NMDA Glutamate Receptor NR1, NR2A and NR2B Expression and NR2B Tyr-1472 Phosphorylation in the Lens

Detailed parallels described between lens fiber cell and neuron morphology, sub-cellular structure, and molecular biology include striking similarities in the ultrastructure of their vesicle transport machinery and the membrane protrusions that occur along the lateral surfaces of both cell types. α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and N-methyl-d-aspartate receptor (NMDA) glutamate receptors (AMPARs/NMDARs) are the predominant receptors in neurons. These receptors have fundamental roles in neuron morphogenesis as well as neuron physiology and dynamic cell signaling, and specifically at dendritic spines. As a result, AMPAR and NMDAR dysregulation underlies several primary neural disorders that have also shown epidemiological associations with cataract. Previously, we demonstrated AMPAR GluA1 and REST (RE-1 silencing transcription factor)-regulated GluA2 subunits are expressed in the lens, and showed C-terminal phospho-tyrosine-GluA2, and striatal-enriched tyrosine phosphatase (STEP), as well as GluA2 Q/R RNA editing in lenses similar to neurons. Here, we demonstrated that REST-regulated NMDAR NR1, NR2A, and NR2B are also expressed in lenses and localize predominantly in fiber cell membranes, consistent with REST transcription factors, as well as miR-124 and other REST gene targets identified in the lens. We also showed NR2B Tyr-1472 phosphorylation occurs in lens. These p-Tyr-GluA2 and p-Tyr-NR2B phosphorylation events are linked with membrane insertion regulated by STEP. We next determined that NR1 transcripts that include exon 5 are produced in lens consistent with Fox-1 RNA binding protein isoforms linked with this alternative splicing event, and shown to be expressed in lens as well as brain. These findings provide further evidence that fundamental neuronal morphogenetic programs, and hallmark neuronal gene expression and modes of regulation, are shared with elongated fiber cells of the lens.