Mating induces gene expression shift in chicken oviduct
Ontology highlight
ABSTRACT: We compared gene expression in oviduct tissues between unmated (control) and mated hen. As spermatozoa are foreign to the female reproductive tract therefore we were interested to look at how spermatozoa survive in the female reproductive tract and keep their fertilization potentiality. To check that we collected tissues from oviduct of both control and mated female chicken and compared if sperm deposition to the oviduct made any gene expression shift related to sperm survival.
Project description:The objective of the present study is to investigate if females have the ability to recognise X or Y chromosome bearing spermatozoa and present a different response to different spermatozoa. In order to investigate if females have the ability to recognise X or Y chromosome bearing spermatozoa and present a different response to different spermatozoa. Samples of semen were sorted for chromosomal sex by M-oM-,M-^Bow cytometric-sorting. Using laparoscopic insemination one oviduct was inseminated with X spermatozoa and the contralateral oviduct was inseminated with Y spermatozoa.This allowed us to obtain samples from the oviduct in contact with X sperm and Y sperm in the the contrlateral oviduct from the same sow. Oviduct pig samples were collected for RNA isolation and hybridization on Affymetrix microarrays. Four biological replicates were performed (n= 4 sows) and a total of 8 arrays were used for microarrays study (4 arrays for oviduct samples in contact with X spermatozoa and 4 arrays for oviduct samples in contact with Y spermatozoa.
Project description:We specifically deleted Mettl3 in the female reproductive tract using the Pgr-Cre driver. To portray the molecular mechanism for Mettl3 function in oviduct, whole oviduct were collected from Mettl3f/f and Mettl3d/d mice on gestational day 3 and subjected to RNA-seq analysis.
Project description:Male-derived accessory gland proteins (Acps) that are transferred to females during mating have profound effects on female reproductive physiology including increased ovulation, mating inhibition, and effects on sperm utilization and storage. The extreme rates of evolution seen in Acps may be driven by sperm competition and sexual conflict, processes which may ultimately drive complex interactions between female- and male-derived molecules and sperm. However, little is known of how gene expression in female reproductive tissues changes in response to the presence of male molecules and sperm. To characterize this response, we conducted parallel genomic and proteomic analyses of gene expression in the reproductive tract of 3-day-old unmated and mated female Drosophila melanogaster. Using DNA microarrays, we identified 539 transcripts that are differentially expressed in unmated vs. mated females and revealed a striking peak in differential expression at 6 hrs postmating and a marked shift from primarily down-regulated to primarily up-regulated transcripts within 3 hrs after mating. Combining two-dimensional gel electrophoresis and liquid chromatography mass spectrometry analyses, we identified 84 differentially expressed proteins at 3 hrs postmating, including proteins which appeared to undergo post-translational modification. Together, our observations define transcriptional and translational response to mating within the female reproductive tract and suggest a bimodal model of postmating gene expression initially correlated with mating and the final stages of female reproductive tract maturation and later with the declining presence of male reproductive molecules and with sperm maintenance and utilization. Experiment Overall Design: Three-day-old mated and unmated females were dissected to remove the lower reproductive tract (upper uterus, sperm-storage organs, and accessory glands). Mated females were dissected either immediately following mating (0 hr) or at 3, 6, or 24 hrs following the termination of mating. Tracts of 12-40 females of like category were pooled and total RNA extracted via a TRIzol-based protocol. Processing and labeling of transcript was performed by the Molecular Biology Core Facility at the Medical College of Georgia. Arrays from mated females at the different timepoints were compared to unmated females.
Project description:Female fertility in mammals requires the iterative remodeling of the entire adult female reproductive tract across the menstrual/estrous cycle. Here we examine global transcriptional dynamics of the mouse oviduct along the anteroposterior axis and across the estrous cycle. Though we observed robust patterns of differential oviduct gene expression along the anteroposterior axis, we found surprisingly few changes in gene expression across the estrous cycle, in marked contrast to other mammals. We speculate that this is an evolutionarily derived state that may reflect the extremely rapid five-day mouse estrous cycle.
