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1.
Am J Hum Genet ; 108(2): 337-345, 2021 02 04.
Artigo em Inglês | MEDLINE | ID: mdl-33434492

RESUMO

Mayer-Rokitansky-Küster-Hauser syndrome (MRKHS) is associated with congenital absence of the uterus, cervix, and the upper part of the vagina; it is a sex-limited trait. Disrupted development of the Müllerian ducts (MD)/Wölffian ducts (WD) through multifactorial mechanisms has been proposed to underlie MRKHS. In this study, exome sequencing (ES) was performed on a Chinese discovery cohort (442 affected subjects and 941 female control subjects) and a replication MRKHS cohort (150 affected subjects of mixed ethnicity from North America, South America, and Europe). Phenotypic follow-up of the female reproductive system was performed on an additional cohort of PAX8-associated congenital hypothyroidism (CH) (n = 5, Chinese). By analyzing 19 candidate genes essential for MD/WD development, we identified 12 likely gene-disrupting (LGD) variants in 7 genes: PAX8 (n = 4), BMP4 (n = 2), BMP7 (n = 2), TBX6 (n = 1), HOXA10 (n = 1), EMX2 (n = 1), and WNT9B (n = 1), while LGD variants in these genes were not detected in control samples (p = 1.27E-06). Interestingly, a sex-limited penetrance with paternal inheritance was observed in multiple families. One additional PAX8 LGD variant from the replication cohort and two missense variants from both cohorts were revealed to cause loss-of-function of the protein. From the PAX8-associated CH cohort, we identified one individual presenting a syndromic condition characterized by CH and MRKHS (CH-MRKHS). Our study demonstrates the comprehensive utilization of knowledge from developmental biology toward elucidating genetic perturbations, i.e., rare pathogenic alleles involving the same loci, contributing to human birth defects.


Assuntos
Transtornos 46, XX do Desenvolvimento Sexual/genética , Anormalidades Congênitas/genética , Ductos Paramesonéfricos/anormalidades , Ductos Paramesonéfricos/crescimento & desenvolvimento , Mutação , Ductos Mesonéfricos/crescimento & desenvolvimento , Adulto , Proteína Morfogenética Óssea 4/genética , Proteína Morfogenética Óssea 7/genética , Códon sem Sentido , Feminino , Estudos de Associação Genética , Pleiotropia Genética , Proteínas Homeobox A10/genética , Proteínas de Homeodomínio/genética , Humanos , Fator de Transcrição PAX8/genética , Herança Paterna , Penetrância , Proteínas com Domínio T/genética , Fatores de Transcrição/genética , Proteínas Wnt/genética , Ductos Mesonéfricos/anormalidades
2.
Differentiation ; 110: 49-63, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31622789

RESUMO

The study of male and female reproductive tract development requires expertise in two separate disciplines, developmental biology and endocrinology. For ease of experimentation and economy, the mouse has been used extensively as a model for human development and pathogenesis, and for the most part similarities in developmental processes and hormone action provide ample justification for the relevance of mouse models for human reproductive tract development. Indeed, there are many examples describing the phenotype of human genetic disorders that have a reasonably comparable phenotype in mice, attesting to the congruence between mouse and human development. However, anatomic, developmental and endocrinologic differences exist between mice and humans that (1) must be appreciated and (2) considered with caution when extrapolating information between all animal models and humans. It is critical that the investigator be aware of both the similarities and differences in organogenesis and hormone action within male and female reproductive tracts so as to focus on those features of mouse models with clear relevance to human development/pathology. This review, written by a team with extensive expertise in the anatomy, developmental biology and endocrinology of both mouse and human urogenital tracts, focusses upon the significant human/mouse differences, and when appropriate voices a cautionary note regarding extrapolation of mouse models for understanding development of human male and female reproductive tracts.


Assuntos
Epitélio/crescimento & desenvolvimento , Genitália Feminina/crescimento & desenvolvimento , Ductos Paramesonéfricos/crescimento & desenvolvimento , Útero/crescimento & desenvolvimento , Animais , Feminino , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Humanos , Camundongos , Organogênese/fisiologia
3.
Genetics ; 213(2): 529-553, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31399485

RESUMO

Fetal mammalian testes secrete Anti-Müllerian hormone (Amh), which inhibits female reproductive tract (Müllerian duct) development. Amh also derives from mature mammalian ovarian follicles, which marks oocyte reserve and characterizes polycystic ovarian syndrome. Zebrafish (Danio rerio) lacks Müllerian ducts and the Amh receptor gene amhr2 but, curiously, retains amh To discover the roles of Amh in the absence of Müllerian ducts and the ancestral receptor gene, we made amh null alleles in zebrafish. Results showed that normal amh prevents female-biased sex ratios. Adult male amh mutants had enormous testes, half of which contained immature oocytes, demonstrating that Amh regulates male germ cell accumulation and inhibits oocyte development or survival. Mutant males formed sperm ducts and some produced a few offspring. Young female mutants laid a few fertile eggs, so they also had functional sex ducts. Older amh mutants accumulated nonvitellogenic follicles in exceedingly large but sterile ovaries, showing that Amh helps control ovarian follicle maturation and proliferation. RNA-sequencing data partitioned juveniles at 21 days postfertilization (dpf) into two groups that each contained mutant and wild-type fish. Group21-1 upregulated ovary genes compared to Group21-2, which were likely developing as males. By 35 dpf, transcriptomes distinguished males from females and, within each sex, mutants from wild types. In adult mutants, ovaries greatly underexpressed granulosa and theca genes, and testes underexpressed Leydig cell genes. These results show that ancestral Amh functions included development of the gonadal soma in ovaries and testes and regulation of gamete proliferation and maturation. A major gap in our understanding is the identity of the gene encoding a zebrafish Amh receptor; we show here that the loss of amhr2 is associated with the breakpoint of a chromosome rearrangement shared among cyprinid fishes.


