Abstract
The cytokine-inducible suppressor of cytokine signalling SOCS1, or JAB, has been shown to be implicated in vitro in the negative regulation of the prolactin-receptor-induced activation of JAK2 and STAT5. Disruption of this gene in vivo resulted in an accelerated mammary gland development. In the present experiment, we assessed the potential impact on the lactation process of the doxycycline-inducible mammary-controlled expression of this gene in transgenic mice. Three transgenic mouse lines that expressed JAB specifically in the mammary gland in a conditional manner following doxycycline treatment were successfully established. The resulting overall expression of JAB was high and ranged from half to four times that of the endogenously expressed homologous gene in the thymus. It was found to be highly heterogeneous in the mammary epithelium, with less than 5% of JAB-expressing cells detected. Phenotypic analysis of these transgenic mice exhibiting doxycycline-induced JAB expression did not reveal any obvious effect on the lactation process. Double immunostaining experiments suggested that JAB expression in vivo did not significantly affect the β-casein gene expression and the STAT5a nuclear localisation. These results do not support a role for JAB in the disruption of the lactation process.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Blau H and Rossi FMV (1999) Tet B or not tet B: advances in tetracycline-inducible gene expression. Proc Natl Acad Sci USA 96: 797–799.
Brisken C, Kaur S, Chavarria TE, Binart N, Sutherland RL, Weinberg RA et al. (1999) Prolactin controls mammary gland development via direct and indirect mechanisms. Dev Biol 210: 96–106.
Chughtai N, Schimchowitsch S, Lebrun JJ and Ali S (2002) Prolactin induces SHP-2 association with Stat5, nuclear translocation and binding to the β-casein gene promoter in mammary cells. J Biol Chem 277: 31107–31114.
De Wet JR, Wood KV, De Luca M, Helinski DR and Subramani S (1987) Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol 7: 725–737.
Dobie KW, Lee M, Fantes JA, Graham E, Clark AJ, Springbett A et al. (1996) Variegated transgene expression in mouse mammary gland is determined by the transgene integration locus. Proc Natl Acad Sci USA 93: 6659–6664.
Endo TA, Masuhara M, Yokouchi M, Suzuki R, Sakamoto H, Mitsui K et al. (1997) A new protein containing and SH2 domain that inhibits JAK kinases. Nature 387: 921–924.
Faerman A, Barash I, Puzis R, Nathan M, Hurwitz DR and Shani M (1995) Dramatic heterogeneity of transgene expression in the mammary gland of lactating mice: a model system to study the synthetic activity of mammary epithelial cells. J Histochem Cytochem 43: 461–470.
Forster K, Helbl V, Lederer T, Urlinger S, Wittenburg N and Hillen W (1999) Tetracycline-inducible expression systems with reduced basal activity in mammalian cells. Nucl Acids Res 27: 708–710.
Gallego MI, Binart N, Robinson GW, Okagaki R, Coschigano KT, Perry J et al. (2001) Prolactin, growth hormone, and epidermal growth factor activate STAT5 in different compartments of mammary tissue and exert different and overlapping developmental effects. Dev Biol 229: 163–175.
Gossen M and Bujard H (1992) Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci USA 89: 5547–5551.
Goupille O, Daniel N, Bignon C, Jolivet G and Djiane J (1997) Prolactin signal transduction to milk protein genes: carboxyterminal part of the prolactin receptor and its tyrosine phosphorylation are not obligatory for JAK2 and STAT5 activation. Mol Cell Endocrinol 127: 155–169.
Gual E, Baron V, Lequoy V and Van Obberghen E (1998) Interaction of janus kinases JAK-1 and JAK-2 with the insulin receptor and the insulin-like growth factor-1 receptor. Endocrinology 139: 884–893.
Harris S, McClenaghan M, Simons JP, Ali S and Clark AJ (1991) Developmental regulation of the sheep β-lactoglobulin gene in the mammary gland of transgenic mice. Dev Gen 12: 299–307.
Hennighausen L, Wall RJ, Tillmann U, Li M and Furth PA (1995) Conditional gene expression in secretory tissues and skin of transgenic mice using the MMTV-LTR and the tetracycline responsive system.J Cell Biochem 59: 463–472.
Hynes NE, Horsch K, Olayioye MA and Badache A (2001) The ErbB receptor tyrosine family as signal integrators. Endocrinol Rel Cancer 8: 151–159.
Hogan B, Constantini F and Lacy E (1986) Manipulating the Mouse Embryo: A Laboratory Manual. Cold Spring Harbor Laboratory Press, New York.
