title: Production of recombinant Japanese eel gonadotropins by baculovirus in silkworm larvae.
abstract: Recombinant follicle-stimulating hormone (reFSH) and luteinizing hormone (reLH) of the Japanese eel Anguilla japonica were produced by baculovirus in silkworm Bombyx mori larvae. cDNAs encoding Japanese eel gonadotropin subunits (i.e., FSH beta, LH beta, and common alpha) were introduced into the baculovirus, which was infected into silkworm larvae after propagation of the recombinant virus in B. mori culture cells. A 100ml solution of pooled hemolymph from silkworm larvae containing reFSH or reLH were obtained from approximately 250 infected larvae. Ten milliliters of hemolymph were applied to Ni-affinity choromatography, and 5.6 and 3.5mg of partially purified reFSH and reLH were obtained, respectively. Using Western blot analysis concentrations of reFSH and reLH in the original hemolymph was estimated to be 2.2 and 1.1mg/ml, respectively. Biological activities of reFSH and reLH were assessed in vitro and in vivo. Purified reFSH and reLH induced eel oocyte maturation in vitro, and administration of hemolymph containing reFSH or reLH induced spermatogenesis in vivo in sexually immature Japanese eel. The present study indicates that a baculovirus-silkworm system could produce large amounts of biologically active recombinant fish gonadotropins for use in investigations in reproductive endocrinology and/or aquaculture of fish.
authors.affiliation: Department of Life Science, International Christian University, Mitaka, Tokyo 181-8585, Japan. makito@icu.ac.jp
authors.forename: Makito
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Youichi
authors.initials: Y
authors.lastname: Hayakawa
authors.forename: Woodong
authors.initials: W
authors.lastname: Park
authors.forename: Akira
authors.initials: A
authors.lastname: Banba
authors.forename: Goro
authors.initials: G
authors.lastname: Yoshizaki
authors.forename: Katsuya
authors.initials: K
authors.lastname: Kumamaru
authors.forename: Hirohiko
authors.initials: H
authors.lastname: Kagawa
authors.forename: Hiroki
authors.initials: H
authors.lastname: Kaki
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Nagaya
authors.forename: Young Chang
authors.initials: YC
authors.lastname: Sohn
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D012831:Bombyx / Q000254:growth & development / Q000378:metabolism*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004353:Drug Evaluation, Preclinical; D004524:Eels / Q000235:genetics*; D005260:Female; D005822:Genetic Vectors / Q000008:administration & dosage; D006062:Gonadotropins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D007814:Larva / Q000378:metabolism; D008297:Male; D008954:Models, Biological; D009865:Oocytes / Q000187:drug effects / Q000502:physiology; D009866:Oogenesis / Q000187:drug effects / Q000502:physiology; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D013091:Spermatogenesis / Q000187:drug effects / Q000502:physiology; D014161:Transduction, Genetic / Q000379:methods
chemical_list: D006062:Gonadotropins; D011994:Recombinant Proteins