Regulation of protein and prostaglandin secretion in polarized primary cultures of caprine uterine epithelial cells

  • G. R. Newton
  • D. W. Weise
  • J. A. Bowen
  • S. Woldesenbet
  • R. C. Burghardt
Cellular Models

Summary

Caprine uterine epithelial (UE) cells were cultured on Matrigel-coated filters. Transmission electron microscopy revealed polarized UE cells characterized by basally located nuclei, apical microvilli, convoluted lateral membranes, and junctional complexes. Domain-specific secretion of prostaglandins and radiolabeled proteins provide further evidence of functional epithelial cell polarity. Two experiments were conducted to evaluate factors controlling prostaglandin E2 (PGE) and prostaglandin F (PGF) secretion. In experiment one, steroid-treated (estradiol, progesterone, or estradiol + progesterone) polarized UE cells were treated with interferon tau (IFNτ) and/or oxytocin (OT). Steroid treatment did not influence PGE or PGF secretion. However, analysis of variance revealed an IFNτ by OT interaction (P<.01) for both PGE and PGF. This interaction was caused by a reduction in PGE and PGF secretion by cultures receiving only IFNτ and the inability of IFNτ to block OT-induced release of PGE or PGF. In experiment 2, polarized UE cells were cultured in progesterone, with or without IFNτ, and sequentially challenged with estradiol and OT. Oxytocin stimulated the release of both PGE and PGF by polarized cUE cells (P<.01) and resulted in an increased accumulation of PGE (OT*domain; P<.01) in the basal compartment. Interferon tau did not influence PGE (P<.1) secretion. However, further analysis revealed that IFNτ reduced PGF secretion and was unable to block OT-induced PGF secretion (IFNτ*OT; P<.05) by polarized UE cells. Therefore, caprine UE cells form polarized monolayers and retain responsiveness to IFNτ and OT in vitro.