Horseman ND, Zhao W, Montecino-Rodriguez E, Tanaka M, Nakashima K, Engle SJ et al. (1997) Defective mammopoiesis, 706 but normal hematopoiesis, in mice with a targeted disruption of the prolactin gene. EMBO J 16: 6926–6935.
Jahn GA, Daniel N, Jolivet G, Belair L, Bole-Feysot C, Kelly PA et al. (1997) In vivo study of prolactin intracellular signalling during lactogenesis in the rat: JAK/STAT pathway is activated by prolactin in the mammary gland but not in the liver. Biol Reprod 57: 894–900.
Jiang W, Zhou L, Breyer B, Feng T, Cheng H, Haydon R et al. (2001) Tetracycline-regulated gene expression mediated by a novel chimeric repressor that recruits histone deacetylases in mammalian cells. J Biol Chem 276: 45168–45174.
Kawazoe Y, Naka T, Fujimoto M, Kohzaki H, Morita Y, Narazaki M et al. (2001) Signal transducer and activator of transcription (STAT)-induced STAT Inhibitor 1 (SSI-1)/suppressor of cytokine signaling 1 (SOCS1) inhibits insulin signal transduction pathway through modulating insulin receptor substrate 1 (IRS-1) phosphorylation. J ExpMed 193: 263–269.
Kile BT and Alexander WS (2001) The suppressors of cytokine signalling (SOCS). Cell Mol Life Sci 58: 1627–1635.
Leuchtenberger S, Perz A, Gatz C and Bartsch JW (2001) Conditional ablation by stringent tetracycline-dependent regulation of barnase in mammalian cells. Nucl Acids Res 29: e76.
Lindeman GJ, Wittlin S, Lada H, Naylor MJ, Santamaria M, Zhang JG et al. (2001) SOCS1 deficiency results in accelerated mammary gland development and rescues lactation in prolactin receptor-deficient mice. Genes Dev 15: 1631–1636.
Liu X, Robinson GW and Henninghausen L (1996) Activation of STAT5a and STAT5b by tyrosine phosphorylation is tightly linked to mammary gland differentiation. Mol Endocrinol 10: 1496–1506.
Liu X, Robinson GW, Wagner KU, Garrett L, Wynshow-Boris A and Henninghausen L (1997) STAT5a is mandatory for adult mammary gland development and lactogenesis. Genes Dev 11: 179–186.
Liu XMI, Gallego GHS, Robinson W and Hennighausen L (1998) Functional release of STAT5a-null mammary tissue through the activation of compensating signals including STAT5b. Cell Growth Diff 9: 795–803.
Lkhider M, Petridou B, Aubourg A and Ollivier-Bousquet M (2001) Prolactin signaling to milk protein secretion but not to gene expression depends on the integrity of the golgi region. J Cell Sci 114: 1883–1891.
Marine JC, Topham DJ, McKay C, Wang D, Parganas E, Stravopodis D et al. (1999) SOCS1 defieciency causes a lymphocyte-dependent perinatal lethality. Cell 98: 609–16.
Matsumoto A, Seki Y, Kubo M, Ohtsuka S, Suzuki A, Hayashi I et al. (1999) Suppression of STAT5 functions in liver, mammary glands, and T cells in cytokine-inducible SH2-containing protein 1 transgenic mice.Mol Cell Biol 19: 6396–6407.
Miyoshi K, Shillingford JM, Smith GH, Grimm SL, Wagner KU, Oka T et al. (2001) Signal transducer and activator of transcription (STAT) 5 controls the proliferation and differentiation of mammary alveolar epithelium. J Cell Biol 155: 531–542.
Morita Y, Naka T, Kawazoe Y, Fujimoto M, Narazaki M, Nakagawa R et al. (2000) Signal transducers and activators of transcription (STAT)-induced STAT inhibitor-1 (SSI-1)/suppressor of cytokine signaling-1 (SOCS-1) suppresses tumor necrosis factor α-induced cell death in fibroblasts. Proc Natl Acad Sci USA 97: 5405–5410.
Ormandy CJ, Camus A, Barra J, Damotte D, Lucas B, Buteau H et al. (1997) Null mutation of the prolactin receptor gene produces multiple reproductive defects in the mouse. Genes Dev 11: 167–178.
Pezet A, Favre H, Kelly PA and Edery M (1999) Inhibition and restoration of prolactin signal transduction by SOCS. J Biol Chem 274: 24497–24502.
