Triphenyltin induced darker body coloration by disrupting melanocortin system and pteridine metabolic pathway in a reef fish, Amphiprion ocellaris.

Triphenyltin (TPT) is a typical persistent organic pollutant whose occurrence in coral reef ecosystems may threaten the survival of reef fishes. In this study, a brightly colored representative reef fish, Amphiprion ocellaris was used to explore the effects of TPT at environmental levels (1, 10, and 100 ng/L) on skin pigment synthesis. After the fish were exposed to TPT for 60 days, the skin became darker, owing to an increase in the relative area of black stripes, a decrease in orange color values while an increase in brown color values, and an increase in the number of melanocytes in the orange part of the skin tissues. To explore the mechanisms by which TPT induces darker body coloration, the enzymatic activity and gene expression levels of the members of melanocortin system that affect melanin synthesis were evaluated. Leptin levels and lepr expression were found to be increased after TPT exposure, which likely contributed to the increase found in pomc expression and α-melanocyte-stimulating hormone (α-MSH) levels. Then Tyr activity and mc1r, tyr, tyrp1, mitf, and dct were upregulated, ultimately increasing melanin levels. Importantly, RT-qPCR results were consistent with the transcriptome analysis of trends in lepr and pomc expression. Because the orange color values decreased, pterin levels and the pteridine metabolic pathway were also evaluated. The results showed that TPT induced BH4 levels and spr, xdh, and gch1 expression associated with pteridine synthesis decreased, ultimately decreasing the colored pterin content (sepiapterin). We conclude that TPT exposure interferes with the melanocortin system and pteridine metabolic pathway to increase melanin and decrease colored pterin levels, leading to darker body coloration in A. ocellaris. Given the importance of body coloration for the survival and reproduction of reef fishes, studies on the effects of pollutants (others alongside TPT) on body coloration are of high priority.

Monotropein Induced Apoptosis and Suppressed Cell Cycle Progression in Colorectal Cancer Cells.

OBJECTIVE: To determine whether monotropein has an anticancer effect and explore its potential mechanisms against colorectal cancer (CRC) through network pharmacology and molecular docking combined with experimental verification. METHODS: Network pharmacology and molecular docking were used to predict potential targets of monotropein against CRC. Cell counting kit assay, plate monoclonal assay and microscopic observation were used to investigate the antiproliferative effects of monotropein on CRC cells HCT116, HT29 and LoVo. Flow cytometry and scratch assay were used to analyze apoptosis and cell cycle, as well as cell migration, respectively in HCT116, HT29, and LoVo cells. Western blotting was used to detect the expression of proteins related to apoptosis, cell cycle, and cell migration, and the expression of proteins key to the Akt pathway. RESULTS: The Gene Ontology and Reactome enrichment analyses indicated that the anticancer potential of monotropein against CRC might be involved in multiple cancer-related signaling pathways. Among these pathways, RAC-beta serine/threonine-protein kinase (Akt1, Akt2), cyclin-dependent kinase 6 (CDK6), matrix metalloproteinase-9 (MMP9), epidermal growth factor receptor (EGFR), cell division control protein 42 homolog (CDC42) were shown as the potential anticancer targets of monotropein against CRC. Molecular docking suggested that monotropein may interact with the 6 targets (Akt1, Akt2, CDK6, MMP9, EGFR, CDC42). Subsequently, cell activity of HCT116, HT29 and LoVo cell lines were significantly suppressed by monotropein (P<0.05). Furthermore, our research revealed that monotropein induced cell apoptosis by inhibiting Bcl-2 and increasing Bax, induced G1-S cycle arrest in colorectal cancer by decreasing the expressions of CyclinD1, CDK4 and CDK6, inhibited cell migration by suppressing the expressions of CDC42 and MMP9 (P<0.05), and might play an anticancer role through Akt signaling pathway. CONCLUSION: Monotropein exerts its antitumor effects primarily by arresting the cell cycle, causing cell apoptosis, and inhibiting cell migration. This indicates a high potential for developing novel medication for treating CRC.

Brain metabolite profiles provide insight into mechanisms for behavior sexual dimorphisms in zebrafish (Danio rerio).

