Abstract
BackgroundRecovering high quality intact RNA from post-mortem tissue is of major concern for gene expression studies in animals and humans. Since the availability of post-mortem tissue is often associated with substantial delay, it is important that we understand the temporal variation in the stability of total RNA and of individual gene transcripts so as to be able to appropriately interpret the data generated from such studies. Hence, the objective of this experiment was to qualitatively and quantitatively assess the integrity of total and messenger RNA extracted from bovine skeletal muscle, subcutaneous adipose tissue and liver stored at 4°C at a range of time points up to 22 days post-mortem. These conditions were designed to mimic the environment prevailing during the transport of beef from the abattoir to retail outlets.ResultsThe 28S and 18S rRNA molecules of total RNA were intact for up to 24 h post-mortem in liver and adipose tissues and up to 8 days post-mortem in skeletal muscle. The mRNA of housekeeping genes (GAPDH and ACTB) and two diet-related genes (RBP5 and SCD) were detectable up to 22 days post-mortem in skeletal muscle. While the mRNA stability of the two housekeeping genes was different in skeletal muscle and liver, they were similar to each other in adipose tissue. After 22 days post-mortem, the relative abundance of RBP5 gene was increased in skeletal muscle and in adipose tissue and decreased in liver. During this period, the relative abundance of SCD gene also increased in skeletal muscle whereas it decreased in both adipose tissue and liver.ConclusionStability of RNA in three tissues (skeletal muscle, subcutaneous adipose tissue and liver) subjected to long-term post-mortem storage at refrigeration temperature indicated that skeletal muscle can be a suitable tissue for recovering biologically useful RNA for gene expression studies even if the tissue is subjected to post-mortem storage for weeks, whereas adipose tissue and liver should be processed within 24 hours post-mortem.
Highlights
Recovering high quality intact RNA from post-mortem tissue is of major concern for gene expression studies in animals and humans
It is a normal practice for beef to be consumed after 2–3 weeks due to the fact that an elaborate meat tenderization process occurs over this time period. This experiment was designed to mimic the real life situation from the slaughtering of the animal to the length of time when meat is made available to consumers in retail outlets. The objective of this experiment was to qualitatively and quantitatively assess the integrity of total and messenger RNA (mRNA) extracted from bovine skeletal muscle, subcutaneous adipose tissue and liver stored at 4°C at a range of time points up to 22 days post-mortem
The present study evaluated the qualitative and quantitative variations in the integrity of total RNA and mRNA of two common housekeeping genes (GAPDH and ACTB) and two diet-related genes (RBP5 and stearoyl-CoA desaturase gene (SCD)) in bovine skeletal muscle, adipose tissue and liver stored at 4°C for up to 22 days post-slaughter
Summary
Recovering high quality intact RNA from post-mortem tissue is of major concern for gene expression studies in animals and humans. The objective of this experiment was to qualitatively and quantitatively assess the integrity of total and messenger RNA extracted from bovine skeletal muscle, subcutaneous adipose tissue and liver stored at 4°C at a range of time points up to 22 days post-mortem. These conditions were designed to mimic the environment prevailing during the transport of beef from the abattoir to retail outlets. Since the post-mortem stability of an mRNA population can not be generalised [6,7], knowledge of tissue specific stability of mRNA transcripts of interest is essential for evaluating the usefulness of postmortem tissues for gene expression studies
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