Postmortem stability of total RNA isolated from rabbit ligament, tendon and cartilage

被引:81
作者
Marchuk, L [1 ]
Sciore, P [1 ]
Reno, C [1 ]
Frank, CB [1 ]
Hart, DA [1 ]
机构
[1] Univ Calgary, Hlth Sci Ctr, Fac Med, McCaig Ctr Joint Injury & Arthrit Res, Calgary, AB T2N 4N1, Canada
来源
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS | 1998年 / 1379卷 / 02期
基金
英国医学研究理事会;
关键词
RNA stability; connective tissue; ligament; tendon; cartilage; postmortem;
D O I
10.1016/S0304-4165(97)00094-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 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 96h postmortem. These findings confirm that it is likely appropriate to use properly stored postmortem dense connective tissues for molecular biological investigations. (C) 1998 Elsevier Science B.V.
引用
收藏
页码:171 / 177
页数:7
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