Journal Mobile Options
Table of Contents
Vol. 182, No. 1, 2006
Issue release date: April 2006
Cells Tissues Organs 2006;182:22–31

Immunohistochemical Detection of Macrophage Migration Inhibitory Factor in Fetal and Adult Bovine Epididymis: Release by the Apocrine Secretion Mode?

Eickhoff R. · Jennemann G. · Hoffbauer G. · Schüring M.-P. · Kaltner H. · Sinowatz F. · Gabius H.-J. · Seitz J.
To view the fulltext, log in and/or choose pay-per-view option

Individual Users: Register with Karger Login Information

Please create your User ID & Password

Contact Information

I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in


Originally defined as a lymphokine inhibiting the random migration of macrophages, the macrophage migration inhibitory factor (MIF) is an important mediator of the host response to infection. Beyond its function as a classical cytokine, MIF is currently portrayed as a multifunctional protein with growth-regulating properties present in organ systems beyond immune cells. In previous studies, we detected substantial amounts of MIF in the rat epididymis and epididymal spermatozoa, where it appears to play a role during post-testicular sperm maturation and the acquisition of fertilization ability. To explore its presence in other species not yet examined in this respect, we extended the range of studies to the bull. Using a polyclonal antibody raised against MIF purified from bovine eye lenses, we detected MIF in the epithelium of the adult bovine epididymis with the basal cells representing a prominently stained cell type. A distinct accumulation of MIF at the apical cell pole of the epithelial cells and in membranous vesicles localized in the lumen of the epididymal duct was obvious. In the fetal bovine epididymis, we also detected MIF in the epithelium, whereas MIF accumulation was evident at the apical cell surface and in apical protrusions. By immunoelectron microscopy of the adult bovine epididymis, we localized MIF in apical protrusions of the epithelial cells and in luminal membrane-bound vesicles that were found in close proximity to sperm cells. Although the precise origin of the MIF-containing vesicles remains to be delineated, our morphological observations support the hypothesis that they become detached from the apical surface of the epididymal epithelial cells. Additionally, an association of MIF with the outer dense fibers of luminal spermatozoa was demonstrated. Data obtained in this study suggest MIF release by an apocrine secretion mode in the bovine epididymis. Furthermore, MIF localized in the basal cells of the epithelium and in the connective tissue could be responsible for regulating the migration of macrophages in order to avoid contact of immune cells with spermatozoa that carry a wide range of potent antigens.

Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.


  1. Agrawal, Y., T. Vanha-Perttula (1988) Electron microscopic study of the secretion process in bovine reproductive organs. J Androl 9: 307–316.
  2. Agarwal, A., A.P. Hoffer (1989) Ultrastructural studies on the development of the blood-epididymis barrier in immature rats. J Androl 10: 425–431.
  3. Andre, S., S. Kojima, N. Yamazaki, C. Fink, H. Kaltner, K. Kayser, H.-J. Gabius (1999) Galectins-1 and -3 and their ligands in tumor biology. Non-uniform properties in cell-surface presentation and modulation of adhesion to matrix glycoproteins for various tumor cell lines, in biodistribution of free and liposome-bound galectins and in their expression by breast and colorectal carcinomas with/without metastatic propensity. J Cancer Res Clin Oncol 125: 461–474.
  4. Aumüller, G., M. Bergmann, J. Seitz (1991) Immunohistochemical distribution of sulfhydryl oxidase in the human testis. Cell Tissue Res 266: 23–28.
  5. Bacher, M., C.N. Metz, T. Calandra, K. Mayer, J. Chesney, M. Lohoff, D. Gemsa, T. Donnelly, R. Bucala (1996) An essential regulatory role for macrophage migration inhibitory factor in T-cell activation. Proc Natl Acad Sci USA 93: 7849–7854.
  6. Bacher, M., A. Meinhardt, H.Y. Lan, W. Mu, C.N. Metz, J.A. Chesney, T. Calandra, D. Gemsa, T. Donnelly, R.C. Atkins, R. Bucala (1997) Migration inhibitory factor expression in experimentally induced endotoxemia. Am J Pathol 150: 235–246.
  7. Bernhagen, J., T. Calandra, R.A. Mitchell, S.B. Martin, K.J. Tracey, W. Voelter, K.R. Manogue, A. Cerami, R. Bucala (1993) MIF is a pituitary-derived cytokine that potentiates lethal endotoxaemia. Nature 365: 756–759.
  8. Bloom, B.R., B. Bennett (1966) Mechanism of a reaction in vitro associated with delayed-type hypersensitivity. Science 153: 80–82.
  9. Bove, S.E., M.G. Petroff, M. Nishibori, J.L. Pate (2000) Macrophage migration inhibitory factor in the bovine corpus luteum: characterization of steady-state messenger ribonucleic acid and immunohistochemical localization. Biol Reprod 62: 879–885.
  10. Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254.
  11. Bronson, R.A., F.M. Fusi (1994) Autoimmunity to sperm antigens. Allergy Clin North Am 14: 773–786.

