Overactivation of hedgehog signaling in the developing Müllerian duct interferes with duct regression in males and causes subfertility

The influence of the hedgehog signaling pathway on reproduction was studied in transgenic mice in which a dominant active allele of the hedgehog signal transducer, smoothened (Smo), was conditionally expressed in the developing Müllerian duct and gonads through recombination mediated by anti-Müllerian hormone receptor 2-cre (Amhr2cre). Previous studies showed that development of the oviduct and uterus are abnormal in femaleAmhr2cre/+SmoM2mice. In the current study, focusing on mutant males, litter size was reduced 53% in crosses with wild-type females. An extra band of undifferentiated tissue extended along each epididymis and vas deferens, a position suggesting derivation from Müllerian ducts that failed to regress fully. Hedgehog signaling was elevated in this tissue, based on mRNA levels of target genes.Amhr2mRNA was dramatically reduced in the uterus of mutant females and in the extra tissue in the tract of mutant males, suggesting that AMHR2 signaling was inadequate for complete Müllerian duct regression. Spermatogenesis and sperm motility were normal, but testis weight was reduced 37% and epididymal sperm number was reduced 36%. The number of sperm recovered from the uteri of wild-type females after mating with mutant males was reduced 78%. This suggested that sperm transport through the male tract was reduced, resulting in fewer sperm in the ejaculate. Consistent with this, mutant males had unusually tortuous vas deferentia with constrictions within the lumen. We concluded that persistence of a relatively undifferentiated remnant of Müllerian tissue is sufficient to cause subtle changes in the male reproductive tract that reduce fertility.

Download Full-text

Proteolysis of cubitus interruptus in Drosophila requires phosphorylation by protein kinase A

Development ◽

10.1242/dev.126.19.4331 ◽

1999 ◽

Vol 126 (19) ◽

pp. 4331-4339 ◽

Cited By ~ 12

Author(s):

M.A. Price ◽

D. Kalderon

Keyword(s):

Protein Kinase ◽

Protein Kinase A ◽

Hedgehog Signaling ◽

Target Genes ◽

Ectopic Expression ◽

Full Length ◽

Mutant Form ◽

Wild Type ◽

Cubitus Interruptus ◽

Kinase A

The Hedgehog signal transduction pathway is involved in diverse patterning events in many organisms. In Drosophila, Hedgehog signaling regulates transcription of target genes by modifying the activity of the DNA-binding protein Cubitus interruptus (Ci). Hedgehog signaling inhibits proteolytic cleavage of full-length Ci (Ci-155) to Ci-75, a form that represses some target genes, and also converts the full-length form to a potent transcriptional activator. Reduction of protein kinase A (PKA) activity also leads to accumulation of full-length Ci and to ectopic expression of Hedgehog target genes, prompting the hypothesis that PKA might normally promote cleavage to Ci-75 by directly phosphorylating Ci-155. Here we show that a mutant form of Ci lacking five potential PKA phosphorylation sites (Ci5m) is not detectably cleaved to Ci-75 in Drosophila embryos. Moreover, changes in PKA activity dramatically altered levels of full-length wild-type Ci in embryos and imaginal discs, but did not significantly alter full-length Ci5m levels. We corroborate these results by showing that Ci5m is more active than wild-type Ci at inducing ectopic transcription of the Hh target gene wingless in embryos and that inhibition of PKA enhances induction of wingless by wild-type Ci but not by Ci5m. We therefore propose that PKA phosphorylation of Ci is required for the proteolysis of Ci-155 to Ci-75 in vivo. We also show that the activity of Ci5m remains Hedgehog responsive if expressed at low levels, providing further evidence that the full-length form of Ci undergoes a Hedgehog-dependent activation step.