The zebrafish (Danio rerio) has historically been a useful model for research in genetics, ecology, biology, toxicology, and neurobehavior. Zebrafish have been demonstrated to have brain sexual dimorphism. However, the sexual dimorphism of zebrafish behavior demands our attention, particularly. To evaluate the behavior and brain sexual dimorphisms in zebrafish, this study assessed sex differences in adult D. rerio in four behavioral domains, including aggression, fear, anxiety, and shoaling, and further compared with metabolites in the brain tissue of females and males. Our findings showed that aggression, fear, anxiety and shoaling behaviors were significantly sexually dimorphic. Interestingly, we also show through a novel data analysis method, that the female zebrafish exhibited significantly increased shoaling behavior when shoaled with male zebrafish groups and, for the first time, we offer evidence that male shoals are beneficial in dramatically alleviating anxiety in zebrafish. In addition, there were significant changes in metabolites in zebrafish brain tissue between the sexes. Furthermore, zebrafish behavioral sexual dimorphism may be associated with brain sexual dimorphism, with significant differences in brain metabolites. Therefore, to prevent the influence or even bias of behavioral sex differences on results, it is suggested that behavioral studies or behavioral-based other relevant investigations consider sexual dimorphism of behavior and brain.

An integrative investigation of developmental toxicities induced by triphenyltin in a larval coral reef fish, Amphiprion ocellaris.

Triphenyltin (TPT) is widely distributed on coastlines, which makes coral reef fish a potential target of TPT pollution. However, the negative effects of TPT on coral reef fish remain poorly understood. Therefore, in the present study, the larval coral reef fish Amphiprion ocellaris was used to investigate the developmental toxicities of TPT at environmentally relevant concentrations (0, 1, 10 and 100 ng/L). After TPT exposure for 14 d, the cumulative mortality increased, and growth was suppressed. In addition, TPT exposure inhibited the development of melanophores and xanthophores and delayed white strip formation, which might be responsible for the disruption of the genes (erbb3b, mitfa, kit, xdh, tyr, oca2, itk and trim33) related to pigmentation. TPT exposure also attenuated ossification of head skeletal elements and the vertebral column and inhibited the expression of genes (bmp2, bmp4 and sp7) related to skeletal development. The observed developmental toxicities on growth, pigmentation and skeleton development might be associated with the disruption of thyroid hormones and the genes related to thyroid hormone regulation (tshß, thrα, thrß, tg, tpo, dio2, and ttr). In addition, TPT exposure interfered with locomotor and shoaling behavior, and the related genes dbh, avp and avpr1aa. Taken together, our results suggest that TPT pollution might threaten the development of one of the most iconic coral reef fish, which might produce disastrous consequences on the health of coral reef ecosystems.

Environmental concentrations of benzophenone-3 disturbed lipid metabolism in the liver of clown anemonefish (Amphiprion ocellaris).

Benzophenone-3 (BP-3) often used as a UV filter in various products and an endocrine disruptor. In this work, we exposed the clown anemonefish to 10 µg/L and 50 µg/L BP-3 for 7 and 14 days. Liver histological, biochemical analysis, and transcriptome sequencing were used to explore the mechanism of the lipid metabolism disorder in the liver of three-month-old clown anemonefish treated with BP-3. The histological and biochemical analysis showed that BP-3 induces morphological changes and lipid droplet accumulation, and the lipid content, lipase, and antioxidant enzyme activity were abnormal. After treatment with 10 µg/L and 50 µg/L BP-3 for 7 days, the transcriptome analysis further demonstrated that the KEGG analysis revealed that the differentially expressed genes (DEGs) were mainly associated with fat digestion and absorption, PPAR signaling pathway, circadian rhythm, and mineral absorption pathways; After 10 µg/L and 50 µg/L of BP-3 exposure for 14 days, the KEGG analysis were mainly associated with circadian rhythm, circadian rhythm-fly, protein processing in the endoplasmic reticulum, and beta-alanine metabolism pathways. Several key genes were involved in the process of liver lipid metabolism, including CD36, APoA-Ⅰ, FABP, LPL, ACS, and PEPCK. The qRT-PCR validation results showed that eight genes (CYP8B1, FABP1, LPL, MGAT, PEPCK, PER1, PSMB4, PSME2) were significantly down-regulated, and the other two genes (Fbxl3, RXR) were significantly up-regulated after 7 days of BP-3 exposure. Similarly, eleven genes (AMPK, ARNTL, Bmal1, CASP3, CYC, CYP2J, CYP2U1, GSK3A, PEPCK, RAC1, RORA) were significantly up-regulated, and the other four genes (NR1D1, PER1, PTGDS, HLF) were significantly down-regulated after 14 days of BP-3 exposure. In conclusion, our results elucidate the physiological and molecular responses to BP-3 exposure in the liver lipid metabolism of clown anemonefish, and these findings reveal that the regulation of lipid metabolism is disturbed when clown anemonefish is exposed to UV filters.

