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Julie Sanderson, Marina Hopes, David C Broadway; HDAC Inhibition in Human Organotypic Retinal Cultures (HORCs) protects against loss of THY-1 mRNA. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1890.
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Histone deacetylase (HDAC) inhibitors have been associated with potential neuroprotective properties in relation to glaucoma. The purpose of these experiments was to investigate the effect of the HDAC inhibitor trichostatin A (TSA) on gene expression in human organotypic retinal cultures (HORCs).
Donor eyes were obtained from the East Anglian Eye Bank within 24 hours post mortem. 4mm diameter paramacular explants were dissected from the retina and cultured in serum free DMEM/Ham F12 medium for 48 hours in the presence or absence of TSA (0.1, 1 or 10µM). LDH release was used to assess cell death. Total RNA levels were assessed by spectrophotometric analysis. Gene expression was assessed by QRT-PCR.
There was no significant change in LDH release from the HORCs following 48 hours' exposure to 0.1 - 10µM TSA indicating that the HDAC inhibitor was not causing toxicity (n=4). During the culture period, there was an approximate 60% decrease in total RNA (n=4). TSA led to a significant amelioration of loss of total RNA (approximately 25% at 10μM; n=4). Analysis of the expression of two housekeeping genes (cytochrome c-1; CYC1 and topoisomerase 1; TOP1) showed no significant change in expression in TSA-treated retina compared with control. The expression of three markers for retinal neurons was also assessed: Thy-1 (THY-1) for retinal ganglion cells (RGCs); recoverin (RCVRN) for photoreceptors and calbindin (CALB) for horizontal cells. There was no significant change in expression of CALB in HORCs treated with TSA compared with control (n=4). RCVRN showed a significant increase in expression of approximately 20% (n=4) and THY-1 an approximate 80% increase (n=4) in expression in HORCs treated with 10µM TSA compared with control at the 48 hour time point.
HDAC inhibition with TSA differentially inhibited loss of gene expression in the cultured human retina. Of the genes investigated, the RGC marker THY-1 showed the greatest protection. Loss of normal gene expression has been shown to be an early event in animal models of glaucoma. The current data may support a potential neuroprotective role for HDAC inhibitors in relation to glaucoma.
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