Biology subjects

Winuthayanon, W.

Publications and source records attributed to Winuthayanon, W..

5 recordsLinked to original sources

bioRxiv2026

Method for modeling oviduct function and impact on embryonic development

Direct experimental analysis of the mammalian oviduct is constrained by limited tissue access and the short lifespan of ex vivo preparations. Extracellular matrix-embedded three-dimensional epithelial organoids provide longer-term in vitro models. However, their inward-facing apical surface and the absence of supporting stromal cells limit physiological studies of the oviduct, including ciliary activity and maternal-embryonic interactions. Here, we provide a step-wise protocol detailing the generation of mouse and human oviductal assembloids in which epithelial cells form an outward-facing (apical-out) layer around a stromal core. Epithelial and stromal cells from adult mouse oviducts or human Fallopian tubes are isolated, expanded separately, and subsequently aggregated in a rotational culture system. The protocol also outlines morphological and immunostaining criteria for confirming cellular organization, whole-mount detection of external cilia, measurement of ciliary beat frequency, and co-culture of mouse assembloids with preimplantation embryos. Mouse and human assembloids retained epithelial and stromal identity and displayed cilia at the accessible outer surface. In a proof-of-concept experiment, embryos co-cultured with the assembloids developed to blastocysts at a rate similar to that of in vivo-derived blastocysts. This reductionist system provides a straightforward and tractable model to investigate oviduct physiology and embryo-maternal communication while allowing direct manipulation and observation of the epithelial interface. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=148 SRC="FIGDIR/small/743297v1_ufig1.gif" ALT="Figure 1"> View larger version (50K): org.highwire.dtl.DTLVardef@1acb462org.highwire.dtl.DTLVardef@17695f8org.highwire.dtl.DTLVardef@6fd6c3org.highwire.dtl.DTLVardef@7cc470_HPS_FORMAT_FIGEXP M_FIG C_FIG SummaryThe protocol for generating mouse and human oviductal assembloids by combining epithelial and stromal cells for studying oviductal function in an in vitro setting.

cell biology↗

bioRxiv2025

Vaginal epithelial estrogen receptor α coordinates glycogen deposition, microbial stability, and pH regulation in mice

Estrogen plays a central role in regulating the vaginal environment, but the specific contribution of epithelial estrogen receptor (ESR1) to microbial and biochemical homeostasis has not been fully defined. In our previous work, we showed that epithelial ESR1 is indispensable for estrogen-induced epithelial proliferation, cornification, and MUC1 expression. Here, using mice with conditional deletion of Esr1 in vaginal epithelial cells, called epithelial Esr1d/d, we extend these findings to demonstrate that epithelial ESR1 also regulates glycogen deposition, luminal pH, and microbial stability. Compared to control littermates, epithelial Esr1d/d mice reduced glycogen abundance, elevated vaginal pH, and a compositional shift in the vaginal microbiome, marked by enrichment of Comamonadaceae and loss of Lactobacillus species, without significant differences in alpha diversity. These changes parallel features of postmenopausal dysbiosis in women. Together, our findings identify epithelial ESR1 as a master regulator of multiple pathways that sustain vaginal homeostasis, integrating epithelial metabolism, barrier function, and host-microbe interactions. This work provides a mechanistic framework to understand postmenopausal vaginal dysbiosis and suggests epithelial estrogen signaling as a potential therapeutic target for genitourinary syndrome of menopause. Significance StatementThe vaginal environment is essential for reproductive and gynecologic health, yet the mechanisms by which estrogen shapes this niche remain incompletely understood. We show that epithelial estrogen receptor (ESR1) regulates glycogen deposition, luminal pH, and microbial composition in the murine vagina. Loss of epithelial ESR1 reduced glycogen and increased luminal pH without altering overall microbial diversity, but shifted community structure toward enrichment of Comamonadaceae, a family associated with neutral to mildly alkaline environments. These findings identify epithelial ESR1 as a key regulator of the metabolic and physicochemical conditions that maintain vaginal microbial balance and provide a mechanistic framework for understanding postmenopausal dysbiosis.

physiology↗

bioRxiv2024

Multi-omics analyses and machine learning prediction of oviductal responses in the presence of gametes and embryos

