Wipawee Winuthayanon

University of Missouri | Mizzou · Department of Obstetrics, Gynecology and Women's Health

Contraception is a practice with extensive and complicated social and scientific histories. From cycle tracking, to the very first prescription contraceptive pill, to now having over-the-counter (OTC) contraceptives on demand, family planning is an aspect of healthcare that has undergone and will continue to undergo several transformations through...

Ciliary action performs a critical role in the oviduct (Fallopian tube) during pregnancy establishment through sperm and egg transport. The disruption of normal ciliary function in the oviduct affects oocyte pick-up and is a contributing factor to female infertility. Estrogen is an important regulator of ciliary action in the oviduct and promotes c...

Pre‐implantation embryonic development occurs in the oviduct during the first few days of pregnancy. The presence of oviductal extracellular vesicles (oEVs, also called oviductosomes) is crucial for pre‐implantation embryonic development in vivo as oEVs often contain molecular transmitters such as proteins. Therefore, evaluating oEV cargo during ea...

The oviduct is the site of fertilization and preimplantation embryo development in mammals. Evidence suggests that gametes alter oviductal gene expression. To delineate the adaptive interactions between the oviduct and gamete/embryo, we performed a multi-omics characterization of oviductal tissues utilizing bulk RNA-sequencing (RNA-seq), single-cel...

The oviduct is the site of fertilization and preimplantation embryo development in mammals. Evidence suggests that gametes alter oviductal gene expression. To delineate the adaptive interactions between the oviduct and gamete/embryo, we performed a multi-omics characterization of oviductal tissues utilizing bulk RNA-sequencing (RNA-seq), single-cel...

The epithelial cell lining of the oviduct plays an important role in oocyte pickup, sperm migration, preimplantation embryo development, and embryo transport. The oviduct epithelial cell layer comprises ciliated and non-ciliated secretory cells. The ciliary function has been shown to support gamete and embryo movement in the oviduct, yet secretory...

The oviduct is the site of fertilization and preimplantation embryo development in mammals. Evidence suggests that gametes alter oviductal gene expression. To delineate the adaptive interactions between the oviduct and gamete/embryo, we performed a multi-omics characterization of oviductal tissues utilizing bulk RNA-sequencing (RNA-seq), single-cel...

Kallikreins (KLKs) are serine peptidases. It was established that Klks are estrogen-target genes in mouse uteri. However, the functional requirement of KLK family in the uterine function during reproduction is unknown. Here we generated a compound deletion of Klk1b3, Klk1b4, Klk1b5, and Klk1 in a mouse model using CRISPR/Cas9 strategy with four sin...

Postnatal development of the uterus involves specification of undifferentiated epithelium into uterine-type epithelium. That specification is regulated by stromal-epithelial interactions as well as intrinsic cell-specific transcription factors and gene regulatory networks. This study utilized mouse genetic models of Esr1 deletion, endometrial epith...

The oviduct comprises 4 main regions: infundibulum (oocyte pick-up), ampulla (fertilization), isthmus (sperm capacitation and reservoir, preimplantation embryonic development), and uterotubal junction (UTJ; sperm and embryo transport). Mounting evidence in livestock and rodents suggest that gametes alter gene expression in secretory and ciliated ep...

Progesterone has been shown to be a potent suppressor of several inflammatory pathways. During pregnancy, progesterone levels increase, allowing for normal pregnancy establishment and maintenance. The dysregulation of progesterone, as well as inflammation, leads to poor pregnancy outcomes. However, it is unclear how progesterone imbalance could imp...

Prostate Specific Antigen (PSA) is a prostate-specific serine protease enzyme that hydrolyzes gel-forming proteins (semenogelins) and changes the semen from gel-like to watery viscosity, a process called semen liquefaction. Highly viscous semen and abnormal liquefaction reduce sperm motility and contribute to infertility. Previously, we showed that...

One of the endogenous estrogens, 17β‐estradiol (E2) is a female steroid hormone secreted from the ovary. It is well established that E2 causes biochemical and histological changes in the uterus. However, it is not completely understood how E2 regulates the oviductal environment in vivo. In this study, we assessed the effect of E2 on each oviductal...

The mammalian oviduct is a dynamic organ where important events such as final maturation of oocytes, transport of gametes, sperm capacitation, fertilization, embryo development, and transport take place. Prostaglandin-endoperoxide synthase 2 (PTGS2), also known as cyclooxygenase 2 (COX-2), is the rate-limiting enzyme in the production of prostaglan...

