Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 88
Filtrar
1.
Hum Mol Genet ; 28(10): 1671-1681, 2019 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-30649340

RESUMO

Hand-Foot-Genital syndrome is a rare condition caused by mutations in the HOXA13 gene and characterized by limb malformations and urogenital defects. While the role of Hoxa13 in limb development has been extensively studied, its function during the development of the urogenital system remains elusive mostly due to the embryonic lethality of Hoxa13 homozygous mutant mice. Using a conditional inactivation strategy, we show that mouse fetuses lacking Hoxa13 function develop megaureters, hydronephrosis and malformations of the uterus, reminiscent of the defects characterizing patients with Hand-Foot-Genital syndrome. Our analysis reveals that Hoxa13 plays a critical role in Müllerian ducts fusion and in ureter remodeling by regulating the elimination of the caudal common nephric duct, eventually preventing the separation from the nephric duct. Our data also reveal a specific role for Hoxa13 in the urogenital sinus, which is in part mediated by Gata3, as well as Hoxa13 requirement for the proper organization of the ureter. Finally, we provide evidence that Hoxa13 provides positional and temporal cues during the development of the lower urogenital system, a sine qua non condition for the proper function of the urinary system.


Assuntos
Anormalidades Múltiplas/genética , Deformidades Congênitas do Pé/genética , Fator de Transcrição GATA3/genética , Deformidades Congênitas da Mão/genética , Proteínas de Homeodomínio/genética , Anormalidades Urogenitais/genética , Sistema Urogenital/fisiopatologia , Anormalidades Múltiplas/fisiopatologia , Animais , Extremidades/crescimento & desenvolvimento , Extremidades/fisiopatologia , Deformidades Congênitas do Pé/fisiopatologia , Deformidades Congênitas da Mão/fisiopatologia , Humanos , Rim/anormalidades , Rim/patologia , Deformidades Congênitas dos Membros/genética , Deformidades Congênitas dos Membros/fisiopatologia , Camundongos , Ductos Paramesonéfricos/fisiopatologia , Mutação , Ureter/anormalidades , Ureter/fisiopatologia , Anormalidades Urogenitais/fisiopatologia , Sistema Urogenital/crescimento & desenvolvimento
2.
Differentiation ; 103: 100-119, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30287094

RESUMO

We have studied the ontogeny of the developing human male and female urogenital tracts from 9 weeks (indifferent stage) to 16 weeks (advanced sex differentiation) of gestation by immunohistochemistry on mid-sagittal sections. Sixteen human fetal pelvises were serial sectioned in the sagittal plane and stained with antibodies to epithelial, muscle, nerve, proliferation and hormone receptor markers. Key findings are: (1) The corpus cavernosum in males and females extends into the glans penis and clitoris, respectively, during the ambisexual stage (9 weeks) and thus appears to be an androgen-independent event. (2) The entire human male (and female) urethra is endodermal in origin based on the presence of FOXA1, KRT 7, uroplakin, and the absence of KRT10 staining. The endoderm of the urethra interfaces with ectodermal epidermis at the site of the urethral meatus. (3) The surface epithelium of the verumontanum is endodermal in origin (FOXA1-positive) with a possible contribution of Pax2-positive epithelial cells implying additional input from the Wolffian duct epithelium. (4) Prostatic ducts arise from the endodermal (FOXA1-positive) urogenital sinus epithelium near the verumontanum. (5) Immunohistochemical staining of mid-sagittal and para-sagittal sections revealed the external anal sphincter, levator ani, bulbospongiosus muscle and the anatomic relationships between these developing skeletal muscles and organs of the male and female reproductive tracts. Future studies of normal human developmental anatomy will lay the foundation for understanding congenital anomalies of the lower urogenital tract.


