Cytological analysis of interference in mouse meiosis.

Abstract

In most eukaryotes, meiotic crossovers (COs) are non-randomly placed along the bivalents, such that the presence of a CO reduces the probability of additional COs nearby. This phenomenon, named CO interference, was originally defined genetically, but can also be analyzed cytologically by studying the chromosomal positions of protein complexes that are involved in CO formation, or by studying the positions of chiasmata. Here we focus on the cytological analysis of interference among protein complexes involved in meiotic recombination and CO formation in the mouse. During the pachytene stage of meiosis, these protein complexes can be visualized as immunofluorescent foci along synaptonemal complexes (SCs), which are linear protein structures that are formed along homologous chromosome pairs (bivalents) during meiotic prophase. We describe how to make cytological preparations that are suitable for the analysis of interference among these foci, and how to estimate the strength of interference among foci, using the gamma distribution as a mathematical model for focus/CO positioning.