Assuntos
Hormônio Antimülleriano/genética , Genitália Feminina/crescimento & desenvolvimento , Processos de Determinação Sexual , Peixe-Zebra/genética , Animais , Feminino , Gônadas/crescimento & desenvolvimento , Ductos Paramesonéfricos/crescimento & desenvolvimento , Folículo Ovariano/crescimento & desenvolvimento , Ovário/crescimento & desenvolvimento , RNA-Seq , Receptores de Peptídeos/genética , Receptores de Fatores de Crescimento Transformadores beta/genética , Peixe-Zebra/crescimento & desenvolvimento
4.
Methods Mol Biol ; 1965: 173-186, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31069675

RESUMO

The reproductive cycle encompasses processes such as sex organ differentiation and development in the early life stages and maturation of the gametes in the adult organism. During the early life stages, critical developmental programming of the endocrine and reproductive systems occurs, and exposure to chemicals during these critical developmental windows can result in impaired reproductive function later in life. It is therefore important to evaluate long-term consequences of early life stage exposure to endocrine-disrupting chemicals. The African clawed frog Xenopus tropicalis has several characteristics that facilitate studies of developmental and reproductive toxicity. Here I present a X. tropicalis life cycle test protocol including study design, exposure regimes, and endpoints for chemical disruption of sex differentiation, gonadal and Müllerian duct development, the thyroxin-regulated metamorphosis, estrogen synthesis (activity of the CYP19 aromatase enzyme), spermatogenesis, oogenesis, puberty and fertility.


Assuntos
Gametogênese/efeitos dos fármacos , Ductos Paramesonéfricos/crescimento & desenvolvimento , Xenopus/crescimento & desenvolvimento , Animais , Embrião não Mamífero/efeitos dos fármacos , Feminino , Estágios do Ciclo de Vida/efeitos dos fármacos , Masculino , Metamorfose Biológica/efeitos dos fármacos , Modelos Animais , Ductos Paramesonéfricos/efeitos dos fármacos , Diferenciação Sexual , Tiroxina/metabolismo , Xenopus/metabolismo
5.
Hum Reprod ; 34(6): 1117-1125, 2019 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-31111890

RESUMO

STUDY QUESTION: Is there an increased prevalence of male microchimerism in women with Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome, as evidence of fetal exposure to blood and anti-Müllerian hormone (AMH) from a (vanished) male co-twin resulting in regression of the Müllerian duct derivatives? SUMMARY ANSWER: Predominant absence of male microchimerism in adult women with MRKH syndrome does not support our hypothesis that intrauterine blood exchange with a (vanished) male co-twin is the pathophysiological mechanism. WHAT IS KNOWN ALREADY: The etiology of MRKH is unclear. Research on the phenotype analogous condition in cattle (freemartinism) has yielded the hypothesis that Müllerian duct development is inhibited by exposure to AMH in utero. In cattle, the male co-twin has been identified as the source for AMH, which is transferred via placental blood exchange. In human twins, a similar exchange of cellular material has been documented by detection of chimerism, but it is unknown whether this has clinical consequences. STUDY DESIGN, SIZE, DURATION: An observational case-control study was performed to compare the presence of male microchimerism in women with MRKH syndrome and control women. Through recruitment via the Dutch patients' association of women with MRKH (comprising 300 members who were informed by email or regular mail), we enrolled 96 patients between January 2017 and July 2017. The control group consisted of 100 women who reported never having been pregnant. PARTICIPANTS/MATERIALS, SETTING, METHODS: After written informed consent, peripheral blood samples were obtained by venipuncture, and genomic DNA was extracted. Male microchimerism was detected by Y-chromosome-specific real-time quantitative PCR, with use of DYS14 marker. Possible other sources for microchimerism, for example older brothers, were evaluated using questionnaire data. MAIN RESULTS AND THE ROLE OF CHANCE: The final analysis included 194 women: 95 women with MRKH syndrome with a mean age of 40.9 years and 99 control women with a mean age of 30.2 years. In total, 54 women (56.8%) were identified as having typical MRKH syndrome, and 41 women (43.2%) were identified as having atypical MRKH syndrome (when extra-genital malformations were present). The prevalence of male microchimerism was significantly higher in the control group than in the MRKH group (17.2% versus 5.3%, P = 0.009). After correcting for age, women in the control group were 5.8 times more likely to have male microchimerism (odds ratio 5.84 (CI 1.59-21.47), P = 0.008). The mean concentration of male microchimerism in the positive samples was 56.0 male genome equivalent per 1 000 000 cells. The prevalence of male microchimerism was similar in women with typical MRKH syndrome and atypical MRKH syndrome (5.6% versus 4.9%, P = 0.884). There were no differences between women with or without microchimerism in occurrence of alternative sources of XY cells, such as older brothers, previous blood transfusion, or history of sexual intercourse. LIMITATIONS, REASON FOR CAUTION: We are not able to draw definitive conclusions regarding the occurrence of AMH exchange during embryologic development in women with MRKH syndrome. Our subject population includes all adult women and therefore is reliant on long-term prevalence of microchimerism. Moreover, we have only tested blood, and, theoretically, the cells may have grafted anywhere in the body during development. It must also be considered that the exchange of AMH may occur without the transfusion of XY cells and therefore cannot be discovered by chimerism detection. WIDER IMPLICATIONS OF THE FINDINGS: This is the first study to test the theory that freemartinism causes the MRKH syndrome in humans. The study aimed to test the presence of male microchimerism in women with MRKH syndrome as a reflection of early fetal exposure to blood and AMH from a male (vanished) co-twin. We found that male microchimerism was only present in 5.3% of the women with MRKH syndrome, a significantly lower percentage than in the control group (17.2%). Our results do not provide evidence for an increased male microchimerism in adult women with MRKH as a product of intrauterine blood exchange. However, the significant difference in favor of the control group is of interest to the ongoing discussion on microchimeric cell transfer and the possible sources of XY cells. STUDY FUNDING/COMPETING INTEREST(S): None. TRIAL REGISTRATION NUMBER: Dutch trial register, NTR5961.


