Concurrent In Situ Hybridization and Antibody Staining in Red Flour Beetle (Tribolium) Embryos

INTRODUCTION

The red flour beetle, Tribolium castaneum, has emerged as an important model system for studying the evolution of development. Studies with Tribolium complement the vast amount of research done with Drosophila. Developmental features that are conserved between Drosophila and Tribolium, such as body segmentation, are achieved by quite different means, and thus comparison of developmental mechanisms between these two insects can address many interesting questions concerning the evolution of morphology and other characters. Most in situ and antibody staining protocols used for Tribolium have been adapted from Drosophila studies, and to date, few antibodies have been raised specifically against Tribolium antigens. Whole-mount in situ hybridization allows detection of mRNA expression patterns, while immunohistochemical staining detects protein expression patterns. This protocol allows both procedures to be performed concurrently in the same Tribolium embryo. Such double staining is particularly useful for comparing the expression pattern of a new gene to that of a known protein.