One of the endogenous estrogens, 17b-estradiol (E2) is a female steroid hormone secreted from the ovary. It is well established that E2 causes biochemical and histological changes in the uterus. The oviduct response to E2 is virtually unknown in an in vivo environment. In this study, we assessed the effect of E2 on each oviductal cell type, using a...

In mammals, the oviduct (or the Fallopian tube in humans) can be divided into the infundibulum (responsible for oocyte pick-up), ampulla (site of fertilization), isthmus (where preimplantation embryos develop), and uterotubal junction (where embryos transit to the uterus). The oviductal fluid, as well as extracellular vesicles produced from the ovi...

In mammals, the oviduct (or the Fallopian tube in humans) can be divided into the infundibulum (responsible for oocyte pick-up), ampulla (site of fertilization), isthmus (where preimplantation embryos develop), and uterotubal junction (where embryos transit to the uterus). The oviductal fluid, as well as extracellular vesicles produced from the ovi...

At birth, all female mice, including those that either lack estrogen receptor α (ERα-knockout) or that express mutated forms of ERα (AF2ERKI), have a hypoplastic uterus. However, uterine growth and development that normally accompanies pubertal maturation does not occur in ERα-knockout or AF2ERKI mice, indicating ERα mediated estrogen signaling is...

Semen liquefaction is a proteolytic process where a gel-like ejaculated semen becomes watery due to the enzymatic activity of prostate-derived serine proteases in the female reproductive tract. The liquefaction process is crucial for the sperm to gain their motility and successful transport to the fertilization site in Fallopian tubes (or oviducts...

Inhibition of the sperm transport process in the female reproductive tract could lead to infertility. We previously showed that a pan-serine protease inhibitor, 4-(2-aminoethyl) benzenesulfonyl fluoride (AEBSF), blocked semen liquefaction in vivo and resulted in a drastic decrease in the number of sperm in the oviduct of female mice. In this study,...

We previously demonstrated that 5’-adenosine monophosphate-activated protein kinase (AMPK) is essential for normal reproductive functions in female mice. Conditional ablation of Prkaa1 and Prkaa2, genes that encode the α1 and α2 catalytic domains of AMPK, resulted in early reproductive senescence, faulty artificial decidualization, uterine inflamma...

Estrogen receptor α (encoded by Esr1) is a crucial nuclear transcription factor for female reproduction and is expressed throughout the female reproductive tract. To assess the function of ESR1 in reproductive tissues without confounding effects from a potential developmental defect arising from global deletion of ESR1, we generated a mouse model i...

The oviduct (known as the fallopian tube in humans) is the site for fertilization and pre-implantation embryo development. Female steroid hormones, estrogen and progesterone, are known to modulate the morphology and function of cells in the oviduct. In this review, we focus on the actions of estrogen and progesterone on secretory, ciliated, and mus...

Pausing of RNA polymerase II (Pol II) during early transcription, mediated by the negative elongation factor (NELF) complex, allows cells to coordinate and appropriately respond to signals by modulating the rate of transcriptional pause release. Promoter proximal enrichment of Pol II occurs at uterine genes relevant to reproductive biology; thus, w...

In the female reproductive tract, the innate immune system is modulated by two sex steroid hormones, estrogen and progesterone. A cyclical wave of neutrophils in the vaginal lumen is triggered by chemokines and correlates with circulating estrogen levels. Classical estrogen signaling in the female reproductive tract is activated through estrogen re...

Multiciliated cells of the airways, brain ventricles, and female reproductive tract provide the motive force for mucociliary clearance, cerebrospinal fluid circulation, and ovum transport. Despite their clear importance to human biology and health, the molecular mechanisms underlying multiciliated cell differentiation are poorly understood. Prior s...

Effects of loss of CEP164 on the ciliary localization of Arl13b and INPP5E in MEFs. Mouse embryonic fibroblasts (MEFs) were prepared from E8.5 CEP164-KO or control embryos and serum-starved for 48 hours to induce primary cilia. MEFs were double-labeled for Arl13b or INPP5E (green) and the ciliary marker acetylated α-tubulin (A-tub). Nuclei were vis...

Primary antibodies used for IF staining. (TIF)

Deletion of CEP164 in multiciliated tissues leads to significant loss of multicilia in the trachea. Tracheal sections from CEP164fl/fl and FOXJ1-Cre;CEP164fl/fl adult mice were immunostained for A-tub (green). Nuclei were detected with DAPI. Scale bar, 100 μm. (TIF)

CEP164 is important for ependymal multiciliated cell maturation. (A) SVZ whole mount preparations from CEP164fl/fl or FOXJ1-Cre;CEP164fl/fl adult mice were immunostained for G-tub (white) and β-catenin (red). β-Catenin demarcates the cell boundaries, and γ-tubulin labels basal bodies that are found in patches in ependymal multiciliated cells. Scale...

