Neurocirugía MVS, Mexico City, 14629, México

    Miguel Hernández-Cerón

  8. Departamento de Neuroanestesiología, Instituto Nacional de Neurología y Neurocirugía MVS, Mexico City, 14269, México

    Cesar Martínez-de los Santos

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  1. Edith González-Guevara
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Contributions

All authors contributed sufficiently for being credited as an author of this article. Conception and design: JCML, JFP and VC. JFP and MHC performed the stereotaxic surgery and the development of the kindling model. EGG and CMS performed the molecular experiments. ELG, ERO, and AL and JB performed the patch clamp experiment and data analysis. FPS performed the enzymatic activity experiment. VC provided technical support. EGG, ELG, and JCML performed data analysis. The first draft of the manuscript was written by JCML and all authors contributed to previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Juan Carlos Martínez-Lazcano.

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This study was conducted in accordance with the institutional guidelines and national regulation (NOM-062-ZOO-1999) from the Guide for the Care and Use of Laboratory Animals (8th edition), and international guiding principles (Council for International Organizations of Medical Sciences, CIOMS)

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González-Guevara, E., Lara-González, E., Rendon-Ochoa, E. et al. Inhibition of the NMDA Currents by Probenecid in Amygdaloid Kindling Epilepsy Model. Mol Neurobiol (2024). https://doi.org/10.1007/s12035-024-03969-0

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