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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Leucocyte population changes in the reproductive tract of the ewe in response to insemination

Jennifer L. Scott A B , Natkunam Ketheesan A and Phillip M. Summers A
+ Author Affiliations
- Author Affiliations

A School of Veterinary and Biomedical Sciences, James Cook University, Solander Drive, Douglas, Qld 4811, Australia.

B Corresponding author. Email: jennifer.scott1@jcu.edu.au

Reproduction, Fertility and Development 18(6) 627-634 https://doi.org/10.1071/RD05165
Submitted: 16 December 2005  Accepted: 21 March 2006   Published: 9 June 2006

Abstract

Leucocyte changes after insemination may affect conceptus implantation, but information regarding leucocyte populations in the ruminant reproductive tract is limited. The present study investigated changes in leucocyte populations and distribution in the ovine reproductive tract following oestrus and insemination. Fifteen ewes were mated with a ram for 1 h and their reproductive tracts collected 3, 6, 18, 24 or 48 h later. Another 15 ewes were used as oestrus controls. Tissues were collected from 10 sites in each reproductive tract and stained with haematoxylin and eosin, Toluidine blue and immunohistochemically using a monoclonal CD68 antibody. Luminal mucus smears were collected from seven sites and stained with a modified Wright’s stain and immunohistochemically. Neutrophils, eosinophils, mast cells and macrophages were identified and quantified, and temporal changes in their distribution within tissues were examined. Neutrophils and macrophages increased significantly (P < 0.05) in posterior cervical and uterine tissues following insemination. In uterine tissues, neutrophils peaked at 6 h after insemination, whereas macrophages peaked at 18–24 h. Mast cells decreased and eosinophils remained constant. Neutrophils increased significantly (P < 0.05) in the cervical and uterine lumen following insemination. In conclusion, leucocyte population changes after insemination vary between different sites in the ovine reproductive tract and may contribute to pregnancy establishment.

Extra keywords: eosinophils, macrophages, mast cells, neutrophils, ovine.


Acknowledgments

The authors thank Mr Chris Coleman and Mr Laurie Reilly, School of Veterinary and Biomedical Sciences, James Cook University, for assistance with the management of animals and advice on immunohistochemical techniques, respectively.


References

Barratt, C. L. R. , Bolton, A. E. , and Cooke, I. D. (1990). Functional significance of white blood cells in the male and female reproductive tract. Hum. Reprod. 5, 639–648.
PubMed | Carson F. L. (1990). ‘Histotechnology: A Self-Instructional Text.’ (ASCP Press: Chicago, IL, USA.)

Cobb, S. P. , and Watson, E. D. (1995). Immunohistochemical study of immune cells in the bovine endometrium at different stages of the oestrous cycle. Res. Vet. Sci. 59, 238–241.
Crossref | GoogleScholarGoogle Scholar | PubMed | Cotran R. S., Kumar V., and Collins T. (1999). ‘Robbins Pathologic Basis of Disease.’ 6th edn. (W. B. Saunders: Philadelphia, PA, USA.)

De, M. , and Wood, G. W. (1990). Influence of oestrogen and progesterone on macrophage distribution in the mouse uterus. J. Endocrinol. 126, 417–424.
PubMed | Drury R. A. B., and Wallington E. A. (1967). ‘Carleton’s Histological Technique.’ 4th edn. (Oxford University Press: New York, NY, USA.)

Frayne, J. , and Stokes, C. R. (1994). MHC class II positive cells and T cells in the equine endometrium throughout the oestrous cycle. Vet. Immunol. Immunopathol. 41, 55–72.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Garfield, R. E. , Bytautiene, E. , Vedernikov, Y. P. , Marshall, J. S. , and Romero, R. (2000). Modulation of rat uterine contractility by mast cells and their mediators. Am. J. Obstet. Gynecol. 183, 118–125.
PubMed |

Kaeoket, K. , Dalin, A.-M. , Magnusson, U. , and Persson, E. (2001). The sow endometrium at different stages of the oestrous cycle: immunohistochemical study on the distribution of SWC3-expressing cells (granulocytes, monocytes and macrophages). J. Vet. Med. A 48, 507–511.
Crossref | GoogleScholarGoogle Scholar |

Kaeoket, K. , Persson, E. , and Dalin, A.-M. (2002). Corrigendum to ‘The sow endometrium at different stages of the oestrous cycle: studies on morphological changes and infiltration by cells of the immune system’. Anim. Reprod. Sci. 73, 89–107.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Kaplanski, G. , Marin, V. , Montero-Julian, F. , Mantovani, A. , and Farnarier, C. (2003). IL-6: a regulator of the transition from neutrophil to monocyte recruitment during inflammation. Trends Immunol. 24, 25–29.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Kaushic, C. , Frauendorf, E. , Rossoll, R. M. , Richardson, J. M. , and Wira, C. R. (1998). Influence of the estrous cycle on the presence and distribution of immune cells in the rat reproductive tract. Am. J. Reprod. Immunol. 39, 209–216.
PubMed |

Kotilainen, T. , Huhtinen, M. , and Katila, T. (1994). Sperm-induced leukocytosis in the equine uterus. Theriogenology 41, 629–636.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Lovell, J. E. , and Getty, R. (1968). Fate of semen in the uterus of the sow: histologic study of endometrium during the 27 h after natural service. Am. J. Vet. Res. 29, 609–625.
PubMed |

Mahajan, S. C. , and Menge, A. C. (1967). Influence of reproductive phase on the inflammatory response and rate of sperm removal in the uterus and oviduct of the cow. Am. J. Vet. Res. 28, 1037–1041.
PubMed |

Matthijs, A. , Engel, B. , and Woelders, H. (2003). Neutrophil recruitment and phagocytosis of boar spermatozoa after artificial insemination of sows, and the effects of inseminate volume, sperm dose and specific additives in the extender. Reproduction 125, 357–367.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Mattner, P. E. (1963). Spermatozoa in the genital tract of the ewe: II. Distribution after coitus. Aust. J. Biol. Sci. 16, 688–694.