Download Full-text

Studies on sex-organ development. Oestrogenic effect on ornithine decarboxylase activity in the differentiating Müllerian ducts and other organs of the chick embryo

Biochemical Journal ◽

10.1042/bj1760143 ◽

1978 ◽

Vol 176 (1) ◽

pp. 143-149 ◽

Cited By ~ 6

Author(s):

C S Teng ◽

C T Teng

Keyword(s):

Enzyme Activity ◽

Ornithine Decarboxylase ◽

Decarboxylase Activity ◽

Mullerian Duct ◽

Stages Of Development ◽

Müllerian Duct ◽

Mullerian Ducts ◽

The Right ◽

The Brain

The activity of ornithine decarboxylase in the differentiating left and right Müllerian ducts was assayed and compared with that in other embryonic organs, i.e. the liver and the brain throughout the stages of development. In general the enzyme activity was high in the early stages and decreased extensively in the late stages of development. Specifically, in the left and righ Müllerian ducts, the enzyme activity was high from day 8 to day 9 of incubation. In the right duct the enzyme activity started to decline on day 9 and then continuously decreased to an almost undetectable value on day 18 of incubation. In the left duct the enzyme activity also decreased slightly from day 9 to day 12; however, it increased from day 13 to day 15 and finally decreased to a constant value from day 18 until hatching. The alteration in enzyme activity in the Müllerian duct as assayed in vitro during development is not due to the effect of the size of the endogenous ornithine pool. When the enzyme activity was subjected to oestrogen stimulation, an increase of 5–10-fold for the left duct and of 5–3-fold for the right duct was observed during the course of development. No such stimulation was observed with the treatment of progesterone. Testosterone consistently caused a 25–30% inhibition of the enzyme activity in the Müllerian duct. Oestrogen slightly stimulated the enzyme activity in the developing liver but inhibited that of the brain. The concentration of the three polyamines measured in the Müllerian duct corresponds to the activity of the enzyme determined.

Download Full-text

SIX1 cooperates with RUNX1 and SMAD4 in cell fate commitment of Müllerian duct epithelium

10.1101/427351 ◽

2018 ◽

Author(s):

Jumpei Terakawa ◽

Vanida A. Serna ◽

Devi Nair ◽

Shigeru Sato ◽

Kiyoshi Kawakami ◽

...

Keyword(s):

Transcription Factor ◽

Transcription Factors ◽

Epithelial Cells ◽

Cell Fate ◽

Reproductive Tract ◽

Mesenchymal Cells ◽

Mullerian Duct ◽

Müllerian Duct ◽

Duct Epithelium ◽

Putative Precursor

AbstractDuring female mammal reproductive tract development, epithelial cells of the lower Müllerian duct are committed to become stratified squamous epithelium of vagina and ectocervix, when the expression of ΔNp63 transcription factor is induced by mesenchymal cells. The absence of ΔNp63 expression leads to adenosis, the putative precursor of vaginal adenocarcinoma. Our previous studies with genetically engineered mouse models have established that fibroblast growth factor (FGF)/mitogen-activated protein kinase (MAPK), bone morphogenetic protein (BMP)/SMAD, and activin A/runt related transcription factor 1 (RUNX1) signaling pathways are independently required for ΔNp63 expression in Müllerian duct epithelium (MDE). Here we report that sine oculis homeobox homolog 1 (SIX1) plays a critical role in the activation of ΔNp63 locus in MDE as a downstream transcription factor of mesenchymal signals. In mouse developing reproductive tract, SIX1 expression was restricted to MDE of the future cervix and vagina. SIX1 expression was totally absent in SMAD4 null MDE and was reduced in RUNX1 null and FGFR2 null MDE, indicating that SIX1 is under the control of vaginal mesenchymal factors, BMP4, activin A and FGF7/10. Furthermore, Six1, Runx1 and Smad4 gene-dose-dependently activated ΔNp63 expression in MDE within vaginal fornix. Using a mouse model of diethylstilbestrol (DES)-associated vaginal adenosis, we found DES action through epithelial estrogen receptor α (ESR1) down-regulates SIX1 and RUNX1 in MDE within the vaginal fornix. This study establishes that the vaginal/ectocervical cell fate of MDE is regulated by a collaboration of multiple transcription factors including SMAD4, SIX1 and RUNX1, and the down-regulation of these key transcription factors leads to vaginal adenosis.Author SummaryIn embryogenesis, differentiation fate of cells is specified through constant communication between neighboring cells. In this study, we investigated the molecular mechanism of epithelial cell fate commitment in the lower female reproductive organs utilizing mouse genetic models. The cell fate of epithelial cells in the uterus, cervix and vagina is directed by signaling from mesenchymal cells. We demonstrated that within the epithelial cells of the developing vagina, signals from mesenchymal cells are integrated into activities of transcription factors including SMAD4, RUNX1 and SIX1, which dose-dependently co-operate in the determination of vaginal epithelial cell fate. Disruption of these processes alters the cell fate from vaginal to uterine epithelium, resulting in a condition called vaginal adenosis, a putative precursor of vaginal adenocarcinoma. Women exposed to diethylstilbestrol (DES) in the womb have about 40 times the risk of developing vaginal adenocarcinoma. We determined that developmental exposure to DES induces vaginal adenosis by repressing SIX1 and RUNX1 through ESR1 in the epithelial cells. This discovery enhances the understanding of how early-life events, such as exposure to endocrine disruptors, causes vaginal adenosis, and thus may contribute to the prevention and therapeutic treatment of idiopathic vaginal adenocarcinoma.

