title: Transcriptomic analysis of human retinal surgical specimens using jouRNAI.
abstract: Retinal detachment (RD) describes a separation of the neurosensory retina from the retinal pigmented epithelium (RPE). The RPE is essential for normal function of the light sensitive neurons, the photoreceptors. Detachment of the retina from the RPE creates a physical gap that is filled with extracellular fluid. RD initiates cellular and molecular adverse events that affect both the neurosensory retina and the RPE since the physiological exchange of ions and metabolites is severely perturbed. The consequence for vision is related to the duration of the detachment since a rapid reapposition of the two tissues results in the restoration of vision (1). The treatment of RD is exclusively surgical. Removal of vitreous gel (vitrectomy) is followed by the removal non essential part of the retina around the detached area to favor retinal detachment. The removed retinal specimens are res nullius (nothing) and consequently normally discarded. To recover RNA from these surgical specimens, we developed the procedure jouRNAl that allows RNA conservation during the transfer from the surgical block to the laboratory. We also standardized a protocol to purify RNA by cesium chloride ultracentrifugation to assure that the purified RNAs are suitable for global gene expression analysis. The quality of the RNA was validated both by RT-PCR and microarray analysis. Analysis of the data shows a simultaneous involvement of inflammation and photoreceptor degeneration during RD.
authors: U968, Institut National de la Santé et de la Recherche Médicale.
authors: Marie-Noëlle
authors: MN
authors: Delyfer
authors: Najate
authors: N
authors: Aït-Ali
authors: Hawa
authors: H
authors: Camara
authors: Emmanuelle
authors: E
authors: Clérin
authors: Jean-François
authors: JF
authors: Korobelnik
authors: José-Alain
authors: JA
authors: Sahel
authors: Thierry
authors: T
authors: Léveillard
mesh_terms: D002586:Cesium / Q000737:chemistry; D002712:Chlorides / Q000737:chemistry; D004587:Electrophoresis, Agar Gel / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D012313:RNA / Q000235:genetics / Q000302:isolation & purification*; D012160:Retina / Q000737:chemistry* / Q000502:physiology; D012163:Retinal Detachment / Q000235:genetics; D014461:Ultracentrifugation / Q000379:methods
chemical_list: D002712:Chlorides; D002586:Cesium; D012313:RNA; C028019:cesium chloride
references: Proc Natl Acad Sci U S A. 1990 Mar;87(5):1663-7
references: 1689846
references: Br J Ophthalmol. 2010 Jun;94(6):678-84
references: 19515646
references: BMC Genomics. 2008 May 05;9:208
references: 18457592
references: PLoS One. 2011;6(12):e28791
references: 22174898
references: Sci Transl Med. 2010 Apr 7;2(26):26ps16
references: 20375363
references: BMC Mol Biol. 2007 Aug 31;8:74
references: 17764561
references: Nucleic Acids Res. 1975 Dec;2(12):2315-28
references: 79997
references: Science. 2010 May 7;328(5979):710-722
references: 20448178
references: Biochemistry. 1974 Jun 4;13(12):2633-7
references: 4831907
references: Proc Natl Acad Sci U S A. 2007 Feb 13;104(7):2425-30
references: 17284607
references: Biochem Biophys Res Commun. 1981 Jan 15;98(1):261-7
references: 7011323