A method for the extraction of high-quality RNA and protein from single small samples of arteries and veins preserved in RNAlater

Abstract

Introduction: The mesenteric vasculature plays an important role in cardiovascular homeostasis, both in terms of arterial resistance and venous capacitance. However, the limited quantity of tissue available and the activity of RNases and proteases during dissection of the vessels make it difficult to study gene and protein expression in this tissue. Methods: The mesenteric arcade was harvested from rats. The mesenteric arteries and veins were separated from each other and from contaminating adipose, nerve, and connective tissue by blunt dissection while submerged in RNAlater (Ambion). Total ribonucleic acid (RNA) and protein were extracted from the same mesenteric artery and vein samples. Results: Good quality RNA and protein were obtained by this method. The method preserved the content of proteins including biglycan, adrenergic receptors, estrogen receptors, and protein kinase Cδ, and also preserved the differential expression of estrogen receptors. Discussion: A method for the isolation of high-quality RNA and protein from the arteries and veins of the same mesenteric sample is described. The content of proteins from a variety of cellular compartments and molecular weights was preserved by this method, as well as differential expression of proteins. This method will facilitate the study of gene and protein expression in the mesenteric vasculature in response to pharmacologic treatments and in disease states.