RESUMO
Background. The immunophenotypes and potential excretory function of human mesonephros are not well studied. Methods. Five mesonephros specimens of human embryos from the 6th to 10th weeks of gestation were stained with immunohistochemical markers. Results. PAX8 was universally expressed in all renal tubules, while α-methyacyl-CoA racemase (AMACAR) was positive in proximal tubules and GATA3 was positive in distal tubular mesonephric structures. At the 8th weeks of gestation, the mesonephric glomeruli were characterized by opened glomerular capillary loops with Periodic Acid Schiff (PAS)-positive glomerular basement membranes and GATA3-positive mesangial-like cells. By the 8th week, proximal tubules showed PAS-positive brush borders, indicating reabsorption capacity, and the proximal tubules also demonstrated positivity with kidney injury molecule-1 (KIM-1), representing tubular response to injury. Conclusion. Our overall findings show detailed phenotypes of the glomerular and tubular structures of the mesonephros and indicate that at the 8th week of gestation, the mesonephros may carry out temporary excretory function before metanephros becomes fully functional.
Assuntos
Glomérulos Renais , Mesonefro , Humanos , Mesonefro/irrigação sanguínea , Mesonefro/química , Túbulos Renais Proximais , RimRESUMO
Mesonephric remnants are embryonic vestiges of the mesonephric (Wolffian) ducts which regress during normal development. These remnants have been uncommonly reported in the female and male reproductive tract as a spectrum of morphologic lesions that can be misdiagnosed as carcinoma. One case of mesonephric remnant hyperplasia of the jejunal mesentery incidentally found in a 47-year-old man is herein reported. This is the first description of mesonephric hyperplasia arisen in the mesentery. The presence of ducts, tubules, and cysts lined by bland, epithelial, cuboidal cells with scant cytoplasm, and diffuse pseudoinfiltrative growth pattern can raise the possibility of neoplasia. Immunohistochemically, mesonephric epithelia have a characteristic staining. CD10 highlights the apical-luminal aspect of the cells. Besides, intense reactivity is showed for high-molecular-weight cytokeratin (CK), CK7, bcl2, and vimentin. The main differential diagnosis includes mesothelial hyperplasia, epithelial mesothelioma, well-differentiated neuroendocrine tumor, and infiltration due to acinar adenocarcinoma of the prostate. However, a detailed microscopic study with the aid of immunohistochemistry helps separate mesonephric remnants from malignant processes. The mesonephric hyperplasia of the mesentery we have reported adds to the spectrum of mesonephric remnants a new location. Familiarity with this lesion is indispensable to avoid overdiagnosis.
Assuntos
Achados Incidentais , Jejuno/patologia , Mesentério/patologia , Mesonefro/metabolismo , Ductos Mesonéfricos/patologia , Biomarcadores/análise , Biópsia , Diagnóstico Diferencial , Humanos , Hiperplasia , Imuno-Histoquímica , Jejuno/química , Jejuno/cirurgia , Masculino , Mesentério/química , Mesentério/cirurgia , Mesonefro/química , Mesonefro/cirurgia , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Ductos Mesonéfricos/química , Ductos Mesonéfricos/cirurgiaRESUMO
When biological specimens are cut into physical sections for three-dimensional (3D) imaging by confocal laser scanning microscopy, the slices may get distorted or ruptured. For subsequent 3D reconstruction, images from different physical sections need to be spatially aligned by optimization of a function composed of a data fidelity term evaluating similarity between the reference and target images, and a regularization term enforcing transformation smoothness. A regularization term evaluating the total variation (TV), which enables the registration algorithm to account for discontinuities in slice deformation (ruptures), while enforcing smoothness on continuously deformed regions, was proposed previously. The function with TV regularization was optimized using a graph-cut (GC) based iterative solution. However, GC may generate visible registration artifacts, which impair the 3D reconstruction. We present an alternative, multilabel TV optimization algorithm, which in the examined samples prevents the artifacts produced by GC. The algorithm is slower than GC but can be sped up several times when implemented in a multiprocessor computing environment. For image pairs with uneven brightness distribution, we introduce a reformulation of the TV-based registration, in which intensity-based data terms are replaced by comparison of salient features in the reference and target images quantified by local image entropies.
