Abstract
Two groups of transgenic rainbow trout (Oncorhynchus mykiss, Walbaum) have been produced and compared. One group harbored the reporter gene of chloramphenicol acetyltransferase (CAT) associated with mouse immunoglobulin (Ig) promoter/enhancer (pUCL-CAT-E). The other group carried the same reporter gene under the control of the cytomegalovirus promoter/enhancer (pCMV-CAT). Slot blot analysis of DNA from blood cells and other tissues from pUCL-CAT-E fish showed variation of copy number between the major tissues but not between red and white blood cells. Southern blot analysis indicated that multiple copies organized in concatemers were incorporated into the genome. The pCMV-CAT fish had a pronounced expression of CAT in both white and red blood cells. In contrast, activity of CAT was found in the white blood cells of all pUCL-CAT-E fish but not in their red blood cells. Expression in white blood cells was found preferentially in sIg+ cells, indicating that B cells are the major expressors. High expression was also found in spleen and kidney, but the activity found in thymocytes was equal to the background level. Analysis of some major tissues showed high white blood cell expression associated with low tissue expression, except that liver (known to contain lymphoid tissue in fish) was higher. Thus the regulatory elements of the Ig gene from mouse induce a tissue-specific expression in fish.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Amemiya, C. T., and Litman, G. W. Complete nucleotide sequence of adn immunoglobulin heavy-chain gene and analysis of immunoglobulin gene organization in a primitive teleost species. Proc Natl Acad Sci USA 87: 811–815, 1990.
Banerji, J., Olson, L., and Schaffner, W. A lymphocyte-specific cellular enhancer is located downstream of the joining region in immunoglobulin heavy chain gene. Cell 33: 729–740, 1983.
Bengtén, E., Leanderson, T., and Pilström, L. Immunoglobulin heavy chain cDNA from the teleost Atlantic cod (Gadus morhua L.): nucleotide sequence of secretory and membrane form show an unusual splicing pattern. Eur J Immunol 21: 3027–3033, 1991.
Chourrout, D, Guyomard, R., and Houdebine, L. M. High efficiency gene transfer in rainbow trout (Salmo gairdneri R.) by microinjection into egg cytoplasm. Aquaculture 51: 143–150, 1986.
Culp, P., Nüsslein-Volhard, C., and Hopkins, N. High efficiency germline transmission of plasmid DNA sequences injected into fertilized zebrafish eggs. Proc Natl Acad Sci USA 88: 7953–7957, 1991.
Dunham, R. A., Eash, J., Askins, J., and Townes, T. M. Transfer of metallothionein-human growth hormone fusion gene into channel catfish. Trans Am Fish Soc 116: 87–91, 1987.
Ellis, A. E., Roberts, R. J., and Tytler, P. The anatomy and physiology of teleosts. In R. J. Roberts (ed.): Fish Pathology, 2nd edn, pp. 13–55, Bailliere Tindal, London, 1989.
Fletcher, G. L., Davies, P. L., and Hew, C. L. Evidence for antifreeze protein gene transfer in Atlantic salmon (Salmo salar). Can J Fish Aquat Sci 45: 352–357, 1988.
Foecking, M. K., and Hofstetter, H. Powerful and versatile enhancer-promoter unit for mammalian expression vectors. Gene 45: 101–105, 1986.
Ghaffari, S. H. and Lobb, C. J. Cloning and sequence analysis of channel catfish heavy chain cDNA indicate phylogenetic diversity within IgM immunoglobulin family. J Immunol 142: 1356–1365, 1989.
Ghaffari, S. H. and Lobb, C. J. Organisation of immunoglobulin heavy chain constant and joining genes in the channel catfish. Mol Immunol 2: 151–159, 1992.
Gillies, S. D., Morrison, S. L., Oi, V. T., and Tonegawa, S. A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell 33: 717–728, 1983.
Gorman, C. M., Moffat, L. F., and Howard, B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol 2: 1044–1051, 1982.
Grosschedl, R. and Baltimore, D. Cell-type specificity of immunoglobulin gene expression is regulated by at least three DNA sequence elements. Cell 41: 885–897, 1985.
Guyomard, R., Chourrout, D., Leroux, C., Houdebine, L. M., and Pourrain, F. Integration and germline transmission of foreign genes microinjected into fertilized trout eggs. Biochimie 71: 857–863, 1989.
Hart, S., Wrathmell, A. B., Harris, J. E., and Grayson, T. H. Gut immunology in fish: a review. Dev Comp Immunol 12: 453–480, 1988.
Hawley, R. G., Schulman, M. J., Murialdo, H., Gibson, D. M., and Hozumi, N. Mutant immunoglobulin genes have repetitive DNA elements inserted into their intervening sequences. Proc Natl Acad Sci USA 79: 7425–7429, 1982.
