Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
권호 표지

Changes in Reproductive Function and White Blood Cell Proliferation Induced in Mice by Injection of a Prolactin-expressing Plasmid into Muscle

  • Lee, Jung-Sun (Department of Life Science, The Catholic University of Korea) ;
  • Yun, Bo-Young (Department of Life Science, The Catholic University of Korea) ;
  • Kim, Sang-Soo (Department of Life Science, Hanyang University) ;
  • Cho, Chunghee (Department of Life Science Gwangju Institute of Science and Technology) ;
  • Yoon, Yong-Dal (Department of Life Science, Hanyang University) ;
  • Cho, Byung-Nam (Department of Life Science, The Catholic University of Korea)
  • Received : 2006.05.18
  • Accepted : 2006.07.24
  • Published : 2006.10.31
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Abstract

Prolactin (PRL) is a pituitary hormone involved in various physiological processes, including lactation, mammary development, and immune function. To further investigate the in vivo and comparative endocrine roles of PRL, mouse PRL cDNA fused to the cytomegalovirus promoter, was introduced into muscle by direct injection. Previously we studied the function of rat PRL using the same protocol. PRL mRNA was detected in the muscle following injection by RT-PCR and subsequent Southern blot analysis. PRL was also detected and Western blot analysis revealed a relatively high level of serum PRL. In the pCMV-mPRL-injected female mice, the estrous cycle was extended, especially in diestrus stage and the uterus thickening that was shown in normal estrous stage was not observed. In the pCMV-mPRL-injected male mice, new blood vessels were first found at 5 weeks of age and fully developed blood vessels were found after 8 weeks in the testis. The number of Leydig cells increased within the testis and the testosterone level in serum was observed high. Finally, the number of white blood cells (WBCs) increased in the pCMV-mPRL-injected mice. The augmentation of WBCs persisted for at least 20 days after injection. When injection was combined with adrenalectomy, there was an even greater increase in number of WBCs, especially lymphocytes. This increase was returned normal by treatment with dexamethansone. Taken together, our data reveal that intramuscularly expressed mouse PRL influences reproductive functions in female, induces formation of new blood vessels in the testis, and augments WBC numbers. Of notice is that the Leydig cell proliferation with increased testosterone was conspicuously observed in the pCMV-mPRL-injected mice. These results also suggest subtle difference in function of PRL between mouse and rat species.

