Molecular characterization of viral infections in vivo

Abstract

Abstract Natural and experimental infections of animals continue to be important as models in studying the pathogenesis and management of infectious diseases. Classically, methods for examining animal models have required extraction of individual tissues for biochemical or histological analysis. Examples of these techniques reviewed elsewhere in this volume include nucleic acid and protein electrophoresis, immunohistochemistry and enzyme linked immunoadsorbent assays (ELISA). Over the past few years, however, we have used a new technology which allows for sensitive and specific detection of nucleic acids and proteins in whole animal sections (WAS) fixed to tape or artificial membranes. The power of WAS technology rests in its unique potential for defining and quantitating the distribution of genes and gene products in an anatomical context. Further, because the entire animal is examined, there is no bias a priori against recognizing novel distributions of genes or gene products. Although we have used WAS primarily for studies in viral pathogenesis, problems in developmental biology, neoplasia, and pharmacology may also be suitable for WAS analysis. WAS would also seem ideal for the investigation of tissue-specific distributions of transcripts and proteins in transgenic animal systems. In this chapter we will discuss our methods for WAS in situ hybridization and protein blotting (Figure 1 ). For illustrations of WAS applications the reader is referred to a recent review (1 ).