Tributyltin exposure disrupted the locomotor activity rhythms in adult zebrafish (Danio rerio) and the mechanism involved.

The fish circadian rhythm system might be an emerging target of tributyltin (TBT), however, the mechanism by which TBT interferes with the circadian rhythm is poorly understood. Therefore, in the present study, zebrafish were used to assess the effects of TBT at environmental concentrations (1 and 10 ng/L) on locomotor activity rhythm. Furthermore, we focused on the visual system to explore the potential mechanism involved. After 90 d of exposure, TBT disturbed the locomotor activity rhythms in zebrafish, which manifested as: (1) low activities and lethargy during the arousing period; (2) inability to fall asleep quickly and peacefully during the period of latency to sleep; and (3) no regular "waves" of locomotor activities during the active period. After TBT exposure, the histological structure of the eyes significantly changed, the boundary between layers became blurred, and the melanin concentrations significantly decreased. Using KEGG and GSEA pathway analyses, the differentially expressed genes in the eyes screened by transcriptomics were significantly enriched in the tyrosine metabolism pathway and retinol metabolism pathway. Furthermore, a decrease in melanin and disruption of retinoic acid were found after TBT exposure, which would affect the reception of phototransduction, and then interfere with the circadian rhythm in fish. The disruption of the circadian rhythm of fish by aquatic pollutants would decrease their ecological adaptability, which should be considered in future research.

Triphenyltin exposure induced abnormal morphological colouration in adult male guppies (Poecilia reticulata).

Fish morphological colouration is essential for their survival and reproduction success; however, it is vulnerable to environmental factors, such as pollutants. Triphenyltin (TPT) is widespread in aquatic ecosystems, and its impacts on fish have been problematic. Therefore, the purpose of this study was to investigate the effects of TPT at environment-related concentrations (0, 1, 10 and 100 ng Sn/L) on morphological colouration in male guppies (Poecilia reticulata). The results showed that TPT exposure affected both orange/red and dark morphological colouration in guppies. The faded orange/red colouration might be related to the decrease of coloured pteridine and Pts (6-Pyruvoyltetrahydropterin Synthase) expression. In addition, TPT exposure induced melanogenesis, however, much melanin was distributed diffusely in the skin and did not seem to form a spot pattern, giving the fish a dull appearance. According to the skin transcriptional profiles, the changes of dark morphological colouration might be related to the changes in genes related to the functions of melanosome components (Gpnmb, Slc45a2 and Tyr), construction (Ap3d1, Fig4, Hps3, Hps5, Lyst, Rabggta, Txndc5 and Vps33a), and transport (Rab27a). Additionally, genes related to the regulation of melanogenesis (Atrn and Pomc) and system effects (Atox1, Atp6ap2, Atp6v1f, Atp6v1h, Rpl24, Rps19 and Rps20) might also be involved in the molecular mechanisms of abnormal morphological colouration induced by TPT. The present study provides crucial data on the molecular basis of abnormal morphological colouration in fish exposed to TPT and underscores the importance of toxicological studies of the effects of pollutants in aquatic environments on fish morphological colouration.

Different action mechanisms of low- and high-level quercetin in the brains of adult zebrafish (Danio rerio).

