1. Abnormally up-regulated cystic fibrosis transmembrane conductance regulator expression and uterine fluid accumulation contribute to Chlamydia trachomatis-induced female infertility 
Qiong He, Ph.D., Lai Ling Tsang, M.Phil., Louis Chukwuemeka Ajonuma, Ph.D., Hsiao Chang Chan, Ph.D. 
Fertility and Sterility 
Volume 93, Issue 8, Pages (May 2010)
DOI: /j.fertnstert
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
The cystic fibrosis transmembrane conductance regulator-mediated short circuit current (Isc) response and uterine fluid formation throughout the estrous cycle. Freshly isolated uterine endometrial epithelia stripped off the serosa and muscular layers from sexually mature Imprinting Control Region mice at different estrous stages (proestrus, estrus, metestrus, and diestrous) were used for Isc measurement. (A) Representative traces of Isc (μA/cm2) response to forskolin (10 μM) added to the apical side of the epithelia obtained from different estrous stages. Arrows indicate the time of forskolin addition. (B) Statistical analysis of forskolin-elicited Isc responses from the four cycle stages. Each point was obtained from at least four individual experiments. Values are means ±SEM of maximal increase in Isc. Forskolin-induced Isc at estrus is significantly higher than that at the diestrous stage (P<.01). (C) Uterine wet weight was examined when the uteri were taken at the estrus and diestrus stages (n = 5, P<.01).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Enhanced cystic fibrosis transmembrane conductance regulator (CFTR) expression, CFTR-mediated uterine anion secretion, and reduced implantation rate with C. trachomatis LPS stimulation. Mice at the initial onset of the diestrus stage were used for intrauterine injection with C. trachomatis lipopolysaccharide (LPS) (1 μg) with saline injection as the control. Endometrial epithelia removed from mice confirmed at the diestrus stage 24 hours after treatment was used for experiments. (A) Representative traces of short circuit current (Isc) (μA/cm2) response to apical addition of forskolin (10 μM) with statistical analysis. Values are means ±SEM of maximal increase in Isc. (B) Statistical analysis showed only C. trachomatis LPS-treated endometria, but not the saline-treated controls, responds to forskolin stimulation with an increase in Isc at the diestrus stage (n = 4, P<.01). (C) Western blot analysis of protein expression of uterine CFTR at 160 kDa with β-tubulin at 55 kDa used as a loading control. (D) Analysis showing increased uterine wet weights in the uteri infected by C. trachomatis LPS compared with that in control uteri (n = 5, P<.05). Pregnant mice were injected on day 3 with 10 μL of C. trachomatis LPS at 100 μg/mL or 10 μL sterile saline in each uterine horn. The effects of C. trachomatis LPS on implantation were determined by comparing the total number of viable embryos in both uterine horns on pregnant day 5 mice. (E) Results represent the means ±SEM of replicate assays (n = 6). Difference in implantation number between saline and C. trachomatis LPS-injected uterine horns was statistically significant (P<.05).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Up-regulated cystic fibrosis transmembrane conductance regulator (CFTR) expression, CFTR-related endometrial ion transport, and uterine fluid accumulation with stimulation of tumor necrosis factor-α (TNF-α) (100 ng/mL) at the diestrus stage. (A) Uteri from mice confirmed at diestrus were collected 24 hours after TNF-α intrauterine injection to detect CFTR messenger RNA (mRNA) expression by reverse transcriptase–polymerase chain reaction (RT-PCR). Relative expression of uterine CFTR mRNA was normalized with the internal marker GAPDH. Experiments were repeated three times. The CFTR mRNA expression in the TNF-α-stimulated uteri was significantly increased compared with control (∗P<.05). (B) Cystic fibrosis transmembrane conductance regulator-mediated short circuit current (Isc) upon stimulation of TNF-α at diestrus. Representative traces of Isc response to apical addition of forskolin (10 μM) in the two groups were shown and compared. Values are means ±SEM of maximal increase in Isc. Statistical analysis showed that TNF-α pretreated endometria, but not the saline-treated controls, responded to forskolin stimulation with an increase in Isc at the diestrus stage (n = 5, ∗∗P<.01). (C) Uterine wet weights were examined after stimulation with TNF-α. Statistical analysis showed that the uterine wet weights were significantly increased after stimulation with TNF-α (n = 6, ∗P<.05).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

5. Figure 4
Up-regulated CFTR expression, ion transport, and abnormal uterine fluid accumulation by estrogen (E) and blastocyst implantation outcome. Pregnant mice were intraperitoneally injected with 0.5 μg of 17β-E2 on day 3, with saline injection as control. (A) The CFTR expression in 17β-E2 or saline pretreated mouse endometrium on pregnant day 4 (the day of blastocyst implantation). (B) Analysis showing increased uterine wet weights in uteri from 17β-E2-treated mice compared with that in control animals (n = 5, ∗∗P<.01). (C) Morphological appearance of 17β-E2 and saline-pretreated mice uterus on pregnant day 5. The control uterus exhibits obvious implantation sites, whereas the 17β-E2 pretreated one filled with fluid without blastocyst implantation. (D) Effect of CFTR up-regulation and abnormal uterine fluid accumulation on blastocyst implantation. The total numbers of viable embryos in uterine horns from 17β-E2 and saline pretreated mice on pregnant day 5 were compared. Results represent the means ±SEM of replicate assays (n = 7). Differences in implantation number between saline and 17β-E2 pretreated mice were highly significant (∗∗∗P<.001).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions