MiRNA Changes in Retinal Ganglion Cells after Optic Nerve Crush and Glaucomatous Damage.

Ben Mead,Alicia Kerr,Naoki Nakaya,Stanislav I Tomarev

doi:10.3390/cells10071564

Ben Mead, Alicia Kerr + Show 2 more

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https://doi.org/10.3390/cells10071564

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Abstract

The purpose of this study was to characterize the miRNA profile of purified retinal ganglion cells (RGC) from healthy and diseased rat retina. Diseased retina includes those after a traumatic optic nerve crush (ONC), and after ocular hypertension/glaucoma. Rats were separated into four groups: healthy/intact, 7 days after laser-induced ocular hypertension, 2 days after traumatic ONC, and 7 days after ONC. RGC were purified from rat retina using microbeads conjugated to CD90.1/Thy1. RNA were sequenced using Next Generation Sequencing. Over 100 miRNA were identified that were significantly different in diseased retina compared to healthy retina. Considerable differences were seen in the miRNA expression of RGC 7 days after ONC, whereas after 2 days, few changes were seen. The miRNA profiles of RGC 7 days after ONC and 7 days after ocular hypertension were similar, but discrete miRNA differences were still seen. Candidate mRNA showing different levels of expression after retinal injury were manipulated in RGC cultures using mimics/AntagomiRs. Of the five candidate miRNA identified and subsequently tested for therapeutic efficacy, miR-194 inhibitor and miR-664-2 inhibitor elicited significant RGC neuroprotection, whereas miR-181a mimic and miR-181d-5p mimic elicited significant RGC neuritogenesis.

Highlights

Retinal ganglion cells (RGC) are the sole projection neurons of the retina to the brain, and their axons makes up the optic nerve
Adult female outbred Sprague-Dawley rats weighing 150 to 200 g (Charles River, Wilmington, MA, USA) and adult 2–3-month-old C57BL/6J mice were maintained in accordance with guidelines described in the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research, using protocols approved by the National Eye Institute Committee on the Use and Care of Animals/Cardiff University’s Biological Standard’s committee
We identified over 100 miRNA whose abundance was significantly (p < 0.001) different in injured retinal ganglion cells (RGC) (Group 2, 3, and 4) in comparison to uninjured RGC (Group 1; Figure 2)

Summary

Introduction

Retinal ganglion cells (RGC) are the sole projection neurons of the retina to the brain, and their axons makes up the optic nerve. They are the one of the cell types most responsible for irreversible vision loss (e.g., glaucoma and traumatic optic neuropathy), and an accessible model used to study spinal cord injury and, to a lesser extent, traumatic brain injury [1]. It is for this reason that RGC have become perhaps one of the most attractive cell types from a research perspective. The optic nerve is the main site of damage, which can come about due to trauma or degeneration

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