Project description:Intensive aquaculture and environmental changes will inevitably lead to hypoxic stress for largemouth bass (Micropterus salmoides). To better understand the hypoxia responds mechanisms of largemouth bass, we compared the miRNA profile in liver under different environmental DO to determine which miRNAs are most affected during hypoxia. A total of 266 miRNAs were identified, and 84 miRNAs were differentially expressed compared with in control group. GO and KEGG analysis indicated that the miRNAs may play important roles in environment information processing. Specifically, we considered the VEGF signaling pathway, Phosphatidylinositol signaling system and MAPK signaling pathway, the results show that, the 13 miRNAs (miR-15b-5p, miR-30a-3p, miR-133a-3p, miR-19d-5p, miR-1288-3p, miR456, miR-96-5p, miR-23a-3p, miR-23b, miR-214, miR-24, miR-20a-3p and miR-2188-5p) involved in these three pathways are significantly down-regulated during hypoxia stress. And 12 target genes of these miRNAs were showed a higher degree of expression. We found the obvious negative correlation between miRNA and their target mRNAs, providing several miRNA-mRNA interaction networks in largemouth bass in response to hypoxia. Although relatively little information is currently available concerning the biological function of miRNAs identified to date, we strongly suggest that miRNAs play an important role in modulating gene expression involved in the physiological response to hypoxic stress in the fish liver.
Project description:Toxaphene (TOX) is an organochlorine pesticide (OCPs) that remains abundant in the environment due to its heavy use in agriculture, its persistence, and its atmospheric transport. In Florida, the north shore of Lake Apopka was heavily used for agriculture decades ago, and TOX remains present in the soil. Wild largemouth bass inhabit the lake, and are potentially exposed to a variety of OCPs. While other OCPs (dieldrin, methoxychlor, p,p’DDE) have been studied for their endocrine disrupting effects in largemouth bass, there are little data available for TOX. Here we performed a 2-month feeding study of male and female largemouth bass with TOX-laced food to mimic the levels found in fish at Lake Apopka. Subsamples of the gonad and liver were collected and preserved in 10% buffered formalin for histopathology and reproductive staging. Plasma levels of vitellogenin (Vtg) testosterone (T) and 17β-estradiol (E2) were quantified and snippets of testis and ovaries were placed into culture to determine ex vivo production of hormones. Lastly, transcriptomics was conducted in both the male testis and liver, as most studies investigating OCPs have analyzed female ovary/liver and data are scarce for OCPs and male bass. Female GSI did not change with TOX but was trending towards a decrease while male GSI did not change with exposure. Female vitellogenin decreased with toxaphene by ~40% but male vitellogenin was not induced. Female and male E2 and T were not different following dietary exposure compared to control bass and this corresponded to ex vivo culture experiments, which revealed no change in E2 and T production from the gonads. Taken together, TOX did not alter the production of E2 and T directly in the gonad tissue. In the testis, transcriptomics revealed that androgen receptor/beta-2-microglobulin signaling was up-regulated while insulin-related pathways were suppressed with TOX. In the liver, the transcriptome analysis revealed an overwhelming suppression in immune-related signaling cascades (e.g. lectin-like receptor and ITSM-Containing Receptor signaling, CD16/CD14 Proinflammatory Monocyte Activation, and CD38/CD3 -> JUN/FOS/NF-kB Signaling in T-cell Proliferation). Ninety percent of the pathways were reduced between 10 to 40% in median network expression. Reduced immune pathways are consistent with other studies investigating OCPs in fish. In the current study, TOX, at the concentrations tested in the diet, did not act as a weak estrogen as many other OCPs do, but rather appeared to be acting as an antiestrogen (or androgen) based upon reduced female Vtg and activation of androgen-receptor mediated pathways in the testis. These data contribute to a comprehensive transcriptomic-physiological framework for TOX and suggests that endocrine disruption may not be the result of direct effects on steroidogenesis in the gonad, but may rather occur at higher levels of the reproductive axis.