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This Article Full Text Full Text (PDF) All Versions of this Article: JOE-09-0315v1 205/1/69    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Google Scholar Articles by Chen, M. J Articles by Chen, T. T PubMed PubMed Citation Articles by Chen, M. J Articles by Chen, T. T

Department of Molecular and Cell Biology, University of Connecticut, 91 North Eagleville Road, U-3125 Storrs, Connecticut 06269, USA

(Correspondence should be addressed to T T Chen; Email: thomas.chen{at}uconn.edu)

(P P Chiou is now at Marine Research Station, Institute of Cellular and Organismal Biology, Academia Sinica, Yilan, Taiwan, ROC)

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 °C was 3.6±0.7, 2.8±0.7, 3.2±0.8, 5.5±0.6, and 6.6±0.6 days respectively. Each single-cell clone contains 60±2 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 (e2r 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 o,p'-dichlorodiphenyltrichloroethane. All together, these results confirm that each of the single-cell 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.