Keywords

Acknowledgement

Supported by : Korea Science and Engineering Foundation

References

  1. Bernton, E. W., Meltzer, M. S., and Holaday, J. W. (1988) Suppression of macrophage activation and lymphocyte function in hypoprolactinemic mice. Science 239, 401−404
  2. Berry, S. J., Coffey, D. S., Walsh, P. C., and Ewing, L. L. (1984) The development of human benign prostatic hyperplasia with age. J. Urol. 132, 474−479
  3. Binart, N., Melaine, N., Pineau, C., Kercret, H., Touzalin, A. M., et al. (2003) Male reproductive function is not affected in prolactin receptor-deficient mice. Endocrinology 144, 3779− 3782 https://doi.org/10.1210/en.2003-0409
  4. Bole-Feysor, C., Goffin, V., Edery, M., Binart, N., and Kelly, P. A. (1998) Prolactn (PRL) and its receptor: actions, signal transduction pathways and phenotypes observed in PRL receptor knockout mice. Endocrine Rev. 19, 225−268 https://doi.org/10.1210/er.19.3.225
  5. Bouchard, B., Ormandy, C. J., Di Santo, J. P., and Kelly, P. A. (1999) Immune system development and function in prolactin receptor-deficient mice. J. Immunol. 163, 576−582
  6. Boutin, J.-M., Jolicoeur, C., Okamura, H., Gagnon, J., Edery, M., et al. (1988) Cloning and gene expression of the rat prolactin receptor, a member of the growth hormone/prolactin receptor gene family. Cell 53, 69−77 https://doi.org/10.1016/0092-8674(88)90488-6
  7. Cho, B. N., McMullen, M. L., Pei, L., Yates, C. J., and Mayo, E. K. (2001) Reproductive deficiencies in the transgenic mice expressing the rat inhibin $\alpha$-subunit gene. Endocrinology 142, 4994−5004 https://doi.org/10.1210/en.142.11.4994
  8. Clapp, C., Martial, J. A., Guzman, R. C., Rentier-Delrue, F., and Weiner, R. I. (1993) The 16-kilodalton N-terminal fragment of human prolactin is a potent inhibitor of angiogenesis. Endocrinology 133, 1292−1299 https://doi.org/10.1210/en.133.3.1292
  9. Clevenger, C. V., Russell, D. H., Appasamy, P. M., and Prystowsky, M. B. (1990) Regulation of IL2-driven Tlymphocyte proliferation by prolactin. Proc. Natl. Acad. Sci. USA 87, 6460−6464
  10. Clevenger, C. V., Altmann, S. W., and Prystowsky, M. B. (1991) Requirement of nuclear prolactin for interleukin-2- stimulated proliferation of T lymphocytes. Science 253, 77−79 https://doi.org/10.1126/science.2063207
  11. Clevenger, C. V., Sillman, A. L., Hanley-Hyde, J., and Prystowsky, M. B. (1992) Requirement for prolactin during cell cycle regulated gene expressed in cloned T-lymphocytes. Endocrinology 130, 3216−3222 https://doi.org/10.1210/en.130.6.3216
  12. Clevenger, C. V., Chang, W. P., Ngo, W., Pasha, T. L. M., Montone, K. T., et al. (1995) Expression of prolactin and prolactin receptor in human breast carcinoma. Evidence for an autocrine/paracrine loop. Am. J. Pathol. 146, 695−705
  13. Danko, I., Williams, P., Herweijer, H., Zhang, G., Latendresses, J. S., et al. (1997) High expression of naked plasmid DNA in muscles of young rodents. Hum. Mol. Genet. 6, 1453−1443
  14. Dombrowicz, D., Sente, B., Closset, J., and Hennen, G. (1992) Dose-dependent effects of human prolactin on the immature hypophysectomized rat testis. Endocrinology 130, 695−700 https://doi.org/10.1210/en.130.2.695
  15. Elias, K. A. and Weiner, R. L. (1984) Direct arterial vascularization of estrogen-induced prolactin-secreting anterior pituitary tumors. Proc. Natl. Acad. Sci. USA 81, 4549−4553
  16. Ferrara, N., Clapp, C., and Weiner, R. (1991) The 16K fragment of prolactin specifically inhibits basal or fibroblast growth factor stimulated growth of capillary endothelial cells. Endocrinology 129, 896−900
  17. Fletcher-Chiappini, S. E., Comptom, M. M., Lavoie, H. A., Day, E. B., and Witorsch, R. J. (1993) Glucocorticoid-prolactin interactions in Nb2 lymphoma cells: antiproliferative versus anticytolytic effects. Proc. Soc. Exp. Biol. Med. 202, 345− 352
  18. Folkman, J. (1995) Angiogenesis inhibitors generated by tumors. Mol. Med. 1, 120−122
  19. Gala, R. R. and Shevach, E. M. (1997) The immunomodulatory effects of a recombinant bovine prolactin (rbPRL) are secondary to endotoxin contamination. FASEB. J. 11, 99A
  20. Ginsberg, E. and Vonderharr, B. K. (1995) Prolactin synthesis and secretion by human breast cancer cells. Cancer Res. 55, 2591−2595
  21. Gunasekar, P. G., Kumaran, B., and Govindarajulu, P. (1988) Prolactin and Leydig cell steroidogenic enzymes in the bonnet monkey (macaca radiata). Int. J. Androl. 11, 53−59
  22. Hammond, G. L., Kontturi, M., Maattala, P., Puukka, M., and Vihko, R. (1977) Serum FSH, LH and prolactin in normal males and patients with prostatic diseases. Clin. Endocrinol. (Oxf) 7, 129−135
  23. Hanahan, D. and Folkman, J. (1996) Pattern and emerging mechanism of the angiogenic switch during tumorigenesis. Cell 86, 353−364
  24. Harper, M. E., Peeling, W. B., Cowley, T., Brownsey, B. G., Philips, M. E. A., et al. (1976) Plasma steroid and protein hormone concentration in patients with prostatic carcinoma, before and during oestrogen therapy. Acta. Endocrinol. (Copenh) 81, 409−426
  25. Horseman, N. D., Zhao, W., Montecino-Rodriquez, E., Tanaka, M., Nakashima, K., et al. (1997) Defective mammopoiesis, but hematopoiesis, in mice with a targeted disruption of the prolactin gene. EMBO J. 16, 6926−6935 https://doi.org/10.1093/emboj/16.23.6926
