| HOME | HELP | CONTACT US | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Center of Marine Biotechnology, University of Maryland Biotechnology Institute, 701 East Pratt Street, Baltimore, Maryland 21202, USA
1 Research Group Endocrinology and Metabolism, Department Biology, Faculty of Sciences, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands
2 Physiology of Growth and Reproduction in Fish, Institute of Marine Research, PO Box 1870 Nordnes, 5817 Bergen, Norway
(Requests for offprints should be addressed to Y Kazeto who is now at Division of Marine Life Sciences Research, Faculty of Fisheries Science Hokkaido University 3-1-1 Minato-cho Hakodate, Hokkaido 041-8611 Japan; Email: kazeto{at}fish.hokudai.al.jp)
(R S Kumar is now at Cellular and Molecular Sciences, Wyeth Biopharma, 1 Burtt Road, Andover, Massachusetts 01810, USA)
Due to the lack of purified, native gonadotropins (GtH) for almost all species of fish, we designed a system for the production of recombinant bioactive luteinizing hormone (LH) and follicle stimulating hormone (FSH) using the channel catfish (Ictalurus punctatus) as a model animal. The strategy was to produce the three subunits composing FSH and LH, i.e. the common 
-subunit (-glycoprotein hormone (-GP)), ß-FSH, and ß-LH subunit, individually in stable recombinant insect cells (S2) with C-terminal His-tag. This expression system was also used to co-express the -subunit without the His-tag with each of the His-tagged ß-subunits. The recombinant S2 cells were capable of secreting FSH and LH heterodimers and -GP in abundance; however, expression of the individual ß-subunits was much less successful. The recombinant GtHs were partially purified from the cell medium by immobilized metal affinity chromatography to ~15% purity with a yield of 7 and 4 mg per liter of medium for FSH and LH respectively. These recombinant GtHs activated their receptors in vitro, enhanced estrogen secretion, up-regulated several steroidogenic enzyme genes in channel catfish ovarian follicles, and increased androgen secretion from African catfish testis. Interestingly, the FSH and LH dose–response curves for each of these biological activities clearly demonstrate differences in their cellular action and physiological roles. This expression system may be an important development for the production of species-specific GtHs so that FSH- and LH-specific mechanisms of actions within the reproductive endocrine processes can finally be examined with homologous, albeit recombinant, hormones.
This article has been cited by other articles:
|
|
 |
|
  P M Lokman, Y Kazeto, Y Ozaki, S Ijiri, R Tosaka, M Kohara, S L Divers, H Matsubara, L G Moore, and S Adachi Effects of reproductive stage, GH, and 11-ketotestosterone on expression of growth differentiation factor-9 in the ovary of the eel, Anguilla australis Reproduction, January 1, 2010; 139(1): 71 - 83. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. F.L. Almeida, G. L. Taranger, B. Norberg, O. Karlsen, J. Bogerd, and R. W. Schulz Photoperiod-Modulated Testis Maturation in Atlantic Cod (Gadus morhua, L.) Biol Reprod, April 1, 2009; 80(4): 631 - 640. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Garcia-Lopez, J. Bogerd, J. C. M. Granneman, W. van Dijk, J. M. Trant, G. L. Taranger, and R. W. Schulz Leydig Cells Express Follicle-Stimulating Hormone Receptors in African Catfish Endocrinology, January 1, 2009; 150(1): 357 - 365. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Kazeto, M. Kohara, T. Miura, C. Miura, S. Yamaguchi, J. M. Trant, S. Adachi, and K. Yamauchi Japanese Eel Follicle-Stimulating Hormone (Fsh) and Luteinizing Hormone (Lh): Production of Biologically Active Recombinant Fsh and Lh by Drosophila S2 Cells and Their Differential Actions on the Reproductive Biology Biol Reprod, November 1, 2008; 79(5): 938 - 946. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | CONTACT US | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
