溫度短暫變化對紡錘體及卵細胞之絕對影響

使用polescopy觀察MII卵子紡錘體顯示溫度之短暫變化亦會造成紡錘體之分解
時間過久將產生不可逆之損害，顯示溫度變化對卵子細胞之絕對影響
溫度變化影響到紡錘體，進一步影響染色體複製合成有絲分裂之正常與否及胚胎細胞之正常分裂

http://humrep.oxfordjournals.org/content/16/11/2374.full

Limited recovery of meiotic spindles in living human oocytes after cooling–rewarming observed using polarized light microscopy

1. Wei-Hua Wang1,2,4, 
2. Li Meng1,2, 
3. Rickard J. Hackett1,2, 
4. Rudolf Odenbourg3and 
5. David L. Keefe1,2,3

+Author Affiliations

1. ¹Division 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. ²Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, New England Medical Center, Boston, MA and
3. ³Marine Biological Laboratory, Woods Hole, MA, USA

• Received March 12, 2001.
• Accepted July 27, 2001.

 

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Abstract

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.

Figure 3.

Spindle images in human oocytes after cooling and rewarming. The oocytes were imaged after being maintained at 37°C or cooled to 33, 28 and 25°C and then rewarmed to 37°C. Images were taken before cooling, 5 and 10 min after cooling and again 10 and20 min after rewarming to 37°C. Arrows indicate the spindles, and arrowheads indicate the first polar body. Original magnification, ×120.