Quercetin is reported to be beneficial to or pose hazards to the health of animals, the inconsistence remains to be recognized and debated. This work was conducted to understand the neuroprotective or neurotoxic properties of quercetin, and investigate the different action mechanisms between low- and high-level quercetin. Therefore, we evaluated brain oxidative stress and monoamine neurotransmitters in adult zebrafish (Danio rerio) after exposure to 1 and 1000 µg/L quercetin. In addition, the brain transcriptional profiles were analyzed to identify genes and pathways that were differentially regulated in the brains. The results of oxidative stress and neurotransmitters suggest that low-level quercetin might be beneficial to nervous system, while high-level quercetin might exert detrimental effects. Furthermore, transcriptional profiles also suggested different toxic mechanisms occurred between low- and high-level quercetin. At 1 µg/L quercetin, enrichment analysis of differently expressed genes (DEGs) revealed that the fanconi anemia pathway might be an important mechanism in neuroprotective effects. At 1000 µg/L quercetin, the up-regulated DEGs were enriched in many Gene Ontology (GO) terms related to neuronal synapses, indicating potential neuroprotective effects; however, enrichment of up-regulated DEGs in GO terms of response to stimulus and the MAPK signaling pathway was also found, which indicated increases of stress. Notably, at 1000 µg/L quercetin, the down-regulated DEGs were enriched in several GO terms related to the proteostasis and the proteasome pathway, indicating impairment of proteasome functions which was involved in neurodegenerative diseases. Moreover, several hub genes involved in the pathology of neurodegenerative diseases were identified by Protein-protein interaction analysis at 1000 µg/L quercetin. Thus, high-level quercetin might pose potential risk inducing neurodegenerative diseases, which should receive more attention in the future. Additionally, our findings may provide awareness to society and researchers about toxicity possibilities of phytochemicals on wildlife and human.

Prognostic Value of Neoantigen Load in Immune Checkpoint Inhibitor Therapy for Cancer.

Immune checkpoint inhibitors (ICIs) have made great progress in the field of tumors and have become a promising direction of tumor treatment. With advancements in genomics and bioinformatics technology, it is possible to individually analyze the neoantigens produced by somatic mutations of each patient. Neoantigen load (NAL), a promising biomarker for predicting the efficacy of ICIs, has been extensively studied. This article reviews the research progress on NAL as a biomarker for predicting the anti-tumor effects of ICI. First, we provide a definition of NAL, and summarize the detection methods, and their relationship with tumor mutation burden. In addition, we describe the common genomic sources of NAL. Finally, we review the predictive value of NAL as a tumor prediction marker based on various clinical studies. This review focuses on the predictive ability of NAL's ICI efficacy against tumors. In melanoma, lung cancer, and gynecological tumors, NAL can be considered a predictor of treatment efficacy. In contrast, the use of NAL for urinary system and liver tumors requires further research. When NAL alone is insufficient to predict efficacy, its combination with other indicators can improve prediction efficiency. Evaluating the response of predictive biomarkers before the treatment initiation is essential for guiding the clinical treatment of cancer. The predictive power of NAL has great potential; however, it needs to be based on more accurate sequencing platforms and technologies.

Significant benefits of pembrolizumab in treating refractory advanced pulmonary sarcomatoid carcinoma: A case report.

BACKGROUND: Pulmonary sarcomatoid carcinoma (PSC), a rare subtype of non-small cell lung cancer (NSCLC), is poorly differentiated and highly aggressive. Treatment is limited, and the prognosis is poor. Pembrolizumab is an anti-programmed death (PD)-1 antibody with good efficacy in NSCLC. Recent studies have demonstrated that PD-ligand 1 (PD-L1) overexpression is common in PSCs, which suggests that anti-PD-L1 treatment is an ideal option. However, the response to pembrolizumab in PSC has not been studied. CASE SUMMARY: We present a PSC case with PD-L1 overexpression that significantly benefited from pembrolizumab. A 73-year-old Chinese male was detected with a right lung lesion. Pathological analysis of the right upper lobectomy confirmed PSC. The PD-L1 test revealed overexpression (TPS: 90%). Multiple metastases occurred 1 mo after surgery, representing stage IV PSC. Neither first-line chemotherapy nor second-line antiangiogenic agents showed any benefit. Radiotherapy (1200 cGy) was administered to relieve chest wall pain. The patient received the PD-1 inhibitor pembrolizumab (100 mg) as third-line therapy; however, because of fever and severe infection, he refused to receive immunotherapy any longer. Thus, only one dose of pembrolizumab was administered. Deep sustained remission of most of the metastases was achieved except for lesions in the right adrenal gland, which first shrank and then progressed. The patient died because of disease progression in the right adrenal gland. He achieved a progression-free survival time of 8 mo and an overall survival time of 9 mo with third-line pembrolizumab. CONCLUSION: Our findings highlight and offer direct evidence of the efficacy of pembrolizumab in PD-L1-overexpressing PSCs. Combined radiotherapy and immunotherapy may enhance treatment efficacy.

