Chapter 7 Meiotic Chromosome Preparation and Protein Labeling

Abstract

Publisher Summary This chapter discusses the preparation of meiotic chromosome and protein labeling. The prophase of the first meiotic division appears an ideal experimental system for the analysis of the functional organization of the nucleus. The chromatin and the nuclear matrix and lamina undergo a series of precisely timed and well-defined structural changes to fulfill the apparently equally well-defined functions of genetic exchange and chromosome segregation. The synaptonemal complexes (SCs) look particularly inviting: these structures—which are formed between homologous chromosomes during meiotic prophase—undergo a series of morphological alterations which closely correlate with the successive rearrangements of chromatin. The chapter describes the steps followed in the assembly of SCs. The isolation of SCs is an essential step for the biochemical analysis of the role of these structures in meiotic chromosome behavior. The preparation of polyclonal and monoclonal anti-SC antibodies is another precondition to be fulfilled for such an analysis. The antibodies can now be used to analyze the assembly and disassembly of SCs by immunocytochemical analysis and the genes coding for SC components can be isolated by screening expression libraries with the available antibodies.