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Toxicity of beta-amyloid in HEK293 cells expressing NR1/NR2A or NR1/NR2B N-methyl-D-aspartate receptor subunits.
Neurochem Int. 2007 May; 50(6):872-80.NI

Abstract

Neurotoxicity induced by beta-amyloid peptide (Abeta) involves glutamate toxicity, resulting from overactivation of N-methyl-D-aspartate (NMDA) receptors and elevation of intracellular calcium. However, the heterogeneity of the NMDA receptors, frequently composed of NR1 and NR2A-D subunits, has been less studied. Thus, we determined the contribution of NMDA receptor subtypes on Abeta(1-40) toxicity in HEK293 cells transiently expressing NR1/NR2A or NR1/NR2B subunits. Analysis of lactate dehydrogenase (LDH) release and trypan blue exclusion revealed an increase in Abeta(1-40) toxicity upon NR1/NR2A expression, compared to NR1/NR2B, indicating loss of plasma membrane integrity. Furthermore, Abeta(1-40) decreased intracellular ATP in cells expressing NR1/NR2A. MK-801 ((+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate), a noncompetitive NMDA receptor antagonist, partially prevented the decrease in cell viability and the energy impairment. These differences were not accounted for by the activation of caspases 2, 3, 8 and 9 or calpains or by DNA fragmentation, excluding the hypothesis of apoptosis. Functional NR1/NR2A and NR1/NR2B receptor subtypes were further evidenced by single-cell calcium imaging. Stimulation of NR1/NR2A receptors with NMDA/glycine revealed an increase in intracellular calcium in cells pre-exposed to Abeta(1-40). Opposite effects were observed upon activation of NR1/NR2B receptors. These results suggest that NR1/NR2A-composed NMDA receptors mediate necrotic cell death in HEK293 cells exposed to Abeta(1-40) through changes in calcium homeostasis.

Authors+Show Affiliations

Domingues A
Institute of Biochemistry, Faculty of Medicine and Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal.
Almeida S
No affiliation info available
da Cruz e Silva EF
No affiliation info available
Oliveira CR
No affiliation info available
Rego AC
No affiliation info available

MeSH

Adenosine TriphosphateAmyloid beta-PeptidesBlotting, WesternCalciumCaspasesCell LineCell SurvivalDNA FragmentationDizocilpine MaleateExcitatory Amino Acid AntagonistsHumansImmunohistochemistryL-Lactate DehydrogenaseReceptors, N-Methyl-D-AspartateStimulation, ChemicalTransfection

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17403555

Citation

Domingues, A, et al. "Toxicity of Beta-amyloid in HEK293 Cells Expressing NR1/NR2A or NR1/NR2B N-methyl-D-aspartate Receptor Subunits." Neurochemistry International, vol. 50, no. 6, 2007, pp. 872-80.
Domingues A, Almeida S, da Cruz e Silva EF, et al. Toxicity of beta-amyloid in HEK293 cells expressing NR1/NR2A or NR1/NR2B N-methyl-D-aspartate receptor subunits. Neurochem Int. 2007;50(6):872-80.
Domingues, A., Almeida, S., da Cruz e Silva, E. F., Oliveira, C. R., & Rego, A. C. (2007). Toxicity of beta-amyloid in HEK293 cells expressing NR1/NR2A or NR1/NR2B N-methyl-D-aspartate receptor subunits. Neurochemistry International, 50(6), 872-80.
Domingues A, et al. Toxicity of Beta-amyloid in HEK293 Cells Expressing NR1/NR2A or NR1/NR2B N-methyl-D-aspartate Receptor Subunits. Neurochem Int. 2007;50(6):872-80. PubMed PMID: 17403555.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Toxicity of beta-amyloid in HEK293 cells expressing NR1/NR2A or NR1/NR2B N-methyl-D-aspartate receptor subunits. AU - Domingues,A, AU - Almeida,S, AU - da Cruz e Silva,E F, AU - Oliveira,C R, AU - Rego,A C, Y1 - 2007/03/07/ PY - 2007/02/28/received PY - 2007/03/01/accepted PY - 2007/4/4/pubmed PY - 2007/7/27/medline PY - 2007/4/4/entrez SP - 872 EP - 80 JF - Neurochemistry international JO - Neurochem Int VL - 50 IS - 6 N2 - Neurotoxicity induced by beta-amyloid peptide (Abeta) involves glutamate toxicity, resulting from overactivation of N-methyl-D-aspartate (NMDA) receptors and elevation of intracellular calcium. However, the heterogeneity of the NMDA receptors, frequently composed of NR1 and NR2A-D subunits, has been less studied. Thus, we determined the contribution of NMDA receptor subtypes on Abeta(1-40) toxicity in HEK293 cells transiently expressing NR1/NR2A or NR1/NR2B subunits. Analysis of lactate dehydrogenase (LDH) release and trypan blue exclusion revealed an increase in Abeta(1-40) toxicity upon NR1/NR2A expression, compared to NR1/NR2B, indicating loss of plasma membrane integrity. Furthermore, Abeta(1-40) decreased intracellular ATP in cells expressing NR1/NR2A. MK-801 ((+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate), a noncompetitive NMDA receptor antagonist, partially prevented the decrease in cell viability and the energy impairment. These differences were not accounted for by the activation of caspases 2, 3, 8 and 9 or calpains or by DNA fragmentation, excluding the hypothesis of apoptosis. Functional NR1/NR2A and NR1/NR2B receptor subtypes were further evidenced by single-cell calcium imaging. Stimulation of NR1/NR2A receptors with NMDA/glycine revealed an increase in intracellular calcium in cells pre-exposed to Abeta(1-40). Opposite effects were observed upon activation of NR1/NR2B receptors. These results suggest that NR1/NR2A-composed NMDA receptors mediate necrotic cell death in HEK293 cells exposed to Abeta(1-40) through changes in calcium homeostasis. SN - 0197-0186 UR - https://www.unboundmedicine.com/medline/citation/17403555/Toxicity_of_beta_amyloid_in_HEK293_cells_expressing_NR1/NR2A_or_NR1/NR2B_N_methyl_D_aspartate_receptor_subunits_ DB - PRIME DP - Unbound Medicine ER -