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Alterations in hepatic gene expression by trivalent chromium in Fundulus heteroclitus.

ABSTRACT: Cr(III) is the dominant toxicant at some Superfund sites within the United States and therefore we are interested in its effects. Cr(III)s mechanisms are not well studied or understood because of its low bioavailability. We have attempted to characterize the effects of Cr(III) on gene expression in Fundulus heteroclitus (mummichog) liver. The NOEC and LOEC were determined at 32 and 64 mg/L, respectively, by measuring growth and mortality after exposing juveniles for 30 days. Secondary adult male exposures were performed at 32 mg/L, livers excised, and RNA extracted. Arrays were probed with cDNA from untreated or Cr(III)-exposed adult fish and gene expression was quantified. Cr(III) at 32 mg/L altered the expression of five genes, including GSTtau, GSTalpha, and ALDH4. Ultimately, we anticipate using this gene expression information to ascertain whether Cr(III) is bioavailable at potentially adverse concentrations in contaminated sites.

SUBMITTER: Roling JA

PROVIDER: S-EPMC1522065 | biostudies-literature | 2006 Jul

REPOSITORIES: biostudies-literature

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Alterations in hepatic gene expression by trivalent chromium in Fundulus heteroclitus.

Roling Jonathan A JA Baldwin William S WS

Marine environmental research 20060407

Cr(III) is the dominant toxicant at some Superfund sites within the United States and therefore we are interested in its effects. Cr(III)s mechanisms are not well studied or understood because of its low bioavailability. We have attempted to characterize the effects of Cr(III) on gene expression in Fundulus heteroclitus (mummichog) liver. The NOEC and LOEC were determined at 32 and 64 mg/L, respectively, by measuring growth and mortality after exposing juveniles for 30 days. Secondary adult male ...[more]

PMID: 16690119

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Project description:The Atlantic killifish (Fundulus heteroclitus) is an environmental sentinel organism used extensively for studies on environmental toxicants and salt (NaCl) homeostasis. Previous research in our laboratory has shown that rapid acclimation of killifish to seawater is mediated by trafficking of CFTR chloride channels from intracellular vesicles to the plasma membrane in the opercular membrane within the first hour in seawater, which enhances chloride secretion into seawater, thereby contributing to salt homeostasis. Acute transition to seawater is also marked by an increase in both mRNA and protein levels of serum glucocorticoid kinase 1 (SGK1) within 15 minutes of transfer. Although the rise in SGK1 in gill and its functional analog, the opercular membrane, after seawater transfer precedes the increase in membrane CFTR, a direct role of SGK1 in elevating membrane CFTR has not been established in vivo. To test the hypothesis that SGK1 mediates the increase in plasma membrane CFTR we designed two functionally different vivo-morpholinos to knock down SGK1 in gill, and developed and validated a vivo-morpholino knock down technique for adult killifish. Injection (intraperitoneal, IP) of the splice blocking SGK1 vivo-morpholino reduced SGK1 mRNA in the gill after transition from fresh to seawater by 66%. The IP injection of the translational blocking and splice blocking vivo-morpholinos reduced gill SGK1 protein abundance in fish transferred from fresh to seawater by 64% and 53%, respectively. Moreover, knock down of SGK1 completely eliminated the seawater induced rise in plasma membrane CFTR, demonstrating that the increase in SGK1 protein is required for the trafficking of CFTR from intracellular vesicles in mitochondrion rich cells to the plasma membrane in the gill during acclimation to seawater. This is the first report of the use of vivo-morpholinos in adult killifish and demonstrates that vivo-morpholinos are a valuable genetic tool for this environmentally relevant model organism.

Dataset Information

Alterations in hepatic gene expression by trivalent chromium in Fundulus heteroclitus.

Publications

Alterations in hepatic gene expression by trivalent chromium in Fundulus heteroclitus.

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