Effects of CEP164 deletion on the ciliary localization of TTBK2 and Arl13b in multiciliated cells. (A) ALId14 MTECs from CEP164fl/fl and FOXJ1-Cre;CEP164fl/fl mice were immunostained for TTBK2 (green) and the ciliary/basal body maker A-tub (red). Nuclei were detected with DAPI (blue). (B) ALId5 MTECs were immunostained for Arl13b (green) and A-tub...

Efficient removal of CEP164 by FOXJ1-Cre-mediated recombination in multiciliated cells in MTEC cultures. (A) MTECs were prepared from CEP164fl/fl and FOXJ1-Cre;CEP164fl/fl mice, fixed at ALId14, and immunostained for FOXJ1 (green) and CEP164 (red). Nuclei were stained using DAPI (blue). ~90% of multiciliated cells in MTEC cultures from FOXJ1-Cre;CE...

Transmission electron microscopy reveals short cilia as well as intact transition fibers and transition zone structures in CEP164-KO multiciliated cells. (A) Structure of multicilia. CP, cilia proper; BP, basal plate; TZ, transition zone, BB, basal body; TF, transition fiber (arrowheads). Scale bar, 100 nm. (B) Elongated cilia were abundant in cros...

Generation of FOXJ1-Cre;CEP164fl/fl mice. (A) Schematic diagram of CEP164 protein structure illustrating the WW domain and the three coiled-coiled (CC) domains. The N-terminal portion of the protein encoded by exon 4 (ex4), which was removed upon Cre-mediated recombination, is depicted. The numbers indicate amino acid positions. (B) Shown are the o...

Aberrant regulation of uterine cell growth can lead to endometrial cancer and infertility. To understand the molecular mechanisms of estrogen-induced uterine cell growth, we removed the estrogen receptor α (Esr1) from mouse uterine stromal cells, where the embryo is implanted during pregnancy. Without ESR1 in neighboring stroma cells, epithelial ce...

In mice, ejaculated semen is deposited in the uterus. After ejaculation, the semen changes consistency from gel-like to watery, a process called liquefaction. In this study, we show how to collect the post-ejaculated semen from the female reproductive tract in a mouse model. First, adult female mice in the estrus stage were housed in a male's cage...

Estrogen (E2) signaling through its nuclear receptor, estrogen receptor α (ERα) increases insulin-like growth factor 1 (IGF1) in the rodent uterus, which then initiates further signals via the IGF1 receptor (IGF1R). Directly administering IGF1 results in similar biological and transcriptional uterine responses. Our studies using global ERα-null mic...

Semen liquefaction changes semen from a gel-like to watery consistency and is required for sperm to gain mobility and swim to the fertilization site in the Fallopian tubes. Kallikrein-related peptidases 3 (KLK3) and other kallikrein-related peptidases from male prostate glands are responsible for semen liquefaction by cleaving gel-forming proteins...

List of primer sequencesπ used in this study. π m and h prefixes refer to mouse and human primers, respectively. (DOCX)

Representative video recording of the semen freshly collected from Wnt7aCre/+;Esr1f/f uteri approximately 8 h after mating at 1000x magnification with a speed of 12 frames/second (n = 3 females). (AVI)

Microarray analysis showing all the transcript levelsπ of aquaporins detected in the uterus of Esr1f/f and Wnt7aCre/+;Esr1f/f female 2 h and 24 h after E2 treatment. π All the transcripts detected in the gene family in the uteri were listed in the table regardless of the fold changes. Transcript levels indicated by the raw signal intensities; cut o...

Representative video recording of the semen freshly collected from control uteri approximately 8 h after mating at 1000x magnification with a speed of 12 frames/second (n = 3 females). (AVI)

Diarylheptanoids from Curcuma comosa (C. comosa), Zingiberaceae family, exhibit diverse estrogenic activities. In this study we investigated the estrogenic activity of a major hydroxyl diarylheptanoid, 7-(3,4 dihydroxyphenyl)-5-hydroxy-1-phenyl-(1E)-1-heptene (compound-092) isolated from C. comosa. The compound elicited different transcriptional ac...

