Electrostimulation improves progenitor cell potential of Muller glial cells.

Abstract

Purpose : We know that retinal tissue can repair itself upon injury and specific cells in the retina possess the ability to fully regeneration without producing scar tissue. The Muller glial cells in zebrafish act as progenitor cells and respond as rescue cells upon injury. Mammals have not spontaneously shown this ability. However, it appears that low current electricity can challenge this fact noninvasively.

Methods : Retinal cells isolated from neonatal B6 mice aged postnatal day 6–7 (P6–7) were dissociated with papain and incubated for 14 days in Dulbecco’s modified Eagle’s medium (DMEM)/F12 medium to yield 95% enriched Muller cell cultures. The cells were seeded on a poly-D-lysine–coated cover glass. Optimal ES conditions established from our earlier work with ramp waveform were applied for 15 minutes and for 1 hour. Gene expression was profiled using RNA sequencing, and progenitor cell markers were quantified 2 days after ES by reverse transcriptase–polymerase chain reaction (qPCR).

Results : We observed upregulation of retinal progenitor cells markers, such as Crx, Chx10, Sox2, and Pax6 in the MCs group that received optimal ES conditions for 1 hour. Upregulation of Wnt7 and CNTF was also observed post ES. KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis of the RNA sequencing data showed upregulation of the Ca²⁺ signaling pathway, glutamatergic synapse, cholinergic synapse, GABAergic synapse, and dopaminergic synapse respectively.

Conclusions : Our results show that ES with ramp waveform changes the genetic landscape and improves the progenitor cell potential of MCs, suggesting the exciting possibility of modifying MC’s regenerative abilities by ES in the future.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.