Download Full-text

Wnt4 coordinates directional cell migration and extension of the Müllerian duct essential for ontogenesis of the female reproductive tract

Human Molecular Genetics ◽

10.1093/hmg/ddv621 ◽

2015 ◽

Vol 25 (6) ◽

pp. 1059-1073 ◽

Cited By ~ 29

Author(s):

Renata Prunskaite-Hyyryläinen ◽

Ilya Skovorodkin ◽

Qi Xu ◽

Ilkka Miinalainen ◽

Jingdong Shan ◽

...

Keyword(s):

Cell Migration ◽

Reproductive Tract ◽

Female Reproductive Tract ◽

Mullerian Duct ◽

Müllerian Duct ◽

Directional Cell Migration

Download Full-text

SIX1 cooperates with RUNX1 and SMAD4 in cell fate commitment of Müllerian duct epithelium

Cell Death and Differentiation ◽

10.1038/s41418-020-0579-z ◽

2020 ◽

Vol 27 (12) ◽

pp. 3307-3320

Author(s):

Jumpei Terakawa ◽

Vanida A. Serna ◽

Devi M. Nair ◽

Shigeru Sato ◽

Kiyoshi Kawakami ◽

...

Keyword(s):

Transcription Factor ◽

Cell Fate ◽

Reproductive Tract ◽

Critical Role ◽

Activin A ◽

Genetically Engineered ◽

Mitogen Activated Protein Kinase ◽

Mullerian Duct ◽

Müllerian Duct ◽

Duct Epithelium

AbstractDuring female mammal reproductive tract development, epithelial cells of the lower Müllerian duct are committed to become stratified squamous epithelium of the vagina and ectocervix, when the expression of ΔNp63 transcription factor is induced by mesenchymal cells. The absence of ΔNp63 expression leads to adenosis, the putative precursor of vaginal adenocarcinoma. Our previous studies with genetically engineered mouse models have established that fibroblast growth factor (FGF)/mitogen-activated protein kinase (MAPK), bone morphogenetic protein (BMP)/SMAD, and activin A/runt-related transcription factor 1 (RUNX1) signaling pathways are independently required for ΔNp63 expression in Müllerian duct epithelium (MDE). Here, we report that sine oculis homeobox homolog 1 (SIX1) plays a critical role in the activation of ΔNp63 locus in MDE as a downstream transcription factor of mesenchymal signals. In the developing mouse reproductive tract, SIX1 expression was restricted to MDE within the future cervix and vagina. SIX1 expression was totally absent in SMAD4 null MDE and was reduced in RUNX1 null and FGFR2 null MDE, indicating that SIX1 is under the control of vaginal mesenchymal factors: BMP4, activin A and FGF7/10. Furthermore, Six1, Runx1, and Smad4 gene-dose-dependently activated ΔNp63 expression in MDE within the vaginal fornix. Using a mouse model of diethylstilbestrol (DES)-associated vaginal adenosis, we found DES action through epithelial estrogen receptor α (ESR1) inhibits activation of ΔNp63 locus in MDE by transcriptionally repressing SIX1 and RUNX1 in the vaginal fornix.