    External Resources

  12. Calandra, T., J. Bernhagen, R.A. Mitchell, R. Bucala (1994) The macrophage is an important and previously unrecognized source of macrophage migration inhibitory factor. J Exp Med 179: 1895–1902.
  13. Calandra, T., J. Bernhagen, C.N. Metz, L.A. Spiegel, M. Bacher, T. Donnelly, A. Cerami, R. Bucala (1995) MIF as a glucocorticoid-induced modulator of cytokine production. Nature 377: 68–71.
  14. Calandra, T., L.A. Spiegel, C.N. Metz, R. Bucala (1998) Macrophage migration inhibitory factor is a critical mediator of the activation of immune cells by exotoxins of Gram-positive bacteria. Proc Natl Acad Sci USA 95: 11383–11388.
  15. Calne, R.Y. (2000) Immunological tolerance – the liver effect. Immunol Rev 174: 280–282.
  16. Choufani, G., R. Ghanooni, C. Decaestecker, K. Delbrouck, P. Simon, M.P. Schuring, Y. Zick, S. Hassid, H.-J. Gabius, R. Kiss (2001) Detection of macrophage migration inhibitory factor (MIF) in human cholesteatomas and functional implications of correlations to recurrence status and to expression of matrix metalloproteinases-3/9, retinoic acid receptor-beta, and anti-apoptotic galectin-3. Laryngoscope 111: 1656–1662.
  17. Cooper, T.G. (1999) Epididymis. Encyclopedia Reprod 2: 1–17.
  18. Cropp, C.S., W.D. Schlaff (1990) Antisperm antibodies. Arch Immunol Ther Exp (Warsz) 38: 31–46.
  19. David, J.R. (1966) Delayed hypersensitivity in vitro: its mediation by cell-free substances formed by lymphoid cell-antigen interaction. Proc Natl Acad Sci USA 56: 72–77.
  20. Eickhoff, R., B. Wilhelm, H. Renneberg, G. Wennemuth, M. Bacher, D. Linder, R. Bucala, J. Seitz, A. Meinhardt (2001) Purification and characterization of macrophage migration inhibitory factor as a secretory protein from rat epididymis: evidences for alternative release and transfer to spermatozoa. Mol Med 7: 27–35.
  21. Eickhoff, R., C. Baldauf, H.W. Koyro, G. Wennemuth, Y. Suga, J. Seitz, R. Henkel, A. Meinhardt (2004) Influence of macrophage migration inhibitory factor (MIF) on the zinc content and redox state of protein-bound sulphydryl groups in rat sperm: indications for a new role of MIF in sperm maturation. Mol Hum Reprod 10: 605–611.
  22. Frenette, G., C. Lessard, R. Sullivan (2002) Selected proteins of ‘prostasome-like particles’ from epididymal cauda fluid are transferred to epididymal caput spermatozoa in bull. Biol Reprod 67: 308–313.
  23. Frenette, G., C. Lessard, E. Madore, M.A. Fortier, R. Sullivan (2003) Aldose reductase and macrophage migration inhibitory factor are associated with epididymosomes and spermatozoa in the bovine epididymis. Biol Reprod 69: 1586–1592.
  24. Frenette, G., C. Legare, F. Saez, R. Sullivan (2005) Macrophage migration inhibitory factor in the human epididymis and semen. Mol Hum Reprod 11: 575–582.
  25. Goyal, H.O. (1985) Morphology of the bovine epididymis. Am J Anat 172: 155–172.
  26. Hoffer, A.P., B.T. Hinton (1984) Morphological evidence for a blood-epididymis barrier and the effects of gossypol on its integrity. Biol Reprod 30: 991–1004.
