Cytokine expression in the gilt oviduct: effects of seminal plasma, spermatozoa and extender after insemination.
Effects of semen components [fresh semen in extender, spermatozoa in extender (Spz), seminal plasma (SP)], or extender alone (Beltsville thawing solution, BTS) on the expression of selected cytokines [interleukin (IL)-1beta, IL-6, IL-10 and transforming growth factor (TGF)-beta1)] as well as the presence of cells positive for CD8 or CD25 were studied in the pig oviduct. In addition, cytokines in SP and oviductal flushings were analyzed. In experiment (Exp) I, groups of gilts were sampled at 5-6h after insemination with SP, Spz, fresh semen in BTS or only BTS (control). In Exp II, gilts were sampled 35-40 h after insemination with SP, Spz, BTS or only catheter insertion (control). Most oviductal flushing samples were positive (> or =detectable limits) for IL-10 and TGF-beta1 but only few for IL-6. The IHC-labelling of IL-6, IL-10 and TGF-beta1 was evident, especially in the epithelial cells of the isthmus and infundibulum as well as in the cells of the regional (mesometrial) lymph node. Cilia of the epithelium were positive for IL-6 (strongest in the infundibulum) and TGF-beta1 (strongest in the isthmus) but negative for IL-10. There were no consistent differences in IHC-labelling of the cytokines in relation to different treatments, except at 35-40 h after insemination (Exp II), when IL-6 was slightly higher in epithelium of the SP group and IL-10 in the infundibular connective tissue was higher in the SP and Spz groups. In the isthmus and infundibulum, there were no differences between animals inseminated with BTS (control) and the semen components for any of the cytokine mRNAs at 5-6h after insemination (Exp I). However, later (35-40 h, Exp II), insemination with SP, Spz and BTS alone appeared to up-regulate TGF-beta1 mRNA expression compared with the control group (without any fluid infused). In all treatment groups, the mRNA level for TGF-beta1 was higher than for IL-1beta, IL-6 and IL-10. Higher mRNA levels of all cytokines were found in the isthmus compared with the infundibulum. Numbers of CD8-positive cells (both in epithelium and connective tissue) appeared higher in the infundibulum compared with the isthmus and were mostly higher shortly (Exp I) after treatment with SP, SPZ and BTS than later (Exp II) in both segments. CD25-positive cells were few and found solely in the sub-epithelial connective tissue. The results indicate that in the porcine oviduct, IL-6, IL-10 and TGF-beta1 are endogenous produced and that TGF-beta1 may have a more important role for immunomodulation than the other cytokines, especially in isthmus. Differences between isthmus and infundibulum in cytokine mRNA expression and in presence of CD8-positive cells indicate different patterns of immune reactivity in the upper and lower parts of the oviduct.
Jiwakanon
J
J
Division of Reproduction, Department of Clinical Sciences, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Berg
M
M
Persson
E
E
Fossum
C
C
Dalin
A-M
AM
D000818:Animals; D016827:CD8 Antigens / Q000032:analysis; D015925:Cryopreservation / Q000662:veterinary; D016207:Cytokines / Q000032:analysis / Q000235:genetics*; D005187:Fallopian Tubes / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D007315:Insemination, Artificial / Q000379:methods / Q000662:veterinary; D016753:Interleukin-10 / Q000032:analysis / Q000235:genetics; D015850:Interleukin-6 / Q000032:analysis / Q000235:genetics; D008297:Male; D012333:RNA, Messenger / Q000032:analysis; D012661:Semen / Q000737:chemistry / Q000502:physiology*; D012662:Semen Preservation / Q000662:veterinary; D012996:Solutions; D013094:Spermatozoa / Q000502:physiology*; D013552:Swine / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000032:analysis / Q000235:genetics
D016827:CD8 Antigens; D016207:Cytokines; D015850:Interleukin-6; D012333:RNA, Messenger; D012996:Solutions; D053773:Transforming Growth Factor beta1; D016753:Interleukin-10