Differential Expression of the Synapse Regulatory Proteins Neurexins in Early Diabetic Retinal Disease

Neurexins (NRXNs), a complex family of synapse regulatory proteins, represent attractive candidate molecular mediators of retinal neuronal dysfunction in early diabetic retinal disease (DRD) given their roles in the central nervous system and recent work suggesting a potential link with diabetes mellitus (DM). As antibodies are unable to distinguish NRXN family members and isoforms, the purpose of this study was to investigate differential expression of individual NRXN transcripts in relevant retinal cell types in early DRD.

RNAscope multiplexed fluorescent in situ hybridization was used to quantify transcripts of NRXNs in mouse and human retina. DM was induced in C57BL/6J mice using streptozotocin (STZ) and the retinal phenotype characterized by electroretinogram (ERG), optical coherence tomography (OCT), and optokinetic tracking (OKT) after 6 weeks. Retinal ganglion cells (RGCs) were generated in vitro by directed differentiation from human embryonic stem cells (hESCs), cultured in normal or high glucose, and NRXN expression assessed by qPCR and Western blot.

All NRXN family members (NRXN1, NRXN2, and NRXN3) were enriched in inner retinal neurons in both human and mouse tissue. We identified decreased Nrxn3 transcripts, specifically the Nrxn3β isoform, in RGCs of diabetic mice at 6 weeks after STZ treatment. Interestingly, Nrxn3α and Nrxn3β were differentially expressed in inner retinal layers. Finally, we confirmed decreased Nrxn3β expression in hESC-RGCs cultured in high glucose in vitro.

Our findings suggest that NRXNs may play cell-type-specific roles in the inner retina and associate decreased Nrxn3β expression in RGCs with inner retinal dysfunction in early DRD.