Bowen Li

Acute administration of tributyltin and trimethyltin modulate glutamate and N-methyl-D-aspartate receptor signaling pathway in Sebastiscus marmoratus

Tributyltin (TBT), widely used as an antifouling biocide, is the most abundant pesticide in coastal environments. Trimethyltin (TMT) is a potent neurotoxicant of a mechanism of action yet to be uncovered. The neurotoxicity of TBT and TMT on the brain of marine fish Sebastiscus marmoratus was investigated in this study. The results showed that TBT and TMT can modulate amino acid neurotransmitters and N-methyl-D-aspartate receptor (NMDAR) signaling pathway in the brain of marine fish in a different manner. TBT did not increase the content of the amino acid neurotransmitters except gamma-aminobutyricd acid (GABA). TMT increased the content of aspartate (Asp), glutamate (Glu) and GABA in a dose-dependent manner. The expression of NADAR and components on its signaling pathway, such as calmodulin, calmodulin-dependent kinase II (CaMKII) and cAMP-response element-binding (CREB) protein was significantly decreased in a dose-dependent manner after TBT exposure. However, the low dose of TMT exposure up-regulate rather than down-regulate the expression of NMDAR and other genes of its pathway. It is suggested that the Glu-NMDAR pathway plays a role in the mechanism for the brain injury in marine fish after TBT or TMT exposure. The alteration of expression of glutamatergic receptor NMDAR and components on its signaling pathway accompanied with the change of total brain transmitter level indicated the importance of glutamatergic system in organotin toxicity.

Tissue-specific and embryonic expression of the retinoid X receptors in Sebastiscus marmoratus

Retinoid X receptors (RXRs) are highly conserved members of the nuclear receptor family and mediate various physiological processes in vertebrates. Most studies on RXRs have concentrated on their structure and function in mammals and their characterization and developmental expression in Danio rerio. However, there is little information concerning the distribution of RXRs in teleost tissues. In the present study, we cloned partial sequences of three RXR subtypes (RXRa, -b, -g) from Sebastiscus marmoratus by RACE PCR and analyzed the phylogeny of the teleost and the tetrapod RXR genes, and identified some inconsistencies with previous studies. The tissue-specific and embryonic expression profiles of each RXR gene were explored using real time quantitative PCR. This analysis demonstrated that these RXRs were expressed in all test tissues indicating their participation in many physiological processes. However, we found a great difference in the distribution of RXRg between teleosts and mammals. Furthermore, we followed expression of the three subtypes through various embryo developmental stages and found that the RXRa orthologues of teleosts might be involved in the development of the anterior hindbrain, tailbud and neural crest and in the formation of the pharynx and fin, that RXRb played ubiquitous roles in fish early development, and that RXRg probably played a role in brain and nervous system development and function.

Exposure of Sebastiscus marmoratus embryos to pyrene results in neurodevelopmental defects and disturbs related mechanisms

Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental contaminants, which are known to be carcinogenic and teratogenic. These compounds cause a range of macroscopic malformations, particularly to the craniofacial apparatus and cardiovascular system during vertebrate development. However, little is known concerning microscopic effects, especially on the sensitive early life stages or on the molecular basis of developmental neurotoxicity. Using the rockfish (Sebastiscus marmoratus), we explored the neurodevelopmental defects caused by early-life exposure to environmentally relevant concentrations of pyrene, a 4-ring PAH. The results showed that pyrene substantially disrupted the cranial innervation pattern and caused deficiency of motor nerves. The expression of a protein associated with axon growth, growth associated protein 43, was decreased in the central nervous system after treatment with pyrene. N-methyl-D-aspartate receptor (NMDAR) plays a vital role in a variety of processes, including neuronal development, synaptic plasticity, and neuronal survival and death. Our results showed that the expression of Ca²⁺/calmodulin dependent kinase II and cAMP-response element-binding, which belong to the NMDAR pathway, were increased in a dose-dependent manner after exposure to pyrene. Acetylcholine, an important neurotransmitter which is known to suppress retinal cells neurite outgrowth, was increased by pyrene exposure. Nitric oxide (NO) acts as an activity-dependent retrograde signal that can coordinate axonal targeting and synaptogenesis during development. The level of NO was decreased in a dose-dependent manner following exposure to pyrene. Taken together, the defects in neurodevelopment and the damage to related mechanisms provided the basis for a better understanding of the neurotoxic effects of pyrene.

