Histological and Histochemical Evaluation of the Effects of Graphene Oxide on Thyroid Follicles and Gas Gland of Japanese Medaka (Oryzias latipes) Larvae.

Graphene oxide (GO) has become a topic of increasing concern for its environmental and health risks. However, studies on the potential toxic effects of GO, especially as an endocrine disrupting chemical (EDC), are very limited. In the present study we have used Japanese medaka fish as a model to assess the endocrine disruption potential of GO by evaluating its toxic and histopathologic effects on thyroid follicles and the gas gland (GG) of medaka larvae. One day post-hatch (dph) starved medaka fries were exposed to GO (2.5, 5.0, 10.0, and 20 mg/L) for 96 h, followed by 6 weeks depuration in a GO-free environment with feeding. Larvae were sacrificed and histopathological evaluation of thyroid follicles and the GG cells were done microscopically. Different sizes of spherical/oval shape thyroid follicles containing PAS positive colloids, surrounded by single-layered squamous/cuboidal epithelium, were found to be scattered predominantly throughout the pharyngeal region near the ventral aorta. We have apparently observed a sex-specific difference in the follicular size and thyrocytes height and a non-linear effect of GO exposure on the larvae on 47th day post hatch (dph). The GG is composed of large uniform epithelial cells with eosinophilic cytoplasm. Like thyroids, our studies on GG cells indicate a sex-specific difference and GO exposure non-linearly reduced the GG cell numbers in males and females as well as in XY and XX genotypes. Our data further confirm that sex effect should be carefully considered while assessing the toxicity of EDCs on the thyroid gland.