Assuntos
Transtornos 46, XX do Desenvolvimento Sexual/genética , Quimerismo , Anormalidades Congênitas/genética , Genes Ligados ao Cromossomo Y/genética , Ductos Paramesonéfricos/anormalidades , Ductos Paramesonéfricos/crescimento & desenvolvimento , Transtornos 46, XX do Desenvolvimento Sexual/sangue , Transtornos 46, XX do Desenvolvimento Sexual/diagnóstico , Adulto , Biomarcadores/análise , Estudos de Casos e Controles , Anormalidades Congênitas/sangue , Anormalidades Congênitas/diagnóstico , Feminino , Humanos , Pessoa de Meia-Idade , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , Adulto Jovem
6.
Reprod Toxicol ; 86: 56-61, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30940504

RESUMO

In female mice, the Müllerian duct develops into the oviduct, uterus and vagina. The fate of epithelia is determined by factors secreted from the mesenchyme. Retinoic acid (RA) and its receptors are present in the mesenchyme of cranial Müllerian duct. RA induces Müllerian duct to uterine epithelial differentiation whereas inhibition of RA receptors induces vaginal epithelial differentiation. Thus, RA signaling in the Müllerian duct is required to promote differentiation of the mesenchyme into the future uterus. Perinatal estrogen exposure induces various abnormalities in Müllerian duct-derived organs. These include a cranial shift of the border among oviduct, uterus and vagina as well as precancerous lesions suppressed by co-treatment with RA and estrogen. Since RA synthesis enzymes and receptors are expressed both in the epithelium and stroma after birth, RA signaling may act in the epithelia to maintain adult epithelial homeostasis and to prevent irreversible lesions induced by perinatal estrogen exposure.


Assuntos
Ductos Paramesonéfricos/crescimento & desenvolvimento , Tretinoína/fisiologia , Útero/crescimento & desenvolvimento , Animais , Estrogênios/metabolismo , Feminino , Humanos , Transdução de Sinais
8.
Differentiation ; 103: 46-65, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30236463

RESUMO

Development of the human female reproductive tract is reviewed from the ambisexual stage to advanced development of the uterine tube, uterine corpus, uterine cervix and vagina at 22 weeks. Historically this topic has been under-represented in the literature, and for the most part is based upon hematoxylin and eosin stained sections. Recent immunohistochemical studies for PAX2 (reactive with Müllerian epithelium) and FOXA1 (reactive with urogenital sinus epithelium and its known pelvic derivatives) shed light on an age-old debate on the derivation of vaginal epithelium supporting the idea that human vaginal epithelium derives solely from urogenital sinus epithelium. Aside for the vagina, most of the female reproductive tract is derived from the Müllerian ducts, which fuse in the midline to form the uterovaginal canal, the precursor of uterine corpus and uterine cervix an important player in vaginal development as well. Epithelial and mesenchymal differentiation markers are described during human female reproductive tract development (keratins, homeobox proteins (HOXA11 and ISL1), steroid receptors (estrogen receptor alpha and progesterone receptor), transcription factors and signaling molecules (TP63 and RUNX1), which are expressed in a temporally and spatially dynamic fashion. The utility of xenografts and epithelial-mesenchymal tissue recombination studies are reviewed.