Assuntos
Algoritmos , Aumento da Imagem/métodos , Mesonefro/ultraestrutura , Microscopia Confocal/métodos , Animais , Artefatos , Galinhas , Embrião de Mamíferos , Embrião não Mamífero , Entropia , Aumento da Imagem/instrumentação , Mesonefro/química , Microtomia/métodos , Inclusão em Parafina , Ratos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Coloração e Rotulagem , TartarugasRESUMO
Although renal regeneration is limited to repair of the proximal tubule in mammals, some bony fish are capable of renal regeneration through nephron neogenesis in the event of renal injury. We previously reported that nephron development in the medaka mesonephros is characterized by four histologically distinct stages, generally referred to as condensed mesenchyme, nephrogenic body, relatively small nephron, and the mature nephron. Developing nephrons are positive for wt1 expression during the first three of these stages. In the present study, we examined the regenerative response to renal injury, artificially induced by the administration of sublethal amounts of gentamicin in adult medaka. Similar to previous reports in other animals, the renal tubular epithelium and the glomerulus of the medaka kidney exhibited severe damage after exposure to this agent. However, kidneys showed substantial recovery after gentamicin administration, and a significant number of developing nephrons appeared 14 days after gentamicin administration (P < 0.01). Similarly, the expression of wt1 in developing nephrons also indicated the early stages of nephrogenesis. These findings show that medaka has the ability to regenerate kidney through nephron neogenesis during adulthood and that wt1 is a suitable marker for detecting nephrogenesis.
Assuntos
Mesonefro/fisiologia , Néfrons/fisiologia , Regeneração/fisiologia , Células-Tronco Adultas/fisiologia , Animais , Biomarcadores , Divisão Celular , Gentamicinas/toxicidade , Masculino , Mesonefro/química , Mesonefro/efeitos dos fármacos , Néfrons/química , Oryzias , Especificidade da Espécie , Proteínas WT1/análiseRESUMO
The spatial and temporal expression patterns of cytokeratins, vimentin, epithelial growth factor (EGF) and transforming growth factor alpha (TGF-alpha), were investigated in the 5-9-week old human mesonephros and metanephros. Vimentin was found in all mesonephric structures, while cytokeratins were seen only in the mesonephric tubules. EGF and TGF-alpha were detected early in all mesonephric structures, and immunoreactivity to both factors decreased in later stages. In the 5-6-week metanephros, vimentin immunoreactivity was found in all structures and later increased in the collecting system and interstitium. In the 5th week, cytokeratins 8 and 19 appeared in the ureteric bud and ampullae, and later showed increasing immunoreactivity in the collecting system and nephrons. The coexpression of intermediate filament proteins in metanephric development is a temporary feature and might be associated with mesenchymal to epithelial transformation of developing nephrons. In adult kidneys, such coexpression is associated with fibrosis or carcinomatous changes. At early stages, immunoreactivity to EGF and TGF-alpha was detected in all metanephric structures and from the 7th week onward, it decreased in differentiating nephrons. EGF and TGF-alpha patterns of appearance indicate their role in induction, proliferation and growth of metanephric structures. Disturbances in that pattern might cause reduction in kidney growth.
