Waterborne exposure to microcystin-leucine arginine induces endocrine disruption and gonadal dysplasia of Pelophylax nigromaculatus tadpoles via the hypothalamic-pituitary-gonadal-liver axis

Abstract

The impact of microcystins on the gonad development and reproduction endocrine in the tadpole stage on amphibians remains unclear. In this study, the tadpoles (Pelophylax nigromaculatus) were exposed to 0, 1, and 10 μg/L of microcystin-leucine arginine (MC-LR) for 60 days to explore the impacts of environmental realistic concentration MC-LR on gonad development and reproduction endocrine, respectively. After MC-LR exposure, the germ cell structure has changed, especially in oocytes. The 10 μg/L MC-LR exposure group showed a significantly diminished gonad somatic index (GSI) in females. However, the sex ratio of tadpoles did not differ significantly. Moreover, gene transcription (figla and nobox) related to ovarian development and genes (sox9 and dmrt1) associated with testicular development were down-regulated after MC-LR exposure. After MC-LR exposure, the gene transcripts encoding gonadotropin-releasing hormone (gnrh1 and gnrh2) were down-regulated in the hypothalamus, while gonadotropins (FSH and LH) levels increased in serum. The transcripts of testosterone synthesis-related genes (star, cyp11a1, 3β-hsd, cyp17a1, and 17β-hsd) were up-regulated in the gonads, and the testosterone (T) concentration increased in serum. However, key gene transcript (cyp19a1) involved in estradiol synthesis was down-regulated and the estradiol (E2) concentration decreased in serum, resulting in the absence of a compensatory mechanism for positive feedback regulation of the hypothalamic-pituitary-gonadal (HPG) axis to maintain E2 levels. The vitellogenin gene (vtg1) transcription level was significantly down-regulated. The E2/T content ratio decreased in MC-LR concentration-dependent manner. Consequently, MC-LR exposure interfered with the hypothalamic-pituitary-gonadal-liver (HPGL) axis in tadpoles, which in turn affects gonadal development, especially the ovaries. Overall, this study provides the initial evidence that MC-LR exerts significant effects on reproductive endocrinology and gonadal development in amphibian tadpoles, highlighting the susceptibility of the tadpole reproductive system to the environmental risks of MC-LR.