Intrinsic redox properties of N-methyl-d-aspartate receptor can determine the developmental expression of excitotoxicity in rat cortical neurons in vitro

Sinor, Jeroo D.; Boeckman, Faye A.; Aizenman, Elias

doi:10.1016/s0006-8993(96)01237-1

Cited by 32 publications

(29 citation statements)

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“…This is the same period of development that we had previously showed to closely coincide with the full expression of sensitivity to NMDAinduced toxicity (Fig. 2 A; Sinor et al, 1997). Hence, the NR1/ NR2B component of the total receptor population in cortical neurons appears to diminish at a time when NMDA receptormediated excitotoxicity becomes fully expressed.…”

Section: Nmda Receptor Subunits Are Functionally Segregated In Maturesupporting

confidence: 79%

“…NMDA receptor-mediated injury was normalized to the nearly complete neuronal death produced by an overnight exposure to 1 mM kainate (Koh and Choi, 1988). As seen previously in our culture system (Sinor et al, 1997) and in others (Choi, 1985), younger cultures were substantially less sensitive to NMDA toxicity than mature cultures, although a considerable amount of cell death could be produced with the higher NMDA concentration in the immature neurons (Fig. 2 A).…”

Section: Developmental Changes In Nmda Receptor-mediated Toxicitysupporting

confidence: 77%

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NMDA and Glutamate Evoke Excitotoxicity at Distinct Cellular Locations in Rat Cortical NeuronsIn Vitro

et al. 2000

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The development of cortical neurons in vivo and in vitro is accompanied by alterations in NMDA receptor subunit expression and concomitant modifications in the pharmacological profile of NMDA-activated ionic currents. For example, we observed that with decreasing NR2B/NR2A subunit expression ratio, the block of NMDA receptor-mediated whole-cell responses by the NR2B-selective antagonist haloperidol was also decreased. In mature cultures (Ͼ22 d in vitro), however, NMDA responses obtained from excised nucleated macropatches, which comprised a large portion of the soma, remained strongly antagonized by haloperidol. These results suggest that in more mature neurons NR1/NR2B receptors appear to be preferentially expressed in the cell body. As predicted from the whole-cell recording pharmacological profile, NMDA-induced toxicity was largely unaffected by haloperidol in mature cultures. However, haloperidol effectively blocked glutamate toxicity in the same cultures, suggesting that the neurotoxic actions of this amino acid were mostly due to the activation of somatic NMDA receptors. In experiments in which the potency of glutamate toxicity was increased by the transport inhibitor L-trans-pyrrolidine-2,4-dicarboxylic acid, the neuroprotective effects of haloperidol were significantly diminished. This was likely because of the fact that glutamate, now toxic at much lower concentrations, was able to reach and activate dendritic receptors under these conditions. These results strongly argue that exogenous glutamate and NMDA normally induce excitotoxicity at distinct cellular locations in mature mixed neuronal cultures and that NR1/NR2B receptors remain an important component in the expression of glutamate, but not NMDA-induced excitotoxicity.

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Section: Nmda Receptor Subunits Are Functionally Segregated In Maturesupporting

confidence: 79%

Section: Developmental Changes In Nmda Receptor-mediated Toxicitysupporting

confidence: 77%

NMDA and Glutamate Evoke Excitotoxicity at Distinct Cellular Locations in Rat Cortical NeuronsIn Vitro

et al. 2000

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“…Hence, mature cortical neurons in culture, which have been shown to express comparatively high levels of NR2A, in conjunction with NR1 and NR2B (42,43), show a more pronounced sensitivity to KCN-mediated NMDA current potentiation than younger neurons, which express primarily NR1/NR2B receptors and less NR2A. This is in stark contrast to the potentiating actions of DTT on native NMDA receptors, which we have recently reported to be substantially larger in immature cultured neurons than in older cells (50).…”

