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

Abstract

One of the endogenous estrogens, 17β-estradiol (E 2 ) is a female steroid hormone secreted from the ovary. It is well established that E 2 causes biochemical and histological changes in the uterus. However, it is not completely understood how E 2 regulates the oviductal environment in vivo. In this study, we assessed the effect of E 2 on each oviductal cell type, using an ovariectomized-hormone-replacement mouse model, single-cell RNA-sequencing (scRNA-seq), in situ hybridization, and celltype-specific deletion in mice. We found that each cell type in the oviduct responded to E 2 distinctively, especially ciliated and secretory epithelial cells. The treatment of exogenous E 2 did not drastically alter the transcriptomic profile from that of endogenous E 2 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 E 2 -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, including epithelial, stromal, and muscle cells, are differentially regulated by E 2 and support gene expression changes, such as growth factors that are required for normal embryo development and transport in mouse models. Furthermore, we have identified cell-specific and region-specific gene markers for targeted studies and functional analysis in vivo. K E Y W O R D Sembryo development, embryo transport, estrogen, insulin-like growth factor 1, oviduct, scRNA-seq 2 of 19 | MCGLADE Et AL.

Progesterone and Inflammatory Response in the Oviduct during Physiological and Pathological Conditions

“…Leukocytes (expressing PTPRC, also known as CD45) are the most abundant in human Fallopian tubes compared to other reproductive tissues. We have re-analyzed previously published scRNA-seq data from human Fallopian tubes [ 3 ] and found the presence of PTPRC + cells ( Figure 2 ) that are consistent with previous reports [ 37 ]. T cell marker CD52 + has also been detected at a high abundancy.…”

“…The oviduct is comprised of multiple cell types, including stromal cells (fibroblasts), ciliated epithelial cells, secretory epithelial cells, endothelial cells, and smooth muscle cells. These cells are tightly regulated by ovarian steroid hormones [ 3 ]. Oviductal epithelial cells directly interact with sperm [ 4 ], eggs [ 5 ], and embryos [ 6 ].…”

“…The Fallopian tube was originally thought to be a sterile environment. However, recent studies show that there are resident immune cells in the Fallopian tube even in the absence of infection [ 3 , 33 , 34 , 35 , 36 , 37 ]. Immunohistochemical, fluorescent-activated cell sorting (FACS), and single-cell RNA-sequencing (scRNA-seq) analyses reveal that immune cells in both innate and adaptive immune responses are present in human Fallopian tubes [ 3 , 33 , 34 , 35 , 36 , 37 ].…”

“…However, recent studies show that there are resident immune cells in the Fallopian tube even in the absence of infection [ 3 , 33 , 34 , 35 , 36 , 37 ]. Immunohistochemical, fluorescent-activated cell sorting (FACS), and single-cell RNA-sequencing (scRNA-seq) analyses reveal that immune cells in both innate and adaptive immune responses are present in human Fallopian tubes [ 3 , 33 , 34 , 35 , 36 , 37 ]. Leukocytes (expressing PTPRC, also known as CD45) are the most abundant in human Fallopian tubes compared to other reproductive tissues.…”

“…T cell marker CD52 + has also been detected at a high abundancy. However, CD14 + monocytes/macrophages [ 3 ] and NCAM1 + (also known as CD56 +) NK cells are present at low levels compared to other immune cell types [ 3 , 35 ]. Recent scRNA-seq data from pooled human Fallopian tubes show that tissue CD69 + resident memory T cells are also detected [ 37 ].…”