ContentThe aim of this study was to localize and evaluate the role of Toll-like receptor 2 (TLR2) in the endometrium and cervix of bitches at different stages of the oestrous cycle and in bitches with pyometra. Sixty-seven nulliparous dogs, ranging in age from 1 to 13 years, were allocated amongst five groups (prooestrus; n = 7, oestrus; n = 10, dioestrus; n = 16, anoestrus; n = 11, pyometra; n = 23). Blood samples were collected for the measurement of progesterone concentration. The mean progesterone concentration was analysed as a parameter for validating the stage of the oestrous cycle in bitches. Tissues collected from uterine horn and cervix were fixed in 4% paraformaldehyde for immunohistochemical examination of TLR2. The expression of TLR2 was assessed semi-quantitatively. No pathological changes were found in the uterine samples of healthy dogs. In bitches with pyometra, the glandular epithelium expressed TLR2 more intensely than the surface epithelium. The expression of TLR2 in the glandular epithelium was also significantly higher in healthy dogs at oestrus, dioestrus and dogs with pyometra compared with anoestrous dogs (p < 0.01). The expression of TLR2 in the stroma was not observed in the group of healthy dogs at all stages. The surface epithelium of cervix in dogs with pyometra expressed TLR2 significantly more intensely than did the stoma, whereas the expression of TLR2 during oestrus and dioestrus was absent in the stroma of cervix. This study provides the first report of immunohistochemical localization of TLR2 in the canine reproductive tract. In the present study, TLR2 was expressed in endometrial epithelium but was absent in the endometrial stroma of healthy dogs at all oestrous cycle stages. These findings suggest differential expression of TLR in endometrial cells. On the other hand, the lack of TLR2 in the stroma of healthy uteri of dogs may predispose to infection from the invading pathogens once the epithelial cells have been destroyed by the pathogens, especially Gram-positive bacteria.
Worldwide heat stress (HS) conditions have a negative impact on dairy cow fertility. However, understanding of the effect of heat stress on endometrial functions is still unclear. The present study aimed to investigate the effects of differential heat exposure conditions on the immune response and prostaglandin biosynthesis of bovine endometrium challenged with bacterial lipopolysaccharide (LPS). Cultures of endometrial cells were grown to confluence at 37 °C (control) and 40.4 °C for 24 h after confluence (short-term heat exposure) and 40.4 °C for 8 days from the beginning of the culture (long-term heat exposure), prior to a challenge by 100 ng/mL LPS for 12 h. LPS altered ALOX12, IL8, IL1B, S100A8, PTGES and AKR1B1 expressions, as well as secretory IL8 and PGF2α. Short-term heat exposure decreased S100A8, IL8 and PGF2α compared with the control temperature, while long-term heat exposure decreased S100A8 and PGF2α. In contrast, HSPA5 expression was not altered by heat exposure or LPS. Indeed, the short-term heat treatment was insufficient for accomplishing the responses of the endometrium to LPS treatment for IL8, S100A8 and PTGES expressions when compared with other temperature conditions. Our findings showed that heat exposure could compromise endometrium immune response and prostaglandin biosynthesis in different ways based on elevated temperature duration, which could reduce subsequent fertility.
This study was performed to monitor estrous patterns and, more importantly, changes in anti-Müllerian hormone (AMH) concentrations during the peri-ovulatory period in deslorelin-induced estrous bitches. Healthy anestrous bitches (n = 4) were used. Estrus and ovulation were monitored after deslorelin implantation. Blood samples were collected for analysis of progesterone, estradiol-17ß and AMH concentrations before implantation (day 0) and on days 6, 8, 10, 12, 14, 16, 18, 20 and 22 after implantation. Six days following treatment, all bitches showed estrus signs. Ovulation took place between days 12 and 15. Circulating AMH concentrations varied among bitches from 0.12 to 3.08 ng/mL. However, no significant differences in AMH levels (mean ± SD) were observed between day 0 and days following post-implantation (p > 0.05). There were no significant correlations between AMH and estradiol or AMH and progesterone (p > 0.05). Ultrasonographically, the number of clearly identifiable ovarian follicles was higher before ovulation and the area of ovaries increased after ovulation (p < 0.05). Except for AMH, changes in vaginal cytology, estradiol-17ß and progesterone levels observed in our study were similar to naturally occurring estrus. Large intra- and inter-individual variation in AMH were observed suggesting that AMH is currently not suitable as a canine fertility marker to monitor ovarian response to deslorelin treatment for estrus induction.
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