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GABA-mediated inhibition of cadmium uptake and accumulation in apples
2022
Li, Yuxing | Li, Yunhao | Cui, Yinglian | Xie, Yuanmei | Shi, Yanjiao | Shang, Yueming | Ma, Fengwang | Zhang, Jing | Li, Cuiying
GABA, a four-carbon non-protein amino acid, plays an important role in animals and plants. We previously found GABA could alleviate alkali stress in apple seedlings. However, its physiological mechanism under heavy metal cadmium (Cd) stress need to be further studied. Thus, we explored its biological role in response to Cd stress. It was verified that 0.5 mM GABA could effectively alleviate Cd toxicity. Using NMT technique, we found that exogenous GABA could significantly reduce the net Cd²⁺ fluxes in apple roots, and Cd content was significantly lower than that in roots under Cd stress. Further analysis indicated exogenous GABA could significantly reduce the expression of genes related to the uptake and transport of Cd in apples under Cd stress. In addition, exogenous GABA could significantly increase the content of amino acids in apple roots under Cd stress. GAD is a key enzyme in GABA synthesis, we obtained transgenic apple roots of overexpression MdGAD1. Compared with the control, transgenic roots accumulated less Cd, maintained lower Cd uptake by roots, and lower expression of related transport genes. These results showed that GABA could effectively alleviate Cd toxicity in apple seedlings and provide a new perspective of GABA to alleviate Cd stress.
Show more [+] Less [-]Carbamazepine induces hepatotoxicity in zebrafish by inhibition of the Wnt/β-catenin signaling pathway
2021
Bai, Zhonghui | Jia, Kun | Chen, Guilan | Liao, Xinjun | Cao, Zigang | Zhao, Yangqi | Zhang, Chunping | Lu, Huiqiang
As drug abuse has become increasingly serious, carbamazepine (CBZ) is discharged into the aquatic environment with municipal sewage, causing potential harm to aquatic organisms. Here, we utilized zebrafish, an aquatic vertebrate model, to comprehensively evaluate the hepatotoxicity of CBZ. The larvae were exposed to 0.07, 0.13, and 0.26 mmol/L CBZ from 72 hpf to 144 hpf, and the adults were exposed to 0.025, 0.05, and 0.1 mmol/L CBZ for 28 days. The substantial changes were observed in the size and histopathology of livers, indicating that CBZ induced severe hepatoxicity in the larvae and adults. Oil red O staining demonstrated CBZ exposure caused severe lipid accumulation in the livers of both larvae and adults. Furthermore, CBZ exposure facilitated hepatocyte apoptosis through TUNEL staining, which was caused by rising ROS content. Subsequently, down-regulation of genes related to the Wnt pathway in exposure groups indicated that CBZ inhibited the development of liver via the Wnt/β-catenin signaling pathway. In conclusion, CBZ induced severe hepatotoxicity by promoting lipid accumulation, generating excessive ROS production, and inhibiting the Wnt/β-catenin signaling pathway in zebrafish. The results reveal the occurrence of CBZ-induced hepatotoxicity in zebrafish and clarify its mechanism of action, which potentially illustrate environmental concerns associated with CBZ exposure.
Show more [+] Less [-]Parental exposure to environmental concentrations of tris(1,3-dichloro-2-propyl)phosphate induces abnormal DNA methylation and behavioral changes in F1 zebrafish larvae
2020
Ding, Xisheng | Sun, Wen | Dai, Lili | Liu, Chunsheng | Sun, Qian | Wang, Jianghua | Zhang, Panwei | Li, Kun | Yu, Liqin
Tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) has been demonstrated to be transferred from parental animals to their offspring. However, whether parental exposure to environmental concentrations of TDCIPP show neurodevelopmental toxicity in the F1 generation and the possible underlying mechanism remain unclear. Therefore, in this study, zebrafish embryos were exposed to environmental concentrations of TDCIPP (3, 30 and 300 ng L⁻¹) for 120 days. The effects of exposure on motor behaviors, neurotransmitter levels, DNA methylation, and gene expression of F1 larvae were investigated. Parental exposure left TDCIPP residues in F1 eggs as well as reduced body length of F1 larvae. Moreover, parental exposure significantly reduced swimming activity in F1 5 dpf larvae, although it did not significantly alter serotonin, dopamine, 3,4-dihydroxyphenylacetic acid, γ-aminobutyrate, and acetylcholine levels. Genes encoding DNA methylation transferases (dnmt3aa and dnmt1) were downregulated in F1 larvae. Reduced representation bisulfite sequencing analysis revealed 446 differentially methylated regions and enriched neuronal cell body Gene Ontology term in F1 generation. Correlation analysis between the expression of genes related to neural cell body and swimming speed indicated that solute carrier family 1 member 2b (slc1a2b) downregulation might be responsible for the inhibition of motor behaviors. Furthermore, bisulfite amplicon sequencing analysis confirmed hypermethylation of the promoter region of slc1a2b in F1 larvae following parental exposure to 300 ng L⁻¹ TDCIPP, which might have led to significant downregulation of gene expression and, in turn, influenced the motor behaviors. These results indicate that parental exposure to environmental concentrations of TDCIPP alters DNA methylation, downregulates gene expressions and, thus inducing developmental neurotoxicity, in F1 larvae.