  26. Jackson, D., Volpert, O. V., Bouck, N., and Linzer, D. I. H. (1994) Stimulation and inhibition of angiogenesis by placental proliferin and proliferin-related protein. Science 266, 1581−1584 https://doi.org/10.1126/science.7527157
  27. Kant, G. J., Bauman, R. A., Anderson, S. M., and Mougey, E. H. (1992) Effects of controllable vs. uncontrollable chronic stress on stress-responsive plasma hormones. Physiol. Behav. 51, 1285−1288
  28. Ko, J.-Y., Ahn, Y.-L., and Cho, B. N. (2003) Angiogenesis and white blood proliferation induced in mice by injection of a prolactin-expressing plasmid into muscle. Mol. Cells 15, 262−270
  29. Lee, D. W. and Markoff, E. (1986) Synthesis and release of glycosylated prolactin by human deciduas in vitro. J. Clin. Endocrinol. Metab. 62, 990−994
  30. Lee, C. W., Lee, C. U., Kim, J. H., Son, B. C., Kim, D. H., et al. (2003) Effects of Aroclor 1254 on the expression of rat placental PRL-family genes. Mol. Cells 15, 114−121
  31. Leong, D. A., Frawley, L. S., and Neill, J. D. (1983) Neuroendocrine control of prolactin secretion. Ann. Rev. Physiol. 45, 109−127
  32. Loudon, A. S., Brinklow, B. R., Gulland, F. D., Boyle. J., and Flint, A. P. (1990) Roles of prolactin and the uterus in the control of luteal regression in the Bennetts's wallaby (Macropus rufogriseus rufogriseus). Reprod. Fertil. Dev. 2, 71−78
  33. Matsuyama, S., Shiota, K., and Takahashi, M. (1990) Possible role of transforming growth factor-$\beta$ as a mediator of luteotropic action of prolactin in rat luteal cell cultures. Endocrinology 127, 1561−1567 https://doi.org/10.1210/endo-127-4-1561
  34. Maurer, R. A. (1980) Dopaminergic inhibition of prolactin synthesis and prolactin messenger RNA accumulation in cultured pituitary cells. J. Biol. Chem. 255, 8092−8097
  35. Maurer, R. A. (1982) Estradiol regulates the transcription of the prolactin gene. J. Biol. Chem. 257, 2133−2136
  36. Montgomery, D. W., LeFevre, J. A., Ulrich, E. D., Adamson, C. R., and Zukoski, C. F. (1990) Identification of prolactin-like proteins synthesized by normal murine lymphocytes. Endocrinology 127, 2601−2603 https://doi.org/10.1210/endo-127-5-2601
  37. Neill, J. (1988) Prolactin secretion and its control; in The Physiology of Reproduction, Knobil, E. and Neill, J. (eds.), pp. 1379−1390, Raven Press, New York
  38. Nevalainen, M. T., Valve, E. M., Ingelton, P. M., Nurmi, M., Martikainen, P. M., et al. (1997) Prolactin and prolactin receptors are expressed and functioning in the human prostate. J. Clin. Invest. 99, 618−627
  39. Olazabal, I., Munoz, J., Ogueta, S., Obregon, E., and Garcia- Ruiz, J. P. (2000) Prolactin (PRL)-PRL receptor system increases cell proliferation involving JNK (c-Jun amino terminal kinase) and AP-1 activation: Inhibition by glucocorticoids. Mol. Endocrinol. 14, 564？575 https://doi.org/10.1210/me.14.4.564
  40. Ormandy, C. J., Camus, A., Barra, J., Damotte, D., Lucas, B., et al. (1997) Null mutation of the prolactin receptor gene produces multiple reproductive defects in the mouse. Genes Dev. 11, 167−178
  41. Pellegrini, I., Lebrun, J.-J., Ali, S., and Kelly, P. A. (1992) Expression of prolactin and its receptor in human lymphoid cells. Mol. Endocrinol. 6, 1023−1031 https://doi.org/10.1210/me.6.7.1023
  42. Raymond, V., Beaulieu, M., Labrie, F., and Boissier, J. (1978) Potent antidopaminergic activity of estradiol at the pituitary level on prolactin release. Science 200, 1173−1175
  43. Russell, D. H., Matrisian, L., Kibler, R., Larson, D. F., Poulos, B., et al. (1984) Prolactin receptors on human lymphocytes and their modulation by cyclosporine. Biochem. Biophys. Res. Commun. 121, 899−906
  44. Saroff, J., Kirdani, R. Y., Chu, M., Wajsman, Z., and Murhpy, G. P. (1980) Measurements of prolactin and androgens in patients with prostatic diseases. Oncology 37, 46−52
  45. Sinha, Y. N. (1995) Structure variants of prolactin: Occurrence and physiological significance. Endocrine Rev. 16, 354−369
  46. Steger, R. W., Chandrashekar, V., Zhao, W., Bartke, A., and Horseman, N. D. (1998) Neuroendocrine and reproductive functions in male mice with targeted disruption of the prolactin gene. Endocrinology 139, 3691−3695
  47. Struman, I., Bentzien, F., Lee, H., Mainfroid, V., D'Angelo, G., et al. (1999) Opposing actions of intact and N-terminal fragments of the human prolactin/growth hormone family members on angiogenesis. Proc. Natl. Acad. Sci. USA 96, 1246− 1251
  48. Walker, A. M., Montgomery, D. W., Saraiya, S., Ho, T. W. C., Garewal, H. S., et al. (1995) Prolactin-immunoglobulin G complexes from human serum act as costimulatory ligands causing proliferation of malignant B lymphocytes. Proc. Natl. Acad. Sci. USA 92, 3278−3282
  49. Wennbo, H., Kindblom, J., Isaksson, O. G., and Tornell, J. (1997) Transgenic mice overexpressing the prolactin gene develop dramatic enlargement of the prostate gland. Endocrinology 138, 4410−4415
  50. Witorsch, R. J., Day, E. B., Lavoie, H. A., Hashemi, N., and Taylor, J. K. (1993) Comparison of glucocorticoid-induced effects in prolactin-dependent and autonomous rat Nb2 lymphoma cells. Proc. Soc. Exp. Biol. Med. 203, 454−460