Rossi FMV, Guicherit OM, Spicher A, Kringstein AM, Fatyol K, Blakely BT et al. (1998) Tetracycline-regulatable factors with distinct dimerization domains allow reversible growth inhibition by p16. Nat Genet 20: 389.
Saad MJA, Carvalho CRO, Thirone ACP and Velloso LA (1996) Insulin induces tyrosine phosphorylation of JAK2 in insulin-sensitive tissues of the intact rat. J Biol Chem 271: 22100–22104.
Schultze N, Burki Y, Lang Y, Certa U and Bluethmann H (1996) Efficient control of gene expression by single step integration of the tetracycline system in transgenic mice. Nat Biotech 14: 499–503.
Shillingford JM, Miyoshi K, Robinson GW, Grimm SL, Rosen JM, Neubauer H et al. (2002) Jak2 is an essential tyrosine kinase involved in pregnancy-mediated development of mammary secretory epithelium. Mol Endocrinol 16: 563–570.
Soulier S, Vilotte JL, Stinnakre MG and Mercier JC (1992) Expression analysis of ruminant α-lactalbumin in transgenic mice: developmental regulation and general location of important cisregulatory elements. FEBS Lett 1–2: 13–18.
Soulier S, Stinnakre MG, Lepourry L, Mercier JC and Vilotte JL (1999) Use of doxycycline-controlled gene expression to reversibly alter milk-protein composition in transgenic mice. Eur J Biochem 260: 533–539.
Tam SP, Lau P, Djiane J, Hilton DJ and Waters MJ (2001) Tissuespecific induction of SOCS gene expression by PRL. Endocrinology 142: 5015–5026.
Teglund S, McKay C, Schuetz E, van Deursen JM, Stravopodis D, Wang D et al. (2002) STAT5a and STAT5b proteins have essential and nonessential, or redundant, roles in cytokine responses. Cell 93: 841–850.
Tomic S, Chughtai N and Ali S (1999) SOCS-1,-2,-3: selective targets and functions downstream of the prolactin receptor. Mol Cell Endocrinol 158: 45–54.
Tonko-Geymayer S, Goupille O, Tonko M, Soratroi C, Yoshimura A, Streuli C et al. (in press) Regulation and function of the cytokine inducible SH-2 domain proteins CIS and SOCS3 in mammary epithelial cells. Mol Endocrinol 16: 1680–1695
Tourkine N, Schindler C, Larose M and Houdebine LM (1995) Activation of STAT factors by prolactin, interferon-gamma, growth hormones and a tyrosine phosphatase inhibitor in rabbit primary mammary epithelial cells. J Biol Chem 270: 20952–20961.
Udy GB, Snell RG, Wilkins RJ, Park SH, Ram PA, Waxman DJ et al. (1997) Requirement of STAT5b for sexual dimorphism of body growth rates and liver gene expression. Proc Natl Acad Sci USA 94: 7239–7244.
Urlinger S, Baron U, Thellmann M, Hasan MT, Bujard H and Hillen W (2000) Exploring the sequence space for tetracyclinedependent transcriptional activators: novel mutations yield expanded range and sensitivity. Proc Natl Acad Sci USA 97: 7963–7968.
Valencik ML and McDonald JA (2001) Codon optimization markedly improves doxycycline regulated gene expression in the mouse heart. Transgenic Res 10: 269–275.
Vilotte JL, Soulier S, Stinnakre MG, Massoud M and Mercier JC (1989) Efficient tissue-specific expression of bovine α-lactalbumin in transgenic mice. Eur J Biochem 186: 43–48.
Yasukawa H, Sasaki A and Yoshimura A (2000) Negative regulation of cytokine signaling pathways. Annu Rev Immunol 18: 143–164.
Zhu Z, Ma B, Homer RJ, Zheng T and Elias JA (2001) Use of the tetracycline-controlled transcriptional silencer (tTS) to eliminate transgene leak in inducible overexpression transgenic mice. J Biol Chem 276: 25222–25229.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Petridou, B., Soulier, S., Besnard, N. et al. Heterogeneous Inducible Mammary-specific Expression of JAB/SOCS1 in Lactating Transgenic Mice is Associated with no Obvious Phenotype, even at the Cellular Level. Transgenic Res 12, 693–706 (2003). https://doi.org/10.1023/B:TRAG.0000005165.85843.2e
Issue Date:
DOI: https://doi.org/10.1023/B:TRAG.0000005165.85843.2e