Abstract
Chromosome segregation is conserved throughout eukaryotes. In most systems, it is solely driven by a spindle machinery that is assembled from microtubules. We have recently discovered that actin filaments that are embedded inside meiotic spindles (spindle actin) are needed for accurate chromosome segregation in mammalian oocytes. To understand the function of spindle actin in oocyte meiosis, we have developed high-resolution and super-resolution live and immunofluorescence microscopy assays that are described in this chapter.
Highlights
Every mammalian life begins with the fertilization of an egg by a sperm cell
For the resulting genetically unique zygote to grow into a healthy offspring, both the egg and sperm should first contain the correct number of chromosomes
Meiotic chromosome segregation is driven by a spindle machinery that is assembled from microtubules and separates the chromosomes in two rounds of cell division [3]
Summary
Every mammalian life begins with the fertilization of an egg by a sperm cell. For the resulting genetically unique zygote to grow into a healthy offspring, both the egg and sperm should first contain the correct number of chromosomes. The rate of egg aneuploidy increases dramatically with advancing maternal age [2] This phenomenon, often referred to as “the maternal age effect,” is highly attributed to errors in meiosis, the specialized form of cell division that generates eggs from oocytes [3]. We recently discovered that the actin cytoskeleton plays a vital role inside the meiotic spindle—actin filaments embedded inside the spindle help to organize microtubules into functional bundles that can accurately separate the chromosomes [6]. This finding constitutes an important safety mechanism in mammalian meiosis that prevents aneuploidy in oocytes and eggs. This chapter discusses in detail the live and immunofluorescence microscopy assays that have for the first time enabled highly resolved visualization and functional analysis of spindle actin in mammalian oocyte meiosis
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