Assuntos
Fator de Crescimento Epidérmico/metabolismo , Queratinas/metabolismo , Rim/embriologia , Fator de Crescimento Transformador alfa/metabolismo , Vimentina/metabolismo , Anticorpos , Fator de Crescimento Epidérmico/análise , Fator de Crescimento Epidérmico/imunologia , Humanos , Imuno-Histoquímica , Queratinas/análise , Queratinas/imunologia , Rim/química , Rim/metabolismo , Mesonefro/química , Mesonefro/metabolismo , Fator de Crescimento Transformador alfa/análise , Fator de Crescimento Transformador alfa/imunologia , Vimentina/análise , Vimentina/imunologiaRESUMO
The aim of this study was to investigate androgen receptor (AR) expression in the developing human urogenital tract. The distribution of AR was examined in paraffin-embedded tissue sections of the lower urogenital tract using 55 human embryos of 8-12 weeks of gestation. Immunohistochemistry was performed for AR detection and gender was determined by polymerized chain reaction. There were no differences in the distribution of AR in male and female embryos at any stage of gestation. AR was present only in the mesenchymal tissues of the urogenital sinus at 8 weeks whilst the epithelium was negative, but after 9 weeks the epithelium also showed progressively more positive staining. In the phallus, AR staining was prominent. There was far less staining in the epithelium of the urethral groove from 8 to 10 weeks, whilst the mesenchyme of the urethral folds showed positive staining. At 11 and 12 weeks, both the urethral groove and folds showed uniform staining. The genital tubercle, genital swelling and bulbourethral gland precusors were also positively stained, although paramesonephric ducts were negative. Staining was observed in the mesonephric duct from 9 weeks. There was an absence of staining in the rectum at all stages of gestation. The expression of AR in an epithelium may be dependent upon the mesenchyme. Mesenchymal-epithelial interactions played an important role in development, as has been described in experimental animals. AR expression could play a part in the growth of the genital organs.
Assuntos
Mesoderma/química , Receptores Androgênicos/análise , Sistema Urogenital/química , Sistema Urogenital/embriologia , Glândulas Bulbouretrais/química , Glândulas Bulbouretrais/embriologia , Epitélio/química , Epitélio/embriologia , Feminino , Identidade de Gênero , Humanos , Imuno-Histoquímica/métodos , Cariotipagem , Masculino , Mesonefro/química , Mesonefro/embriologia , Pênis/química , Pênis/embriologia , Gravidez , Primeiro Trimestre da Gravidez , Uretra/química , Uretra/embriologiaRESUMO
Natriuretic peptides (NPs), a family of structurally related hormones and nitric oxide (NO), generated by nitric oxide synthase (NOS), are believed to be involved in the regulation of fluid balance and sodium homeostasis. Differential expression and regulation of these factors depend on both physiological and pathological conditions. Both NPs and NO act in target organs through the activation of guanylate cyclase (GC) and the generation of guanosine 3',5'-cyclic monophosphate (cGMP), which is considered a common messenger for the action of these factors. The present study was designed to investigate--by histochemical methods--the expression of some NPs (proANP and ANP) and isoforms of NOS (neuronal NOS, nNOS, and inducible NOS, iNOS) in the mesonephros of Rana esculenta in different periods of the year including hibernation, to evaluate possible seasonal changes in their expression. We also studied the enzyme activity of NOS-related nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) and of GC. The experiments were performed on pieces of kidney of R. esculenta collected in their natural environment during active and hibernating life. The study was carried out using immunohistochemical techniques to demonstrate proANP, ANP, and some NOS isoforms. Antigen capture by enzyme linked immunosorbent assay (ELISA) was also performed to determine the presence of NPs in the frog kidney extract. Enzyme histochemistry was used to demonstrate the NOS-related NADPHd activity at light microscopy; GC activity was visualized at the electron microscope, using cerium as capture agent. The application of the immunohistochemical techniques demonstrated that frog mesonephros tubules express different patterns of distribution and/or expression of ANP and NOS during the annual cycle. Comparing the results obtained on active and hibernating frogs has provided interesting data; the NOS/NADPHd and GC activities showed some variations as well. Furthermore, the presence of NPs in the frog kidney extract was evidenced by dose-dependent response in the ELISA. The data suggest that both ANP and NO are intra-renal paracrine and/or autocrine factors which may modulate the adaptations of frog renal functions to seasonal changes through the action of the cGMP generated from GC activity.
