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Hamid Ahmadieh, Bahareh Safdari, Fatemeh Suri, Zahra-Soheila Soheili, Iman Salahshourifar, Mozhgan Rezaeikanavi; NOGO-A Gene Expression in Amniotic Fluid Treated Human RPE Cells. Invest. Ophthalmol. Vis. Sci. 2017;58(8):608.
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© ARVO (1962-2015); The Authors (2016-present)
Using retinal pigment epithelial (RPE) cells as a potential source of retinal neurons has already been reported. Previous studies have shown that human amniotic fluid (HAF) can enhance trans-differentiation of RPE cells into the retinal neurons and progenitor cells. NOGO-A is the longest member of NOGO family, all of which are coded by RTN-4 gene. During neural development, NOGO is expressed mainly by neurons and provides an inhibitory signal for the migration and sprouting of endothelial cells, thereby restricting blood vessel density. NOGO-A is expressed in retinal neurons, being up-regulated after neural lesions. NOGO-A inhibits axon growth and synaptic function in the central nervous system. Suppressing the activity of the NOGO receptor has been shown to enhance neural regeneration and survival. In this study, we examined the expression of NOGO-A in cultivated human RPE cells and investigated the effect of HAF treatment on NOGO-A gene expression.
Primary cultured human RPE cells were established from three unrelated human cadaver globes. RPE cells were cultured in DMEM/F12 supplemented with 10% FBS. Cultures were treated with 30% HAF, obtained from normal fetuses of 14–16 weeks gestational age. Real time PCR analysis was used to determine NOGO-A gene expression. All the experiments were performed in duplicate.
Results showed that NOGO-A gene was expressed in cultured human RPE cells. Comparative analysis also demonstrated that NOGO-A gene expression was increased one hundred times in average in HAF-treated cultures, when compared to FBS-treated controls (P < 0.001).
NOGO-A gene expression was detected in human RPE cells’ culture. The high level of NOGO-A transcripts seems to be a key regulatory factor in HAF-treated RPE cells. This may support previous evidences on neural differentiation of RPE cells after exposure to HAF. Further investigations are necessary to verify the role of NOGO-A in trans-differentiation of RPE cells and recruitment of the RPE cells in retinal regeneration studies.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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