Assuntos
Desenvolvimento Fetal/genética , Imuno-Histoquímica , Uretra/crescimento & desenvolvimento , Sistema Urogenital/crescimento & desenvolvimento , Clitóris/crescimento & desenvolvimento , Clitóris/metabolismo , Epitélio/crescimento & desenvolvimento , Epitélio/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Genitália Feminina/crescimento & desenvolvimento , Fator 3-alfa Nuclear de Hepatócito/genética , Humanos , Queratina-10/genética , Masculino , Fator de Transcrição PAX2/genética , Pênis/crescimento & desenvolvimento , Pênis/metabolismo , Uretra/metabolismo , Sistema Urogenital/metabolismo , Vagina/crescimento & desenvolvimento , Vagina/metabolismo
3.
Differentiation ; 103: 5-13, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30245193

RESUMO

We present a detailed review of fetal development of the male and female human urogenital tract from 8 to 22 weeks gestation at the macroscopic and morphometric levels. Human fetal specimens were sexed based on macroscopic identification of fetal testes or ovaries, Wolffian or Müllerian structures and the presence of the SRY gene in the specimens at or near the indifferent stage (8-9 weeks). Specimens were photographed using a dissecting microscope with transmitted and reflected light. Morphometric measurements were taken of each urogenital organ. During this time period, development of the male and female urogenital tracts proceeded from the indifferent stage to differentiated organs. The kidneys, ureters, and bladder developed identically, irrespective of sex with the same physical dimensions and morphologic appearance. The penis, prostate and testis developed in males and the clitoris, uterus and ovary in females. Androgen-dependent growth certainly influenced size and morphology of the penile urethra and prostate, however, androgen-independent growth also accounted for substantial growth in the fetal urogenital tract including the clitoris.


Assuntos
Diferenciação Celular/genética , Ovário/ultraestrutura , Testículo/ultraestrutura , Sistema Urogenital/ultraestrutura , Feminino , Desenvolvimento Fetal , Feto , Genitália/embriologia , Genitália/crescimento & desenvolvimento , Genitália/ultraestrutura , Humanos , Masculino , Ovário/embriologia , Ovário/crescimento & desenvolvimento , Testículo/embriologia , Testículo/crescimento & desenvolvimento , Sistema Urogenital/crescimento & desenvolvimento
4.
Differentiation ; 103: 24-45, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30224091

RESUMO

This paper provides a detailed compilation of human prostatic development that includes human fetal prostatic gross anatomy, histology, and ontogeny of selected epithelial and mesenchymal differentiation markers and signaling molecules throughout the stages of human prostatic development: (a) pre-bud urogenital sinus (UGS), (b) emergence of solid prostatic epithelial buds from urogenital sinus epithelium (UGE), (c) bud elongation and branching, (d) canalization of the solid epithelial cords, (e) differentiation of luminal and basal epithelial cells, and (f) secretory cytodifferentiation. Additionally, we describe the use of xenografts to assess the actions of androgens and estrogens on human fetal prostatic development. In this regard, we report a new model of de novo DHT-induction of prostatic development from xenografts of human fetal female urethras, which emphasizes the utility of the xenograft approach for investigation of initiation of human prostatic development. These studies raise the possibility of molecular mechanistic studies on human prostatic development through the use of tissue recombinants composed of mutant mouse UGM combined with human fetal prostatic epithelium. Our compilation of human prostatic developmental processes is likely to advance our understanding of the pathogenesis of benign prostatic hyperplasia and prostate cancer as the neoformation of ductal-acinar architecture during normal development is shared during the pathogenesis of benign prostatic hyperplasia and prostate cancer.


Assuntos
Mesoderma/crescimento & desenvolvimento , Próstata/crescimento & desenvolvimento , Neoplasias da Próstata/genética , Sistema Urogenital/crescimento & desenvolvimento , Androgênios/genética , Androgênios/metabolismo , Diferenciação Celular/genética , Células Epiteliais/metabolismo , Estrogênios/genética , Estrogênios/metabolismo , Feminino , Humanos , Masculino , Neoplasias da Próstata/patologia , Uretra/crescimento & desenvolvimento , Sistema Urogenital/metabolismo
6.
Differentiation ; 103: 14-23, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30262218

RESUMO

Recent studies in our lab have utilized three imaging techniques to visualize the developing human fetal urogenital tract in three dimensions: optical projection tomography, scanning electron microscopy and lightsheet fluorescence microscopy. We have applied these technologies to examine changes in morphology and differential gene expression in developing human external genital specimens from the ambisexual stage (<9 weeks fetal age) to well-differentiated male and female organs (>13 weeks fetal age). This work outlines the history and function of each of these three imaging modalities, our methods to prepare specimens for each and the novel findings we have produced thus far. We believe the images in this paper of human fetal urogenital organs produced using lightsheet fluorescence microscopy are the first published to date.