Assuntos
Guanilato Ciclase/metabolismo , Mesonefro/metabolismo , Peptídeos Natriuréticos/análise , Óxido Nítrico Sintase/análise , Periodicidade , Rana esculenta/metabolismo , Animais , Fator Natriurético Atrial/análise , Ensaio de Imunoadsorção Enzimática , Feminino , Hibernação , Técnicas Imunoenzimáticas , Masculino , Mesonefro/química , Mesonefro/ultraestrutura , Óxido Nítrico Sintase Tipo I , Óxido Nítrico Sintase Tipo II , Precursores de Proteínas/análise , Estações do AnoRESUMO
cDNA cloning from chicken embryonic gonad subtracted from tissues of the brain, heart, liver, gizzard, mesonephros, and muscle was performed to identify growth factor genes with expression unique to embryonic ovary and testis. We obtained several cDNA clones encoding known and many unknown genes. We found for the first time that the transforming growth factor beta2 (TGF-beta2) is preferentially expressed in the chicken embryonic ovary and testis. cDNA subtraction cloning with respect to the selective expression of TGF-beta2 in the ovary and testis was further analyzed by reverse transcription-polymerase chain reaction analyses of other embryonic tissues. The ontogeny of TGF-beta2 was evaluated in chicken embryonic ovary and testis. In both testis and ovary, the levels of TGF-beta2 transcripts were high during the early period of embryonic development (E7), gradually decreased until the late embryonic days (E14--E17), and then slightly increased at the last embryonic day (E21). There was no difference in the TGF-beta2 transcripts per RNA between the left and the right ovaries. TGF-beta2 may have a critical role in the regulation of the development of chicken ovarian and testicular germ cells during the embryonic period.
Assuntos
Clonagem Molecular/métodos , Expressão Gênica , Gônadas/embriologia , Gônadas/metabolismo , Ovário/embriologia , Testículo/embriologia , Fator de Crescimento Transformador beta/genética , Animais , Encéfalo/embriologia , Química Encefálica , Embrião de Galinha , DNA Complementar/análise , Feminino , Biblioteca Gênica , Moela das Aves/química , Moela das Aves/embriologia , Coração/embriologia , Fígado/química , Fígado/embriologia , Masculino , Mesonefro/química , Mesonefro/embriologia , Músculos/química , Músculos/embriologia , Miocárdio/química , Ovário/química , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Testículo/química , Fatores de Tempo , Fator de Crescimento Transformador beta2RESUMO
Platelet-derived growth factors (PDGFs) are paracrine factors with roles in mesenchymal-epithelial interactions during normal and pathologic processes. Previously, PDGF and its receptor (PDGFR) have been shown to be present in perinatal, peripubertal, and adult rat testes. The role of PDGF in embryonic testicular cord formation is not known. The hypothesis tested is that PDGFs and PDGFRs are expressed during cord formation and that inhibition of their action influences normal cord formation during embryonic testis development. Embryonic Day (E) 13 gonadal organ cultures were used. Organs were cultured for 3 days and treated daily with vehicle or a PDGFR-specific tyrosine phosphorylation inhibitor (i.e., the tyrphostin AG1295 or AG1296). Vehicle-treated testes formed normal cords, whereas tyrphostin-treated testes formed "swollen cords," a phenomenon characterized by a significant decrease in the number of cords per testis area and increased cord diameter due to fusion of cords. Expression of PDGF and PDGFR in E13, E14, E16, Postnatal Day (P) 0, and P20 testes was examined. Messenger RNAs for PDGF-A and -B and PDGF alpha- and beta-receptors were expressed in isolated testes during all developmental periods examined. Immunoreactivity for PDGF was present throughout the testicular compartment at E14, restricted primarily to testicular cords at E16, and present in cells of the testicular cords with a stronger immunoreactivity in certain interstitial cell types of P0 testis. PDGFR beta-receptor immunoreactivity was primarily localized to the mesonephros of E14 organs and the testicular interstitium of E16 and P0 testes. Tyrphostins did not affect apoptotic cell number in the testis. PDGF had no effect on cell growth in P0 testis cultures. The results show that PDGFs and PDGFRs are expressed in embryonic testis during cord formation in a tissue-specific manner. Inhibition of PDGF actions does not inhibit cord formation but does alter normal cord development and morphology. The observations provide insight into the factors involved in male sex differentiation and embryonic testis development.
