Publications by authors named "Tracy A Sherwood"

7 Publications

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Characterization of the differential expressed genes and transcriptomic pathway analysis in the liver of sub-adult red drum (Sciaenops ocellatus) exposed to Deepwater Horizon chemically dispersed oil.

Ecotoxicol Environ Saf 2021 May 2;214:112098. Epub 2021 Mar 2.

Mote Marine Laboratory, 1600 Ken Thompson Pkwy, Sarasota, FL 34236, USA.

The Deepwater Horizon blowout resulted in the second-largest quantity of chemical dispersants used as a countermeasure for an open water oil spill in the Gulf of Mexico. Of which, the efficacy of dispersant as a mitigation strategy and its toxic effects on aquatic fauna remains controversial. To enhance our understanding of potential sub-lethal effects of exposure to chemically dispersed-oil, sub-adult red drum (Sciaenops ocellatus) were continuously exposed to a Corexit 9500: DWH crude oil chemically enhanced water accommodated fraction (CEWAF) for 3-days and transcriptomic responses were assessed in the liver. Differential expressed gene (DEG) analysis demonstrated that 63 genes were significantly impacted in the CEWAF exposed fish. Of these, 37 were upregulated and 26 downregulated. The upregulated genes were primarily involved in metabolism and oxidative stress, whereas several immune genes were downregulated. Quantitative real-time RT-PCR further confirmed upregulation of cytochrome P450 and glutathione S-transferase, along with downregulation of fucolectin 2 and chemokine C-C motif ligand 20. Ingenuity Pathway Analysis (IPA) predicted 120 pathways significantly altered in the CEWAF exposed red drum. The aryl hydrocarbon receptor pathway was significantly activated, while pathways associated with immune and cellular homeostasis were primarily suppressed. The results of this study indicate that CEWAF exposure significantly affects gene expression and alters signaling of biological pathways important in detoxification, immunity, and normal cellular physiology, which can have potential consequences on organismal fitness.
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http://dx.doi.org/10.1016/j.ecoenv.2021.112098DOI Listing
May 2021

Characterizing transcriptomic responses of southern flounder (Paralichthys lethostigma) chronically exposed to Deepwater Horizon oiled sediments.

Aquat Toxicol 2021 Jan 7;230:105716. Epub 2020 Dec 7.

Mote Marine Laboratory, 1600 Ken Thompson Parkway, Sarasota, FL, 34236, United States.

To obtain a deeper understanding of the transcriptomic responses to oil in southern flounder (Paralichthys lethostigma), we performed quantitative PCR and RNA sequencing on liver and gill tissue after a chronic exposure (35 days) to Deepwater Horizon crude oiled sediment and after a 30-day recovery period. We wanted to understand which specific genes are differentially expressed in liver and gill tissues directly after oiled sediment exposure and with the addition of a recovery period. Furthermore, we wanted to examine specific enriched pathways in these two tissues to determine the impact of exposure with and without a recovery period on biological processes (e.g. immune function). Liver and gill tissues were chosen because they represent two distinct organs that are highly important to consider when examining the impacts of oiled sediment exposure. The liver is the classic detoxification organ, while the gill is in direct contact with sediment in benthic fishes. Examination of these two tissues, therefore, generates a broad understanding of the transcriptomic consequences of oil exposure across an organism. Gene expression for interleukin 8 (il8) and interleukin 1B (il1β) was significantly increased versus control measurements for fish exposed to oiled sediments for 35 days in gill tissue. Hierarchical clustering of gene expression showed that tissue type was the main driver of gene expression (rather than treatment). The inclusion of a 30-day post-exposure recovery period showed a return of il8 and il1β gene expression in the gill to baseline expression levels. However, the recovery period increased the number of differentially expressed genes and significantly affected canonical pathways in both tissue types. Pathways related to cholesterol biosynthesis were significantly suppressed in oil-exposed flounder with a recovery period, but not in the exposed flounder without a recovery period. At the end of the exposure, 17 pathways were significantly affected in the gill, including thyroid hormone metabolism-related pathways, which were the most influenced. Liver tissue from the recovered fish had the greatest number of enriched pathways for any tissue or time point (187). Cellular and humoral immune response pathways were considerably impacted in the liver after the recovery period, suggesting that the immune system was attempting to respond to potential damage caused from the chronic oil exposure. Our results demonstrate that liver and gill tissues from southern flounder were differentially altered by Deepwater Horizon oiled sediment exposure and that a 30-day recovery period after exposure substantially shifted gene expression and canonical pathway profiles.
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http://dx.doi.org/10.1016/j.aquatox.2020.105716DOI Listing
January 2021

Nonlethal Biomarkers of Oxidative Stress in Oiled Sediment Exposed Southern Flounder (): Utility for Field-Base Monitoring Exposure and Potential Recovery.