Assuntos
Desenvolvimento Fetal/genética , Imageamento Tridimensional/métodos , Diferenciação Sexual/genética , Sistema Urogenital/ultraestrutura , Feminino , Humanos , Masculino , Microscopia Eletrônica de Varredura , Sistema Urogenital/crescimento & desenvolvimento
7.
Differentiation ; 103: 66-73, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30236462

RESUMO

The urinary bladder collects urine from the kidneys and stores it until the appropriate moment for voiding. The trigone and ureterovesical junctions are key to bladder function, by allowing one-way passage of urine into the bladder without obstruction. Embryological development of these structures has been studied in multiple animal models as well as humans. In this report we review the existing literature on bladder development and cellular signalling with particular focus on bladder development in humans. The bladder and ureterovesical junction form primarily during the fourth to eighth weeks of gestation, and arise from the primitive urogenital sinus following subdivision of the cloaca. The bladder develops through mesenchymal-epithelial interactions between the endoderm of the urogenital sinus and mesodermal mesenchyme. Key signalling factors in bladder development include shh, TGF-ß, Bmp4, and Fgfr2. A concentration gradient of shh is particularly important in development of bladder musculature, which is vital to bladder function. The ureterovesical junction forms from the interaction between the Wolffian duct and the bladder. The ureteric bud arises from the Wolffian duct and is incorporated into the developing bladder at the trigone. It was previously thought that the trigonal musculature developed primarily from the Wolffian duct, but it has been shown to develop primarily from bladder mesenchyme. Following emergence of the ureters from the Wolffian ducts, extensive epithelial remodelling brings the ureters to their final trigonal positions via vitamin A-induced apoptosis. Perturbation of this process is implicated in clinical obstruction or urine reflux. Congenital malformations include ureteric duplication and bladder exstrophy.


Assuntos
Desenvolvimento Embrionário/genética , Rim/crescimento & desenvolvimento , Bexiga Urinária/crescimento & desenvolvimento , Ductos Mesonéfricos/crescimento & desenvolvimento , Animais , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Masculino , Sistema Urogenital/crescimento & desenvolvimento
8.
Differentiation ; 101: 25-38, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29684807

RESUMO

This study documented, for the first time, the morphological patterns of differentiation of male and female genital organs of Spix cavy (Galea spixii) using histological and ultrastructural analyses, with immuno-localization of steroidogenic enzymes, cytochromes P450 aromatase (P450arom) and 17α-hydroxylase/17, 20-lyase (P450c17), involved in the synthesis of estrogens and androgens respectively throughout fetal sexual development. Undifferentiated gonads of Spix cavy develop into ovaries in females after 25 days of gestation (DG), exhibiting P450arom immunoreactivity. After 25 DG, paramesonephric ducts develop and form oviducts, uterine horns and cranial portion of the vagina. The caudal portion of the vagina originates from the urogenital sinus, and a vaginal closure membrane is present at the end of gestation. Partial channeling of the urethra into the clitoris occurs after 40 DG, but complete channeling never occurs. A preputial meatus emerges near the tip of organ. In males, undifferentiated gonads develop into testes at 25 DG and develop immunoreactivity for P450c17, which is required for androgens synthesis and likely maintenance of mesonephric ducts. Mesonephric ducts develop subsequently, forming the epididymis and ductus deferens. The pelvic urethra develops after 25 DG with channeling into the penis occurring around 30 DG. This is the first morphological study describing the process of sexual differentiation during gestation in a hystricomorph rodent and one of the most comprehensive analyses conducted in any mammal. Male genital organ development follows the general pattern described in other domestic mammals, but does not include formation of the baculum as occurs in mice and rats. In females, clitoral development includes partial canalization by the urethra and development of a preputial meatus. Further studies are required to clarify the mechanisms involved in the differentiative processes described.


