Effect of chilling on the organization of tubulin and chromosomes in rhesus monkey oocytes.
OBJECTIVE
To determine the effects of chilling on the organization and distribution of tubulin and chromosomes in rhesus monkey oocytes.


METHODS
Comparative laboratory study.


METHODS
Academic research laboratory.


METHODS
Eight adult female rhesus monkeys (Macaca mulatta) aged 6-16 years.


METHODS
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.


METHODS
Organization of cytoskeleton and chromosomes.


RESULTS
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.


CONCLUSIONS
We conclude that rhesus monkey oocytes are extremely sensitive to chilling injury. Their successful cryopreservation may require rapid cooling to outpace this injury.
Songsasen
Nucharin
N
Department of Biological Sciences, University of New Orleans, New Orleans, Louisiana, USA.
Yu
Il Jeoung
IJ
Ratterree
Marion S
MS
VandeVoort
Catherine A
CA
Leibo
Stanley P
SP
D000818:Animals; D002875:Chromosomes / Q000648:ultrastructure*; D003080:Cold Temperature; D005260:Female; D008253:Macaca mulatta; D008540:Meiosis; D009865:Oocytes / Q000737:chemistry / Q000648:ultrastructure*; D011108:Polymers; D013997:Time Factors; D014404:Tubulin / Q000032:analysis / Q000648:ultrastructure*
D011108:Polymers; D014404:Tubulin