Assuntos
Genitália Feminina/crescimento & desenvolvimento , Ductos Paramesonéfricos/crescimento & desenvolvimento , Útero/crescimento & desenvolvimento , Vagina/crescimento & desenvolvimento , Feminino , Genitália Feminina/metabolismo , Proteínas de Homeodomínio/genética , Humanos , Proteínas com Homeodomínio LIM/genética , Receptores de Progesterona/genética , Fatores de Transcrição/genética , Proteínas Supressoras de Tumor/genética
9.
Differentiation ; 97: 54-72, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29053991

RESUMO

Human female reproductive tract development rests mostly upon hematoxilyn and eosin stained sections despite recent advances on molecular mechanisms in mouse studies. We report application of immunohistochemical methods to explore the ontogeny of epithelial and mesenchymal differentiation markers (keratins, homobox proteins, steroid receptors), transcription factors and signaling molecules (TP63 and RUNX1) during human female reproductive tract development. Keratins 6, 7, 8, 10, 14 and 19 (KRT6, KRT7, KRT8, KRT10, KRT14, KRT19) were expressed in a temporally and spatially dynamic fashion. The undifferentiated Müllerian duct and uterovaginal canal, lined by simple columnar epithelia, expressed KRT7, KRT8 and KRT19. Glandular derivatives of the Müllerian duct (uterine tube, uterine corpus and endocervix) maintained expression of these keratins, while tissues that undergo stratified squamous differentiation (exocervix and vagina) expressed KRT6, KRT14 and KRT10 during development in an age-dependent fashion. TP63 and RUNX1 were expressed prior to KRT14, as these two transcription factors are known to be upstream from KRT14 in developing Müllerian epithelium. In the vagina, KRT10, a marker of terminal differentiation, appeared after endogenous estrogens transformed the epithelium to a thick glycogenated squamous epithelium. Uroplakin, a protein unique to urothelium, was expressed only in the bladder, urethra and vaginal introitus, but not in the female reproductive tract itself. Mesenchymal differentiation was examined through immunostaining for HOXA11 (expressed in uterine mesenchyme) and ISL1 (expressed in vaginal mesenchyme). A detailed ontogeny of estrogen receptor alpha (ESR1), progesterone receptor (PGR) and the androgen receptor (AR) provides the mechanistic underpinning for the teratogenicity of estrogens, progestins and androgens on female reproductive tract development. Immunohistochemical analysis of differentiation markers and signaling molecules advance our understanding of normal development of the human female reproductive tract. These observations demonstrate remarkable similarities in mouse and human female reproductive tract development, but also highlight some key differences.


Assuntos
Genitália Feminina/crescimento & desenvolvimento , Proteínas de Homeodomínio/genética , Queratinas/genética , Receptores de Esteroides/genética , Reprodução/genética , Animais , Diferenciação Celular/genética , Desenvolvimento Embrionário/genética , Epitélio/crescimento & desenvolvimento , Epitélio/metabolismo , Receptor alfa de Estrogênio/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Genitália Feminina/metabolismo , Humanos , Camundongos , Ductos Paramesonéfricos/crescimento & desenvolvimento , Ductos Paramesonéfricos/metabolismo , Receptores Androgênicos/genética , Receptores de Progesterona , Útero/crescimento & desenvolvimento , Útero/metabolismo , Vagina/crescimento & desenvolvimento , Vagina/metabolismo
10.
Differentiation ; 97: 9-22, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28918284

RESUMO

We present a detailed review of the embryonic and fetal development of the human female reproductive tract utilizing specimens from the 5th through the 22nd gestational week. Hematoxylin and eosin (H&E) as well as immunohistochemical stains were used to study the development of the human uterine tube, endometrium, myometrium, uterine cervix and vagina. Our study revisits and updates the classical reports of Koff (1933) and Bulmer (1957) and presents new data on development of human vaginal epithelium. Koff proposed that the upper 4/5ths of the vagina is derived from Müllerian epithelium and the lower 1/5th derived from urogenital sinus epithelium, while Bulmer proposed that vaginal epithelium derives solely from urogenital sinus epithelium. These conclusions were based entirely upon H&E stained sections. A central player in human vaginal epithelial development is the solid vaginal plate, which arises from the uterovaginal canal (fused Müllerian ducts) cranially and squamous epithelium of urogenital sinus caudally. Since Müllerian and urogenital sinus epithelium cannot be unequivocally identified in H&E stained sections, we used immunostaining for PAX2 (reactive with Müllerian epithelium) and FOXA1 (reactive with urogenital sinus epithelium). By this technique, the PAX2/FOXA1 boundary was located at the extreme caudal aspect of the vaginal plate at 12 weeks. During the ensuing weeks, the PAX2/FOXA1 boundary progressively extended cranially such that by 21 weeks the entire vaginal epithelium was FOXA1-reactive and PAX2-negative. This observation supports Bulmer's proposal that human vaginal epithelium derives solely from urogenital sinus epithelium. Clearly, the development of the human vagina is far more complex than previously envisioned and appears to be distinctly different in many respects from mouse vaginal development.