Download Full-text

Adhesion G-protein-coupled receptor, GPR56, is required for Müllerian duct development in the chick

Journal of Endocrinology ◽

10.1530/joe-19-0419 ◽

2020 ◽

Vol 244 (2) ◽

pp. 395-413 ◽

Cited By ~ 1

Author(s):

Zahida Yesmin Roly ◽

Andrew T Major ◽

Alex Fulcher ◽

Martin A Estermann ◽

Claire E Hirst ◽

...

Keyword(s):

G Protein ◽

Female Reproductive Tract ◽

Mullerian Duct ◽

G Protein Coupled Receptor ◽

Endocrine Control ◽

Müllerian Duct ◽

Collagen Iii ◽

Mullerian Ducts ◽

Duct Development ◽

G Protein Coupled

The embryonic Müllerian ducts give rise to the female reproductive tract (fallopian tubes, uterus and upper vagina in humans, the oviducts in birds). Embryonic Müllerian ducts initially develop in both sexes, but later regress in males under the influence of anti-Müllerian hormone. While the molecular and endocrine control of duct regression in males have been well studied, early development of the ducts in both sexes is less well understood. Here, we describe a novel role for the adhesion G protein-coupled receptor, GPR56, in development of the Müllerian ducts in the chicken embryo. GPR56 is expressed in the ducts of both sexes from early stages. The mRNA is present during the elongation phase of duct formation, and it is restricted to the inner Müllerian duct epithelium. The putative ligand, Collagen III, is abundantly expressed in the Müllerian duct at the same developmental stages. Knockdown of GPR56 expression using in ovo electroporation results in variably truncated ducts, with a loss of expression of both epithelial and mesenchymal markers of duct development. Over-expression of GPR56 in vitro results in enhanced cell proliferation and cell migration. These results show that GPR56 plays an essential role in avian Müllerian duct development through the regulation of duct elongation.

Download Full-text

Pregnancy in uterus didelphys delivered by caesarean section: a case report

International Journal of Reproduction Contraception Obstetrics and Gynecology ◽

10.18203/2320-1770.ijrcog20175048 ◽

2017 ◽

Vol 6 (11) ◽

pp. 5166 ◽

Cited By ~ 1

Author(s):

Devyani Sawai ◽

Susheel Kumar Sharma ◽

Devashish Singh Sawai ◽

Uttkarsha Sawai ◽

Sangeeta Sharma ◽

...

Keyword(s):

Caesarean Section ◽

High Risk Group ◽

Congenital Defects ◽

Mullerian Duct ◽

Müllerian Duct ◽

Uterus Didelphys ◽

Müllerian Duct Anomalies ◽

Mullerian Anomaly ◽

Mullerian Ducts ◽

Mullerian Duct Anomalies

The aim of this study is to report a rare case of pregnancy in uterus didelphys. Mullerian duct anomalies are congenital defects of the female genital system that arise from abnormal embryological development of the Mullerian ducts. A didelphys uterus, also known as double uterus is one of the least common amongst the various Mullerian duct anomalies. It results from complete failure of fusion of Mullerian ducts. There is presence of double uterine bodies with two separate cervices and often double or septate vagina. We report the case in our institute of a pregnancy in the left sided body of a didelphys uterus, delivered by caesarean section. Patients with uterus didelphys belong to high risk group and complications are increased in malformed uterus. Such cases need a meticulous prenatal care. It is a rare Mullerian anomaly and can present with varied obstetrical and gynaecological complications. Prompt and accurate diagnosis of uterine malformations and appropriate surgical intervention are essential to prevent complications.

Download Full-text

272.Decreased expression of oestrogen receptor β in the reproductive tract of pregnant relaxin-deficient (Rlx - / - ) mice

Reproduction Fertility and Development ◽

10.1071/srb04abs272 ◽

2004 ◽

Vol 16 (9) ◽

pp. 272

Author(s):

J. T. McGuane ◽

H. M. Gehring ◽

L. J. Parry

Keyword(s):

