Effects of chilling to 0°C on the morphology of meiotic spindles in human metaphase II oocytes

被引:125
作者
Zenzes, MT [1 ]
Bielecki, R [1 ]
Casper, RF [1 ]
Leibo, SP [1 ]
机构
[1] Univ Toronto, Div Reprod Sci, Toronto, ON, Canada
基金
英国医学研究理事会;
关键词
chilling; human; meiotic spindle; metaphase II; oocyte;
D O I
10.1016/S0015-0282(00)01800-8
中图分类号
R71 [妇产科学];
学科分类号
100211 ;
摘要
Objective: To determine the effects of chilling to 0 degreesC on the meiotic spindle of human metaphase II as observed by optical sectioning microscopy. Design: Laboratory study. Setting: Academic research laboratory in a medical school. Patient(s): Seventy-two women undergoing infertility treatment donated a total of 108 oocytes. Intervention(s): Metaphase II oocytes were stripped of their cumulus cells, cooled directly to 0 degreesC, and for periods of 1 to 10 minutes. They were then fixed at 37 degreesC, stained for immunofluorescence, and microscopically. Main Outcome Measure(s): Morphology of the meiotic spindle in chilled and control oocytes. Result(s): Microscopic evaluations of 46 chilled oocytes revealed various time-dependent changes in microtubules compared to 9 control oocytes. After 1 minute at 0 degreesC, spindle damage was negligible, but in oocytes cooled for 2 or 3 minutes, there was obvious shortening of the spindle and loss of polarity. Cooling to 0 degreesC for 4 to 9 minutes resulted in increasingly more drastic changes; by 10 minutes the spindles had totally disappeared. Despite depolymerization of microtubular tubulin at 0 degreesC, the chromosomes did not become dispersed, but remained anchored even in the absence of spindles. Conclusion(s): Even brief exposure of human oocytes to temperatures near 0 degreesC causes profound alterations of the meiotic spindle. (Fertil Steril (R) 2001;75:769-77. (C) 2001 by American Society for Reproductive Medicine.) dispersed, but remained anchored even in the absence of spindles.
引用
收藏
页码:769 / 777
页数:9
相关论文
共 54 条
[1]   THE INFLUENCE OF SLOW AND ULTRA-RAPID FREEZING ON THE ORGANIZATION OF THE MEIOTIC SPINDLE OF THE MOUSE OOCYTE [J].
AIGNER, S ;
VANDERELST, J ;
SIEBZEHNRUBL, E ;
WILDT, L ;
LANG, N ;
VANSTEIRTEGHEM, AC .
HUMAN REPRODUCTION, 1992, 7 (06) :857-864
[2]   PRELIMINARY-RESULTS OF CRYOCONSERVATION OF HUMAN OOCYTES [J].
ALHASANI, S ;
DIEDRICH, K ;
VANDERVEN, H ;
KREBS, D .
GEBURTSHILFE UND FRAUENHEILKUNDE, 1986, 46 (09) :643-644
[3]   The effect of temperature fluctuations on the cytoskeletal organisation and chromosomal constitution of the human oocyte [J].
Almeida, PA ;
Bolton, VN .
ZYGOTE, 1995, 3 (04) :357-365
[4]   EFFECTS OF COOLING AND REWARMING ON THE MEIOTIC SPINDLE AND CHROMOSOMES OF IN VITRO-MATURED BOVINE OOCYTES [J].
AMAN, RR ;
PARKS, JE .
BIOLOGY OF REPRODUCTION, 1994, 50 (01) :103-110
[5]  
ANGELL RR, 1991, HUM GENET, V86, P383
[6]   EVALUATION OF THE SPINDLE APPARATUS OF IN-VITRO MATURED HUMAN OOCYTES FOLLOWING CRYOPRESERVATION [J].
BAKA, SG ;
TOTH, TL ;
VEECK, LL ;
JONES, HW ;
MUASHER, SJ ;
LANZENDORF, SE .
HUMAN REPRODUCTION, 1995, 10 (07) :1816-1820
[7]  
Battaglia DE, 1996, HUM REPROD, V11, P2217
[8]   Cryopreservation of human oocytes: A review of current problems and perspectives [J].
Bernard, A ;
Fuller, BJ .
HUMAN REPRODUCTION UPDATE, 1996, 2 (03) :193-207
[9]   In vitro blastocyst formation of human oocytes obtained from unstimulated and stimulated cycles after vitrification at various maturational stages [J].
Chung, HM ;
Hong, SW ;
Lim, JM ;
Lee, SH ;
Cha, WT ;
Ko, JJ ;
Han, SY ;
Choi, DH ;
Cha, KY .
FERTILITY AND STERILITY, 2000, 73 (03) :545-551
[10]   CYTOGENETIC ANALYSIS OF HUMAN OOCYTES AND EMBRYOS IN AN INVITRO FERTILIZATION PROGRAM [J].
EDIRISINGHE, WR ;
MURCH, AR ;
YOVICH, JL .
HUMAN REPRODUCTION, 1992, 7 (02) :230-236