Diabetes changes the levels of ionotropic glutamate receptors in the rat retina.

Purpose: Diabetic retinopathy (DR) is a leading cause of vision loss and blindness among adults between the age 20 to 74. Changes in ionotropic glutamate receptor subunit composition can affect retinal glutamatergic neurotransmission and, therefore, contribute to visual impairment. The purpose of this study was to investigate whether diabetes leads to changes in ionotropic glutamate receptor subunit expression at the protein and mRNA level in the rat retina.

Elevated glucose changes the expression of ionotropic glutamate receptor subunits and impairs calcium homeostasis in retinal neural cells.

Purpose: Altered glutamatergic neurotransmission and calcium homeostasis may contribute to retinal neural cell dysfunction and apoptosis in diabetic retinopathy (DR). The purpose of this study was to determine the effect of high glucose on the protein content of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and kainate glutamate receptor subunits, particularly the GluR2 subunit, because it controls Ca2+ permeability of AMPA receptor-associated channels. The effect of high glucose on the concentration of cytosolic free calcium ([Ca2+]i) was also investigated.

High glucose and diabetes increase the release of [3H]-D-aspartate in retinal cell cultures and in rat retinas.

Several evidences suggest that glutamate may be involved in retinal neurodegeneration in diabetic retinopathy (DR). For that reason, we investigated whether high glucose or diabetes affect the accumulation and the release of [(3)H]-D-aspartate, which was used as a marker of the glutamate transmitter pool. The accumulation of [(3)H]-D-aspartate did not change in cultured retinal neural cells treated with high glucose (30 mM) for 7 days.

Development of chick retina cells in culture: cobalt entry through AMPA receptors and expression of GluR4 AMPA receptor subunit.

The functionality of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors in chick embryo retina cells during development in vitro was studied by using Co(2+) uptake, and these data were correlated with the expression of the AMPA receptor subunit GluR4. We found that at 5 h in vitro only a small number of cells took up Co(2+) upon stimulation with 100 microM kainate or other AMPA receptor agonists, in the presence of cyclothiazide (CTZ), to inhibit desensitisation. The number of cells sensitive to kainate increased from 5 h in vitro to 3 days in vitro (DIV), and remained relatively constant until 14 DIV.

Expression of AMPA/kainate receptors during development of chick embryo retina cells: in vitro versus in vivo studies.

The activity and the subunit expression of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA)/kainate ionotropic glutamate receptors were studied in retina cells developing in chick embryos and in retina cells cultured as retinospheroids, at the same stages of development. In the retinospheroids, the activity of the AMPA/kainate receptors was monitored by following the changes in the intracellular free calcium concentration ([Ca(2+)](i)), in response to AMPA, kainate or to L-glutamate, and the expression of the receptor subunits GluR1, GluR2/3, GluR4 and GluR6/7 was determined in the retinospheroids and in chick retinas by immunodetection using polyclonal antibodies. The changes in [Ca(2+)](i) in response to 400 microM kainate increased from 5h in vitro to 3 days, and remained constant until day 14, whereas the [Ca(2+)](i) in response to 500 microM L-glutamate or 400 microM AMPA increased from 5h in vitro to 3 days, and thereafter decreased slightly until day 14.

Expression of functional N-methyl-D-aspartate receptors during development of chick embryo retina cells: in vitro versus in vivo studies.

The N-methyl-D-aspartate (NMDA) ionotropic glutamate receptors were studied in retina cells developing in chick embryos and in retina cells cultured as retinospheroids, at the same stages of development. In the retinospheroids, the activity of the NMDA receptors was followed by monitoring the changes in the intracellular free calcium concentration ([Ca2+](i)), in response to NMDA or to L-glutamate. The expression of the subunits NMDAR1, NMDAR2A/B and NMDAR2C in the retinospheroids and in chick retinas were determined by Western blot analyses.