Abstract
The whey acidic protein has been found in milk of mice, rats, rabbits and camels, and its gene is expressed specifically in mammary tissue at late pregnancy and throughout lactation. A characteristic of whey acidic protein is the ‘four-disulfide-core’ signature which is also present in proteins involved in organ development. We have generated six lines of transgenic pigs which carry a mouse whey acidic protein transgene and express it at high levels in their mammary glands. Transgenic sows from three lines could not produce sufficient quantities of milk to support normal development of healthy offspring. This phenotype appears to be similar, if not identical, to themilchlos phenotype exhibited by mice expressing whey acidic protein transgenes. Mammary tissue from post-partummilchlos sows had an immature histological appearance, which was distinct from that observed during normal development or involution. Expression of the whey acidic protein transgene was found in mammary tissue from sexually immature pigs frommilchlos lines, but not in sows from lines that appeared to lactate normally. We suggest that precocious synthesis of whey acidic protein impairs mammary development and function. Impaired mammary development due to inappropriate timing of whey acidic protein expression is consistent with the notion that proteins with the ‘four-disulfide-core’ signature participate in tissue formation.
Similar content being viewed by others
References
Araki, K., Kuroki, J., Ito, O., Kuwada, M. and Tachibana, S. (1989) Novel peptide inhibitor (SPAI) of Na+, K+-ATPase from porcine intestine.Biochem. Biophys. Res. Commun. 164, 496–502.
Archibald, A.L., McClenaghan, M., Hornsey, V., Simons, J.P. and Clark, A.J. (1990) High-level expression of biologically active human α1-antitrypsin in the milk of transgenic mice.Proc. Natl Acad. Sci. USA 87, 5178–82.
Bayna, E.M. and Rosen, J.M. (1990) Tissue-specific, high level expression of the rat whey acidic protein gene in transgenic mice.Nucl. Acids Res.,18, 2977–85.
Burdon, T., Sankaran, L., Wall, R.J., Spencer, M. and Hennighausen, L. (1991a) Expression of a whey acidic protein transgene during mammary development: evidence for different mechanisms of regulation during pregnancy and lactation.J. Biol. Chem. 256, 6909–14.
Burdon, T., Wall, R.J., Shamay, A., Smith, G.H. and Hennighausen, L. (1991b) Overexpression of an endogeous milk protein gene in transgenic mice is associated with impaired mammary development and a milchlos phenotype.Mech. Dev. 36, 67–74.
Chomczynski, P. and Sacchi, N. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenolchloroform extraction.Anal. Biochem. 162, 156–9.
Dear, T.N. and Kefford, R.F. (1991) The WDNM1 gene product is a novel member of the ‘four-disulfide core’ family of proteins.Biochem. Biophys. Res. Commun. 176, 247–54.
Ebert, K.M., Selgrath, J.P., DiTullio, P., Denman, J., Smith, T.E., Memon, M.A., Schindler, J.E., Monastersky, G.M., Vitale, J.A. and Gordon, K. (1991) Transgenic production of a variant of human tissue-type plasminogen activator in goat milk: generation of transgenic goats and analysis of expression.Biotechnology 9, 835–838.
Farmer, S.J., Fliss, A.E. and Simmen, R.C.M. (1990) Complementary DNA cloning and regulation of expression of the messenger RNA encoding a pregnancy-associated porcine uterine protein related to human antileukoproteinase.Mol. Endocrinol. 4, 1095–104.
Franco, B., Guioli, S., Pragliola, A., Incerti, B., Bardoni, B., Tonlorenzi, R., Carrozzo, R., Maestrini, E., Pieratti, M., Taillon-Miller, P., Brown, C.J., Willard, H.F., Lawrence, C., Persico, M.G., Camerino, G. and Ballabio, A. (1991) A gene deleted in Kallmann’s syndrome shares homology with neural cell adhesion and axonal path-finding molecules.Nature 353, 529–36.
Gertler, A., Cohen, N. and Maoz, A. (1983) Human growth hormone but not ovine or bovine growth hormones exhibits galactopoietic prolactin-like activity in organ culture from bovine lactating mammary gland.Mol. Cell. Endocrinol. 33, 169–82.
Gordon, K., Lee, E., Vitale, J.A., Smith, A.E., Westphal, H. and Hennighausen, L. (1987) Production of human tissue plasminogen activator in mouse milkBio/Technology 5, 1183–7.
Hennighausen, L.G. and Sippel, A.E. (1982) The mouse whey acidic protein is a novel member of the family of ‘four-disulfide core’ proteins.Nucl. Acids Res. 10, 2677–84.
Hobbs, A.A., Richards, D.A., Kessler, D.J., and Rosen, J.M. (1982) Complex hormonal regulation of rat casein gene expression.J. Biol. Chem. 257, 3598–605.
