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N-Methyl-D-Aspartate Receptor Subunit NR3A in the Retina: Developmental Expression, Cellular Localization, and Functional Aspects

¹From the Department of Biology and Biotechnology Research Institute, Hong Kong University of Science and Technology, Hong Kong, China; the ²Department of Experimental Ophthalmology, University Eye Hospital, Tübingen, Germany; the ⁴Natural and Medical Sciences Institute, Reutlingen, Germany; the ³CNS Research Institute, Brigham and Women’s Hospital, Boston, Massachusetts; and the ⁵Program in Neuroscience, Harvard Medical School, Boston, Massachusetts.

PURPOSE. Recently, a novel N-methyl-D-aspartate receptor (NMDAR) subunit, NR3A, has been discovered in the brain and shown to decrease NMDAR activity by modulating the calcium permeability of the receptor channel. The insertion of NR3A within the NMDAR complex may thus alter NMDAR properties and play a crucial role during processes of neuronal development and degeneration. The present study is the first to investigate the expression and cellular localization of NR3A on the protein level in the retina and to elucidate its putative functional roles within the retinal circuitry.

METHODS. The expression of NR3A in the retina was analyzed by reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry, and Western blot analysis. Functional aspects of NR3A in the retina were addressed by measuring the NMDA-induced increase in intracellular calcium, [Ca²⁺]_i, in retinal cells prepared from wild-type (NR3A^+/+) and NR3A knockout (NR3A^+/-, and NR3A^-/-) mice.

RESULTS. NR3A protein expression was initially observed in the first postnatal week and was predominantly localized to cell bodies in the ganglion cell layer. In older animals, two bands of NR3A immunoreactivity were additionally observed in the inner plexiform layer. NMDA-evoked [Ca²⁺]_i responses were found to be significantly greater in retinal cells in NR3A^-/- mice than in wild-type retinas.

CONCLUSIONS. The data indicate that NR3A is specifically expressed in the inner retina and may modulate NMDAR-mediated calcium influx and thus [Ca²⁺]_i levels in retinal ganglion cells and amacrine cells.

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