Human Reproduction, Vol. 16, No. 11, 2374-2378,
November 2001
© 2001 European Society of Human Reproduction and Embryology
Limited recovery of meiotic spindles in living human oocytes after cooling–rewarming observed using polarized light microscopy
1 Division of Reproductive Medicine and Infertility, Department of Obstetrics and Gynecology, Women and Infants Hospital of Rhode Island, Brown University School of Medicine, Providence, RI, 2 Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, New England Medical Center, Boston, MA and 3 Marine Biological Laboratory, Woods Hole, MA, USA
BACKGROUND: Spindles are formed from microtubules and are exquisitely sensitive to changes in temperature. An orientation-independent polarized light microscope, the Polscope, can be used to image spindles in living oocytes allowing analysis of spindle kinetics in the living state. This study examined the effects of cooling on spindle disassembly in living human oocytes and spindle recovery after rewarming. METHODS: Oocytes were imaged continuously with the Polscope during cooling and rewarming. The quantity of microtubules in the spindles was measured by its birefringence using the Polscope. RESULTS: Spindles had completely disassembled by 5 min after cooling and recovered by 20 min after rewarming to 37°C if rewarming started soon after the oocyte's temperature dropped to room temperature. However, when oocytes were cooled and kept at 33, 28 or 25°C for 10 min and then warmed, it was found that warming allowed 5/5, 2/5 and 0/5 oocytes of the spindles to recover respectively. CONCLUSIONS: These results indicate that human meiotic spindles are exquisitely sensitive to alterations in temperature. The maintenance of temperature at 37°C during in-vitro manipulation is important for spindle integrity and, therefore, is likely to be important for normal fertilization and subsequent embryo development.
Key words: Cooling/human oocytes/polarization microscope/spindle
4 To whom correspondence should be addressed at: IVF laboratory, Department of Obstetrics and Genealogy, Women and Infants Hospital of Rhode Island, 1 Blackstone Place, 1st Floor, Providence RI 02903, USA. E-mail: wwang{at}wihri.org
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