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-cell RNA-sequencing (scRNA-seq), and proteomics collected from distal and proximal at various stages after mating in mice. We observed robust region-specific transcriptional signatures. Specifically, the presence of sperm induces genes involved in pro-inflammatory responses in the proximal region at 0.5 days post-coitus (dpc). Genes involved in inflammatory responses were produced specifically by secretory epithelial cells in the oviduct. At 1.5 and 2.5 dpc, genes involved in pyruvate and glycolysis were enriched in the proximal region, potentially providing metabolic support for developing embryos. Abundant proteins in the oviductal fluid were differentially observed between naturally fertilized and superovulated samples. RNA-seq data were used to identify transcription factors predicted to influence protein abundance in the proteomic data via a novel machine learning model based on transformers of integrating transcriptomics and proteomics data. The transformers identified influential transcription factors and correlated predictive protein expressions in alignment with the in vivo-derived data. Lastly, we found some differences between inflammatory responses in sperm-exposed mouse oviducts compared to hydrosalpinx fallopian tubes from patients. In conclusion, our multi-omics characterization and subsequent in vivo confirmation of proteins/RNAs indicate that the oviduct is adaptive and responsive to the presence of sperm and embryos in a spatiotemporal manner. Significance StatementWe conducted a detailed molecular study of how the oviduct changes its gene expression and protein production in response to sperm and embryos after mating in mice. We found that the oviduct has distinct molecular signatures in different regions - upper versus lower regions. Shortly after mating, inflammatory responses are turned on in the lower regions due to the presence of sperm. A day later, metabolic genes ramp up in the lower regions, likely to provide nutrients for the developing embryos. Overall, this multi-omics study revealed that the oviduct dynamically adapts its molecular makeup over time and space to accommodate and support sperm, eggs and embryos.

cell biology↗

bioRxiv2022

Sperm-mediated regulation of region-specific responses in the oviduct during establishment of pregnancy in mice

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 epithelial cells of the oviduct. To elucidate whether adaptive interactions between the oviduct and gamete/embryo exist, we performed bulk RNA-sequencing on oviductal tissues collected from infundibulum+ampulla (IA) or isthmus+UTJ (IU) at various developmental stages (0.5, 1.5, 2.5-, and 3.5-days post coitus (dpc)) in mice. Samples were also collected during days 0.5, 1.5, 2.5, and 3.5 of pseudopregnancy (dpp). We found a strong region (IA vs. IU)-specific expression of large clusters of genes. The transition from 0.5 dpc to other pregnancy timepoints induces large sets of differentially expressed genes (DEGs) in pregnancy and pseudopregnancy in both IA and IU regions. Specifically, genes involved in pro-inflammatory responses were detected in both IU and IA regions. The presence of sperm at 0.5 dpc induces DEGs involved in pro-inflammatory responses in the IU region with an enrichment of biological processes for inflammatory cytokines, macrophage, and neutrophil recruitment. Additionally, DEGs are enriched in mitogen-activated protein kinase (MAPK) pathways along with genes in the Dusp family, Map3k8, Il1b, and Il1r2, among others. However, at 1.5 dpc we observed a strong shift to an anti-inflammatory condition in the IU region. These observations were absent in 0.5 and 1.5 dpp, suggesting that the DEGs observed for those inflammatory responses during pregnancy were likely induced by the presence of sperm. scRNA-seq analysis revealed that the inflammatory responsive genes were likely produced by secretory epithelial cells, compared to other cell types in the oviduct. In addition, multiple DEGs involved in pyruvate and glycolysis were enriched in the IU region, which could provide metabolic support for developing embryos. Lastly, we have also identified that there were cells that express immune markers in the oviduct, indicating that the oviduct is an immuno-dynamic tissue. In conclusion, our findings indicate that the oviduct is adaptive and responsive to the presence of sperm and embryos in a spatiotemporal manner. In this report, we intend to disseminate our findings on the transcriptional profiles during different stages of pregnancy. The complete study and validation at the protein level are currently underway and will be updated as soon as the data are available.

cell biology↗

bioRxiv2020

Cell-type specific analysis of physiological action of estrogen in mouse oviducts

One of the endogenous estrogens, 17{beta}-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 an ovariectomized-hormone-replacement mouse model, single cell RNA-sequencing (scRNA-seq), in situ hybridization, and cell-type-specific deletion in mice. We found that each cell type in the oviduct responded to E2 distinctively, especially ciliated and secretory epithelial cells. The treatment of exogenous E2 did not drastically alter the transcriptomic profile from that of endogenous E2 produced during estrus. Moreover, we have identified and validated genes of interest in our datasets that may be used as cell- and region-specific markers in the oviduct. Insulin-like growth factor 1 (Igf1) was characterized as an E2-target gene in the mouse oviduct and was also expressed in human Fallopian tubes. Deletion of Igf1 in progesterone receptor (Pgr)-expressing cells resulted in female subfertility, partially due to an embryo developmental defect and embryo retention within the oviduct. In summary, we have shown that oviductal cell types are differentially regulated by E2 and support gene expression changes that are required for normal embryo development and transport in mouse models.

cell biology↗