Assuntos
Fator de Crescimento Derivado de Plaquetas/antagonistas & inibidores , Fator de Crescimento Derivado de Plaquetas/fisiologia , Túbulos Seminíferos/embriologia , Testículo/embriologia , Animais , Apoptose , Becaplermina , Inibidores Enzimáticos/farmacologia , Feminino , Expressão Gênica , Masculino , Mesonefro/química , Técnicas de Cultura de Órgãos , Fator de Crescimento Derivado de Plaquetas/análise , Fator de Crescimento Derivado de Plaquetas/genética , Gravidez , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-sis , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Receptor alfa de Fator de Crescimento Derivado de Plaquetas/análise , Receptor alfa de Fator de Crescimento Derivado de Plaquetas/genética , Receptor beta de Fator de Crescimento Derivado de Plaquetas/análise , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genética , Receptores do Fator de Crescimento Derivado de Plaquetas/análise , Receptores do Fator de Crescimento Derivado de Plaquetas/genética , Testículo/química , Testículo/efeitos dos fármacos , Tirfostinas/farmacologiaRESUMO
Proximal tubules (PT) in 7-10-day old chick mesonephros were examined histochemically to evaluate their structural and functional properties related to the absorption capacity of the epithelium and its possible alterations leading to cystically dilated tubules (CDT). Alkaline phosphatase activity at the apical cell membrane was demonstrated in varying intensities in large PT. A similar heterogeneity was also detected in the expression of proteoglycans (Lewis(x) antigen) localized in the apical part of the epithelial cell membrane but not in the basolateral membrane parts (beta-catenin, Na(+),K(+)-ATPase). In analogy, the ability to accumulate trypan blue was found in the same cell population. We hypothesize that epithelial cells in proximal tubules of nephrons with a defective apical cell membrane cause reduced fluid absorption and subsequent overfilling and dilation of the tubules.
Assuntos
Túbulos Renais Proximais/metabolismo , Mesonefro/metabolismo , Fosfatase Alcalina/análise , Fosfatase Alcalina/metabolismo , Animais , Membrana Celular/química , Membrana Celular/metabolismo , Embrião de Galinha , Citoplasma/ultraestrutura , Células Epiteliais/química , Células Epiteliais/metabolismo , Técnica Indireta de Fluorescência para Anticorpo , Túbulos Renais Proximais/química , Túbulos Renais Proximais/embriologia , Antígenos CD15/análise , Antígenos CD15/metabolismo , Mesonefro/química , Mesonefro/embriologia , Organogênese/fisiologia , Azul Tripano/metabolismoRESUMO
Alpha-fetoprotein (AFP) is the major serum protein during development. AFP is one of the earliest proteins to be synthesised by the embryonic liver. The synthesis of AFP decreases dramatically after birth and only trace amounts are expressed in the adult liver. The tissue distribution of AFP in early human embryogenesis has not been defined. We have studied the expression pattern of AFP mRNA in human and mouse embryos by in situ hybridisation. In humans, AFP is expressed in the hepatic diverticulum at 26 d postovulation as it differentiates from the foregut endoderm (i.e. in the most primitive hepatocytes). It is also expressed in the endoderm of the gastrointestinal tract and in the yolk sac at this age. AFP is subsequently expressed in the mesonephros and transiently in the developing pancreas. In the mouse, no expression of AFP was observed in the mesonephros but other sites of expression were similar. Thus AFP has a distinct temporospatial expression pattern during the embryonic period and this differs between human and mouse species. It is interesting that AFP is expressed by tumours such as primitive gastrointestinal, renal cell and pancreatic tumours as well as those of hepatocyte origin. This distribution reflects the sites of AFP expression during development.