Quercetin affects shoaling and anxiety behaviors in zebrafish: Involvement of neuroinflammation and neuron apoptosis.

Quercetin, a potential fish food supplement, has been reported to process many beneficial properties. However, some negative effects of quercetin have been observed, which pointed out necessity for additional studies to evaluate its safety. Therefore, the present study investigated effects of quercetin (0.01, 0.1, 1, 10, 100 and 1000 µg/L) on shoaling and anxiety behaviors through novel tank tests in zebrafish (Danio rerio). Furthermore, oxidative stress, neuroinflammation and apoptosis in the brains were examined to learn more about mechanisms of action related to quercetin. The results showed that quercetin at the lower concentrations exerted beneficial effects on shoaling and anxiety behaviors. On the contrary, when quercetin was up to 1000 µg/L, it exerted detrimental effects shown as decreases of movement and increases of anxiety behaviors. Generally, U-shaped responses of antioxidant enzyme activities (superoxide dismutase and catalase), and inversed U-shaped responses of inflammatory mediators (cyclooxygenase-2) and cytokines (interleukin-1ß, interleukin-6, interleukin-10, and tumor necrosis factor α) to quercetin treatment were found in the brains. In addition, quercetin at the lower concentrations attenuated cell apoptosis, while even more apoptosis was found at the 1000 µg/L quercetin group. In conclusion, quercetin could exert beneficial or detrimental effects on the shoaling and anxiety behaviors depending on the treatment concentrations, and the underlying mechanisms are potentially associated with neuroinflammation and neuron apoptosis.

Correction to: The Effects of 50 nm Unmodified Nano-ZnO on Lipid Metabolism and Semen Quality in Male Mice.

The original version of this article unfortunately contained a mistake. The correct title should be "The Effects of 50 nm Unmodified Nano-ZnO on Lipid Metabolism and Semen Quality in Male Mice". The original article has been corrected.

The Effects of 50 nm Unmodified Nano-ZnO on Lipid Metabolism and Semen Quality in Male Mice.

Fifty male mice were exposed to 50 nm unmodified nano-ZnO through intragastric administration for 90 days to detect the long-term effects of unmodified nano-ZnO in mice. Results showed that the blood glucose, serum follicle stimulating hormone, luteinizing hormone, testosterone, and estradiol were significantly decreased (p < 0.05). The serum triglyceride, total cholesterol, and low-density lipoprotein were significantly increased (p < 0.05). The semen quality of the 160 mg/kg·bw group were significantly lowered (p < 0.05). The liver and testis catalase and CuZn-SOD activities were significantly elevated (p < 0.05). The abilities of •OH inhibition in the livers and testes of the 160 mg/kg·bw group were significantly lowered (p < 0.05). The liver and testis MDA levels of the 160 mg/kg·bw group were significantly elevated (p < 0.05). Results indicate that exposure of nano-ZnO could induce lipid metabolism disorder, hyperlipidemia, and reproductive toxicity to male mice through oxidative injury.

Sub-lethal effects of the triazole fungicide propiconazole on zebrafish (Danio rerio) development, oxidative respiration, and larval locomotor activity.

Propiconazole is a triazole fungicide used in agriculture. Via run-off, it can enter the aquatic environment, and can adversely affect organisms. However, data are scarce on how propiconazole may affect early developmental life stages of fish. The objectives of this study were to evaluate the potential sub-lethal effects of propiconazole during zebrafish development. Wildtype zebrafish (ABTu strain) embryos and larvae were exposed to propiconazole (0.1-100 µM) for up to 150 hours post fertilization (hpf) depending upon the endpoint measured. Propiconazole decreased survival and induced hypopigmentation in fish at 100 µM compared to the water and solvent controls. Pericardial edema was also noted in embryos and larvae (beginning at 2-3 dpf) exposed to 100 µM propiconazole. To visualize the effects of propiconazole on the circulatory system in more detail, we exposed transgenic zebrafish (globin-LCR:eGFP) to the fungicide. Hematopoietic changes were observed within 48 h of exposure to 100 µM, and localization of blood cells in the cardic region became diffuse, indicating pooling of blood in the pericardial region. We measured oxidative respiration in embryos as sufficient ATP is needed for development. Exposure to 100 µM propiconazole (~6-30 hpf) reduced basal respiration (~50%), oligomycin-induced ATP linked respiration (~70%), proton leak (~30%), and non-mitochondrial respiration (~50%), indicating compromised mitochondrial bioenergetics. A Visual Motor Response (VMR) test was used to measure dark photokinesis behavior in larval fish exposed to propiconazole for a 6-day period. Larval fish exposed to the highest concentration in the assay (10 µM) showed evidence of hypoactivity. This study demonstrates that propiconazole can induce hypopigmentation in zebrafish, disrupt mitochondrial bioenergetics, and can alter locomotor activity. However, these sub-lethal responses were observed at concentrations above what is typically detected in the environment.

