NDLI: Age-related gene response of human corneal endothelium to oxidative stress and DNA damage.

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Age-related gene response of human corneal endothelium to oxidative stress and DNA damage.

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Age-related gene response of human corneal endothelium to oxidative stress and DNA damage.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Harris, Deshea L. Zhu, Cheng C. Joyce, Nancy C.
Description	Country affiliation: United States Author Affiliation: Joyce NC ( Schepens Eye Research Institute, Harvard Medical School, Boston, Massachusetts 02114, USA. nancy.joyce@schepens.harvard.edu)
Abstract	PURPOSE: Nuclear oxidative DNA damage increases with age in human corneal endothelial cells (HCECs) and contributes to their decreased proliferative capacity. These studies investigated whether HCECs respond to this damage by upregulating their expression of oxidative stress and DNA damage-signaling genes in an age-dependent manner. METHODS: HCECs were dissected from the corneas of young (30 years and younger) and older (50 years and older) donors. Total RNA was isolated and reverse-transcribed. Oxidative stress and DNA damage-signaling gene expression were analyzed using commercial PCR-based microarrays. Western blot analyses were conducted on selected proteins to verify the microarray results. Nuclear DNA damage foci were detected in the endothelium of ex vivo corneas by immunostaining for H2AX-Ser139. RESULTS: Four of 84 genes showed a statistically significant age-related difference in the expression of oxidative stress-related genes; however, Western blot analysis demonstrated an age-related increase in only 2 (cytoglobin and GPX-1) of 11 proteins tested. No age-related differences were detected in the expression of DNA damage-signaling genes. Western blot analysis of seven DNA damage-related proteins verified this finding. Intense nuclear staining of DNA damage foci was observed in nuclei within the central endothelium of older donors. Central endothelium from young donors consistently showed a low level of positive staining. CONCLUSIONS: HCECs respond to age-related increases in oxidative nuclear DNA damage by forming DNA damage repair foci; however, they do not vigorously defend against or repair this damage by upregulating the expression of multiple oxidative stress or DNA damage-signaling genes.
ISSN	01460404
e-ISSN	15525783
DOI	10.1167/iovs.10-6492
Journal	Investigative Opthalmology & Visual Science
Issue Number	3
Volume Number	52
Language	English
Publisher	Association for Research in Vision and Ophthalmology
Publisher Date	2011-03-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Aging Physiology Dna Damage Genetics Endothelium, Corneal Metabolism Eye Proteins Gene Expression Regulation Oxidative Stress Adolescent Blotting, Western Child, Preschool Histones Oligonucleotide Array Sequence Analysis Polymerase Chain Reaction Tissue Donors Research Support, N.i.h., Extramural Discipline Ophthalmology
Content Type	Text
Resource Type	Article
Subject	Ophthalmology Sensory Systems Cellular and Molecular Neuroscience

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