The influence of cooling on the organization of the meiotic spindle of the mouse oocyte.

The effect of cooling on the organization of the microtubule system of the mouse oocyte has been investigated. Cooling to 25, 18 or 4 degrees C for varying periods of time resulted in a progressive disassembly of the spindle and the dispersal of the chromosomes. The extent of the changes observed was greater at lower temperatures and with longer periods of exposure. Transfer of oocytes from either 37 or 4 to 24 degrees C resulted in the rapid and transient appearance of polar asters and of multiple cytoplasmic asters associated with the pericentriolar material present at the spindle poles and in the cytocortex of the oocyte. This transient aster formation appeared to be driven by the elevated levels of free tubulin released during spindle disassembly. An apparent reversal of many of the changes induced by low temperature was observed in many but not all oocytes on their restoration to 37 degrees C for 1 h. These results have implications for our understanding of microtubule organization in the oocyte, and for the handling of oocytes during IVF and GIFT therapeutic procedures and during the cryopreservation of oocytes.