Key words

uterus epithelial oxytocin interferon tau prostaglandins polarity caprine 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Akinlosoto, B. A.; Diehl, J. R.; Gimenez, T. Sparing effects of intrauterine treatment with prostaglandin E2 on luteal function in gilts. Prostaglandins 32:291–302; 1986.CrossRefGoogle Scholar
  2. 2.
    Bazer, F. W. Uterine protein secretions: relationship to development of the conceptus. J. Anim. Sci. 41:1276–1282; 1975.Google Scholar
  3. 3.
    Bazer, F. W. Mediators of maternal recognition of pregnancy in mammals. Proc. Soc. Exp. Biol. Med. 18:453–460; 1992.Google Scholar
  4. 4.
    Bazer, F. W.; First, N. L. Pregnancy and parturition. J. Anim. Sci. (Suppl. 1) 57:425–460; 1984.Google Scholar
  5. 5.
    Bazer, F. W.; Spencer, T. E.; Ott, T. L. Placental interferons. Am. J. Reprod. Immunol. 35:297–308; 1996.PubMedGoogle Scholar
  6. 6.
    Bazer, F. W.; Thatcher, W. W. Theory of maternal recognition of pregnancy in swine based on estrogen controlled endocrine versus exocrine secretion of prostaglandin F by the uterine endometrium. Prostaglandins 14:397–401; 1977.PubMedCrossRefGoogle Scholar
  7. 7.
    Betteridge, K. J.; Randall, G. C. B.; Eaglesom, M. D., et al. The influence of PGF secretion in cattle. I. Concentrations of 15-keto-13,14-dihydro-prostaglandin and progesterone in peripheral blood recipients of transferred embryos. Anim. Reprod. Sci. 7:195–206; 1984.CrossRefGoogle Scholar
  8. 8.
    Bowen, J. A.; Newton, G. R.; Weise, D. W., et al. Characterization of a polarized procine uterine epithelial model system. Biol. Reprod. 55:613–619; 1996.PubMedCrossRefGoogle Scholar
  9. 9.
    Cereijido, M.; Gonzalez-Mariscal, L.; Contreras, R. G., et al. The making of tight junctions. J. Cell Sci. 17(Suppl.):127–132; 1993.Google Scholar
  10. 10.
    Cherny, R. A.; Findlay, J. K. Separation and culture of ovine endometrial epithelial and stromal cells: evidence of morphological and functional polarity. Biol. Reprod. 43:241–250; 1990.PubMedCrossRefGoogle Scholar
  11. 11.
    Cunha, G. R.; Bigsby, R. M.; Cooke, P. S., et al. Stromal-epithelial interactions in adult organs. Cell. Differ. 17:137–148; 1985.PubMedCrossRefGoogle Scholar
  12. 12.
    Danet-Desnoyers, G.; Wetzels, C.; Thatcher, W. W. Natural and recombinant bovine interferon tau regulate basal and oxytocin-induced secretion of prostaglandins F2 alpha and E2 by epithelial cells and stromal cells in the endometrium. Reprod. Fertil. Dev. 6:193–202; 1994.PubMedCrossRefGoogle Scholar
  13. 13.
    Enders, A. C. Contributions of comparative studies to understanding mechanisms of implantation. In: Glasser, S. R.; Mulholland, J.; Psychoyos, A., ed. Cellular aspects of implantation in ruminants. New York: Plenum Press; 1994:11–16.Google Scholar
  14. 14.
    Fitz, T. A.; Hoyer, P. B.; Niswender, G. D. Interaction of prostaglandins with subpopulations of ovine luteal cells. I. Stimulatory effects of prostaglandins E1, E2, and I2. Prostaglandins 28:119–126; 1984.PubMedCrossRefGoogle Scholar
  15. 15.
    Fortier, M. A.; Guibault, L. A.; Grasso, F. Specific properties of epithelial and stromal cells from the endometrium of cows. J. Reprod. Fertil. 83:239–248; 1988.PubMedCrossRefGoogle Scholar
  16. 16.
    Glasser, S. R.; Julian, J.; Decker, G. L., et al. Development of morphological and functional polarity in primary cultures of immature rat uterine epithelial cells. J. Cell Biol. 107:2409–2423; 1988.PubMedCrossRefGoogle Scholar
  17. 17.
    Gross, T. S.; Thatcher, W. W.; Hansen, P. J., et al. Presence of an intra-cellular inhibitor of prostaglandin synthesis during early pregnancy in the cow. Prostaglandins 35:359–378; 1988.PubMedCrossRefGoogle Scholar
  18. 18.
    Guillomot, M. Cellular aspects of implantation in ruminants. In: Glasser, S. R.; Mulholland, J.; Psychoyos, A., eds. Cellular aspects of implantation in ruminants. New York: Plenum Press; 1994:41–56.Google Scholar
  19. 19.
    Hagstrom, H. G.; Hahlin, M.; Bennegard-Eden, B., et al. Regulation of corpus luteum function in early human pregnancy. Fertil. Steril. 95:81–86; 1996.Google Scholar
  20. 20.
    Helmer, S. D.; Gross, T. S.; Hansen, P. J., et al. Bovine trophoblast protein-1 complex alters endometrial protein and prostaglandin synthesis and induces an intracellular inhibitor of prostaglandin synthesis in vitro. J. Reprod. Fertil. 87:421–433; 1989.PubMedCrossRefGoogle Scholar
  21. 21.
    Henderson, K. M.; Scaramuzzi, R. J.; Baird, D. T. Simultaneous infusion of prostaglandin E2 antagonizes the luteolytic action of prostaglandin F in vivo. J. Endocrinol. 72:379–386; 1977.PubMedGoogle Scholar
  22. 22.
    Hoffman, L. H.; Olson, G. E. Crystalline inclusions in embryonic and maternal cells. In: van Blerkom, J.; Motta, P. M., eds. Ultrastructure of reproduction. Boston, MA: Martinus Nijhoff Publishers; 1984:235–246.Google Scholar
  23. 23.
    Kazimi, M.; Amann, J. F.; Keisler, D. H., et al. A progesterone-modulated, low molecular-weight protein from the uterus of the sheep is associated with crystalline inclusion bodies in uterine epithelium and embryonic trophectoderm. Biol. Reprod. 43:80–96; 1990.CrossRefGoogle Scholar
  24. 24.
    King, G. J.; Atkinson, B. A.; Robertson, H. A. Implantation and early placentation in domestic ungulates. J. Reprod. Fertil. (Suppl.) 31:17–30; 1982.Google Scholar
  25. 25.