Assuntos
Desenvolvimento Embrionário e Fetal/fisiologia , Fígado/embriologia , RNA Mensageiro/análise , Saco Vitelino/química , alfa-Fetoproteínas/genética , Animais , Idade Gestacional , Humanos , Hibridização In Situ/métodos , Fígado/química , Mesonefro/química , Camundongos , Pâncreas/química , Pâncreas/embriologiaRESUMO
The Wilms' tumour suppressor gene WT1 encodes a zinc-finger transcription factor which is essential for the development of kidney, gonads, spleen and adrenals. WT1-null embryos lack all of these viscerae and they also show a thin ventricular myocardium and unexpectedly die from cardiac failure between 13 and 15 days post coitum. We studied the localization of the WT1 protein in chick and quail embryos between stages HH18 and HH35. In early embryos, WT1 protein was located in specific areas of the coelomic mesothelium adjacent to the nephric ducts, the myocardium or the primordia of the endodermal organs (gut, liver and lungs). These mesothelial areas also showed localized expression of Slug, a zinc-finger transcription factor involved in epithelial-mesenchymal transitions. WT1+ mesenchymal cells were always found below the immunoreactive mesothelial areas, either forming a narrow band on the surface of the endodermal organs (gut, liver and lungs) or migrating throughout the mesodermal organs (mesonephros, metanephros, gonads, spleen and heart). In the developing heart, the invasion of WTI+ cells started at stage HH26, and all the ventricular myocardium was pervaded by these cells, presumably derived from the epicardium, at HH30. We suggest that WT1 is not required for the epithelial-mesenchymal transition of the coelomic mesothelium, but it might be a marker of the mesothelial-derived cells, where this protein would be acting as a repressor of the differentiation.
Assuntos
Proteínas de Ligação a DNA/análise , Mesonefro/química , Pericárdio/química , Fatores de Transcrição/análise , Animais , Especificidade de Anticorpos , Embrião de Galinha , Galinhas , Proteínas de Ligação a DNA/imunologia , Epitélio/química , Feto/química , Feto/embriologia , Gônadas/química , Gônadas/embriologia , Mesonefro/embriologia , Pericárdio/embriologia , Codorniz , Fatores de Transcrição da Família Snail , Baço/química , Baço/embriologia , Fatores de Transcrição/imunologia , Proteínas WT1RESUMO
Recent studies with avian embryos and murine embryonic stem cells have suggested that hematopoietic cells are derived from hemangioblasts, the common precursors of hematopoietic and endothelial cells. We molecularly cloned podocalyxin-like protein 1 (PCLP1) as a novel surface marker for endothelial-like cells in the aorta-gonad-mesonephros (AGM) region of mouse embryos, where long-term repopulating hematopoietic stem cells (LTR-HSCs) are known to arise. PCLP1+ CD45 cells in the AGM region incorporated acetylated low-density lipoprotein and produced both hematopoietic and endothelial cells when cocultured with OP9 stromal cells. Moreover, multiple lineages of hematopoietic cells were generated in vivo when PCLP1 +CD45-cells were injected into neonatal liver of busulfan-treated mice. Thus, PCLP1 can be used to separate hemangioblasts that give rise to LTR-HSCs.