Assuntos
Ovário/crescimento & desenvolvimento , Diferenciação Sexual/fisiologia , Testículo/crescimento & desenvolvimento , Uretra/crescimento & desenvolvimento , Sistema Urogenital/crescimento & desenvolvimento , Animais , Feminino , Gônadas/crescimento & desenvolvimento , Cobaias , Masculino , Ductos Mesonéfricos/crescimento & desenvolvimento
9.
PLoS One ; 13(1): e0191224, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29351342

RESUMO

Congenital anomalies of the kidney and urinary tract (CAKUT) are the most common cause (40-50%) of chronic kidney disease (CKD) in children. About 40 monogenic causes of CAKUT have so far been discovered. To date less than 20% of CAKUT cases can be explained by mutations in these 40 genes. To identify additional monogenic causes of CAKUT, we performed whole exome sequencing (WES) and homozygosity mapping (HM) in a patient with CAKUT from Indian origin and consanguineous descent. We identified a homozygous missense mutation (c.1336C>T, p.Arg446Cys) in the gene Von Willebrand factor A domain containing 2 (VWA2). With immunohistochemistry studies on kidneys of newborn (P1) mice, we show that Vwa2 and Fraser extracellular matrix complex subunit 1 (Fras1) co-localize in the nephrogenic zone of the renal cortex. We identified a pronounced expression of Vwa2 in the basement membrane of the ureteric bud (UB) and derivatives of the metanephric mesenchyme (MM). By applying in vitro assays, we demonstrate that the Arg446Cys mutation decreases translocation of monomeric VWA2 protein and increases translocation of aggregated VWA2 protein into the extracellular space. This is potentially due to the additional, unpaired cysteine residue in the mutated protein that is used for intermolecular disulfide bond formation. VWA2 is a known, direct interactor of FRAS1 of the Fraser-Complex (FC). FC-encoding genes and interacting proteins have previously been implicated in the pathogenesis of syndromic and/or isolated CAKUT phenotypes in humans. VWA2 therefore constitutes a very strong candidate in the search for novel CAKUT-causing genes. Our results from in vitro experiments indicate a dose-dependent neomorphic effect of the Arg446Cys homozygous mutation in VWA2.


Assuntos
Biomarcadores Tumorais/genética , Síndrome de Fraser/genética , Mutação de Sentido Incorreto , Anormalidades Urogenitais/genética , Refluxo Vesicoureteral/genética , Sequência de Aminoácidos , Substituição de Aminoácidos , Animais , Animais Recém-Nascidos , Biomarcadores Tumorais/química , Proteínas de Ligação ao Cálcio , Criança , Consanguinidade , Sequência Conservada , Éxons , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Homozigoto , Humanos , Masculino , Camundongos , Modelos Animais , Modelos Moleculares , Linhagem , Homologia de Sequência de Aminoácidos , Sistema Urogenital/crescimento & desenvolvimento , Sistema Urogenital/metabolismo
10.
Kidney Int ; 91(2): 338-351, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-27914708

RESUMO

Urea transporters (UT) are a family of transmembrane urea-selective channel proteins expressed in multiple tissues and play an important role in the urine concentrating mechanism of the mammalian kidney. UT inhibitors have diuretic activity and could be developed as novel diuretics. To determine if functional deficiency of all UTs in all tissues causes physiological abnormality, we established a novel mouse model in which all UTs were knocked out by deleting an 87 kb of DNA fragment containing most parts of Slc14a1 and Slc14a2 genes. Western blot analysis and immunofluorescence confirmed that there is no expression of urea transporter in these all-UT-knockout mice. Daily urine output was nearly 3.5-fold higher, with significantly lower urine osmolality in all-UT-knockout mice than that in wild-type mice. All-UT-knockout mice were not able to increase urinary urea concentration and osmolality after water deprivation, acute urea loading, or high protein intake. A computational model that simulated UT-knockout mouse models identified the individual contribution of each UT in urine concentrating mechanism. Knocking out all UTs also decreased the blood pressure and promoted the maturation of the male reproductive system. Thus, functional deficiency of all UTs caused a urea-selective urine-concentrating defect with little physiological abnormality in extrarenal organs.