Environ Sci Technol 2019 12 5;53(24):14734-14743. Epub 2019 Dec 5.

Mote Marine Laboratory , 1600 Ken Thompson Parkway , Sarasota , Florida 34236 , United States.

The Deepwater Horizon (DWH) blowout resulted in the deposition of toxic polycyclic aromatic hydrocarbons (PAHs), in the coastal sediments of the Gulf of Mexico. The immediate effects on an ecosystem from an oil spill are clearly recognizable, however the long-term chronic effects and recovery after a spill are still not well understood. Current methodologies for biomonitoring wild populations are invasive and mostly lethal. Here, two potential nonlethal biomonitoring tools for the assessment of PAH toxicity and induced biological alterations in the field, were identified using laboratory-validated methods. In this study, subadult southern flounder () were chronically exposed to DWH surrogate oiled sediments for 35 days; a subset of these exposed flounder were then provided a clean nonexposure period to ascertain the utility of selected biomarkers to monitor recovery post exposure. After chronic exposure, there was an increase in gene expression of cytochrome P450 1A but not glutathione -transferase. There was also a notable imbalance of oxidants to antioxidants, measured as reduced glutathione, oxidized glutathione, and their ratio in the blood. Evidence of subsequent oxidative damage due to chronic exposure was found through lipid peroxidation and DNA damage assessments of liver, gill, and blood.
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http://dx.doi.org/10.1021/acs.est.9b05930DOI Listing
December 2019

De novo assembly and transcriptome dataset of liver, testis and head kidney from red drum ().

Data Brief 2019 Feb 11;22:934-939. Epub 2019 Jan 11.

Mote Marine Laboratory, 1600 Ken Thompson Pkwy, Sarasota, FL 34235, USA.

Red drum () is an estuarine Sciaenid with high commercial value and recreational demand. During the past 50 years, overfishing has caused declines in the population that resulted in the development of red drum commercial and stock enhancement aquaculture fisheries. Despite the potential high economic value in both wild and aquaculture commercial fisheries the availability of transcriptomic data for red drum in public databases is limited. The data here represents the transcriptome profiles of three tissues: liver, testis and head kidney from red drum. The data was generated using Illumina high-throughput RNA sequencing, Trinity for de novo assembly and Blast2GO for annotation. Six individual libraries were pooled for sequencing of the transcriptome and the raw fastq reads have been deposited in the NCBI-SRA database (accession number SRP11690).
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http://dx.doi.org/10.1016/j.dib.2019.01.011DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6362861PMC
February 2019

Linkage Mapping and Comparative Genomics of Red Drum () Using Next-Generation Sequencing.

G3 (Bethesda) 2017 03 10;7(3):843-850. Epub 2017 Mar 10.

Marine Genomics Laboratory, Department of Life Sciences, Texas A&M University, Corpus Christi, Texas 78412.

Developments in next-generation sequencing allow genotyping of thousands of genetic markers across hundreds of individuals in a cost-effective manner. Because of this, it is now possible to rapidly produce dense genetic linkage maps for nonmodel species. Here, we report a dense genetic linkage map for red drum, a marine fish species of considerable economic importance in the southeastern United States and elsewhere. We used a prior microsatellite-based linkage map as a framework and incorporated 1794 haplotyped contigs derived from high-throughput, reduced representation DNA sequencing to produce a linkage map containing 1794 haplotyped restriction-site associated DNA (RAD) contigs, 437 anonymous microsatellites, and 44 expressed sequence-tag-linked microsatellites (EST-SSRs). A total of 274 candidate genes, identified from transcripts from a preliminary hydrocarbon exposure study, were localized to specific chromosomes, using a shared synteny approach. The linkage map will be a useful resource for red drum commercial and restoration aquaculture, and for better understanding and managing populations of red drum in the wild.
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http://dx.doi.org/10.1534/g3.116.036350DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5345714PMC
March 2017

Identification of transcription start sites and preferential expression of select CB2 transcripts in mouse and human B lymphocytes.

J Neuroimmune Pharmacol 2009 Dec 16;4(4):476-88. Epub 2009 Sep 16.

Department of Molecular Medicine, School of Basic Biomedical Sciences, College of Medicine, University of South Florida, 12901 Bruce B Downs Blvd., Tampa, FL 33612, USA.

Marijuana cannabinoids, the endocannabinoids, and cannabinoid cell receptors have been shown to play important roles in immune regulation particularly as potent modulators of anti-inflammatory cytokines. The predominant cannabinoid receptor involved in this immune regulation is cannabinoid receptor 2 (CB(2)), which is predominantly expressed in B lymphocytes. However, the promoter region and mechanisms of CB(2) gene regulation are unknown in this immune cell type. Utilizing a combination of bioinformatics, 5' rapid amplification of cDNA ends (5' RACE), real-time reverse transcription-polymerase chain reaction, DNA sequencing, and luciferase reporter assays, we show that human B cells express one CB(2) transcript while mouse B cells express three CB(2) transcripts, with specific transcript selection occurring during B cell activation by lipopolysaccharide. Alignment of our sequenced RACE products to either the mouse or human genome, along with the GenBank submitted mRNA sequences, revealed that the transcripts we isolated contained previously unidentified transcriptional start sites (TSS). In addition, expression construct testing of the genomic region containing the TSSs of the mouse CB(2) exon 1 transcripts showed an eightfold increase of promoter activity over baseline. These data show for the first time that human B cells use only one TSS for CB(2) while mouse B cells use multiple TSSs and that the mouse TSSs are in a genomic area with promoter activity, thus suggesting the location of the gene promoter region. Defining these TSSs also provides clues to the various gene regulatory factors involved in the expression of CB(2) during B cell activation.
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http://dx.doi.org/10.1007/s11481-009-9169-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2843092PMC
December 2009

Delta-9 tetrahydrocannabinol (THC) inhibits lytic replication of gamma oncogenic herpesviruses in vitro.

BMC Med 2004 Sep 15;2:34. Epub 2004 Sep 15.

Department of Medical Microbiology and Immunology, MDC Box 10, University of South Florida, and the H. Lee Moffitt Cancer Center, 12901 Bruce B. Downs Blvd, Tampa, FL 33612-4799, USA.

Background: The major psychoactive cannabinoid compound of marijuana, delta-9 tetrahydrocannabinol (THC), has been shown to modulate immune responses and lymphocyte function. After primary infection the viral DNA genome of gamma herpesviruses persists in lymphoid cell nuclei in a latent episomal circular form. In response to extracellular signals, the latent virus can be activated, which leads to production of infectious virus progeny. Therefore, we evaluated the potential effects of THC on gamma herpesvirus replication.

Methods: Tissue cultures infected with various gamma herpesviruses were cultured in the presence of increasing concentrations of THC and the amount of viral DNA or infectious virus yield was compared to those of control cultures. The effect of THC on Kaposi's Sarcoma Associated Herpesvirus (KSHV) and Epstein-Barr virus (EBV) replication was measured by the Gardella method and replication of herpesvirus saimiri (HVS) of monkeys, murine gamma herpesvirus 68 (MHV 68), and herpes simplex type 1 (HSV-1) was measured by yield reduction assays. Inhibition of the immediate early ORF 50 gene promoter activity was measured by the dual luciferase method.

Results: Micromolar concentrations of THC inhibit KSHV and EBV reactivation in virus infected/immortalized B cells. THC also strongly inhibits lytic replication of MHV 68 and HVS in vitro. Importantly, concentrations of THC that inhibit virus replication of gamma herpesviruses have no effect on cell growth or HSV-1 replication, indicating selectivity. THC was shown to selectively inhibit the immediate early ORF 50 gene promoter of KSHV and MHV 68.

Conclusions: THC specifically targets viral and/or cellular mechanisms required for replication and possibly shared by these gamma herpesviruses, and the endocannabinoid system is possibly involved in regulating gamma herpesvirus latency and lytic replication. The immediate early gene ORF 50 promoter activity was specifically inhibited by THC. These studies may also provide the foundation for the development of antiviral strategies utilizing non-psychoactive derivatives of THC.
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http://dx.doi.org/10.1186/1741-7015-2-34DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC521080PMC
September 2004