Postmortem stability of total RNA isolated from rabbit ligament, tendon and cartilage.
The stability of RNA, particularly mRNA, in tissues is under complex regulation. Most studies to date have focused on very cellular tissues and not connective tissues such as ligaments, tendons and cartilage. As the availability of such tissues for transplantation or research purposes is frequently delayed following death, it is important to determine whether RNA stability in such tissues is influenced by time postmortem. To approach this question, skeletally mature NZW rabbits were used to investigate RNA integrity over time in dense, hypocellular connective tissues and in several hypercellular organ tissues such as brain, kidney, liver and lung. Samples were analyzed at varying intervals postmortem with respect to rRNA integrity by agarose gel electrophoresis and ethidium bromide staining and mRNA integrity by Northern blot analysis and RT-PCR. No degradation of rRNA or loss in integrity of mRNA for genes of low and high copy number was observed up to 96 h postmortem. These findings confirm that it is likely appropriate to use properly stored postmortem dense connective tissues for molecular biological investigations.
Marchuk
L
L
McCaig Centre for Joint Injury and Arthritis Research, Faculty of Medicine, University of Calgary, Alberta, Canada.
Sciore
P
P
Reno
C
C
Frank
C B
CB
Hart
D A
DA
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism; D002356:Cartilage / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D004355:Drug Stability; D005260:Female; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D066298:In Vitro Techniques; D007668:Kidney / Q000378:metabolism; D008022:Ligaments / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011180:Postmortem Changes; D012313:RNA / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011817:Rabbits; D013710:Tendons / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000235:genetics
D017931:DNA Primers; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D012313:RNA; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases