Analysis of the Zebrafish Proteome during Embryonic Development

Lucitt, Margaret B.; Price, Thomas S.; Pizarro, Angel; Wu, Weichen; Yocum, Anastasia K.; Seiler, Christoph; Pack, Michael; Blair, Ian A.; FitzGerald, Garret A.; Großer, Tilo

doi:10.1074/mcp.m700382-mcp200

Cited by 118 publications

(105 citation statements)

References 63 publications

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“…To show conservation of the injury process across vertebrate species and discover potential biomarkers indicative of outcome, we performed the first analysis of the zebrafish serum plasma proteome. Prior proteomic studies in the zebrafish have focused on the embryo and adult liver (13,29). Our analysis revealed an absence of albumin, the major mammalian serum transport protein, consistent with recent genome analyses (30); apolipoproteins and vitellogenin may serve as alternative carriers for hydrophobic compounds in zebrafish blood (31).…”

Section: Discussionsupporting

confidence: 87%

PGE2-regulated wnt signaling and N -acetylcysteine are synergistically hepatoprotective in zebrafish acetaminophen injury

North

Babu²,

Vedder³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

140

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Acetaminophen (APAP) toxicity is the most common drug-induced cause of acute liver failure in the United States. The only available treatment, N-acetylcysteine (NAC), has a limited time window of efficacy, indicating a need for additional therapeutic options. Zebrafish have emerged as a powerful tool for drug discovery. Here, we developed a clinically relevant zebrafish model of APAP toxicity. APAP depleted glutathione stores, elevated aminotransferase levels, increased apoptosis, and caused dose-dependent hepatocyte necrosis. These outcomes were limited by NAC and conserved in zebrafish embryos. In a targeted embryonic chemical screen, prostaglandin E2 (PGE2) was identified as a potential therapeutic agent; in the adult, PGE2 similarly decreased APAP-associated toxicity. Significantly, when combined with NAC, PGE2 extended the time window for a successful intervention, synergistically reducing apoptosis, improving liver enzymes, and preventing death. Use of a wnt reporter zebrafish line and chemical genetic epistasis showed that the effects of PGE2 are mediated through the wnt signaling pathway. Zebrafish can be used as a clinically relevant toxicological model amenable to the identification of additional therapeutics and biomarkers of APAP injury; our data suggest combinatorial PGE2 and NAC treatment would be beneficial for patients with APAP-induced liver damage.acetaminophen liver toxicity | chemical screen A cetaminophen (N-acetyl-p-aminophenol; APAP) is a commonly used analgesic and antipyretic. Although safe at therapeutic doses, accidental or suicidal drug overdose can cause dosedependent liver damage. APAP is the most common cause for liver transplantation for toxin-induced fulminant hepatic failure and results in more than 300 deaths annually in the United States (1). The only antidote in clinical use is N-acetylcysteine (NAC), which reduces mortality by ∼20-28% (2); however, the time interval of effective intervention after ingestion is typically <12 h, and delayed or prolonged treatment can negatively impact clinical outcome and survival (3). Therapeutic options for hepatic failure are limited to best supportive care and liver transplantation.APAP toxicity results from a hepatotoxic metabolite, N-acetylp-benzoquinone imine (NAPQI), produced by the cytochrome P450 enzymes CYP1A2, 2E1, and 3A4 (4). At therapeutic doses, NAPQI is efficiently inactivated in the liver by glutathione (GSH) conjugation (5). At toxic doses, excess production of NAPQI depletes hepatic GSH. Unconjugated NAPQI causes dysfunction of critical liver proteins, oxidative stress, and mitochondrial damage (6-8). NAC is a true antidote and limits damage by repleting GSH. Clinical efficacy of NAC treatment was shown for patients presenting after APAP ingestion; however, no conclusive clinical trials were conducted to elucidate its efficacy and optimal treatment window. Therapeutic benefits have been shown in mammalian models for other antioxidants (9, 10) that function to restore GSH levels. Compounds that support liver recovery from AP...

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Section: Discussionsupporting

confidence: 87%

PGE2-regulated wnt signaling and N -acetylcysteine are synergistically hepatoprotective in zebrafish acetaminophen injury

North

Babu²,

Vedder³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

140

View full text Add to dashboard Cite

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“…The PRIDE data sets of the brain were acquired on a MALDI TOF/TOF (Applied Biosystems), 37 the fin data with a LTQ ion trap mass spectrometer (Thermo Fisher), 36 and the 3 dpf embryos also on a Thermo Finnigan liquid ion trap mass spectrometer. 38 The p values were calculated as in Figure 4. * p value < 0.05. database searches directly.…”