Assuntos
Fator 3-alfa Nuclear de Hepatócito/genética , Ductos Paramesonéfricos/crescimento & desenvolvimento , Fator de Transcrição PAX2/genética , Vagina/crescimento & desenvolvimento , Animais , Desenvolvimento Embrionário/genética , Epitélio/crescimento & desenvolvimento , Epitélio/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Camundongos , Ductos Paramesonéfricos/metabolismo , Reprodução/genética , Vagina/metabolismo
11.
J Pediatr Surg ; 52(10): 1656-1660, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28599968

RESUMO

BACKGROUND/AIM: Anti-Müllerian hormone (AMH), also called Müllerian inhibiting substance (MIS), is glycoprotein hormone secreted by the fetal Sertoli cells to regulate regression of the Müllerian ducts, the anlagen of the uterus, fallopian tubes, and upper vagina. After its existence was predicted in 1946 and its isolation and purification in the 1970's, a huge amount of information has been gathered on its molecular biology and function in the last 30-40years. Once thought to be a locally acting factor in the male fetus during sexual differentiation, it is now recognized as an endocrine hormone present in both sexes and with functions throughout life. One of the remaining controversies is the possible role of AMH during fetal testicular descent. In the human with aberrant AMH function, the boy has cryptorchidism with persistent Müllerian duct syndrome (PMDS), where the testes are often intraabdominal and on an abnormally long gubernacular cord. By contrast, in rodent models knockout of the AMH gene does not cause cryptorchidism. METHODS/RESULTS: In this review we examined the evidence in the literature for and against a role for AMH in testicular descent and considered the implications of the different anatomy of the gubernacular cord in rodents versus children. CONCLUSION: We conclude that AMH may have a role in shortening the gubernacular cord in humans which is concealed in rodent models by differences in anatomy of the gubernacular cord in rodents. The controversy could be resolved by re-examination of the gubernacular cord in boys with PMDS and mice with AMHKO. TYPE OF STUDY: Review. LEVEL OF EVIDENCE: V.


Assuntos
Hormônio Antimülleriano/metabolismo , Criptorquidismo/metabolismo , Inibidores do Crescimento/metabolismo , Ductos Paramesonéfricos/metabolismo , Animais , Transtorno 46,XY do Desenvolvimento Sexual , Tubas Uterinas/metabolismo , Feminino , Humanos , Masculino , Camundongos , Ductos Paramesonéfricos/crescimento & desenvolvimento , Células de Sertoli/metabolismo
12.
J Appl Toxicol ; 36(12): 1639-1650, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27143402

RESUMO

The Larval Amphibian Growth and Development Assay (LAGDA) is a globally harmonized test guideline developed by the U.S. Environmental Protection Agency in collaboration with Japan's Ministry of the Environment. The LAGDA was designed to evaluate apical effects of chronic chemical exposure on growth, thyroid-mediated amphibian metamorphosis and reproductive development. During the validation phase, two well-characterized endocrine-disrupting chemicals were tested to evaluate the performance of the initial assay design: xenoestrogen 4-tert-octylphenol (tOP) and xenoandrogen 17ß-trenbolone (TB). Xenopus laevis embryos were exposed, in flow-through conditions, to tOP (nominal concentrations: 0.0, 6.25, 12.5, 25 and 50 µg l-1 ) or TB (nominal concentrations: 0.0, 12.5, 25, 50 and 100 ng l-1 ) until 8 weeks post-metamorphosis, at which time growth measurements were taken, and histopathology assessments were made of the gonads, reproductive ducts, liver and kidneys. There were no effects on growth in either study and no signs of overt toxicity, sex reversal or gonad dysgenesis. Exposure to tOP caused a treatment-related decrease in circulating thyroxine and an increase in thyroid follicular cell hypertrophy and hyperplasia (25 and 50 µg l-1 ) during metamorphosis. Müllerian duct development was affected after exposure to both chemicals; tOP exposure caused dose-dependent maturation of oviducts in both male and female frogs, whereas TB exposure caused accelerated Müllerian duct regression in males and complete regression in >50% of the females in the 100 ng l-1 treatment. Based on these results, the LAGDA performed adequately to evaluate apical effects of chronic exposure to two endocrine-active compounds and is the first standardized amphibian multiple life stage toxicity test to date. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.


Assuntos
Embrião não Mamífero/efeitos dos fármacos , Disruptores Endócrinos/toxicidade , Monitoramento Ambiental/métodos , Metamorfose Biológica/efeitos dos fármacos , Fenóis/toxicidade , Acetato de Trembolona/toxicidade , Animais , Bioensaio , Relação Dose-Resposta a Droga , Feminino , Larva , Masculino , Ductos Paramesonéfricos/efeitos dos fármacos , Ductos Paramesonéfricos/embriologia , Ductos Paramesonéfricos/crescimento & desenvolvimento , Glândula Tireoide/efeitos dos fármacos , Glândula Tireoide/embriologia , Glândula Tireoide/crescimento & desenvolvimento , Xenopus laevis
13.
Sci Rep ; 6: 23037, 2016 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-26964900

