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United States Department of Agriculture

Agricultural Research Service

Research Project: IDENTIFYING BIOCHEMICAL PATHWAYS USING GENETICALLY MODIFIED TROUT (UNIVERSITY OF CT)

Location: Cool and Cold Water Aquaculture Research

Title: Development and characterization of five rainbow trout pituitary single-cell clone lines capable of producing pituitary hormones

Authors
item Chen, Maria -
item Chiou, Pinwen Peter -
item Liao, Yi-Hsian -
item Lin, Chen-Mean -
item Chen, Thomas -

Submitted to: Journal of Endocrinology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 19, 2010
Publication Date: August 6, 2010
Citation: Chen, M., Chiou, P., Liao, Y., Lin, C., Chen, T.T. 2010. Development and characterization of five rainbow trout pituitary single-cell clone lines capable of producing pituitary hormones. Journal of Endocrinology. 205:69-78.

Interpretive Summary: Pituitary hormones are important in regulating growth, metabolism, reproduction and many other important physiological functions in fishes. Thorough understanding of how these hormones are produced in the pituitary gland is necessary for manipulating the levels of these hormones in aquaculture. Unfortunately lacking permanent pituitary cell lines from fish has hampered the progress in this area. To overcome this handicap, we have established five single-cell clones from rainbow trout pituitary glands. These cell lines were observed to be capable of expressing genes known to play critical roles in fundamental physiological processes such as growth and reproduction. These cell lines will enable fish endocrinologists and aquaculture researchers to develop strategies to increase the efficiency of fish production.

Technical Abstract: Five single-cell clone lines (mRTP1B, mRTP1E, mRTP1F, mRTP1K, and mRTP2A) have been developed from adult rainbow trout pituitary glands. These cell lines have been maintained in a CO2-independent medium supplemented with 10% fetal bovine serum (FBS) for more than 150 passages. At about 150 passages, the doubling time of each single-cell clone in a CO2-independent medium supplemented with 10% FBS at 20 8C was 3.6G0.7, 2.8G0.7, 3.2G0.8, 5.5G0.6, and 6.6G0.6 days respectively. Each single-cell clone contains 60G2 chromosomes, which is within the range of the 2N chromosome numbers reported for rainbow trout. Reverse transcription-PCR analysis revealed that in addition to expressing gh, prolactin (prl), and estradiol (E2) receptor a (e2ra or esr1) genes, each single-cell clone line also expressed other pituitary-specific genes such as tsh, gonadotropin 1 (gth-1 or fshb), gonadotropin 2 (gth-2 or lhb), somatolactin (sl or smtl), proopiomelanocortin-B (pomcb), and corticosteroid receptor (cr or nr3c1). Immunocytochemical analysis showed that all the five single-cell clones produced both Gh and Prl. Furthermore, the expression of gh and prl genes in the single-cell clone lines is responsive to induction by E2, dexamethasone, and op'-dichlorodiphenyltrichloroethane. All together, these results confirm that each of the singlecell clones was derived from rainbow trout pituitary glands. These single-cell clone lines not only can be used to study factors that regulate the expression of pituitary hormone genes, but can also be developed as a rapid screening system for identifying environmental endocrine disruptors.

Last Modified: 10/20/2014