Assuntos
Aorta/embriologia , Endotélio Vascular/embriologia , Proteínas Fetais/análise , Gônadas/embriologia , Glicoproteínas de Membrana/análise , Mesonefro/química , Peptídeos/fisiologia , Células-Tronco/química , Sequência de Aminoácidos , Animais , Aorta/química , Biomarcadores , Diferenciação Celular/efeitos dos fármacos , Linhagem da Célula , Células Cultivadas , Mapeamento Cromossômico , Técnicas de Cocultura , Endotélio Vascular/citologia , Idade Gestacional , Gônadas/química , Substâncias de Crescimento/farmacologia , Transplante de Células-Tronco Hematopoéticas , Células-Tronco Hematopoéticas/química , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/efeitos dos fármacos , Antígenos Comuns de Leucócito/análise , Lipoproteínas LDL/metabolismo , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Dados de Sequência Molecular , Oncostatina M , Peptídeos/farmacologia , Células-Tronco/classificação , Células-Tronco/efeitos dos fármacosRESUMO
We previously reported that the mouse GATA-2 gene is regulated by two alternative promoters (Minegishi et al, J Biol Chem, 273:3625, 1998). Although the more proximal IG (general) promoter is active in almost all GATA-2-expressing cells, the distal IS (specific) promoter activity was selectively detected in hematopoietic tissues but not in other mesodermal tissues. We report here in vivo analysis of the GATA-2 locus and its regulatory characteristics in hematopoietic tissues of transgenic mice. Transgenes containing 6 or 7 kbp of sequence flanking the 5' end of the IS first exon direct expression of beta-galactosidase or green fluorescent protein (GFP) reporter genes specifically to the para-aortic splanchnopleura, aorta-gonads, and mesonephros (AGM) region, and in the neural tissues. In situ hybridization analysis showed that reporter gene expression specifically recapitulates the endogenous expression profile of GATA-2 in these tissues. The flk-1, CD34, c-kit, and CD45 antigens were identified in the GFP-positive cells from the AGM region and fetal liver, indicating that GATA-2 is expressed in immature hematopoietic cells. Deletion of 3.5 kbp from the 5' end of the 6.0 kbp IS promoter construct, including one of the DNase I hypersensitive sites, completely abolished hematopoietic expression. These experiments describe an early developmental GATA-2 hematopoietic enhancer located between 6.0 and 2.5 kbp 5' to the IS exon.
Assuntos
Proteínas de Ligação a DNA/genética , Embrião de Mamíferos/metabolismo , Expressão Gênica , Hematopoese , Fatores de Transcrição/genética , Animais , Desoxirribonuclease I/metabolismo , Fator de Transcrição GATA2 , Proteínas de Fluorescência Verde , Hibridização In Situ , Fígado/química , Fígado/embriologia , Proteínas Luminescentes/análise , Proteínas Luminescentes/genética , Mesentério/química , Mesentério/embriologia , Mesonefro/química , Camundongos , Camundongos Transgênicos , Tecido Nervoso/química , Tecido Nervoso/embriologia , Glomos Para-Aórticos/química , Pleura/química , Pleura/embriologia , RNA Mensageiro/análise , Baço/química , beta-Galactosidase/genéticaRESUMO
The aim of this study was to investigate stem cell factor and c-kit gene expression and protein localization in the mesonephros and ovary of sheep fetuses at different days of gestation, using RNA in situ hybridization and immunohistochemical procedures. At days 24 and 26 of gestation, stem cell factor mRNA and protein were present in cells throughout the developing gonad and mesonephros. From day 28 to day 40 of gestation, stem cell factor mRNA and protein became increasingly localized to the cortical region of the ovary, where most germ cells were present as actively proliferating oogonia. From day 40 to day 90 of gestation, stem cell factor mRNA and protein localization were confined mainly to the ovarian cortex. Moreover, within the cortical region, stem cell factor mRNA was low or absent where follicles were first forming and highest in the outer ovarian cortex, where germ cells were undergoing mitosis or the early stages of meiosis. In contrast, stem cell factor protein was present in newly forming follicles, as well as in mitotic and meiotic germ cells, which is consistent with the presence of both membrane-bound and soluble forms of this ligand. However, by day 90 of gestation, both stem cell factor mRNA and protein were observed in the granulosa cells of most (> 90%) primordial follicles. C-kit mRNA and protein were observed from day 24 of gestation in both germ cells and somatic cells but, with increasing gestational age, preferentially in germ cells (for example, pre-meiotic germ cells and both isolated oocytes and follicle-enclosed oocytes). C-kit protein, but not mRNA, was also observed in germ cells that were undergoing meiosis. The results indicate that the cells containing stem cell factor mRNA within the ovary up to day 90 of gestation originated from the gonadal blastema and from cells that migrated from the mesonephros before day 28 of gestation. Since stem cell factor mRNA was absent in both mesonephric cells migrating after day 28 of gestation and in regions where follicles were first forming, it is suggested that these later migrating mesonephric cells are the progenitors of the granulosa cells in the first forming follicles. In conclusion, during follicle formation, c-kit mRNA is localized to germ cells whereas c-kit, together with stem cell factor protein, is localized to both germ cells and somatic cells, consistent with the hypothesis that the presence of this receptor-ligand pair is essential to prevent apoptosis.