Long-Term Effects of Unmodified 50 nm ZnO in Mice.

Nanometer zinc oxide (nano-ZnO) is widely used in many kinds of fields. However, information about the toxicity and toxic mechanism of nano-ZnO is limited. The aims of this study were to investigate the long-term toxic effects of unmodified 50 nm ZnO administered by gavage in mice. After 90 days, hematological parameters, hepatic and renal functions, and oxidative and anti-oxidative status were measured. Pathological damages in livers, kidneys, and other tissues were also examined by hematoxylin and eosin (H&E) staining. The results showed that oral nano-ZnO exposure induced anemia and damages to liver and kidney, influenced the antioxidant system, and impacted functions of liver and kidney in mice after a 90-day exposure. The main cause for oxidative stress in vivo induced by nano-ZnO might be hydroxyl free radical. The lowest observed adverse effect level (LOAEL) was 40 mg/kg·bw, and the livers, kidneys, lungs, pancreas, and gastrointestinal tracts are the target organs.

Molecular characterization of p38 MAPK from blunt snout bream (Megalobrama amblycephala) and its expression after ammonia stress, and lipopolysaccharide and bacterial challenge.

p38 mitogen-activated protein kinase (MAPK) is an important protein which plays a key role in regulating the innate immunity, so exploring its molecular characterization is helpful in understanding the resistance against microbial infections in cultured fish. Here, a full-length cDNA of p38 MAPK was cloned from liver of blunt snout bream (Megalobrama amblycephala) which covered 2419 bp with an open reading frame of 1086 bp encoding 361 amino acids. p38 MAPK contained the characteristic structures of Thr-Gly-Tyr (TGY) motif and substrate binding site Ala-Thr-Arg-Trp (ATRW), which are conserved in MAPK family. To investigate p38 MAPK functions, two in vivo experiments were carried out to examine its expression following ammonia exposure and bacterial challenge. Also, an in vitro experiment was conducted to assess the role of p38 MAPK in inflammation of primary hepatocytes induced by lipopolysaccharide (LPS). The results showed the ubiquitous expression of p38 MAPK in all the tested tissues with varying levels. p38 MAPK mRNA expression was significantly up-regulated by ammonia stress and Aeromonas hydrophila challenge, and altered in a time-dependent manner. Moreover, the results indicated that the inflammatory response induced by LPS in hepatocytes is p38 MAPK dependent as knockdown of p38 MAPK using siRNA technology depressed the expression of IL-1ß and IL-6. The findings in this study showed that p38 MAPK has anti-stress property, and plays key role in protection against bacterial infection and inflammation in blunt snout bream.

Triphenyltin exposure affects mating behaviors and attractiveness to females during mating in male guppies (Poecilia reticulata).

The impacts of triphenyltin (TPT) on ecological health have been of great concern due to their widespread use and ubiquity in aquatic ecosystems. However, little is known about the effects of TPT on the reproductive behaviors of fishes. Therefore, the present study was conducted to investigate the effects of TPT at environmentally relevant concentrations (0, 1 and 10 ng Sn/L) on the mating behaviors and the attractiveness to females during mating in male guppies (Poecilia reticulata). The results showed that TPT exposure disturbed the mating behaviors; the TPT-exposed male fish performed more sneaking attempts, but no changes in sigmoid courtship were displayed. The increases in sneaking attempts might be related to increases in testosterone levels induced by TPT exposure. In the context of a competing male, the TPT-exposed males were less attractive to females during mating. The decreases in attractiveness might be related to decreases in carotenoid-based coloration, shown as decreases in caudal fin redness values and skin carotenoid contents. In addition, TPT-induced total antioxidant capacities, the activities of superoxide dismutase and catalase, and the contents of malondialdehyde in liver and intestinal tissues indicated increases in oxidative stress. Both oxidative stress and coloration are linked to carotenoids. Thus, we speculated that the TPT-exposed males might use carotenoids to cope with increases in oxidative stress at the expense of carotenoid-based coloration. The disruption of mating behaviors and the decrease in attractiveness to females in male fish could result in reproductive failure. The present study underscores the importance of using behavioral tests as a sensitive tool in assessing the impact of pollutants present in aquatic environments.

