Oocyte Polarization Is Coupled to the Chromosomal Bouquet, a Conserved Polarized Nuclear Configuration in Meiosis

Abstract

The source of symmetry breaking in vertebrate oocytes is unknown. Animal—vegetal oocyte polarity is established by the Balbiani body (Bb), a conserved structure found in all animals examined that contains an aggregate of specific mRNAs, proteins, and organelles. The Bb specifies the oocyte vegetal pole, which is key to forming the embryonic body axes as well as the germline in most vertebrates. How Bb formation is regulated and how its asymmetric position is established are unknown. Using quantitative image analysis, we trace oocyte symmetry breaking in zebrafish to a nuclear asymmetry at the onset of meiosis called the chromosomal bouquet. The bouquet is a universal feature of meiosis where all telomeres cluster to one pole on the nuclear envelope, facilitating chromosomal pairing and meiotic recombination. We show that Bb precursor components first localize with the centrosome to the cytoplasm adjacent to the telomere cluster of the bouquet. They then aggregate around the centrosome in a specialized nuclear cleft that we identified, assembling the early Bb. We show that the bouquet nuclear events and the cytoplasmic Bb precursor localization are mechanistically coordinated by microtubules. Thus the animal—vegetal axis of the oocyte is aligned to the nuclear axis of the bouquet. We further show that the symmetry breaking events lay upstream to the only known regulator of Bb formation, the Bucky ball protein. Our findings link two universal features of oogenesis, the Bb and the chromosomal bouquet, to oocyte polarization. We propose that a meiotic—vegetal center couples meiosis and oocyte patterning. Our findings reveal a novel mode of cellular polarization in meiotic cells whereby cellular and nuclear polarity are aligned. We further reveal that in zygotene nests, intercellular cytoplasmic bridges remain between oocytes and that the position of the cytoplasmic bridge coincides with the location of the centrosome meiotic—vegetal organizing center. These results suggest that centrosome positioning is set by the last mitotic oogonial division plane. Thus, oocytes are polarized in two steps: first, mitotic divisions preset the centrosome with no obvious polarization yet, then the meiotic—vegetal center forms at zygotene bouquet stages, when symmetry is, in effect, broken. In most vertebrates, an early event in egg development involves the establishment of the so-called animal—vegetal axis; this sets up the embryonic body axes and contributes to germ-line specification, and therefore, is key to embryonic development. The animal—vegetal axis is established during oogenesis by the Balbiani body (Bb), an aggregate of specific mRNAs, proteins, and mitochondria, which forms adjacent to the nucleus and ultimately defines one pole of the oocyte, the vegetal pole. Despite its universal conservation, how the Bb forms and how its position is determined is unknown. Here, we show that Bb formation is initiated at the onset of meiosis, and its position coincides with a previously known meiotic polarized nuclear configuration, the chromosomal bouquet, which gathers the chromosome ends, the telomeres, asymmetrically on the nuclear membrane to assist in homologous chromosome pairing. We reveal that a global cellular organizer functioning via microtubules generates the bouquet and aggregates the Bb precursors asymmetrically towards the centrosome. We determined that these events lie functionally upstream to the Bb regulator Bucky ball. Further upstream, we found that the centrosome appears prepositioned by an intercellular cytoplasmic bridge derived from the last presumptive cell division plane of the premeiotic oogonial cell. Thus, oocyte polarity and the chromosomal bouquet are linked through a common cellular polarization mechanism.