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Identification of ovarian toxicity is very important for safety assessment of drugs and other environmental chemicals. The detection of interference with ovarian function is very hard without a thorough understanding of the normal ovarian morphology based on reproductive physiology. The focus of the present study was therefore a practical analysis in each stage of the estrous cycles using ovaries obtained from 143 rats demonstrating normal cycling. Transversely dissected maximum areas in the ovaries were examined microscopically for the two major features, follicles and corpora lutea (CL). Classification of growing follicles was in reference to Pedersen and Peters (1968), and functionally divided into follicular stimulating hormone (FSH)-independent and dependent categories. The former, small and medium-sized follicles, respectively primordial/primary and preantral follicles, could be readily detected by immunohistochemical staining for proliferating cell nuclear antigen (PCNA). The large antral and Graafian follicles and large sized atretic follicles showed sequential changes depending on the estrous cycle stage. CL could be divided into currently and previously formed examples. Currently formed CL underwent remarkable changes in their appearance with the cycle, reflecting ovulation and progesterone production. Thus morphological analysis that is synchronized the large antral follicle changes with recently formed CL ones allows the ovary to be classified into the each estrous cycle stage. Morphological deviation from any synchronized combination provides a first pointer of ovarian toxicity. PCNA immunohistochemical staining is also useful to detect small follicles.

Morphological characterization of the ovary under normal cycling in rats and its viewpoints of ovarian toxicity detection