Quantifying growing versus non-growing ovarian follicles in the mouse

We found that, by itself, pyknotic nuclei quantification resulted in high numbers of false positives and false negatives

Bahar Uslu

2017

Scholarcy highlights

  • A standard histomorphometric approach has been used for nearly 40 years that identifies atretic follicles by counting the number of pyknotic granulosa cells in the largest follicle cross-section
  • By taking the number of mitotic GC into account, we have developed an improved method of identifying ovarian follicles that are growing versus those that are not growing, and in some cases that have believably committed to atresia
  • Prior analysis tested the hypothesis that the ovaries of animals heterozygous for FX premutation would have a lower primordial ‘reserve’ of follicles in FXPM mice compared to WT controls, as is postulated to occur in the human FX primary ovarian insufficiency condition
  • A detailed evaluation of all pyknotic and mitotic figures in WT ovarian follicles and those from FX 130-repeat harboring mice mice showed that identification of at least one mitotic figure in the largest cross section, and at most, one additional adjacent section, accurately identifies those follicles that are growing
  • In the absence of mitotic figures, pyknotic nuclei are likely to indicate that the follicle has committed irreversibly to atresia
  • While evaluation using the ‘historical’ criteria can be used to generate a reasonable estimate of total and dying follicles, the example of the FXPM animal shows that exceptions where significant mis-identification of follicle life and death can take place
  • Application of this strategy to human ovarian specimens of sufficient histological quality should lead to more accurate determination(s) of the impact of genetic or environmental factors upon follicle growth and survival in women

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