6 Caught in the Act: Deducing Meiotic Function from Protein Immunolocalization

Abstract

Meiotic division comprises a complex series of events, many of which are unique in the life cycle of the organism. The process utilizes both proteins that participate in normal mitotic cell cycle progression and DNA damage repair and proteins expressed only during meiosis. Until recently, few meiotic protein participants had been identified and characterized, but several recent developments have changed this situation. Proteins can be selected for study based on their cDNA sequence and similarity to known proteins with "suspicious" repair/recombination or cell cycle activity and antibodies against these proteins applied to meiotic nuclei to test for activity. With the development of gene sequence data bases from many organisms, similarity to a known protein need not be based on the same or even a closely related species. Potential interactions between two or more proteins can be identified and involvement in a common process inferred based on antibody colocalization. The gene sequence can be disrupted and the effect on meiotic progression directly examined. Previously identified structures, the synaptonemal complex (SC) and both early and late recombination nodules (RNs), provide structural and temporal landmarks that assist in inferring meiotic activity of the protein being studied. Mammalian meiosis is especially attractive for these kinds of studies since spermatocyte and oocyte nuclei are large with distinct nuclear organelles and since meiosis is highly protracted, occurring over a period of several days. In this chapter, an approach to the study of mammalian meiosis based on use of specific antibodies is outlined and methods of coupling this approach to other techniques, such as targeted gene disruption or chromosome aberrations, are described. Some of the proteins already identified as participants in meiotic prophase are reviewed and their presumed functions discussed.