Assuntos
Rim/metabolismo , Proteínas de Membrana Transportadoras/deficiência , Ureia/metabolismo , Animais , Comportamento Animal , Pressão Sanguínea , Simulação por Computador , Proteínas Alimentares/metabolismo , Feminino , Genótipo , Capacidade de Concentração Renal , Masculino , Proteínas de Membrana Transportadoras/genética , Camundongos Endogâmicos C57BL , Camundongos Knockout , Modelos Biológicos , Concentração Osmolar , Fenótipo , Eliminação Renal , Reprodução , Fatores de Tempo , Ureia/urina , Micção , Sistema Urogenital/crescimento & desenvolvimento , Sistema Urogenital/metabolismo , Privação de Água , Transportadores de Ureia
11.
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
12.
Environ Toxicol ; 31(12): 1740-1750, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26443714

RESUMO

Substances that mimic endogenous hormones may alter the cell signaling that govern prostate development and predispose it to developing lesions in adult and senile life. Bisphenol A is able to mimic estrogens, and studies have demonstrated that low levels of exposure to this compound have caused alterations during prostate development. The aim of this study was to describe the prostate development in both male and female neonatal gerbils in normal conditions and under exposure to BPA during intrauterine life, and also to analyze whether the effects of intrauterine exposure to BPA remain in adulthood. Morphological, stereological, three-dimensional reconstruction, and immunohistochemical methods were employed. The results demonstrated that in 1-day-old normal gerbils, the female paraurethral glands and the male ventral lobe are morphologically similar, although its tissue components-epithelial buds (EB), periurethral mesenchyme (PeM), paraurethral mesenchyme (PaM) or ventral mesenchymal pad (VMP), and smooth muscle (SM)-have presented different immunolabeling pattern for androgen receptor (AR), and for proliferating cell nuclear antigen (PCNA). Moreover, we observed a differential response of male and female prostate to intrauterine BPA exposure. In 1-day-old males, the intrauterine exposure to BPA caused a decrease of AR-positive cells in the PeM and SM, and a decrease of the proliferative status in the EB. In contrast, no morphological alterations were observed in ventral prostate of adult males. In 1-day-old females, BPA exposure promoted an increase of estrogen receptor alpha (ERα) positive cells in PeM and PaM, a decrease of AR-positive cells in EB and PeM, besides a reduction of cell proliferation in EB. Additionally, the adult female prostate of BPA-exposed animals presented an increase of AR- and PCNA-positive cells. These results suggest that the prostate of female gerbils were more susceptible to the intrauterine BPA effects, since they became more proliferative in adult life. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1740-1750, 2016.


Assuntos
Compostos Benzidrílicos/toxicidade , Disruptores Endócrinos/toxicidade , Fenóis/toxicidade , Sistema Urogenital/efeitos dos fármacos , Fatores Etários , Animais , Animais Recém-Nascidos , Proliferação de Células/efeitos dos fármacos , Epitélio/efeitos dos fármacos , Epitélio/metabolismo , Receptor alfa de Estrogênio/metabolismo , Feminino , Gerbillinae , Masculino , Exposição Materna/efeitos adversos , Mesoderma/efeitos dos fármacos , Mesoderma/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Próstata/citologia , Próstata/efeitos dos fármacos , Próstata/embriologia , Próstata/crescimento & desenvolvimento , Receptores Androgênicos/metabolismo , Fatores Sexuais , Sistema Urogenital/citologia , Sistema Urogenital/embriologia , Sistema Urogenital/crescimento & desenvolvimento
13.
Sex Dev ; 9(4): 216-28, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26406875