RESUMO

Congenital reproductive tract anomalies could impair fertility. Female and male reproductive tracts are developed from Müllerian ducts and Wolffian ducts, respectively, involving initiation, elongation and differentiation. Genetic basis solely for distal reproductive tract development is largely unknown. Lhfpl2 (lipoma HMGIC fusion partner-like 2) encodes a tetra-transmembrane protein with unknown functions. It is expressed in follicle cells of ovary and epithelial cells of reproductive tracts. A spontaneous point mutation of Lhfpl2 (LHFPL2(G102E)) leads to infertility in 100% female mice, which have normal ovarian development, ovulation, uterine development, and uterine response to exogenous estrogen stimulation, but abnormal upper longitudinal vaginal septum and lower vaginal agenesis. Infertility is also observed in ~70% mutant males, which have normal mating behavior and sperm counts, but abnormal distal vas deferens convolution resulting in complete and incomplete blockage of reproductive tract in infertile and fertile males, respectively. On embryonic day 15.5, mutant Müllerian ducts and Wolffian ducts have elongated but their duct tips are enlarged and fail to merge with the urogenital sinus. These findings provide a novel function of LHFPL2 and a novel genetic basis for distal reproductive tract development; they also emphasize the importance of an additional merging phase for proper reproductive tract development.


Assuntos
Genitália/crescimento & desenvolvimento , Genitália/metabolismo , Perda Auditiva Neurossensorial/metabolismo , Infertilidade Feminina/genética , Reprodução/genética , Animais , Feminino , Perda Auditiva Neurossensorial/genética , Infertilidade Feminina/patologia , Masculino , Camundongos , Ductos Paramesonéfricos/crescimento & desenvolvimento , Ductos Paramesonéfricos/metabolismo , Ovário/crescimento & desenvolvimento , Ovário/metabolismo , Mutação Puntual , Diferenciação Sexual/genética , Sistema Urogenital/crescimento & desenvolvimento , Sistema Urogenital/metabolismo , Sistema Urogenital/patologia , Ductos Mesonéfricos/crescimento & desenvolvimento , Ductos Mesonéfricos/metabolismo
14.
Differentiation ; 87(1-2): 23-31, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24433705

RESUMO

Marsupials differ from eutherian mammals in their reproductive strategy of delivering a highly altricial young after a short gestation. The young, with its undeveloped organ systems completes its development post-natally, usually within a pouch. The young is dependent on milk with a composition that varies through lactation to support its growth and changing needs as it matures over a lengthy period. Gonadal differentiation occurs after birth, providing a unique opportunity to examine the effects of hormonal manipulations on its sexual differentiation of the highly accessible young. In marsupials a difference in the migration of the urinary ducts around the genital ducts from eutherian mammals results in the unique tammar reproductive tract which has three vaginae and two cervices, and two distinctly separate uteri. In the tammar wallaby, a small member of the kangaroo family, we showed that virilisation of the Wolffian duct, prostate and phallus depends on an alternate androgen pathway, which has now been shown to be important for virilisation in humans. Through hormonal manipulations over differing time periods we have achieved sex reversal of both ovaries and testes, germ cells, genital ducts, prostate and phallus. Whilst we understand many of the mechanisms behind sexual differentiation there are still many lessons to be learned from understanding how sex reversal is achieved by using a model such as the tammar wallaby. This will help guide investigations into the major questions of how and why sex determination is achieved in other species. This review discusses the control and development of the marsupial urogenital system, largely drawn from our studies in the tammar wallaby and our ability to manipulate this system to induce sex reversal.


Assuntos
Androgênios/metabolismo , Marsupiais/crescimento & desenvolvimento , Diferenciação Sexual/genética , Sistema Urogenital/crescimento & desenvolvimento , Animais , Feminino , Gônadas/crescimento & desenvolvimento , Masculino , Marsupiais/genética , Ductos Paramesonéfricos/crescimento & desenvolvimento , Testículo/crescimento & desenvolvimento
15.
Int J Dev Biol ; 58(9): 663-8, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25896202

RESUMO

The Müllerian duct gives rise to female reproductive organs, such as the oviduct and uterus. During gestation, the Wolffian duct, which generates male reproductive organs and the kidney, is formed, and the Müllerian duct then elongates caudally along the preformed Wolffian duct. Anatomical separation of these two ducts in chick embryos demonstrated that the Wolffian duct is required for Müllerian duct formation. Likewise, a few reports supported this notion in mice, including studies on Wnt9b mutant mice and Wolffian duct-specific Lhx1 deletion. However, anatomical ablation of the Wolffian duct has not been established in mice. In this study, we addressed the importance of the interaction between these two reproductive ducts, by generating mice that specifically expressed a diphtheria toxin subunit in the Wolffian duct. While this genetic ablation of the Wolffian duct resulted in kidney hypoplasia/agenesis in both male and female mutant mice, the female mutant mice lacked the uterus, which is derived from the Müllerian duct. At mid-gestation, the Müllerian duct was truncated at the level where the mutant Wolffian duct was prematurely terminated, meaning that Müllerian duct elongation was dependent on the preformed Wolffian duct. However, Wnt9b expression in the Wolffian duct and the resultant canonical Wnt activity, as well as Lhx1 expression, were not affected in the mutant mice. These results suggest that the Wolffian duct regulates Müllerian duct elongation by currently unidentified mechanisms that are independent of canonical Wnt signaling or Lhx1 expression.