Transcriptional Activity ◽

Reproductive Tract ◽

Peptide Hormone ◽

Mrna Levels ◽

Wild Type ◽

Taqman Probes ◽

Pregnant Mice ◽

Thermal Cycler ◽

Deficient Mice ◽

Ovariectomised Rats

The peptide hormone relaxin (RLX) is reported to directly affect uterine oestrogen receptors (ERs) in the rat (1). Treatment of immature ovariectomised rats with porcine RLX causes a decrease in uterine ERβ mRNA levels within 6 h. However, RLX has no effect on ERα expression. As both ERβ1 and ERβ2 inhibit ER-mediated transcriptional activity, this RLX-induced downregulation in ERβ could be a prerequisite for oestrogen to exert its effects on target tissues. The aim of the current study was to use relaxin-deficient (Rlx–/–) pregnant mice to investigate if relaxin deficiency results in alterations in either ERβ or ERα mRNA expression in reproductive tissues. Cervix and vagina tissues were obtained from adult C57/Blk6J wild-type mice at five stages of gestation (Days 7.5, 10.5, 14.5, 17.5, 18.5 pc) and Rlx–/– littermates on Days 7.5, 14.5 and 18.5 pc. Q-PCR with TaqMan probes in the Opticon 2 thermal cycler (MJ Research, GeneWorks) was used to quantify ERα and ERα gene expression. ERα mRNA levels were significantly (P < 0.05; ANOVA) increased in the cervix/vagina on Days 17.5 and 18.5 pc in Rlx+/+ mice. The increase in ERα in Rlx+/+ mice was negatively correlated with a significant decrease in ERβ expression from Day 14.5 pc. In contrast, there was no decrease in ERβ expression in the cervix/vagina of Rlx–/– mice; ERβ mRNA levels were significantly (P < 0.05) higher compared to Rlx+/+ mice on Days 14.5 or 18.5 pc. However, there was no corresponding reduction in ERα expression in the cervix/vagina of the Rlx–/– mice, so that ERα mRNA levels were still elevated at term despite the maintenance of high ERα expression. In summary, these data show changes in ERα expression in the cervix/vagina of relaxin-deficient mice, which may subsequently affect ERα-mediated transcriptional activity. (1) Pillai et al. (2002) Biol. Reprod. 67, 1919–1926.

Download Full-text

Reduced Levels of ATF-2 Predispose Mice to Mammary Tumors

Molecular and Cellular Biology ◽

10.1128/mcb.01579-06 ◽

2006 ◽

Vol 27 (5) ◽

pp. 1730-1744 ◽

Cited By ~ 60

Author(s):

Toshio Maekawa ◽

Toshie Shinagawa ◽

Yuji Sano ◽

Takahiko Sakuma ◽

Shintaro Nomura ◽

...

Keyword(s):

Cell Density ◽

Target Genes ◽

High Cell Density ◽

Critical Role ◽

Susceptibility Gene ◽

Mammary Tumors ◽

Mrna Levels ◽

High Cell ◽

Wild Type ◽

Induced Apoptosis

ABSTRACT Transcription factor ATF-2 is a nuclear target of stress-activated protein kinases, such as p38, which are activated by various extracellular stresses, including UV light. Here, we show that ATF-2 plays a critical role in hypoxia- and high-cell-density-induced apoptosis and the development of mammary tumors. Compared to wild-type cells, Atf-2 −/− mouse embryonic fibroblasts (MEFs) were more resistant to hypoxia- and anisomycin-induced apoptosis but remained equally susceptible to other stresses, including UV. Atf-2 −/− and Atf-2 +/− MEFs could not express a group of genes, such as Gadd45α, whose overexpression can induce apoptosis, in response to hypoxia. Atf-2 −/− MEFs also had a higher saturation density than wild-type cells and expressed lower levels of Maspin, the breast cancer tumor suppressor, which is also known to enhance cellular sensitivity to apoptotic stimuli. Atf-2 −/− MEFs underwent a lower degree of apoptosis at high cell density than wild-type cells. Atf-2 +/− mice were highly prone to mammary tumors that expressed reduced levels of Gadd45α and Maspin. The ATF-2 mRNA levels in human breast cancers were lower than those in normal breast tissue. Thus, ATF-2 acts as a tumor susceptibility gene of mammary tumors, at least partly, by activating a group of target genes, including Maspin and Gadd45α.

Download Full-text