Molecular Targets of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) within the Zebrafish Ovary: Insights into TCDD-induced Endocrine Disruption and Reproductive Toxicity

TCDD is a reproductive toxicant and endocrine disruptor, yet the mechanisms by which it causes these reproductive alterations are not fully understood. In order to provide additional insight into the molecular mechanisms that underlie TCDD\u27s reproductive toxicity, we assessed TCDD-induced transcriptional changes in the ovary as they relate to previously described impacts on serum estradiol concentrations and altered follicular development in zebrafish. In silico computational approaches were used to correlate candidate regulatory motifs with observed changes in gene expression. Our data suggest that TCDD inhibits follicle maturation via attenuated gonadotropin responsiveness and/or depressed estradiol biosynthesis, and that interference of estrogen-regulated signal transduction may also contribute to TCDD\u27s impacts on follicular development. TCDD may also alter ovarian function by disrupting various signaling pathways such as glucose and lipid metabolism, and regulation of transcription. Furthermore, events downstream from initial TCDD molecular-targets likely contribute to ovarian toxicity following chronic exposure to TCDD. Data presented here provide further insight into the mechanisms by which TCDD disrupts follicular development and reproduction in fish, and can be used to formulate new hypotheses regarding previously documented ovarian toxicity

Persistent Adverse Effects on Health and Reproduction Caused by Exposure of Zebrafish to 2,3,7,8-Tetrachlorodibenzo-p-dioxin During Early Development and Gonad Differentiation

Little is understood regarding the impacts of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure during early development on the health, survival, and reproductive capability of adults. Here we use zebrafish to determine whether early life stage exposure to TCDD induces toxicity in adult zebrafish and their offspring. Zebrafish were exposed to graded concentrations of TCDD (0–400 pg/ml) via waterborne exposure for 1 h/week from 0 to 7 weeks of age. The heart and swim bladder were identified as being most sensitive to TCDD exposure during early development. Subtle developmental toxic responses collectively impaired survival, and only zebrafish in the 0, 25, and 50 pg TCDD/ml groups survived to adulthood. Surviving fish exhibited TCDD toxicity in craniofacial structures (i.e., operculum and jaw), heart, swim bladder, and ovary. Exposure to 25 pg TCDD/ml impaired egg production (40% of control), fertility (90% of control), and gamete quality. TCDD-treated males contributed more than females to impaired reproductive capacity. Transgenerational effects were also discovered in that offspring from parents exposed to TCDD during early life stages showed a 25% increase in mortality compared with the F1 of dimethyl sulfoxide fish, reduced egg production (30–50% of control) and fertility (96% of control). Thus, adverse effects resulting from TCDD exposure during early life stages for one generation of zebrafish were sufficient to cause adverse health and reproductive effects on a second generation of zebrafish. In the environment, transgenerational effects such as these may contribute to population declines for the most TCDD sensitive fish species

Developmental toxicity of low generation PAMAM dendrimers in zebrafish

Biological molecules and intracellular structures operate at the nanoscale; therefore, development of nanomedicines shows great promise for the treatment of disease by using targeted drug delivery and gene therapies. PAMAM dendrimers, which are highly branched polymers with low polydispersity and high functionality, provide an ideal architecture for construction of effective drug carriers, gene transfer devices and imaging of biological systems. For example, dendrimers bioconjugated with selective ligands such as Arg-Gly-Asp (RGD) would theoretically target cells that contain integrin receptors and show potential for use as drug delivery devices. While RGD-conjugated dendrimers are generally considered not to be cytotoxic, there currently exists little information on the risks that such materials pose to human health. In an effort to compliment and extend the knowledge gleaned from cell culture assays, we have used the zebrafish embryo as a rapid, medium throughput, cost-effective whole-animal model to provide a more comprehensive and predictive developmental toxicity screen for nanomaterials such as PAMAM dendrimers. Using the zebrafish embryo, we have assessed the developmental toxicity of low generation (G3.5 and G4) PAMAM dendrimers, as well as RGD-conjugated forms for comparison. Our results demonstrate that G4 dendrimers, which have amino functional groups, are toxic and attenuate growth and development of zebrafish embryos at sublethal concentrations; however, G3.5 dendrimers, with carboxylic acid terminal functional groups, are not toxic to zebrafish embryos. Furthermore, RGD-conjugated G4 dendrimers are less potent in causing embryo toxicity than G4 dendrimers. RGD-conjugated G3.5 dendrimers do not elicit toxicity at the highest concentrations tested and warrant further study for use as a drug delivery device. © 2007 Elsevier Inc. All rights reserved.Link_to_subscribed_fulltex

Potential Roles of Arnt2 in Zebrafish Larval Development

The aryl hydrocarbon receptor nuclear translocator (ARNT) is a basic helix-loop-helix–PAS heterodimeric transcription factor that dimerizes with other basic helix-loop-helix–PAS proteins to mediate biological responses. The function of ARNT2 is poorly understood. Here we provide an initial characterization of the zebrafish arnt2 null (arnt2−/−) mutant to identify functions of Arnt2 during development. Arnt2−/− mutant zebrafish develop normally until 120 hours postfertilization (hpf) when morphological changes and functional deficits occur. The C-start escape response initiated by either touch or startle stimuli is absent in the mutants. Brain ventricle size is markedly increased at 120 hpf. Heart ventricles are enlarged, with decreased ventricle wall thickness. A cardiac arrhythmia, characterized by missing beats, is also observed in the mutants. This is associated with bradycardia in arnt2−/− larvae. Dilated liver sinusoids merge abnormally to form an extensive, labyrinth-like network of vascular channels. External appearance of arnt2−/− larvae at 120 hpf is indistinguishable from wild type except that the swim bladder is not inflated. The arnt2−/− mutants are not debilitated when phenotypic effects are first detected at 120 hpf that culminate in mortality, 4 days later around 216 hpf. Gross morphological assessment of the development of forebrain, midbrain, and hindbrain regions, neuromasts and Mauthner neurons, inner ear semicircular canals and otoliths, primary motor neurons, trigeminal ganglia, and trunk skeletal muscles, before or when the arnt2−/− phenotype was observed, failed to demonstrate a difference from wild type. The only effect in arnt2−/− larvae that occurred before 120 hpf was a decrease in expression of sim1, an Arnt2 dimerization partner, in the hypothalamus and ventral thalamus at 72 hpf. Further research is needed to determine if the primary functions of Arnt2 occur during the larval stage, when the phenotype is observed, or earlier in development