  27. Howards, S.S., S.J. Jessee, A.L. Johnson (1976) Micropuncture studies of the blood-seminiferous tubule barrier. Biol Reprod 14: 264–269.
  28. Hudson, J.D., M.A. Shoaibi, R. Maestro, A. Carnero, G.J. Hannon, D.H. Beach (1999) A proinflammatory cytokine inhibits p53 tumor suppressor activity. J Exp Med 190: 1375–1382.
  29. Kaltner, H., K. Seyrek, A. Heck, F. Sinowatz, H.J. Gabius (2002) Galectin-1 and galectin-3 in fetal development of bovine respiratory and digestive tracts. Comparison of cell type-specific expression profiles and subcellular localization. Cell Tissue Res 307: 35–46.
  30. Kaunisto, K., S. Parkkila, T. Tammela, L. Ronnberg, H. Rajaniemi (1990) Immunohistochemical localization of carbonic anhydrase isoenzymes in the human male reproductive tract. Histochemistry 94: 381–386.
  31. Kirchhoff, C. (1994) A major messenger ribonucleic acid of the rodent epididymis encodes a small glycosylphosphatidylinositol-anchored lymphocyte surface antigen. Biol Reprod 50: 896–902.
  32. Kirchhoff, C., G. Hale (1996) Cell-to-cell trans fer of glycosylphosphatidylinositol-anchored membrane proteins during sperm maturation. Mol Hum Reprod 2: 177–184.
  33. Kleemann, R., A. Hausser, G. Geiger, R. Mischke, A. Burger-Kentischer, O. Flieger, F.J. Johannes, T. Roger, T. Calandra, A. Kapurniotu, M. Grell, D. Finkelmeier, H. Brunner, J. Bernhagen (2000) Intracellular action of the cytokine MIF to modulate AP-1 activity and the cell cycle through Jab1. Nature 408: 211–216.
  34. Legendre, H., C. Decaestecker, N. Nagy, A. Hendlisz, M.P. Schuring, I. Salmon, H.-J. Gabius, J.C. Pector, R. Kiss (2003) Prognostic values of galectin-3 and the macrophage migration inhibitory factor (MIF) in human colorectal cancers. Mod Pathol 16: 491–504.
  35. Manin, M., P. Lecher, A. Martinez, S. Tournadre, C. Jean (1995) Exportation of mouse vas deferens protein, a protein without a signal peptide, from mouse vas deferens epithelium: a model of apocrine secretion. Biol Reprod 52: 50–62.
  36. Meinertz, H., L. Linnet, H. Wolf, T. Hjort (1991) Antisperm antibodies on epididymal spermatozoa. Am J Reprod Immunol 25: 158–162.
  37. Meinhardt, A., M. Bacher, J.R. McFarlane, C.N. Metz, J. Seitz, M.P. Hedger, D.M. de Kretser, R. Bucala (1996) Macrophage migration inhibitory factor production by Leydig cells: evidence for a role in the regulation of testicular function. Endocrinology 137: 5090–5095.
  38. Meinhardt, A., M. Bacher, M.K. O’Bryan, J.R. McFarlane, C. Mallidis, C. Lehmann, C.N. Metz, D.M. de Kretser, R. Bucala, M.P. Hedger (1999) A switch in the cellular localization of macrophage migration inhibitory factor in the rat testis after ethane dimethane sulfonate treatment. J Cell Sci 112: 1337–1344.
  39. Rosengren, E., R. Bucala, P. Aman, L. Jacobsson, G. Odh, C.N. Metz, H. Rorsman (1996) The immunoregulatory mediator macrophage migration inhibitory factor (MIF) catalyzes a tautomerization reaction. Mol Med 2: 143–149.
