Localization of Tumor Necrosis Factor (TNF) in the Rat and Bovine Ovary Using Immunocytochemistry and Cell Blot: Evidence for Granulosal Production

  • K. F. Roby
  • P. F. Terranova


A protein fraction recently isolated from the bovine ovary has in vitro activity similar to tumor necrosis factor-α (TNF) and was thus hypothesized to be a tumor necrosis-like factor (TNLF) (1). In another related study, rabbit corpora lutea (CL) incubated in vitro in the presence of lipopolysaccharide (LPS) secrete a substance with bioactivity similar to TNF (2). In addition, macrophages which produce TNF (3) are present within the ovary. To investigate the cellular sources of TNF in the ovary, TNF was localized in bovine and rat ovaries using polyclonal antibodies to human recombinant TNFα with a Biotin-StreptAvidin immunostaining technique. Granulosal cells were examined by cell blot technique to ascertain the presence and/or secretion of TNF using a similar Biotin-StreptAvidin immunostaining technique. This study reveals the presence of immunoreactive TNF (I-TNF) in the bovine and rat ovary localized primarily to three regions: the CL, the antral layer of granulosal cells in small antral and large preovulatory follicles, and throughout the granulosal layer of atretic follicles. Cell blot analyses indicate the presence and secretion of TNF by bovine and rat granulosa cells in vitro.


Granulosal Cell Follicular Fluid Antral Follicle Preantral Follicle Corpus Luteum 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Roby KF, Terranova PF. Inhibition of endothelial cell growth by a tumor necrosis-like fraction from the bovine ovary. Biol Reprod (submitted).Google Scholar
  2. 2.
    Bagavandos P, Kunkel SL, Wiggins RC, Keyes PL. Tumor necrosis factor a (TNF a) production and localization of macrophages and T lymphocytes in the rabbit corpus luteum. Endocrinology 1988; 122:1185–7.CrossRefGoogle Scholar
  3. 3.
    Old LJ. Tumor necrosis factor (TNF). Science 1985; 230:630–2.PubMedCrossRefGoogle Scholar
  4. 4.
    Kendall ME, Hymer WC. Cell blotting: a new approach to quantify hormone secretion from individual rat pituitary cells. Endocrinology 1987; 121:2260–2.PubMedCrossRefGoogle Scholar
  5. 5.
    Mannel DN, Moore RN, Mergenhagen SE. Macrophages as a source of tumoricidal activity (tumor-necrotizing factor). Infect Immun 1980; 30:523–30.PubMedGoogle Scholar
  6. 6.
    Byskov AG. Atresia. In: Midgley AR, Sadler WA, eds. Ovarian follicular development and function. New York: Raven Press, 1979.Google Scholar
  7. 7.
    Hay MF, Cran DG, Moor RM. Structural changes occurring during atresia in sheep ovarian follicles. Cell Tissue Res 1976; 169:515–29.PubMedCrossRefGoogle Scholar
  8. 8.
    Hay MF, Moor RM, Cran DG, Dott HM. Regeneration of atretic sheep ovarian follicles in vitro. J Reprod Fertil 1979; 55:195–207.PubMedCrossRefGoogle Scholar
  9. 9.
    Peluso JJ, England-Charlesworth C, Bolender DL, Steger RW. Ultrastructural alterations with the initiation of follicular atresia. Cell Tissue Res 1980; 211:105–15.PubMedCrossRefGoogle Scholar
  10. 10.
    Saidapur SK. Follicular atresia in the ovaries of nonmammalian vertebrates. Int Rev Cytol 1978; 54:225–44.PubMedCrossRefGoogle Scholar
  11. 11.
    Braw RH, Tsafriri A. Effect of PMSG on follicular atresia in the immature rat ovary. J Reprod Fertil 1980; 59:267–72.PubMedCrossRefGoogle Scholar
  12. 12.
    Christensen AK, Gillim SW. The correlation of fine structure and function in steroid-secreting cells, with emphasis on those of the gonads. In: McKerns KW, ed. The gonads. New York: Appleton-Century-Crofts, 1969.Google Scholar
  13. 13.
    Sato N, Goto T, Haranaka K, et al. Actions of tumor necrosis factor on cultured vascular endothelial cells: morphologic modulation, growth inhibition and cytotoxicity. J Natl Cancer Inst 1986; 76:1113–21.PubMedGoogle Scholar
  14. 14.
    Greenwald GS, Terranova PF. Follicular selection and its control. In: Knobil E, Neil JD, eds. The physiology of reproduction. New York: Raven Press, 1988.Google Scholar
  15. 15.
    Nawroth PP, Stem DM. Modulation of endothelial cell hemostatic properties by tumor necrosis factor. J Exp Med 1986; 163:740–5.PubMedCrossRefGoogle Scholar
  16. 16.
    Gamble JR, Harlan JM, Klebanoff S J, Vadas MA. Stimulation of adherence of neutrophils to umbilical vein endothelium by human recombinant tumor necrosis factor. Proc Natl Acad Sci USA 1985; 82:8667–71.PubMedCrossRefGoogle Scholar
  17. 17.
    Dayer JM, Beutler B, Cerami A. Cachectin/tumornecrosis factor stimulates collagenase and prostaglandin E2 production by human synovial cells and dermal fibroblasts. J Exp Med 1985; 162:2163–8.PubMedCrossRefGoogle Scholar
  18. 18.
    Lobel BL, Levy E. Enzymatic correlates of development, secretory function and regression of follicles and corpora lutea in the bovine ovary. Acta Endocrinol (Copenh) 1968;59(suppl 132):7–63.Google Scholar
  19. 19.
    Hepburn A, Boeynaems J, Fiers W, Dumont JE. Modulation of tumor necrosis factor cytotoxicity in L929 cells by bacterial toxins, hydrocortisone and inhibitors of arachidonic acid metabolism. Biochem Biophys Res Commun 1987; 149:815–22.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • K. F. Roby
    • 1
  • P. F. Terranova
    • 1
  1. 1.Department of PhysiologyUniversity of Kansas Medical CenterKansas CityUSA

Personalised recommendations