Mattner, P. E. (1966). Formation and retention of the spermatozoan reservoir in the cervix of the ruminant. Nature 212, 1479–1480.


Mattner, P. E. (1968). The distribution of spermatozoa and leukocytes in the female genital tract in goats and cattle. J. Reprod. Fertil. 17, 253–261.
PubMed |

Mattner, P. E. (1969). Differential leukocyte responses to spermatozoa in the cervix and the uterus in ewes. J. Reprod. Fertil. 18, 297–303.
PubMed |

McDonald, L. E. , Black, W. G. , McNutt, S. H. , and Casida, L. E. (1952). The response of the rabbit uterus to instillation of semen at different phases of the estrous cycle. Am. J. Vet. Res. 13, 419–424.
PubMed |

McMaster, M. T. , Newton, R. C. , Dey, S. K. , and Andrews, G. K. (1992). Activation and distribution of inflammatory cells in the mouse uterus during the preimplantation period. J. Immunol. 148, 1699–1705.
PubMed |

Mitchell, S. E. , Robinson, J. J. , King, M. E. , McKelvey, W. A. C. , and Williams, L. M. (2002). Interleukin 8 in the cervix of non-pregnant ewes. Reproduction 124, 409–416.
Crossref | GoogleScholarGoogle Scholar | PubMed |

O’Leary, S. , Jasper, M. J. , Warnes, G. M. , Armstrong, D. T. , and Robertson, S. A. (2004). Seminal plasma regulates endometrial cytokine expression, leukocyte recruitment and embryo development in the pig. Reproduction 128, 237–247.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Pandya, I. J. , and Cohen, J. (1985). The leukocytic reaction of the human uterine cervix to spermatozoa. Fertil. Steril. 43, 417–421.
PubMed |

Robertson, S. A. (2002). Finding solutions: semen and maternal immune tolerance. Reprod. Fertil. Dev. 14(Suppl.), 60.


Robertson, S. A. , and Sharkey, D. J. (2001). The role of semen in the induction of maternal immune tolerance to pregnancy. Immunology 13, 243–254.


Robertson, S. A. , Mau, V. J. , Tremellen, K. P. , and Seamark, R. (1996). Role of high molecular weight seminal vesicle proteins in eliciting the uterine inflammatory response to semen in mice. J. Reprod. Fertil. 107, 265–277.
PubMed |

Robertson, S. A. , Mau, V. J. , Hudson, S. N. , and Tremellen, K. P. (1997). Cytokine–leukocyte networks and the establishment of pregnancy. Am. J. Reprod. Immunol. 37, 438–442.
PubMed |

Rozeboom, K. J. , Troedsson, M. H. T. , Molitor, T. W. , and Crabo, B. G. (1999). The effect of spermatozoa and seminal plasma on leukocyte migration into the uterus of gilts. J. Anim. Sci. 77, 2201–2206.
PubMed |

Sanford, T. R. , De, M. , and Wood, G. W. (1992). Expression of colony-stimulating factors and inflammatory cytokines in the uterus of CD1 mice during Days 1 to 3 of pregnancy. J. Reprod. Fertil. 94, 213–220.
PubMed |

Starkey, P. M. , Clover, L. M. , and Rees, M. C. P. (1991). Variation during the menstrual cycle of immune cell populations in the human endometrium. Eur. J. Obstet. Gynecol. Reprod. Biol. 39, 203–207.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Summerfield, N. J. , and Watson, E. D. (1998). Endometrial macrophage populations in genitally normal mares at oestrus and dioestrus and in mares susceptible to endometritis. Equine Vet. J. 30, 79–81.
PubMed |

Tekin, S. , and Hansen, P. J. (2004). Regulation of numbers of macrophages in the endometrium of the sheep by systemic effects of pregnancy, local presence of the conceptus, and progesterone. Am. J. Reprod. Immunol. 51, 56–62.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Thompson, L. A. , Barratt, C. L. R. , Bolton, A. E. , and Cooke, I. D. (1992). The leukocytic reaction of the human uterine cervix. Am. J. Reprod. Immunol. 28, 85–89.
PubMed |

Tibbetts, T. A. , Conneely, O. M. , and O’Malley, B. W. (1999). Progesterone via its receptor antagonizes the pro-inflammatory activity of estrogen in the mouse uterus. Biol. Reprod. 60, 1158–1165.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Tremellen, K. P. , Seamark, R. F. , and Robertson, S. A. (1998). Seminal transforming growth factor beta-1 stimulates granulocyte-macrophage colony-stimulating factor production and inflammatory cell recruitment in the murine uterus. Biol. Reprod. 58, 1217–1225.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Troedsson, M. H. T. , Liu, I. K. M. , and Crabo, B. G. (1998). Sperm transport and survival in the mare: a review. Theriogenology 50, 807–818.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Troedsson, M. H. T. , Loset, K. , Alghamdi, A. M. , Dahms, B. , and Crabo, B. G. (2001). Interaction between equine semen and the endometrium: the inflammatory response to semen. Anim. Reprod. Sci. 68, 273–278.
Crossref | GoogleScholarGoogle Scholar | PubMed |