Project description:Male-derived accessory gland proteins (Acps) that are transferred to females during mating have profound effects on female reproductive physiology including increased ovulation, mating inhibition, and effects on sperm utilization and storage. The extreme rates of evolution seen in Acps may be driven by sperm competition and sexual conflict, processes which may ultimately drive complex interactions between female- and male-derived molecules and sperm. However, little is known of how gene expression in female reproductive tissues changes in response to the presence of male molecules and sperm. To characterize this response, we conducted parallel genomic and proteomic analyses of gene expression in the reproductive tract of 3-day-old unmated and mated female Drosophila melanogaster. Using DNA microarrays, we identified 539 transcripts that are differentially expressed in unmated vs. mated females and revealed a striking peak in differential expression at 6 hrs postmating and a marked shift from primarily down-regulated to primarily up-regulated transcripts within 3 hrs after mating. Combining two-dimensional gel electrophoresis and liquid chromatography mass spectrometry analyses, we identified 84 differentially expressed proteins at 3 hrs postmating, including proteins which appeared to undergo post-translational modification. Together, our observations define transcriptional and translational response to mating within the female reproductive tract and suggest a bimodal model of postmating gene expression initially correlated with mating and the final stages of female reproductive tract maturation and later with the declining presence of male reproductive molecules and with sperm maintenance and utilization. Keywords: keywords: reproduction, reproductive tract, accessory gland proteins, sperm, timecourse
Project description:Estrogen receptor α (ERα) is a nuclear transcription factor crucial for the female reproductive function. We previously reported that mice lacking epithelial ERα in the epithelial cells of female reproductive tract (Wnt7aCre+;Esr1fl/fl, conditional knockout or cKO) were infertile, in part, due to an implantation defect. To determine if oviductal dysfunction also contributed to their infertility, we examined the fertilization rates and embryo development in vivo during the first few days of pregnancy. At 0.5 days post coitum (dpc), cKO females had significantly fewer zygotes than wild type control littermates (WT). At 1.5 dpc, cKO females had no 2-cell embryos at all; only dead oocytes or embryos and empty zona pellucidas were observed. These results indicate that lack of ERα in the oviductal epithelium resulted in alterations in the oviductal microenvironment that were detrimental to the embryos. Microarray analysis revealed dramatic differences in gene expression between cKO and WT oviducts collected at 0.5 dpc and significant but less dramatic differences at 1.5 dpc. These findings indicate that signaling via epithelial ERα is essential to generate an oviductal milieu supportive of fertilization and embryo development.and may have implications for infertility in women. The oviduct samples were collected from female reproductive tract epithelial ER α knockout (cKO) and wild type control littermates (WT) at days 1 and 2 of pregnancy (0.5 and 1.5 days post coitum). Each experimental group contains 4 replicates. Gene expression analysis was conducted by using Agilent Whole Mouse Genome 4�44 multiplex format oligo arrays (no. 014868; Agilent Technologies, Santa Clara, CA)
Project description:Proteins play an important role in many reproductive functions such as sperm maturation and physiology, sperm transit in the female genital tract or sperm-oocyte interaction. However, little information concerning reproductive features is available in the case of aquatic animals. The present study aims to characterize the proteome of both spermatozoa and seminal plasma of bottlenose dolphins (Tursiops truncatus) as a model organism for cetaceans. Ejaculate samples were obtained voluntarily from two trained dolphins housed in an aquarium. Spermatozoa and seminal plasma were analyzed by means of proteomic analyses using a LC-MS/MS and a list with the gene symbols corresponding to each protein was submitted to DAVID database. A total of 423 and 307 proteins were identified in spermatozoa and seminal plasma, respectively, and the samples shared 111 proteins. Furthermore, 70 proteins were identified as involved in reproductive processes, 39 in spermatozoa, and 31 in seminal plasma. The top-5 most abundant proteins were also identified in these samples: AKAP3, ODF2, TUBB, GST3, ROPN1 for spermatozoa and CST11, LTF, ALB, HSP90B1, PIGR for seminal plasma. In conclusion, this proteomic study provides the first characterization of proteomic expression patterns in cetacean sperm and seminal plasma, opening a window to future research on the discovery of biomarkers, analysis of conservation capacity or additional applications in the field of assisted reproduction technologies.
Project description:The aim of the study was to assess the cellular composition and transcriptional changes at single-cell resolution of the female reproductive tract across the four stages of estrus cycle in young adult mice (ovary, oviduct, uterus, cervix and vagina, with spleen as a comparison) and compared this with similar analyses of decidualization (uterus) and ageing (ovary, oviduct, uterus, cervix, vagina and spleen). Vaginal smears were used to stage the estrus cycle in young mice. To collect the decidualized uteri mice were mated and uterine tissue was collected from pregnant mice at day 5.5 after mating. Tissues of aged mice were collected at the age of 18 months. All tissues were collected and digested using a collagenase and trypsin protocol to obtain single cell suspension. Libraries were prepared using the 10x scRNA-seq protocol.
Project description:Sperms being foreign to the female are to be promptly eliminated by the female local immune defense. However, avoiding the local immune defense sperm can be stored for lenthy period in the oviductal sperm reservoir. It is currently unknown whether oviductal sperm reservoirs changes their gene expression to tolerate the spermatozoa after mating or sperm free seminal plasma infusion. Therefore this was tested using Swedish Landrace pigs in this study using cDNA microarray. We used 12 sows seperated into three groups- either oestrus sows were inseminated with 50 ml BTS (control, n=4) or mated with boars (treatment 1, n=4) or inseminated with sperm-free seminal plasma (treatment 2, n=4). The utero-tubal junction was retrieved within 24 h of treatment by operation.
Project description:The seminal vesicles are male accessory sex glands that contribute the bulk of the seminal plasma in which mammalian spermatozoa are bathed during ejaculation. In addition to helping convey sperm through the ejaculatory duct, seminal vesicle secretions support sperm survival after ejaculation and influence the female reproductive tract to promote receptivity to pregnancy. Despite its biological importance, there is limited understanding of seminal vesicle fluid (SVF) composition with only 75 proteins having been identified in publicly available proteomic analyses. To address this limitation, here we report new preparative methodology involving sequential solubilization of mouse SVF in guanidine hydrochloride, prior to acetone precipitation and analysis by label-free quantitative mass spectrometry. This strategy identified 126 proteins, including 83 previously undetected in SVF. Members of the seminal vesicle secretory protein family were the most abundant, accounting for 79% of all peptide spectrum matches. Functional analysis of SVF proteins identified inflammation and formation of the vaginal plug as the two most prominent biological processes. Notable additional processes included modulation of sperm function and regulation of the female reproductive tract immune environment. Together, these findings provide a robust methodological framework for future SVF studies and identify novel proteins with the potential to influence both male and female reproductive health.