Abstract
Endometritis is one of the leading causes of infertility in the cattle and buffalo and innate immune mechanism plays an important role in clearing the infection. In this regard, endometrial expression and function of Toll Like Receptors (TLR) are focus of investigation in the recent years. In this study, we report the transcriptional profiles of TLR4 and 5 in the buffalo endometrium during the follicular, early, mid and late luteal phases of estrous cycle and 'subclinical and clinical endometritis' and also at true anestrus (n = 10 for each stage) using RT-PCR and qRT-PCR as they are the ligands for the lipopolysaccharide and flagellin components of E.coli, the most common cause of postpartum endometritis. We found a significant positive correlation between TLR4 and 5 in all the groups (r = 0.696–0.803; P < 0.05) except late luteal phase (r = 0.522; P > 0.05). Chi-square analysis showed that the qualitative expression of endometrial TLR4 and 5 transcripts was significantly associated with the phase of estrous cycle and also with uterine infection (P < 0.05). Further, using true anestrus category as a calibrator group, relative quantitation of TLR4 and 5 revealed that the transcriptional expression of TLR4 and 5 genes were highly upregulated (24.6–83.3 folds) during endometritis conditions and moderately upregulated during mid-luteal phase (6.8–16.2) of the estrous cycle (P < 0.05). The results suggested a role of progesterone in the expression of TLR4 and 5.
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References
Azawi OI (2008) Postpartum uterine infection in cattle – review. Anim Reprod Sci 105:187
Azawi OI (2010) Uterine infection in buffalo cows– review. Buffalo Bull 29:154
Bicalho RC, Machado VS, Bicalho ML, Gilbert RO, Teixeira AG, Caixeta LS, Pereira RV (2010) Molecular and epidemiological characterization of bovine intrauterine Escherichia coli. J Dairy Sci 93:5818
Chapwanya A, Meade KG, Doherty ML, Callanan JJ, Mee JF, O’Farrelly C (2009) Histopathological and molecular evaluation of Holstein–Friesian cows postpartum: towards an improved understanding of uterine innate immunity. Theriogenology 71:1396
Chapwanya A, Meade KG, Doherty ML, Callanan JJ, O’Farrelly C (2013) Endometrial epithelial cells are potent producers of tracheal antimicrobial peptide and serum amyloid A3 gene expression in response to E.coli stimulation. Vet Immunol Immunopathol 151:157
Cronin GJ, Turner LM, Goetze L, Bryant EC, Sheldon IM (2012) Toll Like Receptor 4 and MYD88- dependent signaling mechanisms of the Innate immune system are essential for the response to Lipopolysaccharide by epithelial and stromal cells of the Bovine Endometrium. Biol Reprod 86:51
Davies D, Meade KG, Herath S, Eckersall PD, Gonzalez D, White JO, Conlan RS, O’Farrelly C, Sheldon IM (2008) Toll-like receptor and antimicrobial peptide expression in the bovine endometrium. Reprod Biol Endocrinol 6:53
Fischer C, Drillich M, Odau S, Heuwieser W, Einspanier R, Gabler C (2010) Selected pro-inflammatory factor transcripts in bovine endometrial epithelial cells regulated during the oestrous cycle and elevated in case of subclinical or clinical endometritis. Reprod Fertil Dev 22:818
Heranjal DD, Sheth AR, Wadadekar KB, Desai R, Rao SS (1979) Serum gonadotrophins and prolactin in anoestrus buffaloes. Indian J Dairy Sci 32:383
Jagalan P, Das GK, Sunilkumar BV, Kumar R, Khan FA, Meur SK (2010) Cyclical changes in collagen concentration in relation to growth and development of buffalo corpus luteum. Vet Res Commun 34:511
Kawai T, Akira S (2010) The role of pattern-recognition receptors in innate immunity-Update on Toll-like receptors. Nat Immunol 11:373
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real time quantitative PCR and the 2(−ΔΔ CT) method. Methods 25:4028
Loyi T, Kumar H, Nandi S, Mathapati BS, Patra MK, Pattnaik B (2012) Differential expression of pro-inflammatory cytokines in endometrial tissue of buffaloes with clinical and sub-clinical endometritis. Res Vet Sci 94:336
Pande (2011) Studies on ovarian function in relation to uterine infection in buffaloes. MVSc Thesis. Indian Veterinary Research Institute, Izatnagar, India
Paul WE (2011) Bridging innate and adaptive immunity. Cell 147:1212
Price CJ, Cronin J, Sheldon IM (2012) Toll-Like receptor expression and function in the COV434 Granulosa cell line. Am J Reprod Immunol 68:205
Sheldon IM, Roberts MH (2010) Toll-like receptor 4 mediates the response of epithelial and stromal cells to lipopolysaccharide in the endometrium. PLoS One 5:e12906
Sheldon IM, Cronin J, Donofrio G, Schuberth HJ (2009) Defining postpartum uterine disease and the mechanisms of infection and immunity in the female reproductive tract in cattle. Biol Reprod 81:1025
Sheldon IM, Rycroft AN, Dogan B, Craven M, Bromfield JJ, Chandler A, Roberts MH, Price SB, Gilbert RO, Simpson KW (2010) Specific strains of Escherichia coli are pathogenic for the endometrium of cattle and cause pelvic inflammatory disease in cattle and mice. PLoS One 5:e9192
Singh J, Nanda AS, Adams GP (2000) The reproductive pattern and efficiency of female buffaloes. Anim Reprod Sci 60:593
Usmani RH, Ahmad N, Shafiq P, Mirza MA (2001) Effect of subclinical uterine infections on cervical and uterine involution, estrus activity and fertility in postpartum buffaloes. Theriogenology 55:563
Vahanan MB, Raj GD, Pawar RMC, Gopinath VP, Raja A, Thangavelu A (2008) Expression profile of Toll Like Receptor in a range of water buffalo tissues (Bubalus bubalis). Vet Immunol Immunopathol 126:149
Williams EJ, Fischer DP, Pfeiffer DU, England GE, Noakes DE, Dobson H, Sheldon IM (2005) Clinical evaluation of postpartum vaginal mucus reflects uterine bacterial infection and the immune response in cattle. Theriogenology 63:102
Williams EJ, Sibley K, Miller NA, Fishwick J, Herath S, Deborah, England CW, Dobson H, Sheldon IM (2008) The effects of Escherichia coli lipopolysaccharide and TNF alpha on ovarian function. Am J Reprod Immunol 60:462
Yunhe F, Liu B, Xiaosheng F, Liu Z, Liang D, Li F, Li D, Yongguo C, Feng S, Zang N, Yang Z (2013) Lipopolysaccharide increases Toll-like receptor 4 and downstream Toll-Like Receptor signaling molecules expression in bovine endometrial epithelial cells. Vet Immunol Immunopathol 151:20
Acknowledgments
The authors thank Dr. Senthil, Scientist, High Security Animal Disease Laboratory (HSADL), IVRI campus, Bhopal for reading histopathology slides and the Director, IVRI for providing Institutional Fellowship during the study period.
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Ajevar, G., Muthu, S., Sarkar, M. et al. Transcriptional profile of endometrial TLR4 and 5 genes during the estrous cycle and uterine infection in the buffalo (Bubalus bubalis). Vet Res Commun 38, 171–176 (2014). https://doi.org/10.1007/s11259-014-9594-1
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DOI: https://doi.org/10.1007/s11259-014-9594-1