Differential gene expression in the brain of Sebastiscus marmoratus in response to exposure to polychlorinated biphenyls (PCBs)

Effects of exposure to polychlorinated biphenyls (PCBs) on Sebastiscus marmoratus were investigated using a suppression subtractive hybridization method. A total of 108 gene sequences were identified as having the potential for being differentially expressed, and 45 could be identified with homologous database sequences. Functions with which they were associated included long-term potentiation and neurotransmitter release, neuroendocrine, mitosis and cell proliferation, energy-related metabolism, general metabolism, signal protein, hemopoiesis system, immune system, and structure. The expression of 17 of these genes was analyzed in the brain using real time fluorescent quantitative PCR. The present study provided a basis for studying the response of fish to PCB exposure and allowed the characterization of new potential neurotoxicol biomarkers of PCB contamination in seawater.

Hexabromocyclododecane exposure induces cardiac hypertrophy and arrhythmia by inhibiting miR-1 expression via up-regulation of the homeobox gene Nkx2.5.

Hexabromocyclododecane (HBCD) is one of the most widely used brominated flame retardants. Although studies have reported that HBCD can cause a wide range of toxic effects on animals including humans, limited information can be found about its cardiac toxicity. In the present study, zebrafish embryos were exposed to HBCD at low concentrations of 0, 2, 20 and 200 nM. The results showed that HBCD exposure could induce cardiac hypertrophy and increased deposition of collagen. In addition, disordered calcium (Ca(2+)) handling was observed in H9C2 rat cardiomyocyte cells exposed to HBCD. Using small RNA sequencing and real-time quantitative PCR, HBCD exposure was shown to induce significant changes in the miRNA expression profile associated with the cardiovascular system. Further findings indicated that miR-1, which was depressed by Nkx2.5, might play a fundamental role in mediating cardiac hypertrophy and arrhythmia via its target genes Mef2a and Irx5 after HBCD treatment. HBCD exposure induced an arrhythmogenic disorder, which was triggered by the imbalance of Ryr2, Serca2a and Ncx1 expression, inducing Ca(2+) overload in the sarcoplasmic reticulum and high Ca(2+)-ATPase activities in the H9C2 cells.

Modulation of the DNA repair system and ATR-p53 mediated apoptosis is relevant for tributyltin-induced genotoxic effects in human hepatoma G2 cells

The toxic effects of tributyltin (TBT) have been extensively documented in several types of cells, but the molecular mechanisms related to the genotoxic effects of TBT have still not been fully elucidated. Our study showed that exposure of human hepatoma G2 cells to 1-4 μmol/L TBT for 3 hr caused severe DNA damage in a concentration-dependent manner. Moreover, the expression levels of key DNA damage sensor genes such as the replication factor C, proliferating cell nuclear antigen and poly (ADP-ribose) polymerase-1 were inhabited in a concentration-dependent manner. We further demonstrated that TBT induced cell apoptosis via the p53-mediated pathway, which was most likely activated by the ataxia telangiectasia mutated and rad-3 related (ATR) protein kinase. The results also showed that cytochrome c, caspase-3, caspase-8, caspase-9, and the B-cell lymphoma 2 were involved in this process. Taken together, we demonstrated for the first time that the inhibition of the DNA repair system might be more responsible for TBT-induced genotoxic effects in cells. Then the generated DNA damage induced by TBT initiated ATR-p53-mediated apoptosis.

The concentration-dependent induction of cell death by trimethyltin chloride in rat liver epithelial IAR20 cells

In this study we intended to evaluate the cytotoxic and genotoxic effects of trimethyltin chloride (TMT), one of the most widely used organometallic compounds, on liver cells with the rat liver epithelial cell line IAR20. The results showed that TMT significantly inhibited cell growth in a concentration-dependent manner and caused an increase in DNA damage. Additionally, Hoechst 33342 staining and flow cytometry detected increases in apoptotic and necrotic cells. Western blots demonstrated that the Bcl-2 family of proteins was involved in the apoptotic process, but p53 was not concerned.