Journal
HUMAN REPRODUCTION
Volume 16, Issue 11, Pages 2374-2378Publisher
OXFORD UNIV PRESS
DOI: 10.1093/humrep/16.11.2374
Keywords
cooling; human oocytes; polarization microscope; spindle
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Funding
- PHS HHS [K08 1099] Funding Source: Medline
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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 degreesC 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 degreesC 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 degreesC during in-vitro manipulation is important for spindle integrity and,:therefore, is likely to be important for normal fertilization and subsequent embryo development.
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