Assuntos
Embrião de Mamíferos/metabolismo , Proteínas com Homeodomínio LIM/metabolismo , Ductos Paramesonéfricos/crescimento & desenvolvimento , Organogênese , Fatores de Transcrição/metabolismo , Útero/metabolismo , Proteínas Wnt/metabolismo , Ductos Mesonéfricos/crescimento & desenvolvimento , Animais , Embrião de Mamíferos/citologia , Feminino , Técnicas Imunoenzimáticas , Hibridização In Situ , Proteínas com Homeodomínio LIM/genética , Masculino , Camundongos , Camundongos Knockout , Ductos Paramesonéfricos/embriologia , Ductos Paramesonéfricos/metabolismo , Transdução de Sinais , Fatores de Transcrição/genética , Útero/embriologia , Proteínas Wnt/genética , Ductos Mesonéfricos/embriologia , Ductos Mesonéfricos/metabolismo
16.
Endocrinology ; 154(10): 3931-6, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24025226

RESUMO

Müllerian inhibiting substance (MIS, also known as anti-Müllerian hormone), is a key factor of male sex differentiation in vertebrates. In amniotes, it is responsible for Müllerian duct regression in male embryos. In fish, despite the absence of Müllerian ducts, MIS is produced and controls germ cell proliferation during gonad differentiation. Here we show for the first time the presence of MIS in an amphibian species, Pleurodeles waltl. This is very astonishing because in caudate amphibians, Müllerian ducts do not regress in males. Phylogenetic analysis of MIS P. waltl ortholog revealed that the deduced protein segregates with MIS from other vertebrates and is clearly separated from other TGF-ß family members. In larvae, MIS mRNA was expressed at higher levels in the developing testes than in the ovaries. In the testis, MIS mRNA expression was located within the lobules that contain Sertoli cells. Besides, expression of MIS was modified in the case of sex reversal: it increased after masculinizing heat treatment and decreased after estradiol feminizing exposure. In addition to the data obtained recently in the fish medaka, our results suggest that the role of MIS on Müllerian ducts occurred secondarily during the course of evolution.


Assuntos
Proteínas de Anfíbios/metabolismo , Hormônio Antimülleriano/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Ovário/metabolismo , Pleurodeles/fisiologia , Testículo/metabolismo , Proteínas de Anfíbios/biossíntese , Proteínas de Anfíbios/química , Proteínas de Anfíbios/genética , Animais , Hormônio Antimülleriano/biossíntese , Hormônio Antimülleriano/química , Hormônio Antimülleriano/genética , Feminino , Hibridização In Situ , Larva/crescimento & desenvolvimento , Larva/metabolismo , Fígado/crescimento & desenvolvimento , Fígado/metabolismo , Masculino , Metamorfose Biológica , Ductos Paramesonéfricos/crescimento & desenvolvimento , Ductos Paramesonéfricos/metabolismo , Técnicas de Cultura de Órgãos , Ovário/crescimento & desenvolvimento , Filogenia , Estrutura Terciária de Proteína , RNA Mensageiro/metabolismo , Células de Sertoli/citologia , Células de Sertoli/metabolismo , Diferenciação Sexual , Testículo/citologia , Testículo/crescimento & desenvolvimento
17.
Mol Endocrinol ; 27(9): 1442-54, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23904126

RESUMO

Müllerian duct differentiation and development into the female reproductive tract is essential for fertility, but mechanisms regulating these processes are poorly understood. WNT signaling is critical for proper development of the female reproductive tract as evident by the phenotypes of Wnt4, Wnt5a, Wnt7a, and ß-catenin (Ctnnb1) mutant mice. Here we extend these findings by determining the effects of constitutive CTNNB1 activation within the mesenchyme of the developing Müllerian duct and its differentiated derivatives. This was accomplished by crossing Amhr2-Cre knock-in mice with Ctnnb1 exon (ex) 3(f/f) mice. Amhr2-Cre(Δ/+); Ctnnb1 ex3(f/+) females did not form an oviduct, had smaller uteri, endometrial gland defects, and were infertile. At the cellular level, stabilization of CTNNB1 in the mesenchyme caused alterations within the epithelium, including less proliferation, delayed uterine gland formation, and induction of an epithelial-mesenchymal transition (EMT) event. This EMT event is observed before birth and is complete within 5 days after birth. Misexpression of estrogen receptor α in the epithelia correlated with the EMT before birth, but not after. These studies indicate that regulated CTNNB1 in mesenchyme is important for epithelial cell differentiation during female reproductive tract development.


