OBJECTIVE: To determine the effects of chilling on the organization and distribution of tubulin and chromosomes in rhesus monkey oocytes. DESIGN: Comparative laboratory study. SETTING: Academic research laboratory. ANIMAL(S): Eight adult female rhesus monkeys (Macaca mulatta) aged 6-16 years. INTERVENTION(S): A total of 171 oocytes retrieved from eight rhesus monkeys were separated into nine groups. One group of control oocytes was held at 37 degrees C during the experiment. Four groups of oocytes were rapidly cooled to 0 degrees C and held for 1, 5, 10, or 30 minutes and then fixed and stained. Four other groups of oocytes were cooled to 0 degrees C, held for 1, 5, 10, or 30 minutes, warmed and incubated at 37 degrees C for 60 minutes, and then fixed and stained. MAIN OUTCOME MEASURE(S): Organization of cytoskeleton and chromosomes. RESULT(S): Exposure of rhesus oocytes to 0 degrees C for only 1 minute resulted in complete depolymerization of tubulin. Incubation of chilled oocytes at 37 degrees C for 60 minutes caused partial restoration of tubulin, although most oocytes exhibited abnormal alignment of chromosomes and disorganized meiotic spindles. CONCLUSION(S): We conclude that rhesus monkey oocytes are extremely sensitive to chilling injury. Their successful cryopreservation may require rapid cooling to outpace this injury.
OBJECTIVE: To determine the effects of chilling on the organization and distribution of tubulin and chromosomes in rhesus monkey oocytes. DESIGN: Comparative laboratory study. SETTING: Academic research laboratory. ANIMAL(S): Eight adult female rhesus monkeys (Macaca mulatta) aged 6-16 years. INTERVENTION(S): A total of 171 oocytes retrieved from eight rhesus monkeys were separated into nine groups. One group of control oocytes was held at 37 degrees C during the experiment. Four groups of oocytes were rapidly cooled to 0 degrees C and held for 1, 5, 10, or 30 minutes and then fixed and stained. Four other groups of oocytes were cooled to 0 degrees C, held for 1, 5, 10, or 30 minutes, warmed and incubated at 37 degrees C for 60 minutes, and then fixed and stained. MAIN OUTCOME MEASURE(S): Organization of cytoskeleton and chromosomes. RESULT(S): Exposure of rhesus oocytes to 0 degrees C for only 1 minute resulted in complete depolymerization of tubulin. Incubation of chilled oocytes at 37 degrees C for 60 minutes caused partial restoration of tubulin, although most oocytes exhibited abnormal alignment of chromosomes and disorganized meiotic spindles. CONCLUSION(S): We conclude that rhesus monkey oocytes are extremely sensitive to chilling injury. Their successful cryopreservation may require rapid cooling to outpace this injury.
Authors: Jens O M Karlsson; Abdelmoneim I Younis; Anthony W S Chan; Kenneth G Gould; Ali Eroglu Journal: Mol Reprod Dev Date: 2009-04 Impact factor: 2.609