Kokubu, F., Litman, R., Shamblott, M. J., Hinds, K., and Litman, G. W. Diverse organization of immunoglobulin VH gene loci in a primitive vertebrate. EMBO J 7: 3413–3422, 1988.
Lee, M. A., Bengtén, E., Daggfeldt, A., Rytting, A.-S. and Pilström, L. Characterisation of rainbow trout cDNAs encoding a secreted and membrane-bound Ig heavy chain and the genomic intron upstream of the first constant exon. Mol Immunol 30: 641–648, 1993.
Litman, G. W., Berger, L., Murphy, K., Litman, R., Hinds, K., and Erickson, B. W. Immunoglobulin VH gene structure and diversity in Heterodontus, a phylogenetically primitive shark. Proc Natl Acad Sci USA 82: 2082–2086, 1985.
Lu, J. K., Chen, T. T., Chrisman, C. L., Andrisani, O. M., and Dixon, J. E. Integration, expression, and germ-line transmision of foreign growth hormone genes in medaka (Oryzias latipes). Mol Mar Biol Biotech 1: 366–375, 1992.
Mårtensson, I. L. and Leanderson, T. Transient gene expression in untransformed lymphocytes. Eur J Immunol 17: 1499–1502, 1987.
Matsunaga, T., Chen, T., and Törmänen, V. Characterization of a complete immunoglobulin heavy-chain variable region germ-line gene of rainbow trout. Proc Natl Acad Sci USA 87: 7767–7771, 1990.
Mercola, M., Goverman, J., Mirell, C., and Calame, K. Immunoglobulin heavy-chain enhancer requires one or more tissue-specific factors. Science 227: 266–270, 1985.
Miller, N. W., Bly, J. E., van Ginkel, F., Ellsaesser, C. F., and Clem, L. W. Phylogeny of lymphocyte heterogeneity: Identification and separation of functionally distinct subpopulations of channel catfish lymphocytes with monoclonal antibodies. Dev Comp Immunol 11: 739–747, 1987.
Miller, A. E., Ennist, D. L., Ozato, K., and Westphal, H. Activation of immunoglobulin control elements in transgenic mice. Immunogenetics 35: 24–32, 1992.
Parslow, T. G., Blair, D. L., Murphy, W. J., and Granner, D. K. Structure of the 5′ ends of immunoglobulin genes: a novel conserved sequence. Proc Natl Acad Sci USA 81: 2650–2654, 1984.
Reik, W., Williams, G., Barton, S., Norris, M., Neuberger, M., and Surani, M. A. Provision of the immunoglobulin heavy chain enhancer downstream of a test gene is sufficient to confer lymphoid specific expression in transgenic mice. Eur J Immunol 17: 465–469, 1987.
St. Louis-Cormier, E. A., Osterland, C. K., and Anderson, P. D. Evidence for a cutaneous secretory immune system in rainbow trout (Salmo gairdner). Dev Comp Immunol 8: 71–80, 1984.
Shears, M. A., Fletcher, G. L., Hew, C. L., Gauthier, S., and Davies, P. L. Transfer, expression and stable inheritance of antifreeze protein genes in Atlantic salmon (Salmo salar). Mol Mar Biol Biotech 1: 58–63, 1991.
Storb, U., O'Brien, R. L., McMullen, M. D., Gollahon, K. A., and Brinster, R. L. High expression of cloned immunoglobulin κ gene in transgenic mice is restricted to B lymphoytes. Nature 310: 238–241, 1984.
Stuart, G. W., McMurray, J. V., and Westerfield, M. Replication, integration and stable germline transmission of foreign sequences injected into early zebrafish embryos. Development 103: 403–412, 1988.
Tewari, R., Michard-Vanhée, C., Perrot, E., and Chourrout, D. Mendelian transmission, structure and expression of transgenes following their injection into the cytoplasm of trout eggs. Transgenic Research 1: 250–260, 1992.
Thuvander, A., Fossum, C., and Lorenzen, N. Monoclonal antibodies to salmonid immunoglobulin. characterization and applicability in immunoassays. Dev Comp Immunol 14: 415–423, 1990.
Wilson, M. R., Middleton, D., and Warr, G. W. Immunoglobulin VH genes of the goldfish, Carassius auratus: a re-examination. Mol Immunol 28: 449–457, 1991.
Wilson, M. R. and Warr, G. W. Fish immunoglobulins and the genes that encode them.In M. Faisal and F. M. Hetrick (eds.): Annu Rev Fish Diseases, vol 2, pp. 201–221, Permagon Press, New York, 1992.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Michard-Vanhée, C., Chourrout, D., Strömberg, S. et al. Lymphocyte expression in transgenic trout by mouse immunoglobulin promoter/enhancer. Immunogenetics 40, 1–8 (1994). https://doi.org/10.1007/BF00163958
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00163958