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Human RNA integrity after postmortem retinal tissue recovery.

Authors :
Montanini L
Ferrari S
Crafa P
Ghirardini S
Ponzin D
Orsoni JG
Mora P
Source :
Ophthalmic genetics [Ophthalmic Genet] 2013 Mar-Jun; Vol. 34 (1-2), pp. 27-31. Date of Electronic Publication: 2012 Sep 06.
Publication Year :
2013

Abstract

Purpose: To assess the parameters for postmortem retinal tissue recovery and processing that affect the quality of RNA extracted from the retina/retinal pigment epithelium (RPE) complex.<br />Methods: RNA was extracted from retina/RPE samples. The RNA quality was determined based on qualitative/quantitative measurements made with a Bioanalyzer (Agilent) and on the expression of a long retinal gene (RPE65). After a pilot analysis on rats, ocular RNA was extracted from human donor eyeballs (group A) explanted according to conventional procedures for cornea transplantation. In a second experiment, another group of human donor eyeballs (group B) were processed in a much shorter time. The postmortem interval (T) comprised two periods: T1, the time between a donor's death and enucleation, and T2, the time between eyeball explantation and immersion of the excised retina/RPE sample in preservative solution (T = T1 + T2).<br />Results: A short T2 was correlated with good quality of RNA extracted from the retina/RPE complex (p = 0.043) and successful expression of a tissue-specific gene (p = 0.007). No other parameter appeared to influence RNA quality.<br />Conclusions: The time between eyeball explantation and immersion of the retina/RPE sample in preservative solution was the chief parameter affecting the quality of RNA extracted from the retina/RPE complex.

Details

Language :
English
ISSN :
1744-5094
Volume :
34
Issue :
1-2
Database :
MEDLINE
Journal :
Ophthalmic genetics
Publication Type :
Academic Journal
Accession number :
22950451
Full Text :
https://doi.org/10.3109/13816810.2012.720342