title: Human RNA integrity after postmortem retinal tissue recovery.
abstract: OBJECTIVE
  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.
  
  
  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).
  
  
  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.
  
  
  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.
authors.affiliation: Institute of Ophthalmology, University of Parma, Parma, Italy. luisa.montanini@unipr.it
authors.forename: Luisa
authors.initials: L
authors.lastname: Montanini
authors.forename: Stefano
authors.initials: S
authors.lastname: Ferrari
authors.forename: Pellegrino
authors.initials: P
authors.lastname: Crafa
authors.forename: Stella
authors.initials: S
authors.lastname: Ghirardini
authors.forename: Diego
authors.initials: D
authors.lastname: Ponzin
authors.forename: Jelka G
authors.initials: JG
authors.lastname: Orsoni
authors.forename: Paolo
authors.initials: P
authors.lastname: Mora
mesh_terms: D000328:Adult; D000368:Aged; D000818:Animals; D005260:Female; D015870:Gene Expression; D006801:Humans; D008297:Male; D008875:Middle Aged; D009926:Organ Preservation; D016133:Polymerase Chain Reaction; D011180:Postmortem Changes; D012313:RNA / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000737:chemistry*; D055213:Retinal Pigment Epithelium / Q000737:chemistry*; D014019:Tissue Donors; D009927:Tissue and Organ Procurement; D019745:cis-trans-Isomerases / Q000235:genetics; D000099178:Retinoid Isomerohydrolase
chemical_list: D012313:RNA; D012333:RNA, Messenger; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase