Differential Expression of Immune Response Genes in Steller Sea Lions (Eumetopias jubatus): An Indicator of Ecosystem Health?

Bowen, Lizabeth; Aldridge, Brian; Beckmen, Kimberlee B.; Gelatt, Tom; Rea, Lorrie D.; Burek, Kathy A.; Pitcher, Ken; Stott, Jeffrey L

doi:10.1007/s10393-006-0021-0

Cited by 13 publications

(7 citation statements)

References 21 publications

(30 reference statements)

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“…Association of IL-10 transcription with chronic disease has also been documented in humans (Rigopoulou et al 2005). DRB A component of the major histocompatibility complex, the DRB class II gene, is responsible for the binding and presentation of processed antigenn to T H lymphocytes, thereby facilitating the initiation of an immune response (Goldsby et al 2003, Bowen et al 2006b). Up-regulation of major histocompatibility complex (MHC) genes has been positively correlated with parasite load (Wegner et al 2006), whereas down-regulation of MHC has been associated with contaminant exposure (Dong et al 1997).…”

Section: Il-10mentioning

confidence: 99%

Variations of transcript profiles between sea otters Enhydra lutris from Prince William Sound, Alaska, and clinically normal reference otters

Miles¹,

Bowen²,

Ballachey³

et al. 2012

Mar. Ecol. Prog. Ser.

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Section: Il-10mentioning

confidence: 99%

Variations of transcript profiles between sea otters Enhydra lutris from Prince William Sound, Alaska, and clinically normal reference otters

Miles¹,

Bowen²,

Ballachey³

et al. 2012

Mar. Ecol. Prog. Ser.

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“…Amplifications were performed in an ABI 7300 (Applied Biosystems) under the following conditions: 2 minutes at 50°C, followed by 15 minutes at 95°C, and 35 cycles of 94°C for 30 seconds, 58°C for 30 seconds, and 72°C for 30 seconds, with a final extension step of 72°C for 10 minutes. Reaction specificity was monitored by melting curve analysis using a final data acquisition phase of 60 cycles of 65°C for 30 seconds and verified by direct sequencing of randomly selected amplicons (Bowen et al, 2006). Gene expression was analyzed by relative quantitation, using the comparative C T (cycle threshold) method; values are expressed relative to a calibrator (weakest signal of the normalized values) (Bowen et al, 2006).…”

Section: Quantitative Pcrmentioning

confidence: 99%

“…Reaction specificity was monitored by melting curve analysis using a final data acquisition phase of 60 cycles of 65°C for 30 seconds and verified by direct sequencing of randomly selected amplicons (Bowen et al, 2006). Gene expression was analyzed by relative quantitation, using the comparative C T (cycle threshold) method; values are expressed relative to a calibrator (weakest signal of the normalized values) (Bowen et al, 2006). Amplification efficiencies of S9 and the other genes of interest (GOI) were determined using six dilutions of cDNA preparations (run in triplicate).…”

Section: Quantitative Pcrmentioning

confidence: 99%

Differential Gene Expression Induced by Exposure of Captive Mink to Fuel Oil: A Model for the Sea Otter

et al. 2007

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Free-ranging sea otters are subject to hydrocarbon exposure from a variety of sources, both natural and anthropogenic. Effects of direct exposure to unrefined crude oil, such as that associated with the Exxon Valdez oil spill, are readily apparent. However, the impact of subtle but pathophysiologically relevant concentrations of crude oil on sea otters is difficult to assess. The present study was directed at developing a model for assessing the impact of low concentrations of fuel oil on sea otters. Quantitative PCR was used to identify differential gene expression in American mink that were exposed to low concentrations of bunker C fuel oil. A total of 23 genes, representing 10 different physiological systems, were analyzed for perturbation. Six genes with immunological relevance were differentially expressed in oil-fed mink. Interleukin-18 (IL-18), IL-10, inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), and complement cytolysis inhibitor (CLI) were down-regulated while IL-2 was up-regulated. Expression of two additional genes was affected; heat shock protein 70 (HSP70) was up-regulated and thyroid hormone receptor (THR) was down-regulated. While the significance of each perturbation is not immediately evident, we identified differential expression of genes that would be consistent with the presence of immune system-modifying and endocrine-disrupting compounds in fuel oil. Application of this approach to identify effects of petroleum contamination on sea otters should be possible following expansion of this mink model to identify a greater number of affected genes in peripheral blood leukocytes.

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“…A component of the major histocompatibility complex, the DRB class II gene, is responsible for the binding and presentation of processed antigen to T H lymphocytes, thereby facilitating the initiation of an immune response (Goldsby et al 2003;Bowen et al 2006). Up-regulation of MHC genes has been positively correlated with parasite load (Wegner et al 2006), whereas down-regulation of MHC has been associated with contaminant exposure (Dong et al 1997).…”

Section: Drbmentioning

confidence: 99%

Gene transcription in sea otters (Enhydra lutris); development of a diagnostic tool for sea otter and ecosystem health

Bowen

Miles

Murray

et al. 2011

Molecular Ecology Resources

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Gene transcription analysis for diagnosing or monitoring wildlife health requires the ability to distinguish pathophysiological change from natural variation. Herein, we describe methodology for the development of quantitative real-time polymerase chain reaction (qPCR) assays to measure differential transcript levels of multiple immune function genes in the sea otter (Enhydra lutris); sea otter-specific qPCR primer sequences for the genes of interest are defined. We establish a 'reference' range of transcripts for each gene in a group of clinically healthy captive and free-ranging sea otters. The 10 genes of interest represent multiple physiological systems that play a role in immuno-modulation, inflammation, cell protection, tumour suppression, cellular stress response, xenobiotic metabolizing enzymes, antioxidant enzymes and cell-cell adhesion. The cycle threshold (C(T)) measures for most genes were normally distributed; the complement cytolysis inhibitor was the exception. The relative enumeration of multiple gene transcripts in simple peripheral blood samples expands the diagnostic capability currently available to assess the health of sea otters in situ and provides a better understanding of the state of their environment.

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Differential Expression of Immune Response Genes in Steller Sea Lions (Eumetopias jubatus): An Indicator of Ecosystem Health?

Cited by 13 publications

References 21 publications

Variations of transcript profiles between sea otters Enhydra lutris from Prince William Sound, Alaska, and clinically normal reference otters

Variations of transcript profiles between sea otters Enhydra lutris from Prince William Sound, Alaska, and clinically normal reference otters

Differential Gene Expression Induced by Exposure of Captive Mink to Fuel Oil: A Model for the Sea Otter

Gene transcription in sea otters (Enhydra lutris); development of a diagnostic tool for sea otter and ecosystem health

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