Development and characterization of five rainbow trout pituitary single-cell clone lines capable of producing pituitary hormones.
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 CO(2)-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 CO(2)-independent medium supplemented with 10% FBS at 20 degrees 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 (E(2)) receptor alpha (e2ralpha 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 E(2), 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.
Chen
Maria J
MJ
Department of Molecular and Cell Biology, University of Connecticut, U-3125 Storrs, Connecticut 06269, USA.
Chiou
Pinwen Peter
PP
Liao
Yu-Hsian
YH
Lin
Chun-Mean
CM
Chen
Thomas T
TT
D000818:Animals; D002460:Cell Line; D003907:Dexamethasone / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D006065:Gonadotropins, Pituitary / Q000378:metabolism; D013006:Growth Hormone / Q000378:metabolism; D023421:Models, Animal; D017686:Oncorhynchus mykiss / Q000378:metabolism*; D010902:Pituitary Gland / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D010907:Pituitary Hormones / Q000378:metabolism*; D011333:Pro-Opiomelanocortin / Q000378:metabolism; D011388:Prolactin / Q000378:metabolism; D011987:Receptors, Steroid / Q000378:metabolism; D013972:Thyrotropin / Q000378:metabolism
D006065:Gonadotropins, Pituitary; D010907:Pituitary Hormones; D011987:Receptors, Steroid; D004958:Estradiol; D011333:Pro-Opiomelanocortin; D003907:Dexamethasone; D011388:Prolactin; D013972:Thyrotropin; D013006:Growth Hormone