title: Production and characterization of recombinant Manchurian trout thyrotropin.
abstract: Thyrotropin (thyroid-stimulating hormone, TSH), a heterodimeric glycoprotein hormone produced in the pituitary, stimulates the thyroid gland and release of thyroid hormones. In contrast to a well-known efficacy of recombinant mammalian TSHs, there is no report about the production of teleost recombinant TSH and its biological activity. In this study, we report the production of a single-chain recombinant TSH (mtTSH) of Manchurian trout (Brachymystax lenok), by baculovirus in silkworm (Bombyx mori) larvae. The mtTSH was produced in silkworm larvae and characterized as a form of N-linked glycosylation. The cAMP signaling system in transiently transfected COS-7 cells revealed that the mtTSH was recognized by their cognate receptors, salmon TSHα and TSHβ receptors, but not LH receptor. The thyrotropic potency of the mtTSH was examined by rainbow trout basibranchial tissues containing thyroid follicles. The height of follicle epithelial cells was significantly increased by treatments of mtTSH in vivo and in vitro. In conclusion, the present study suggests that the mtTSH produced by baculovirus-silkworm larvae is a biologically active recombinant TSH.
authors.affiliation: Department of Marine Molecular Biotechnology, Gangneung-Wonju National University, Gangneung, 210-702, Republic of Korea.
authors.forename: Jihye
authors.initials: J
authors.lastname: Shin
authors.forename: Mi Ae
authors.initials: MA
authors.lastname: Kim
authors.forename: Makito
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Young Chang
authors.initials: YC
authors.lastname: Sohn
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D016367:Baculoviridae / Q000378:metabolism; D019149:Bioreactors; D015153:Blotting, Western; D012831:Bombyx / Q000821:virology; D019556:COS Cells; D002522:Chlorocebus aethiops; D000242:Cyclic AMP / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000096:biosynthesis; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells / Q000187:drug effects; D006031:Glycosylation; D013006:Growth Hormone / Q000096:biosynthesis* / Q000378:metabolism / Q000494:pharmacology; D007814:Larva / Q000821:virology; D017686:Oncorhynchus mykiss / Q000378:metabolism; D013972:Thyrotropin / Q000096:biosynthesis* / Q000378:metabolism / Q000494:pharmacology; D014337:Trout / Q000235:genetics*
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D013972:Thyrotropin; D013006:Growth Hormone; D000242:Cyclic AMP