RESUMO

The mammalian prostate is a compact structure in humans but multi-lobed in mice. In humans and mice, FOXA1 and SOX9 play pivotal roles in prostate morphogenesis, but few other species have been examined. We examined FOXA1 and SOX9 in the marsupial tammar wallaby, Macropus eugenii, which has a segmented prostate more similar to human than to mouse. In males, prostatic budding in the urogenital epithelium (UGE) was initiated by day 24 postpartum (pp), but in the female the UGE remained smooth and had begun forming the marsupial vaginal structures. FOXA1 was upregulated in the male urogenital sinus (UGS) by day 51 pp, whilst in the female UGS FOXA1 remained basal. FOXA1 was localised in the UGE in both sexes between day 20 and 80 pp. SOX9 was upregulated in the male UGS at day 21-30 pp and remained high until day 51-60 pp. SOX9 protein was localised in the distal tips of prostatic buds which were highly proliferative. The persistent upregulation of the transcription factors SOX9 and FOXA1 after the initial peak and fall of androgen levels suggest that in the tammar, as in other mammals, these factors are required to sustain prostate differentiation, development and proliferation as androgen levels return to basal levels.


Assuntos
Expressão Gênica , Fator 3-alfa Nuclear de Hepatócito/genética , Macropodidae/metabolismo , Próstata/crescimento & desenvolvimento , Fatores de Transcrição SOX9/genética , Sistema Urogenital/crescimento & desenvolvimento , Envelhecimento , Animais , Feminino , Fator 3-alfa Nuclear de Hepatócito/análise , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Reação em Cadeia da Polimerase/veterinária , Antígeno Nuclear de Célula em Proliferação/análise , Próstata/metabolismo , Fatores de Transcrição SOX9/análise , Caracteres Sexuais , Sistema Urogenital/química , Sistema Urogenital/metabolismo , Vagina/crescimento & desenvolvimento
14.
Cell Rep ; 9(6): 2180-91, 2014 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-25497095

RESUMO

Maf1 is a conserved repressor of RNA polymerase (Pol) III transcription; however, its physiological role in the context of a multicellular organism is not well understood. Here, we show that C. elegans MAFR-1 is functionally orthologous to human Maf1, represses the expression of both RNA Pol III and Pol II transcripts, and mediates organismal fecundity and lipid homeostasis. MAFR-1 impacts lipid transport by modulating intestinal expression of the vitellogenin family of proteins, resulting in cell-nonautonomous defects in the developing reproductive system. MAFR-1 levels inversely correlate with stored intestinal lipids, in part by influencing the expression of the lipogenesis enzymes fasn-1/FASN and pod-2/ACC1. Animals fed a high carbohydrate diet exhibit reduced mafr-1 expression and mutations in the insulin signaling pathway genes daf-18/PTEN and daf-16/FoxO abrogate the lipid storage defects associated with deregulated mafr-1 expression. Our results reveal physiological roles for mafr-1 in regulating organismal lipid homeostasis, which ensure reproductive success.


Assuntos
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/metabolismo , Fertilidade , Metabolismo dos Lipídeos , Proteínas Repressoras/metabolismo , Acetiltransferases/genética , Acetiltransferases/metabolismo , Animais , Caenorhabditis elegans/genética , Caenorhabditis elegans/fisiologia , Proteínas de Caenorhabditis elegans/genética , Ácido Graxo Sintase Tipo I/genética , Ácido Graxo Sintase Tipo I/metabolismo , Homeostase , Mucosa Intestinal/metabolismo , RNA Polimerase I/genética , RNA Polimerase I/metabolismo , RNA Polimerase III/genética , RNA Polimerase III/metabolismo , Proteínas Repressoras/genética , Sistema Urogenital/crescimento & desenvolvimento , Sistema Urogenital/metabolismo , Sistema Urogenital/fisiologia , Vitelogeninas/genética , Vitelogeninas/metabolismo
15.
Dev Biol ; 387(1): 64-72, 2014 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-24394376