    Koji, T.; Chedid, M.; Rubin, J. S., et al. Progesterone-dependent expression of keratinocyte growth factor mRNA in stromal cells of primate endometrium: keratinocyte growth factor as a progestomedin. J. Cell Biol. 125:393–401; 1994.PubMedCrossRefGoogle Scholar
  26. 26.
    Labarca, C.; Paigen, K. A simple, rapid and sensitive DNA assay procedure. Anal. Biochem. 102:344–352; 1980.PubMedCrossRefGoogle Scholar
  27. 27.
    Lukaszewska, J.; Hansel, W. Corpus luteum maintenance during early pregnancy in the cow. J. Reprod. Fertil. 59:445–452; 1980.Google Scholar
  28. 28.
    McCracken, J. A.; Schramm, W.; Okulicz, W. C. Hormone receptor control of pulsatile secretion of PGF from the ovine uterus during luteolysis and its abrogation in early pregnancy. Anim. Reprod. Sci. 7:31–55; 1984.CrossRefGoogle Scholar
  29. 29.
    Meyer, M. D.; Desnoyers, G. D.; Oldick, B., et al. Treatment with recombinant bovine interferon-τ in utero attenuates secretion of prostaglandin F from cultured endometrial epithelial cells. J. Dairy Sci. 79:1375–1384; 1996.PubMedCrossRefGoogle Scholar
  30. 30.
    Nakao, K.; Meyer, C. J.; Noda, Y. Progesterone-specific protein crystals in the endometrium: an electron microscopic study. Am. J. Obstet. Gynecol. 111:1034–1038; 1971.Google Scholar
  31. 31.
    Ott, T. L.; Mirando, M. A.; Davis, M. A., et al. Effects of ovine conceptus secretory proteins and progesterone on oxytocin-stimulated endometrial prostaglandin production and inositol phosphate turnover in ovariectomized ewes. J. Reprod. Fertil. 95:19–29; 1992.PubMedCrossRefGoogle Scholar
  32. 32.
    Pekonen, F.; Nyman, T.; Rutanen, E. M. Differential expression of keratinocyte growth factor and its receptor in the human uterus. Mol. Cell. Endocrinol. 43:43–49; 1993.CrossRefGoogle Scholar
  33. 33.
    Reid, L. M. Defining hormone and matrix requirements for differentiated epithelia. In: Pollard, J. W.; Walker, J. M., eds. Methods in molecular biology. Vol. 3. Animal culture. Totowa, NJ: Humana Press; 1990:237–276.Google Scholar
  34. 34.
    Reynolds, L. P.; Stigler, J.; Hoyner, G. L., et al. Effects of PGE1 or PGE2 on PGF-induced luteolysis in non-bred ewes. Prostaglandins 21:957–972; 1981.PubMedCrossRefGoogle Scholar
  35. 35.
    Roberts, R. M.; Cross, J. C.; Leaman, D. W. Interferons as hormones of pregnancy. Endocr. Rev. 13:432–452; 1992.PubMedCrossRefGoogle Scholar
  36. 36.
    SAS Institute. SAS user’s guide. Cary, NC; 1985.Google Scholar
  37. 37.
    Silvia, W. J.; Lewis, G. S.; McCracken, J. S., et al. Hormonal regulation of uterine secretion of prostaglandin F in ruminants. Biol. Reprod. 45:655–663; 1991.PubMedCrossRefGoogle Scholar
  38. 38.
    Silvia, W. J.; Niswender, G. D. Maintenance of the corpus luteum of early pregnancy in the ewe. III. Differences between pregnant and nonpregnant ewes in luteal responsiveness to prostaglandin F. J. Anim. Sci. 59:746–753; 1984.PubMedGoogle Scholar
  39. 39.
    Silvia, W. J.; Ottobre, J. S.; Inskeep, E. K. Concentrations of prostaglandins E2, F, and 6-keto-prostaglandin F in the utero-ovarian venous plasma of nonpregnant and early pregnant ewes. Biol. Reprod. 30:936–944; 1984.PubMedCrossRefGoogle Scholar
  40. 40.
    Smith, S. K.; Kelly, R. W. Prostaglandins and the establishment of pregnancy. In: Chapman, M.; Grudizinskas, G.; Chard, T., eds. Implantation: biological and clinical aspects. Berlin, Germany: Springer-Verlag; 1988:147–160.Google Scholar
  41. 41.
    Spencer, T. E.; Bazer, F. W. Temporal and spacial alterations in uterine estrogen receptor and progesterone receptor gene expression during the estrous cycle and early pregnancy in the ewe. Biol. Reprod. 53:1527–1543; 1995.PubMedCrossRefGoogle Scholar
  42. 42.
    Spencer, T. E.; Becker, W. C.; George, P., et al. Ovine interferon-τ regulates expression of endometrial receptors for estrogen and oxytocin but not progesterone. Biol. Reprod. 53:732–735; 1995.PubMedCrossRefGoogle Scholar
  43. 43.
    Thatcher, W. W.; Bazer, F. W.; Sharp, D. C., et al. Interrelationships between uterus and conceptus to maintain corpus luteum function in early pregnancy: sheep, cattle, pigs, horses. J. Anim. Sci. 62 (Suppl. 2):25–46; 1986.PubMedGoogle Scholar
  44. 44.
    Thibodeux, J. K.; Myers, M. W.; Roussel, J. D., et al. Intrauterine infusion of prostaglandin E2 and subsequent luteal function in cattle. Prostaglandins 44:531–538; 1983.CrossRefGoogle Scholar
  45. 45.
    Weise, D. W.; Newton, G. R.; Emesih, G. C. Effect of day of the estrous cycle or pregnancy on protein secretion by caprine endometrial tissues. Biol. Reprod. 49:522–527; 1993.PubMedCrossRefGoogle Scholar
  46. 46.
    Wongo, E. O.; Wooding, F. B. P.; Heap, R. B. The role of trophoblast binucleate cells in implantation in the goat: a quantitative study. Placenta 11:381–394; 1990.CrossRefGoogle Scholar
  47. 47.
    Wathes, D. C.; Hamon, M. Localization of oestradiol, progesterone and oxytocin receptors in the uterus during the oestrous cycle and early pregnancy of the ewe. J. Endocrinol. 138:479–491; 1993.PubMedCrossRefGoogle Scholar

Copyright information

© Society for In Vitro Biology 1998

Authors and Affiliations

  • G. R. Newton
    • 1
  • D. W. Weise
    • 1
  • J. A. Bowen
    • 2
  • S. Woldesenbet
    • 1
  • R. C. Burghardt
    • 2
  1. 1.Cooperative Agricultural Research CenterPrairie View A&M UniversityPrairie View
  2. 2.Department of Veterinary Anatomy and Public HealthTexas A&M UniversityCollege Station

Personalised recommendations