Newly fertilized embryos spend the first few days within the oviduct and are transported to the uterus where they implant onto the uterine wall. An implantation of the embryo before reaching the uterus could result in ectopic pregnancy and lead to maternal death. Estrogen is necessary for embryo transport in mammals; however, the mechanism involved...

Animal oviducts and human Fallopian tubes are a part of the female reproductive tract that hosts fertilization and pre-implantation development of the embryo. With an increasing understanding of roles of the oviduct at the cellular and molecular levels, current research signifies the importance of the oviduct on naturally conceived fertilization an...

Estrogen receptor alpha (ERα) is a critical player in development and function of the female reproductive system. Perturbations in ERα response can affect wide-ranging aspects of health in humans as well as in livestock and wildlife. Because of its long-known and broad impact, ERα mechanisms of action continue to be the focus on cutting-edge resear...

The coordinated functions of the ovary and reproductive tract are critical to mammalian fertility. Receptors for ovarian steroids within the uterus and ovary are central mediators of ovulation, implantation, and gestation, thus understanding of cellular localization and mechanisms involved in their functions in these tissues is essential knowledge...

Concerns regarding potential endocrine disrupting chemicals (EDCs) have led to a need for methods to evaluate candidate estrogenic chemicals. Our previous evaluations of two such EDCs revealed a response similar to that of estradiol (E₂) at 2 hours, but a less robust response at 24 hours, similar to the short-acting estrogen, estriol (E<...

Estrogens exert their activity through estrogen receptor alpha (ERalpha) to stimulate hypertrophy and hyperplasia in the uterus. A uterine epithelial ERα conditional knockout mouse model (Wnt7a(Cre+);Esr1(f/f) or cKO) demonstrated that ER&alpha in the epithelial cells was dispensable for an initial uterine proliferative response to 17beta-estradiol...

Estrogen Receptor α (ERα) interacts with DNA, directly, or indirectly via other transcription factors, referred to as "tethering". Evidence for tethering is based on in vitro studies and a widely used "KIKO" mouse model containing mutations that prevent direct estrogen response element (ERE) DNA-binding. KIKO mice are infertile, due in part to the...

Background: Diarylheptanoid (D3) isolated from the medicinal plant, Curcuma comosa, has estrogenic activity. Objective: We aimed to elucidate the mechanism(s) of D3 action and compare it with that of 17β-estradiol (E2) using both in vitro and in vivo uterine models. Methods: We used human uterine (Ishikawa) cells to determine the estrogenic action...

Estrogen receptor a (ERa) is a crucial nuclear transcription factor for female reproduction. ERa is expressed throughout the female reproductive tract, including the ovary, oviduct and uterus. We hypothesized that selective deletion of ERa from the oviduct and uterus would lead to female infertility and lack of response to estrogen (E) in the uteru...

Mammalian fertilization and preimplantation embryo development occur in the oviduct within a microenvironment strictly controlled by steroid hormones. Changes in oviductal cytology in each cell compartment are the most apparent during the estrous stage of the ovarian cycle, immediately prior to fertilization. The goal of this study was to elucidate...

The mouse uterus is a sensitive biological system in which to assess the mechanisms of estrogen response. Biological, biochemical and genomic events initiated by estrogens have been well-characterized and described, allowing assessment of chemicals with suspected estrogenic activity by comparison to the signature responses of proven estrogens. Some...

Female fertility requires estrogen to specifically stimulate estrogen receptor α (ERα)-dependent growth of the uterine epithelium in adult mice, while immature females show proliferation in both stroma and epithelium. To address the relative roles of ERα in mediating estrogen action in uterine epithelium versus stroma, a uterine epithelial-specific...

Diarylheptanoids isolated from Curcuma comosa Roxb. have been recently identified as phyto estrogens. However, the mechanism underlying their actions has not yet been identified. We characterized the estrogenic activity of three active naturally occurring diarylheptanoids both in vitro and in vivo. We characterized mechanisms of estrogenic action o...

Xeno-oestrogens are man-made compounds that have oestrogenic-like activity, and exposure to xeno-oestrogens in critical periods of development and during adulthood may adversely affect both the male and female reproductive systems and fertility. For example, some studies suggest semen quality is declining globally due to xeno-oestrogen exposure, al...

Curcuma comosa Roxb. has traditionally been used as a dietary supplement for health promotion in peri- and postmenopausal women in Thailand. We investigated the estrogenic activity of 7 naturally occurring diarylheptanoids from the extracts of C. comosa both in vitro and in vivo. A yeast recombinant system containing human estrogen receptor alpha,...