RESUMO

Defects of the ventral body wall are prevalent birth anomalies marked by deficiencies in body wall closure, hypoplasia of the abdominal musculature and multiple malformations across a gamut of organs. However, the mechanisms underlying ventral body wall defects remain elusive. Here, we investigated the role of Wnt signaling in ventral body wall development by inactivating Wls or ß-catenin in murine abdominal ectoderm. The loss of Wls in the ventral epithelium, which blocks the secretion of Wnt proteins, resulted in dysgenesis of ventral musculature and genito-urinary tract during embryonic development. Molecular analyses revealed that the dermis and myogenic differentiation in the underlying mesenchymal progenitor cells was perturbed by the loss of ectodermal Wls. The activity of the Wnt-Pitx2 axis was impaired in the ventral mesenchyme of the mutant body wall, which partially accounted for the defects in ventral musculature formation. In contrast, epithelial depletion of ß-catenin or Wnt5a did not resemble the body wall defects in the ectodermal Wls mutant. These findings indicate that ectodermal Wnt signaling instructs the underlying mesodermal specification and abdominal musculature formation during ventral body wall development, adding evidence to the theory that ectoderm-mesenchyme signaling is a potential unifying mechanism for the origin of ventral body wall defects.


Assuntos
Abdome/embriologia , Peptídeos e Proteínas de Sinalização Intracelular/fisiologia , Desenvolvimento Muscular/genética , Receptores Acoplados a Proteínas G/fisiologia , Via de Sinalização Wnt/genética , beta Catenina/fisiologia , Abdome/crescimento & desenvolvimento , Animais , Padronização Corporal/genética , Diferenciação Celular/genética , Ectoderma/embriologia , Ectoderma/crescimento & desenvolvimento , Ectoderma/metabolismo , Embrião de Mamíferos/embriologia , Embrião de Mamíferos/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/genética , Peptídeos e Proteínas de Sinalização Intracelular/genética , Mesoderma/embriologia , Mesoderma/crescimento & desenvolvimento , Mesoderma/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores Acoplados a Proteínas G/genética , Fatores de Transcrição/genética , Sistema Urogenital/embriologia , Sistema Urogenital/crescimento & desenvolvimento , Proteínas Wnt/deficiência , Proteínas Wnt/genética , Proteínas Wnt/metabolismo , Proteína Wnt-5a , beta Catenina/genética , Proteína Homeobox PITX2
16.
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
17.
J Vis Exp ; (76)2013 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-23852031

RESUMO

Progress in prostate cancer research is severely limited by the availability of human-derived and hormone-naïve model systems, which limit our ability to understand genetic and molecular events underlying prostate disease initiation. Toward developing better model systems for studying human prostate carcinogenesis, we and others have taken advantage of the unique pro-prostatic inductive potential of embryonic rodent prostate stroma, termed urogenital sinus mesenchyme (UGSM). When recombined with certain pluripotent cell populations such as embryonic stem cells, UGSM induces the formation of normal human prostate epithelia in a testosterone-dependent manner. Such a human model system can be used to investigate and experimentally test the ability of candidate prostate cancer susceptibility genes at an accelerated pace compared to typical rodent transgenic studies. Since Human embryonic stem cells (hESCs) can be genetically modified in culture using inducible gene expression or siRNA knock-down vectors prior to tissue recombination, such a model facilitates testing the functional consequences of genes, or combinations of genes, which are thought to promote or prevent carcinogenesis. The technique of isolating pure populations of UGSM cells, however, is challenging and learning often requires someone with previous expertise to personally teach. Moreover, inoculation of cell mixtures under the renal capsule of an immunocompromised host can be technically challenging. Here we outline and illustrate proper isolation of UGSM from rodent embryos and renal capsule implantation of tissue mixtures to form human prostate epithelium. Such an approach, at its current stage, requires in vivo xenografting of embryonic stem cells; future applications could potentially include in vitro gland formation or the use of induced pluripotent stem cell populations (iPSCs).