Assuntos
Diferenciação Celular , Células Epiteliais/patologia , Mesoderma/metabolismo , Ductos Paramesonéfricos/crescimento & desenvolvimento , Ductos Paramesonéfricos/metabolismo , Útero/crescimento & desenvolvimento , Útero/metabolismo , beta Catenina/metabolismo , Animais , Animais Recém-Nascidos , Neoplasias do Endométrio/patologia , Endométrio/anormalidades , Endométrio/crescimento & desenvolvimento , Endométrio/patologia , Transição Epitelial-Mesenquimal , Epitélio/crescimento & desenvolvimento , Epitélio/patologia , Feminino , Infertilidade Feminina/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Ductos Paramesonéfricos/patologia , Miométrio/anormalidades , Miométrio/crescimento & desenvolvimento , Miométrio/patologia , Estabilidade Proteica , Sarcoma/patologia , Células Estromais/patologia , Útero/patologia
18.
J Reprod Med ; 58(11-12): 517-28, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24568047

RESUMO

OBJECTIVE: To determine whether the process of vitrification affects the development of the female Müllerian duct. STUDY DESIGN: We examined the difference in morphology and gene expression of the developing uteri of 30 female mice from transferred vitrified embryos and 30 female mice from transferred fresh embryos. RESULTS: By employing histology, the data showed that there were no significant differences between the 2 groups of the same age. With the use of reverse transcription polymerase chain reaction and Western blotting, the data showed that there was no significant change in the expression of Wnt genes (Wnt4, Wnt5a, Wnt7a), beta-catenin/TCF target genes, and homeobox A10 (HOXA10) gene during uterine development in the vitrified group as compared with the control group. CONCLUSION: These data suggest that vitrifying preimplantation embryos may have no effects on morphology and gene expression of the uterus of offspring.


Assuntos
Blastocisto/fisiologia , Criopreservação/veterinária , Útero/crescimento & desenvolvimento , beta Catenina/análise , Animais , Transferência Embrionária/veterinária , Feminino , Expressão Gênica , Proteínas Homeobox A10 , Proteínas de Homeodomínio/análise , Proteínas de Homeodomínio/genética , Camundongos , Ductos Paramesonéfricos/crescimento & desenvolvimento , Gravidez , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Útero/metabolismo , Proteínas Wnt/análise , Proteínas Wnt/genética , beta Catenina/genética
19.
Ann Endocrinol (Paris) ; 71(2): 83-8, 2010 Mar.
Artigo em Francês | MEDLINE | ID: mdl-20137781

RESUMO

Müllerian inhibiting substance (MIS) has been discovered by Alfred Jost at the beginning of the fifties. MIS is a glycoprotein belonging to the TGF-beta family. Its various functions differ between males and females and according to the age of the individual. In male, the protein is synthesized by Sertoli's cells and induces the disappearance of Müllerian's ducts, the development of the male genital tract. Its role in adult males remains quite unknown. In female, the protein is secreted by granulosa cells and plays a role during folliculogenesis as it regulates the initial and cyclic recruitment of ovarian follicles. MIS is also a good marker of follicular reserve and ovarian function. Therefore, it plays a role in different areas such as assisted medical reproduction and oncology. This protein represents a potential major diagnosis as well as prognostic tool in reproduction.


Assuntos
Hormônio Antimülleriano/fisiologia , Ductos Paramesonéfricos/crescimento & desenvolvimento , Folículo Ovariano/fisiologia , Hormônio Antimülleriano/sangue , Hormônio Antimülleriano/genética , Feminino , Genitália Masculina/crescimento & desenvolvimento , Genitália Masculina/fisiologia , Células da Granulosa/fisiologia , Humanos , Masculino , Células de Sertoli/fisiologia , Diferenciação Sexual/fisiologia
20.
Dev Biol ; 334(2): 429-36, 2009 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-19664614

RESUMO

In developing mammalian males, conversion of the Wolffian ducts into the epididymides and vasa deferentia depends on androgen secretion by the testes, whereas in females these ducts remain in a vestigial form or regress. However, there is continuing uncertainty whether the androgen needs to be delivered locally, either by diffusion from the adjacent testis or, by secretion into the lumen of the duct, or whether circulating androgens maintain and virilize the Wolffian ducts. To resolve this uncertainty, we transplanted either day 0-2 or day 8-9 post-partum testes beneath the flank skin of three groups of neonatal (days 0-1) female tammar wallabies, where they developed and secreted physiological levels of hormones. The Wolffian ducts of all these females were retained and had formed extensive epididymides when examined at days 25, 34 and 87 after birth. In the two older groups of females, sampled after the time of prostatic bud formation, the urogenital sinus was virilized and there was extensive prostatic development similar to that of normal males of the same age, showing that androgen secretion had occurred. Virilization of the Wolffian ducts occurred during an early but short-lived window of sensitivity. This study provides the first clear evidence that under physiological conditions virilization can be mediated by circulating androgen.


Assuntos
Androgênios/fisiologia , Diferenciação Sexual/fisiologia , Testículo/metabolismo , Sistema Urogenital/crescimento & desenvolvimento , Virilismo/etiologia , Ductos Mesonéfricos/crescimento & desenvolvimento , Androgênios/sangue , Androgênios/metabolismo , Animais , Animais Recém-Nascidos , Epididimo/crescimento & desenvolvimento , Feminino , Macropodidae , Masculino , Morfogênese , Ductos Paramesonéfricos/crescimento & desenvolvimento , Próstata/crescimento & desenvolvimento , Tela Subcutânea , Testículo/transplante , Fatores de Tempo , Transplante Heterólogo , Virilismo/fisiopatologia
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