Glutamate Receptor Inhibition in Cortical Cultures

Mixed, astrocyte-only and neuron-only cortical cultures were prepared from dissociated rat E18 cortical tissue [29 (link)]. The effect of glutamate receptor inhibition was investigated by adding 10 μM MK801 to the cultures 4 h before incubation with conditioned media. Immunocytochemical assessment was performed as described previously [28 (link)] using antibodies against MAP2 (Synaptic Systems 188004; 1:2000), GFAP (CST 3670; 1:1000), tyrosine hydroxylase (abcam ab112; 1:500), GluN1 (Millipore Ab9864; 1:200), GluN3α (Millipore 07-356; 1:200), GABA Aα1 (abcam ab33299; 1:250) and GABA B1 (abcam ab55051; 1:250).

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Scott H., Phillips T.J., Stuart G.C., Rogers M.F., Steinkraus B.R., Grant S, & Case C.P. (2018). Preeclamptic placentae release factors that damage neurons: implications for foetal programming of disease. Neuronal Signaling, 2(4), NS20180139.

Publication 2018

ab112 Ab9864 ab33299 ab55051

Antibodies Astrocyte Conditioned media Cortical Gaba Gfap Glutamate receptor Inhibition Map2 Mk801 Neuron Tissue Tyrosine hydroxylase

Corresponding Organization : Cardiff University

Other organizations : John Radcliffe Hospital, University of Oxford

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Variable analysis

independent variables

Addition of 10 μM MK801 (glutamate receptor inhibitor) to the cultures 4 h before incubation with conditioned media

dependent variables

Immunocytochemical assessment of the expression of MAP2 (neuronal marker), GFAP (astrocyte marker), tyrosine hydroxylase (dopaminergic neuron marker), GluN1 (NMDA receptor subunit), GluN3α (NMDA receptor subunit), GABA Aα1 (GABAA receptor subunit), and GABA B1 (GABAB receptor subunit)

control variables

Dissociated rat E18 cortical tissue used to prepare mixed, astrocyte-only, and neuron-only cortical cultures
Immunocytochemical assessment procedures described previously [28]

Annotations

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