Section: Discussionmentioning

confidence: 67%

Subunit-specific Interactions of Cyanide with the N-Methyl-d-aspartate Receptor

Arden

Sinor

Potthoff

et al. 1998

Journal of Biological Chemistry

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Cyanide is a mitochondrial poison that adversely affects cellular respiration by inhibiting the reoxidation of cytochrome a 3 by molecular oxygen, thereby obstructing the electron transport chain and oxidative phosphorylation (1). Cytochrome a 3 and cytochrome a form the cytochrome c oxidase complex, which is the terminal enzyme in the electron transport chain. Transferring electrons to oxygen, cytochrome c oxidase is the cellular respiratory component responsible for the critical need for oxygen. Thus, cyanide intoxication in cells is akin to oxygen starvation. In neuronal systems, cyanide treatment has been a widely used model of hypoxia (2-4), particularly in relation to excitotoxic processes. For example, cyanide can raise extracellular glutamate levels (5, 6), increase glutamate-triggered intracellular Ca 2ϩ elevations in neurons (7,8), and potentiate glutamate toxicity (7, 9). Moreover, specific inhibitors of Nmethyl-D-aspartate (NMDA) 1 receptors can inhibit cyanide-induced Ca 2ϩ influx in neurons (7, 10, 11) as well as neurotoxicity (12-14).Interestingly, a direct interaction between cyanide and the NMDA receptor has recently been demonstrated. Cyanide treatment of cultured rat hippocampal or cerebellar neurons potentiated NMDA-induced physiological responses, including single channel activity in excised outside-out membrane patches (15, 16). However, the precise site of action of cyanide at the receptor remained to be elucidated. In the present investigations we have evaluated the possible interaction of KCN on the NMDA receptor redox modulatory sites (17). Via these sites, disulfide-reducing agents such as dithiothreitol (DTT; see Refs. 17 and 18), dihydrolipoic acid (19), or tris(carboxyethyl)phosphine (20) enhance NMDA receptor-mediated physiological responses, whereas thiol-containing oxidants (21, 22), reactive quinones (23, 24), or oxygen-derived free radicals (25,26) can reverse the effects of reductants or depress native responses. Cyanide has well established properties as a disulfide reducing agent in many preparations (27)(28)(29)(30), and thus an effect on the NMDA thiol-sensitive sites would not be surprising. Nonetheless, the experiments described here demonstrate that cyanide can be used to distinguish between different NMDA receptor subtypes by producing either a potentiation or a depression of the physiological responses mediated by this ligand-gated ion channel. EXPERIMENTAL PROCEDURESTissue Culture-Tissue culture and all common reagents were purchased from Sigma, excluding the following: iron-supplemented bovine calf serum, HyClone Laboratories (Logan, UT), and minimal essential medium, Life Technologies, Inc. Chinese hamster ovary cells (CHO-K1; ATTC CCL61) were grown in Ham's F-12 nutrient medium with 10% fetal bovine serum, and 1 mM glutamine (CHO media) in 50-or 200-ml flasks at 37°C in 5% CO 2 . Cells were passaged at a 1:10 dilution at 80% confluency, approximately every 2 days, no more than 30 times. Cerebral cortices were obtained from E-16 Sprague-Dawley C-D rats and dissocia...

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“…Instead, P19 neurons exhibit profound ⌬⌿ m dissipation compared with neurons from younger, less vulnerable animals. These developmental changes contrast with excitotoxic vulnerability seen in cultured embryonic hippocampal neurons in which the onset of vulnerability occurs after 10 -14 d in vitro (Frandsen and Schousboe, 1990;Keilhoff and Erdo, 1991) through progressively increasing expression of the NMDA receptor (Mattson et al, 1991;Xia et al, 1995;Sinor et al, 1997).…”

Section: Discussionmentioning

confidence: 73%

Mitochondrial Nitric Oxide Mediates Decreased Vulnerability of Hippocampal Neurons from Immature Animals to NMDA

Marks¹,

Boriboun²,

Wang³

2005

J. Neurosci.

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Intrinsic redox properties of N-methyl-d-aspartate receptor can determine the developmental expression of excitotoxicity in rat cortical neurons in vitro

Cited by 32 publications

References 29 publications

NMDA and Glutamate Evoke Excitotoxicity at Distinct Cellular Locations in Rat Cortical NeuronsIn Vitro

NMDA and Glutamate Evoke Excitotoxicity at Distinct Cellular Locations in Rat Cortical NeuronsIn Vitro

Subunit-specific Interactions of Cyanide with the N-Methyl-d-aspartate Receptor

Mitochondrial Nitric Oxide Mediates Decreased Vulnerability of Hippocampal Neurons from Immature Animals to NMDA

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