  40. Rüsse, I., F. Sinowatz (1991) Lehrbuch der Embryologie der Haustiere. Berlin, Paul Parey.
  41. Seitz, J., C. Keppler, U. Rausch, G. Aumüller (1990) Immunohistochemistry of secretory transglutaminase from rodent prostate. Histochemistry 93: 525–530.
  42. Steinhoff, M., W. Eicheler, P.M. Holterhus, U. Rausch, J. Seitz, G. Aumüller (1994) Hormonally induced changes in apocrine secretion of transglutaminase in the rat dorsal prostate and coagulating gland. Eur J Cell Biol 65: 49–59.
  43. Sullivan, R., F. Saez, J. Girouard, G. Frenette (2005) Role of exosomes in sperm maturation during the transit along the male reproductive tract. Blood Cells Mol Dis 35: 1–10.
  44. Sun, H.W., J. Bernhagen, R. Bucala, E. Lolis (1996) Crystal structure at 2.6-A resolution of human macrophage migration inhibitory factor. Proc Natl Acad Sci USA 93: 5191–5196.
  45. Suzuki, M., H. Sugimoto, A. Nakagawa, I. Tanaka, J. Nishihira, M. Sakai (1996) Crystal structure of the macrophage migration inhibitory factor from rat liver. Nat Struct Biol 3: 259–266.
  46. Wada, S., S. Fujimoto, Y. Mizue, J. Nishihira (1997) Macrophage migration inhibitory factor in the human ovary: presence in the follicular fluids and production by granulosa cells. Biochem Mol Biol Int 41: 805–814.
  47. Wada, S., T. Kudo, M. Kudo, N. Sakuragi, H. Hareyama, J. Nishihira, S. Fujimoto (1999) Induction of macrophage migration inhibitory factor in human ovary by human chorionic gonadotrophin. Hum Reprod 14: 395–399.
  48. Waeber, G., T. Calandra, R. Roduit, J.A. Haefliger, C. Bonny, N. Thompson, B. Thorens, E. Temler, A. Meinhardt, M. Bacher, C.N. Metz, P. Nicod, R. Bucala (1997) Insulin secretion is regulated by the glucose-dependent production of islet beta cell macrophage migration inhibitory factor. Proc Natl Acad Sci USA 94: 4782–4787.
  49. Weiser, W.Y., P.A. Temple, J.S. Witek-Giannotti, H.G. Remold, S.C. Clark, J.R. David (1989) Molecular cloning of a cDNA encoding a human macrophage migration inhibitory factor. Proc Natl Acad Sci USA 86: 7522–7526.
  50. Wilhelm, B., C. Keppler, G. Hoffbauer, F. Lottspeich, D. Linder, A. Meinhardt, G. Aumüller, J. Seitz (1998) Cytoplasmic carbonic anhydrase II of rat coagulating gland is secreted via the apocrine export mode. J Histochem Cytochem 46: 505–511.
  51. Zeng, F.Y., W.Y. Weiser, H. Kratzin, B. Stahl, M. Karas, H.J. Gabius (1993) The major binding protein of the interferon antagonist sarcolectin in human placenta is a macrophage migration inhibitory factor. Arch Biochem Biophys 303: 74–80.
  52. Zeng, F.Y., V. Gerke, H.J. Gabius (1994a) Characterization of the macrophage migration inhibitory factor-binding site of sarcolectin and its relationship to human serum albumin. Biochem Biophys Res Commun 200: 89–94.
  53. Zeng, F.Y., H. Kratzin, H.J. Gabius (1994b) Migration inhibitory factor-binding sarcolectin from human placenta is indistinguishable from a subfraction of human serum albumin. Biol Chem Hoppe Seyler 375: 393–399.

Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50