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Extramural Research

Grantee Research Project Results

NCER Grantee Research Project Results

Transforming Growth Factor-ß (Tgf-ß) in Teleost Fish: Development of a Quantitative Assay for mRNA Expression and Application to a Field Study of Fish Health

EPA Grant Number: U915209
Title: Transforming Growth Factor-ß (Tgf-ß) in Teleost Fish: Development of a Quantitative Assay for mRNA Expression and Application to a Field Study of Fish Health
Investigators: Harms, Craig A.
Institution: North Carolina State University
EPA Project Officer: Boddie, Georgette
Project Period: January 1, 1997 through January 1, 2000
Project Amount: $102,000
RFA: STAR Graduate Fellowships (1997)
Research Category: Fellowship - Molecular Biology/Genetics , Academic Fellowships , Biology/Life Sciences

Description:

Objective:

The general hypothesis of this research project are to: (1) determine whether cytokines homologous to those characterized in mammals are present in fish; and (2) assess whether cytokine production will provide a powerful tool for monitoring the immune status in a broad range of fish species in response to toxicologic insults and infectious agents. Research is focused on transforming growth factor-ß (TGF-ß), a cytokine which plays an important role in immunity (being primarily immunosuppressive), development, cell proliferation, and carcinogenesis.

Approach:

A TGF-ß was isolated and cloned from hybrid striped bass (Morone saxatilis x Morone. chrysops) anterior kidney mononuclear cells. This isolate contains an open reading frame of 1,146 bases coding for a 382 amino acid protein most similar to rainbow trout TGF-ß (57.3% percent and 78.6% percent identity with precursor and active protein respectively) and rat TGF-ß1 (41.1% percent and 68.8% percent identity with precursor and active protein respectively). Consensus primers were designed, which amplify—by polymerase chain reaction (PCR)Ca TGF-ß segment from at least 11 species of teleost fish comprising 8eight taxonomic families in 5five orders. A reverse transcription quantitative competitive polymerase chain reaction (RT-qcPCR) assay was devised to measure TGF-ß mRNA expression in teleost fish. The RT-qcPCR assay was used to determine TGF-ß transcription levels from mononuclear cells of different lymphoid tissues of hybrid striped bass. The assay was then used to compare TGF-ß mRNA expression with macrophage bacteriocidal activity in hybrid striped bass in response to an immunomodulator and in white perch (Morone americana) in a field study.

Supplemental Keywords:

fellowship, teleost fish, polymerase chain reaction, PCR, reverse transcription quantitative competitive polymerase chain reaction, RT-qcPCR, cytokine production, cell proliferation.

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The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.

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