Assuntos
Células-Tronco Embrionárias/citologia , Epitélio/crescimento & desenvolvimento , Mesoderma/crescimento & desenvolvimento , Próstata/crescimento & desenvolvimento , Transplante Heterólogo/métodos , Sistema Urogenital/crescimento & desenvolvimento , Animais , Células-Tronco Embrionárias/transplante , Feminino , Humanos , Masculino , Mesoderma/citologia , Camundongos , Camundongos Nus , Gravidez , Próstata/citologia , Sistema Urogenital/citologia
18.
J Clin Res Pediatr Endocrinol ; 4(2): 51-60, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22672860

RESUMO

Endocrine disruptors are substances commonly encountered in every setting and condition in the modern world. It is virtually impossible to avoid the contact with these chemical compounds in our daily life. Molecules defined as endocrine disruptors constitute an extremely heterogeneous group and include synthetic chemicals used as industrial solvents/lubricants and their by-products. Natural chemicals found in human and animal food (phytoestrogens) also act as endocrine disruptors. Different from adults, children are not exposed only to chemical toxins in the environment but may also be exposed during their intrauterine life. Hundreds of toxic substances, which include neuro-immune and endocrine toxic chemical components that may influence the critical steps of hormonal, neurological and immunological development, may affect the fetus via the placental cord and these substances may be excreted in the meconium. Children and especially newborns are more sensitive to environmental toxins compared to adults. Metabolic pathways are immature, especially in the first months of life. The ability of the newborn to metabolize, detoxify and eliminate many toxins is different from that of the adults. Although exposures occur during fetal or neonatal period, their effects may sometimes be observed in later years. Further studies are needed to clarify the effects of these substances on the endocrine system and to provide evidence for preventive measures.


Assuntos
Disruptores Endócrinos/farmacologia , Desenvolvimento Fetal/efeitos dos fármacos , Feto/efeitos dos fármacos , Sistema Urogenital/efeitos dos fármacos , Disruptores Endócrinos/metabolismo , Feminino , Feto/metabolismo , Humanos , Recém-Nascido , Exposição Materna , Troca Materno-Fetal , Gravidez , Sistema Urogenital/embriologia , Sistema Urogenital/crescimento & desenvolvimento
19.
Methods Mol Biol ; 886: 223-39, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22639265

RESUMO

Studies into the molecular basis of morphogenesis frequently begin with investigations into gene expression across time and cell type in that organ. One of the most anatomically informative approaches to such studies is the use of in situ hybridization, either of intact or histologically sectioned tissues. Here, we describe the optimization of this approach for use in the temporal and spatial analysis of gene expression in the urogenital system, from embryonic development to the postnatal period. The methods described are applicable for high throughput analysis of large gene sets. As such, ISH has become a powerful technique for gene expression profiling and is valuable for the validation of profiling analyses performed using other approaches such as microarrays.


Assuntos
Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica no Desenvolvimento , Hibridização In Situ/métodos , RNA Mensageiro/genética , Sistema Urogenital/crescimento & desenvolvimento , Animais , Rim/embriologia , Rim/crescimento & desenvolvimento , Rim/metabolismo , Camundongos , RNA Mensageiro/isolamento & purificação , Sistema Urogenital/embriologia , Sistema Urogenital/metabolismo
20.
Methods Mol Biol ; 886: 185-201, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22639262

RESUMO

The Genitourinary Development Molecular Atlas Project (GUDMAP) aims to document gene expression across time and space in the developing urogenital system of the mouse, and to provide access to a variety of relevant practical and educational resources. Data come from microarray gene expression profiling (from laser-dissected and FACS-sorted samples) and in situ hybridization at both low (whole-mount) and high (section) resolutions. Data are annotated to a published, high-resolution anatomical ontology and can be accessed using a variety of search interfaces. Here, we explain how to run typical queries on the database, by gene or anatomical location, how to view data, how to perform complex queries, and how to submit data.


Assuntos
Bases de Dados Genéticas , Regulação da Expressão Gênica no Desenvolvimento , Sistema Urogenital/crescimento & desenvolvimento , Sistema Urogenital/metabolismo , Animais , Feminino , Perfilação da Expressão Gênica , Hibridização In Situ , Masculino , Camundongos , Ferramenta de Busca , Software
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...