Show more [+] Less [-]In vivo and In vitro neurochemical-based assessments of wastewater effluents from the Maumee River area of concern
2016
Arini, Adeline | Cavallin, Jenna E. | Berninger, Jason P. | Marfil-Vega, Ruth | Mills, Marc | Villeneuve, Daniel L. | Basu, Niladri
Wastewater treatment plant (WWTP) effluents contain potentially neuroactive chemicals though few methods are available to screen for the presence of such agents. Here, two parallel approaches (in vivo and in vitro) were used to assess WWTP exposure-related changes to neurochemistry. First, fathead minnows (FHM, Pimephales promelas) were caged for four days along a WWTP discharge zone into the Maumee River (Ohio, USA). Grab water samples were collected and extracts obtained for the detection of alkylphenols, bisphenol A (BPA) and steroid hormones. Second, the extracts were then used as a source of in vitro exposure to brain tissues from FHM and four additional species relevant to the Great Lakes ecosystem (rainbow trout (RT), river otter (RO), bald eagle (BE) and human (HU)). The ability of the wastewater (in vivo) or extracts (in vitro) to interact with enzymes (monoamine oxidase (MAO) and glutamine synthetase (GS)) and receptors (dopamine (D2) and N-methyl-d-aspartate receptor (NMDA)) involved in dopamine and glutamate-dependent neurotransmission were examined on brain homogenates. In vivo exposure of FHM led to significant decreases of NMDA receptor binding in females (24–42%), and increases of MAO activity in males (2.8- to 3.2-fold). In vitro, alkylphenol-targeted extracts significantly inhibited D2 (66% in FHM) and NMDA (24–54% in HU and RT) receptor binding, and induced MAO activity in RT, RO, and BE brains. Steroid hormone-targeted extracts inhibited GS activity in all species except FHM. BPA-targeted extracts caused a MAO inhibition in FHM, RT and BE brains. Using both in vivo and in vitro approaches, this study shows that WWTP effluents contain agents that can interact with neurochemicals important in reproduction and other neurological functions. Additional work is needed to better resolve in vitro to in vivo extrapolations (IVIVE) as well as cross-species differences.
Show more [+] Less [-]The anti-estrogenicity of chronic exposure to semicarbazide in female Japanese flounders (Paralichthys olivaceus), and its potential mechanisms
2018
Yue, Zonghao | Yu, Miao | Zhao, Haifeng | Wang, Jun | Zhang, Xiaona | Tian, Hua | Wang, Wei | Ru, Shaoguo
This study investigated the anti-estrogenic effects of chronic exposure to a new marine pollutant, semicarbazide (SMC; 1, 10, and 100μg/L), in female Paralichthys olivaceus, as well as the associated mechanism. After 130days of exposure, plasma 17β-estradiol and testosterone concentrations, and hepatic estrogen receptors, vitellogenin, and choriogenin mRNA levels decreased significantly in SMC-exposed groups. Moreover, down-regulation of genes in the hypothalamic-pituitary-gonadal axis, including gonadotropin-releasing hormone, gonadotropic hormones and their receptors, the steroidogenic acute regulatory protein, 17α-hydroxylase, 17β-hydroxysteroid dehydrogenase, and cytochrome P450 19A, was observed after SMC exposure. Furthermore, the kisspeptin/g protein-coupled receptor 54 (kiss/gpr54) system and gamma-aminobutyric acid-ergic (GABAergic) system were also affected by SMC: SMC significantly down-regulated mRNA expression of kiss2, gpr54, and the GABA synthesis enzyme gad67. Our results demonstrated for the first time that environmentally relevant concentrations of SMC exerted anti-estrogenicity in female flounders, providing theoretical support for ecological risk assessments of SMC in marine environments.
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