Section: ■ Discussionmentioning

confidence: 99%

Identifying Proteins in Zebrafish Embryos Using Spectral Libraries Generated from Dissected Adult Organs and Tissues

et al. 2014

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Spectral libraries provide a sensitive and accurate method for identifying peptides from tandem mass spectra, complementary to searching genome-derived databases or sequencing de novo. Their application requires comprehensive libraries including peptides from low-abundant proteins. Here we describe a method for constructing such libraries using biological differentiation to "fractionate" the proteome by harvesting adult organs and tissues and build comprehensive libraries for identifying proteins in zebrafish (Danio rerio) embryos and larvae (an important and widely used model system). Hierarchical clustering using direct comparison of spectra was used to prioritize organ selection. The resulting and publicly available library covers 14 164 proteins, significantly improved the number of peptide-spectrum matches in zebrafish developmental stages, and can be used on data from different instruments and laboratories. The library contains information on tissue and organ expression of these proteins and is also applicable for adult experiments. The approach itself is not limited to zebrafish but would work for any model system.

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“…S1). Proteins isolated from washed platelets were separated on a gel and proteolyzed (33) in the presence of the stable isotope-labeled internal standards (Table S1). The peptide was quantitated in the unacetylated and acetylated states (Table S1) using liquid chromatography tandem MS (Fig.…”

Section: Methodsmentioning

confidence: 99%

Differential impairment of aspirin-dependent platelet cyclooxygenase acetylation by nonsteroidal antiinflammatory drugs

Fries

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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The cardiovascular safety of nonsteroidal antiinflammatory drugs (NSAIDs) may be influenced by interactions with antiplatelet doses of aspirin. We sought to quantitate precisely the propensity of commonly consumed NSAIDs-ibuprofen, naproxen, and celecoxib-to cause a drug-drug interaction with aspirin in vivo by measuring the target engagement of aspirin directly by MS. We developed a novel assay of cyclooxygenase-1 (COX-1) acetylation in platelets isolated from volunteers who were administered aspirin and used conventional and microfluidic assays to evaluate platelet function. Although ibuprofen, naproxen, and celecoxib all had the potential to compete with the access of aspirin to the substrate binding channel of COX-1 in vitro, exposure of volunteers to a single therapeutic dose of each NSAID followed by 325 mg aspirin revealed a potent drug-drug interaction between ibuprofen and aspirin and between naproxen and aspirin but not between celecoxib and aspirin. The imprecision of estimates of aspirin consumption and the differential impact on the ability of aspirin to inactivate platelet COX-1 will confound head-to-head comparisons of distinct NSAIDs in ongoing clinical studies designed to measure their cardiovascular risk.aspirin | acetylation | cyclooxygenase | MS | nonsteroidal antiinflammatory drugs C hronic pain, most commonly inflammatory musculoskeletal pain, afflicts hundreds of millions worldwide (1). Nonsteroidal antiinflammatory drugs (NSAIDs) consumed chronically or intermittently remain the mainstay of therapy for inflammationassociated pain. These agents inhibit cyclooxygenase (COX)-1 and COX-2, thereby reducing the production of inflammatory prostanoids, lipid mediators that lower the activation threshold of nociceptors and sensory neurons. The prevalence of chronic pain rises in the elderly, coinciding with an increase in concomitant disease (1), which complicates drug treatment. Pain management in patients with preexisting cardiovascular disease is a particular challenge because of the cardiovascular adverse effects of NSAIDs and the risk of drug-drug interactions that might undermine the antiplatelet effects of aspirin prescribed for cardioprotection (2).Although NSAIDs relieve pain effectively, they can cause serious renal and cardiovascular complications by inhibiting COXdependent prostanoids with homeostatic functions (3, 4). All NSAIDs have the potential to elevate blood pressure and may cause heart failure. COX-2-selective NSAIDs, which were developed to reduce gastrointestinal toxicity, additionally raise the rate of myocardial infraction and stroke (5, 6), affecting ∼1-2% of patients exposed per year (3,4). This adverse drug reaction may have caused thousands of deaths in the general population. Traditional NSAIDs (tNSAIDs) have also been associated with cardiovascular events, but although pharmacoepidemiological studies and metaanalyses of randomized, controlled trials suggest that not all tNSAIDs carry the same risk, there is considerable heterogeneity across studies in the comparative ...

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Analysis of the Zebrafish Proteome during Embryonic Development

Cited by 118 publications

References 63 publications

PGE2-regulated wnt signaling and N -acetylcysteine are synergistically hepatoprotective in zebrafish acetaminophen injury

PGE2-regulated wnt signaling and N -acetylcysteine are synergistically hepatoprotective in zebrafish acetaminophen injury

Identifying Proteins in Zebrafish Embryos Using Spectral Libraries Generated from Dissected Adult Organs and Tissues

Differential impairment of aspirin-dependent platelet cyclooxygenase acetylation by nonsteroidal antiinflammatory drugs

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