Three-dimensional quantitative analysis of chromosomes in the oocytes of aging mice during meiosis I in vitro

The mechanism of senescence is very complicated and can involve formation of chromosome abnormalities and a decline in female fertility. In this study, 3-D visualization of fluorescently labeled chromosomes in oocytes from aging and pubertal mice during in vitro maturation was done with a two-photon laser scanning microscope. Differences between aging and pubertal groups at various maturation stages were analyzed quantitatively in terms of chromosomal morphology, shape, and spatial arrangement. Compared with the pubertal group, the chromosomal morphology of oocytes from aging mice changed: both the mean volume and the mean surface area of chromosomes increased by approximately 20% (P < 0.05) at prometaphase and metaphase of meiosis I (considered to be the weakly condensed folded form of the chromosomes). Furthermore, at these stages, the shape of the chromosomal array became rounder (roundness factor increased by approximately 10%; P < 0.001) and the adhesion among chromosomes became more severe (P < 0.001) at approximately the same stages. Additionally, trends over time for both chromosomal morphology and shape were quite distinct between oocytes from aging and pubertal mice. Interestingly, trends for mean distance were similar; therefore, aging did not seem to influence chromosome movement toward the metaphase plate. These morphologic results should be useful to study age-related degradation of oocyte quality and to interpret results derived from molecular biology.