Chlamydia trachomatis (C. trachomatis) is the most prevalent sexually transmitted bacteria with devastating reproductive consequences that lead to tubal factor infertility (TFI). Recent studies have implicated apoptosis – associated speck – like protein containing a caspase recruitment domain (ASC) as an adaptor of inflammasomes that stimulate IL – 1β and IL – 18 secretion, pro – inflammatory cytokines with critical functions in host defense against a variety of pathogens. Therefore, for the first time, we are reporting the use of ASC-/- mice in a mouse model of Chlamydia infection that might provide some information on the role of inflammasomes in the pathogenesis of Chlamydia infection. In this study, wild type (WT) and ASC-/- mice were infected with Chlamydia. Infectivity, pathology of the upper genital tract (UGT), and, fertility were evaluated. In addition, expression of ASC – dependent inflammasomes and the activation of immune cells within the genital tract (GT) were studied. Results showed that Chlamydia infectivity in ASC-/- mice was significantly higher (p-/- mice which, when compared to infected WT mice, was exhibited by decrease in average number of pups and percent pregnancy. There was also severe UGT damage in ASC-/- mice compared to WT mice, correlating with the higher number of hydrosalpinx observed on the UGT of Chlamydia infected ASC-/- mice. Furthermore, IL – 1β and IL – 18 production as well as immune cell activation were down regulated in the GT of Chlamydia infected ASC-/- mice. This finding indicates that in absence of ASC, host innate and adaptive immunity is impaired. Results imply that ASC plays a protective role in the mucosal immunity against GT Chlamydia infection.

Reproductive success depends on a functional oviduct for gamete storage, maturation, fertilization, and early-conceptus development. The ovarian-derived sex steroids estradiol and progesterone are known to affect functionality of the oviduct. Advances in microarray and NanoString technology allow for gene expression analysis to increase understanding of processes critical for fertility. Studies were conducted to investigate mechanisms regulating oviductal function in cattle and mice by using the Bovine Gene 1.0 ST array and the Mouse Gene 430-2.0 arrays (Affymetrix Inc., CA), respectively. For the first study, oviducts were collected from heifers assigned to luteal or follicular phase groups. In the second study oviducts were collected from immature mice with a global deletion of estrogen receptor-1 (ESR1) and their wild-type littermates at 23 days of age or 48 hr after treatment with 5 IU of PMSG. Following microarray hybridization, the resulting datasets were analyzed using Partek Genomics Suite 6.6 (Partek Inc., MO). The results of the first two studies illustrated a dynamic hormonal regulation of the oviductal epithelium and revealed the identity of novel genes affecting fertility in cattle and gave us insights into the genes regulated by estrogen and ESR1 in mice. Many genes identified as differentially regulated are believed to play an integral role in the regulation of oviductal inflammation. Therefore, the objective of the third study was to test the hypothesis that intraperitoneal administration of E. Coli-derived lipopolysaccharide induces the expression of inflammatory mRNAs in the mouse oviduct. Mice were treated with 0, 2 μg or 10 μg of LPS from E. Coli. and killed 24 h later. Oviducts were collected for determination of inflammatory gene expression by a targeted NanoString approach using the nCounter GX Mouse Inflammation Kit (NanoString Technologies, Wa). Results indicate that systemic treatment with LPS induces inflammation in the oviducts of mice and provides evidence of a repeatable animal model of oviductal inflammation. Overall, data from these studies extends our knowledge of the mechanisms regulating oviductal functions and immune response, as well as identified target molecules and processes to improve production animal and human fertility.