Abstract:
:AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) excitotoxicity was examined in cultured neocortical neurons using the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to measure cell viability. Neurons were exposed to AMPA at different culture periods during development of the neurons. In order to describe the pharmacology of AMPA-mediated toxicity, several glutamate receptor antagonists were used: MK-801, NS 394, NBQX, GYKI 52466, GYKI 53405 and GYKI 53655. Increased excitotoxicity was observed when cortical neurons cultured for 5, 8 and 12 days in vitro (DIV) were exposed to a high concentration of AMPA (500 microM) for 6 h. However, only at DIV 12 was part of the toxicity mediated directly through AMPA receptors since 10 microM MK-801 blocked all AMPA toxicity at DIV 5 and 8, but only some of the AMPA response at DIV 12. This indicated that NMDA receptors were being activated, causing some of the observed toxicity. The high dose of AMPA was not sufficient to damage all neurons since 59% remained viable after exposure to AMPA even for neurons that were cultured for 12 DIV. Since it is known that both glutamate and AMPA activate AMPA receptors with a fast and rapidly desensitizing response, this could explain the relatively low toxicity produced by 500 microM AMPA. This was investigated by blocking AMPA receptor desensitization with cyclothiazide. Using a lower concentration (25 microM) of AMPA, addition of 50 microM cyclothiazide increased the AMPA induced excitotoxicity in cultured cortical neurons at all DIV except for DIV 2. This combination of AMPA + cyclothiazide yielded 77% cell death for DIV 12 cultures. In contrast to the results observed with 500 microM AMPA, the neurotoxicity mediated directly by AMPA receptors when desensitization was blocked was seen as early as 5 DIV since 10 microM MK-801 did not completely block the response whereas 10 microM NBQX did. The 2,3-benzodiazepine GYKI compounds, which have been reported to be selective non-competitive AMPA receptor antagonists, were here observed to block the AMPA toxicity with the following rank order: GYKI 53655 > GYKI 52466 > or = GYKI 53405, which is in agreement with their published potencies.
journal_name
Neurochem Intjournal_title
Neurochemistry internationalauthors
Jensen JB,Schousboe A,Pickering DSdoi
10.1016/s0197-0186(97)00130-7subject
Has Abstractpub_date
1998-05-01 00:00:00pages
505-13issue
5-6eissn
0197-0186issn
1872-9754pii
S0197-0186(97)00130-7journal_volume
32pub_type
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