Oocyte meiotic-stage-specific differences in spindle depolymerization in response to temperature changes monitored with polarized field microscopy and immunocytochemistry

Abstract

Objective: To compare the polymerization status of mouse oocyte spindles exposed to various temperatures at various stages of meiosis.

Design: Experimental animal study.

Setting: University animal laboratory.

Animal(s): CF1 mice.

Intervention(s): Immature oocytes matured to metaphase I (MI), telophase I (TI), and metaphase II (MII) were incubated at 37 °C (control), room temperature (RT), or 4 °C for 0, 10, 30, and 60 minutes. Spindle analysis subsequently was performed using polarized field microscopy and immunocytochemistry. Spindles of TI and MII oocytes that underwent vitrification and warming were analyzed also by immunocytochemistry.

Main outcome measure(s): Detection of polymerized meiotic spindles.

Result(s): At RT, and after 60 minutes at 4 °C, a significant time-dependent decrease in the percentage of polymerized meiotic spindles was observed in MI and MII oocytes, but not in TI oocytes. The polymerization of TI spindles at 4 °C was similar to that of TI spindles at 4 °C that underwent vitrification and warming.

Conclusion(s): Significant differences in the microtubule dynamics of MI, TI, and MII oocytes incubated at different temperatures were observed. In particular, meiotic spindles in TI oocytes exhibited less depolymerization than did metaphase spindles.