Steroid-dependent regulation of the oviduct: A cross-species transcriptomal analysis

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

Differential Expression of mRNA Encoding Cytokines and Chemokines in the Reproductive Tract after Infection of Mice with \u3cem\u3eChlamydia trachomatis\u3c/em\u3e

Infection with Chlamydia trachomatis targets epithelial cells within the genital tract which respond by secreting chemokines and cytokines. Persistent inflammation can lead to fibrosis, tubal infertility and/or ectopic pregnancy; many infections are asymptomatic. Most studies have investigated the inflammatory response in the initial stages of infection, less is known about the later stages of infection, especially with a low, potentially asymptomatic, bacterial load. Our objective was to determine the inflammatory mediators involved in clearance of low-grade infection and the potential involvement in chronic inflammation. Six to eight week old C3H/HeJ mice were pretreated with 2.5 mg medroxyprogesterone acetate on day -10 and -3 before infection. Mice (n=3 for 28 d, n=3 for 35 d) were infected with 5 × 102 inclusion-forming units of C. trachomatis, serovar D; vaginal cultures were obtained weekly to monitor infection. Control mice (n=3 for 28 d, n=3 for 35 d) were sham infected. Mice were killed on day 28 (experiment 1) and day 35 (experiment 2) post-infection and vaginal tissue, uterine horns and oviducts collected for analysis of mRNAs encoding inflammatory cytokines and chemokines. Total RNA was isolated and a superarray analysis performed using mouse Cytokines and Chemokines PCR arrays (Qiagen, Valencia, CA). Statistical differences in gene expression were determined using a paired Students t-test. At 28 days after infection, the expression of mRNA encoding 6, 35 and 3 inflammatory genes differed from controls in vaginal, uterine and oviductal tissues, respectively (P \u3c 0.05). At 35 days after infection, the expression of mRNA encoding 16, 38 and 14 inflammatory genes differed from controls in vaginal, uterine and oviductal tissues, respectively (P \u3c 0.05). Understanding the mechanisms involved in the inflammatory response at later stages of infection should aid in the development of treatment options that minimize the development of asymptomatic, chronic inflammation-induced infertility

Estrogen Receptor Alpha (ESR1)-Dependent Regulation of the Mouse Oviductal Transcriptome

Estrogen receptor-α (ESR1) is an important transcriptional regulator in the mammalian oviduct, however ESR1-dependent regulation of the transcriptome of this organ is not well defined, especially at the genomic level. The objective of this study was therefore to investigate estradiol- and ESR1-dependent regulation of the transcriptome of the oviduct using transgenic mice, both with (ESR1KO) and without (wild-type, WT) a global deletion of ESR1. Oviducts were collected from ESR1KO and WT littermates at 23 days of age, or ESR1KO and WT mice were treated with 5 IU PMSG to stimulate follicular development and the production of ovarian estradiol, and the oviducts collected 48 h later. RNA extracted from whole oviducts was hybridized to Affymetrix Genechip Mouse Genome 430–2.0 arrays (n = 3 arrays per genotype and treatment) or reverse transcribed to cDNA for analysis of the expression of selected mRNAs by real-time PCR. Following microarray analysis, a statistical two-way ANOVA and pairwise comparison (LSD test) revealed 2428 differentially expressed transcripts (DEG’s, P \u3c 0.01). Genotype affected the expression of 2215 genes, treatment (PMSG) affected the expression of 465 genes, and genotype x treatment affected the expression of 438 genes. With the goal of determining estradiol/ESR1-regulated function, gene ontology (GO) and bioinformatic pathway analyses were performed on DEG’s in the oviducts of PMSG-treated ESR1KO versus PMSG-treated WT mice. Significantly enriched GO molecular function categories included binding and catalytic activity. Significantly enriched GO cellular component categories indicated the extracellular region. Significantly enriched GO biological process categories involved a single organism, modulation of a measurable attribute and developmental processes. Bioinformatic analysis revealed ESR1-regulation of the immune response within the oviduct as the primary canonical pathway. In summary, a transcriptomal profile of estradiol- and ESR1-regulated gene expression and related bioinformatic analysis is presented to increase our understanding of how estradiol/ESR1 affects function of the oviduct, and to identify genes that may be proven as important regulators of fertility in the future

UK Ag Equine Programs 2015 Calendar: A 12-Month Planning Calendar for the Care and Use of Your Horses

The information in this calendar is provided to aid owners in planning for the care and use of their horses. When necessary, information is discussed in the month prior to application to allow horse owners adequate time to plan for activities such as weed control, soil testing, and vaccinations. Contact your local veterinarian for health-related issues and your county extension agent for further information. Phone numbers are listed at the end of the calendar

UK Ag Equine Programs 2016 Calendar: A 12-Month Planning Calendar for the Care and Use of Your Horses

The information in this calendar is provided to aid owners in planning for the care and use of their horses for the whole year. When necessary, information is discussed in the month prior to application to allow horse owners adequate time to plan for activities such as weed control, soil and feed testing, vaccinations, etc. Contact your local veterinarian for health-related issues and your county extension ANR, 4-H, or FCS agent for further information. County office phone numbers are listed at the end of the calendar

UK Ag Equine Programs 2014 Calendar: A 12-Month Planning Calendar for the Care and Use of Your Horses

The information in this calendar is provided to aid owners in planning for the care and use of their horses. When necessary, information is discussed in the month prior to application to allow horse owners adequate time to plan for activities such as weed control, soil testing, and vaccinations. Contact your local veterinarian for health-related issues and your county extension agent for further information. Phone numbers are listed at the end of the calendar

Principal component analysis (PCA) of the microarray-derived transcriptomal results for oviducts collected from ESR1KO mice and WT littermates at 23 days of age, or treated with 5 IU PMSG at 23 days of age and collected 48 h later.

<p>Red: ESR1KO, Blue: WT, Green: PMSG-treated ESR1KO, Purple: PMSG-treated WT.</p

Most highly significant canonical pathways identified in the oviducts of PMSG-treated ESR1KO versus PMSG-treated WT identified using QIAGEN’S Ingenuity Pathway Analysis.

<p>Most highly significant canonical pathways identified in the oviducts of PMSG-treated ESR1KO versus PMSG-treated WT identified using QIAGEN’S Ingenuity Pathway Analysis.</p

Gene Ontology (GO) analysis with Molecular Function, Cellular Component and Biological Processes categories.

<p>Pie chart shows the distribution of the DEG’s in the oviducts of PMSG-treated ESR1KO versus PMSG-treated WT mice that were matched to A) a Molecular Function, B) a Cellular Component, and C) a Biological Process, using GO.</p