In Silico Computational Transcriptomics Reveals Novel Endocrine Disruptors in Largemouth Bass ( Micropterus salmoides).

In recent years, decreases in fish populations have been attributed, in part, to the effect of environmental chemicals on ovarian development. To understand the underlying molecular events we developed a dynamic model of ovary development linking gene transcription to key physiological end points, such as gonadosomatic index (GSI), plasma levels of estradiol (E2) and vitellogenin (VTG), in largemouth bass ( Micropterus salmoides). We were able to identify specific clusters of genes, which are affected at different stages of ovarian development. A subnetwork was identified that closely linked gene expression and physiological end points and by interrogating the Comparative Toxicogenomic Database (CTD), quercetin and tretinoin (ATRA) were identified as two potential candidates that may perturb this system. Predictions were validated by investigation of reproductive associated transcripts using qPCR in ovary and in the liver of both male and female largemouth bass treated after a single injection of quercetin and tretinoin (10 and 100 µg/kg). Both compounds were found to significantly alter the expression of some of these genes. Our findings support the use of omics and online repositories for identification of novel, yet untested, compounds. This is the first study of a dynamic model that links gene expression patterns across stages of ovarian development.

Transcriptome analyses of sex differential gene expression in brains of rare minnow (Gobiocypris rarus) and effects of tributyltin exposure.

RNA-sequencing was used to identify sex-biased gene expression in brains of rare minnow (Gobiocypris rarus) by comparing transcriptomic profiles between females and males. Furthermore, transcriptomic responses to 10 ng/L tributyltin (TBT) in both male and female brains were also investigated to understand whether TBT affects the identified sex-biased genes. Differentially expressed genes (DEGs) were identified using the IDEG6 web tool. In this article, we presented male- and female-biased DEGs, and up-regulated and down-regulated DEGs after TBT exposure. The raw reads data supporting the present analyses has been deposited in NCBI Sequence Read Archive (SRA, http://www.ncbi.nlm.nih.gov/Traces/sra) with accession number PRJNA376634. The data presented in this article are related to the research article entitled "Transcriptomic analyses of sexual dimorphism of rare minnow (G. rarus) brains and effects of tributyltin exposure" (doi: 10.1016/j.ecoenv.2018.02.049).

Transcriptomic analyses of tributyltin-induced sexual dimorphisms in rare minnow (Gobiocypris rarus) brains.

The brain of fish displays sexual dimorphisms and exhibits remarkable sexual plasticity throughout their life span. Although reproductive toxicity of tributyltin (TBT) in fish is well documented in fish, it remains unknown whether TBT interrupts sexual dimorphisms of fish brains. In this work, brain transcriptomic profiles of rare minnow (Gobiocypris rarus) was characterized and sex-biased genes were identified using RNA sequencing. Functional annotation and enrichment analysis were performed to reveal differences of gene products and pathways between the brains of male and female fish. Furthermore, transcriptomic responses of male and female brains to TBT at 10 ng/L were also investigated to understand effects of TBT on brain sexual dimorphisms. Only 345 male-biased and 273 female-biased genes were found in the brains. However, significant female-biased pathways of circadian rhythm and phototransduction were identified in the brains by enrichment analysis. Interestingly, following TBT exposure in the female fish, the circadian rhythm pathway was significantly disrupted based on enrichment analysis, while in the male fish, the phototransduction pathway was significantly disrupted. In the female fish, expression of genes (Per, Cry, Rev-Erb α, Ror, Dec and CK1δ/ε) in the circadian rhythm pathway was down-regulated after TBT exposure; while in the male fish, expression of genes (Rec, GNAT1_2, GNGT1, Rh/opsin, PDE and Arr) in the phototransduction pathway was up-regulated after TBT exposure. Overall, our results not only provide key data on the molecular basis of brain sexual dimorphisms in fish, but also offer valuable resources for investigating molecular mechanisms by which environmental chemicals might influence brain sexual plasticity.