Kroczek, R.A. and Siebert, E. (1990) Optimization of northern analysis by vacuum blotting RNA transfer visualization and ultraviolet fixation.Anal. Biochem.,184, 90–5.
Lee, K.F., DeMayo, F.J., Aitee, S.H. and Rosen, J.M. (1988) Tissue-specific expression of the rat β-casein gene in transgenic mice.Nucl. Acids Res. 16, 1027–41.
Legouis, R., Hardelin, J.-P., Levilliers, J., Claverie, J.-M., Compain, S., Wunderle, V., Millasseau, P., LePaslier, D., Cohen, D., Caterina, D., Bougueleret, L., Delemarre-Van de Waal, H., Lutfalla, G., Weissenbach, J. and Petit, C. (1991) The candidate gene for the x-linked Kallmann syndrome encodes a protein related to adhesion molecules.Cell 67, 423–35.
Maschio, A., Brickell, P.M., Kioussis, D., Mellor, A.L., Katz, D. and Craig, R.K. (1991) Transgenic mice carrying the guinea pig α-lactalbumin gene, and normal mice, express milk protein genes in their sabaceous glands during lactation.Biochem. J. 275, 459–67.
McKnight, R.A., Burdon, T., Pursel, V.G., Shamay, A., Wall, R.J. and Hennighausen, L. (1992) The whey acidic protein. In Dickson and Lippman, eds,Breast Cancer: Molecular and Cellular Biology. Boston: Kluwer, Academic Publishers, (in press)
Meade, H., Gates, L., Lacy, E. and Lonberg, N. (1990) Bovine αS1-casein gene sequences direct high level expression of active human urokinase in mouse milk.Bio/Technology 8, 443–6.
Pittius, C.W., Sankaran, S., Topper, Y.J. and Hennighausen, L. (1988) Comparison of the regulation of the whey acidic protein gene to a hybrid gene containing the whey acidic protein gene promoter in transgenic mice.Mol. Endocrinol. 2, 1027–32.
Rahm, S., Geyer, H. and Neuburger, D. (1985) Histologische und histochemische Untersuchungen zur Involution der Schweinemilchdrüse.Zbl. Vet. Med. A.31, 776–91.
Shamay, A., Zeelon, E., Ghez, Z., Cohen, N., Mackinlay, A.G. and Gertler, A. (1987) Inhibition of casein and fat synthesis and α-lactlabumin secretion by progesterone in explants from bovine lactating mammary glands.J. Endocrinol. 113, 81–8.
Shamay, A., Salinas, S., Pursel, V.G., McKnight, R.A., Alexander, L., Beattie, C., Hennighausen, L. and Wall, R.J. (1991) Production of the mouse whey acidic protein in transgenic pigs during lactation.J. An. Sci. 69, 4552–62.
Shamay, A., Pursel, V.G., Wall, R.J. and Hennighausen, L. (1992) Induction of lactogenesis in transgenic virgin pigs: evidence for gene and integration site specific hormonal regulation.Mol. Endocrinol.,6, 191–7.
Simons, J.P., McClenaghan, M. and Clark, A.J. (1987) Alteration of the quality of milk by expression of sheep β-lactoglobulin in transgenic mice.Nature 328, 530–2.
Topper, Y.J. and Freeman, C.S. (1980) Multiple hormone interactions in the developmental biology of the mammary gland.Physiol. Rev. 60, 1049–156.
Vilotte, J.-L., Soulier, S., Sinnakre, M.-G., Massoud, M. and Mercier, J.-C. (1989) Efficient tissue-specific expression of bovine α-lactalbumin in transgenic mice.Eur. J. Biochem. 186, 43–8.
Wall, R.J., Pursel, V.G., Shamay, A., McKnight, R.A., Pittius, C.W. and Hennighausen, L. (1991) High-level synthesis of a heterologous milk protein in the mammary glands of swine.Proc. Natl Acad. Sci. USA 88, 1701–5.
Wiedow, O., Schroder, J.M., Gregory, H., Young, J.A. and Christophers, E. (1990) Elafin: an elastase-specific inhibitor of human skin. Purification, characterization, and complete amino-acid sequence.J. Biol. Chem. 265, 14791–5.
Wright, G., Carver, A., Cottom, D., Reeves, D., Scott, A., Simons, P., Wilmut, I., Garner and Colman, A. (1991) High level expression of active human alpha-1-antitrypsin in the milk of transgenic sheep.Bio/Technology 9, 830–834.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Shamay, A., Pursel, V.G., Wilkinson, E. et al. Expression of the whey acidic protein in transgenic pigs impairs mammary development. Transgenic Research 1, 124–132 (1992). https://doi.org/10.1007/BF02528777
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02528777