Assuntos
Desenvolvimento Embrionário e Fetal , Ovário/embriologia , Proteínas Proto-Oncogênicas c-kit/genética , Ovinos/embriologia , Fator de Células-Tronco/genética , Animais , Feminino , Expressão Gênica , Idade Gestacional , Imuno-Histoquímica , Hibridização In Situ , Mesonefro/química , Ovário/química , Proteínas Proto-Oncogênicas c-kit/análise , RNA Mensageiro/análise , Ovinos/metabolismo , Fator de Células-Tronco/análiseRESUMO
AIMS: Adnexal tumours of probable Wolffian origin are rare low-grade malignant neoplasms that have been previously described in the broad ligament, ovaries and retroperitoneum of females. All are characterized by small, bland epithelial cells growing in a diffuse, trabecular, or tubular pattern. The majority of the cases reported have pursued a benign clinical course. However, recurrences and distant metastases have been described. We present a case of a male adnexal tumour of probable Wolffian origin occurring in the left seminal vesicle of a 29-year-old man with 23 years of follow-up. RESULTS: The diagnosis is supported by immunohistochemical and electron microscopic findings: The tumour cells were immunoreactive for cytokeratin and vimentin while smooth muscle antigen and S100 protein were uniformly negative. By electron microscopy cells were arranged in an acinar pattern and surrounded flocculent, electron-dense material. Individual cells contained numerous dense bodies and free ribrosomes. The patient had recurrences at 14 and 23 years after initial diagnosis. CONCLUSION: This is the first report of this entity in a male. The literature on this unusual tumour is reviewed and the clinicopathological, immunohistochemical and ultrastructural features are described. The differential diagnosis of this seemingly indolent tumour is discussed with genitourinary tumours having a more aggressive clinical course.
Assuntos
Doenças dos Anexos/patologia , Neoplasias dos Genitais Masculinos/patologia , Mesonefro/patologia , Glândulas Seminais/patologia , Adulto , Biomarcadores Tumorais/análise , Feminino , Neoplasias dos Genitais Masculinos/química , Neoplasias dos Genitais Masculinos/ultraestrutura , Humanos , Técnicas Imunoenzimáticas , Masculino , Mesonefro/química , Mesonefro/ultraestrutura , Microscopia Eletrônica , Recidiva Local de Neoplasia/patologia , Glândulas Seminais/química , Glândulas Seminais/ultraestrutura , Neoplasias da Bexiga Urinária/química , Neoplasias da Bexiga Urinária/secundárioRESUMO
The mammalian female reproductive system arises from the uniform paramesonephric duct. The molecular mechanisms that establish differential development along this axis are unknown. We determined the pattern and timing of genes of the Hoxa axis in the development of the Müllerian tract. Hoxa-9, Hoxa-10, Hoxa-11, and Hoxa-13 are all expressed along the length of the paramesonephric duct in the embryonic mouse. After birth, a spatial Hox axis is established, corresponding to the postnatal differentiation of this organ system in the mouse. Hoxa-9 is expressed in the fallopian tubes, Hoxa-10 in the uterus, Hoxa-11 in the uterus and uterine cervix, and Hoxa-13 in the upper vagina. This expression pattern follows the paradigm of spatial colinearity but is a novel exception to temporal colinearity that has been considered typical of Hox genes. These genes remain expressed in the adult mouse and are expressed in the same pattern in the human. The female reproductive system undergoes dramatic structural and functional changes during the estrous cycle and in pregnancy, retaining a high degree of developmental plasticity. The late establishment of a Hox axis and persistent expression of Hox genes in the adult may play an important role in preserving this plasticity.
Assuntos
Expressão Gênica , Genitália Feminina/química , Transativadores/genética , Adulto , Envelhecimento , Animais , Tubas Uterinas/química , Tubas Uterinas/embriologia , Tubas Uterinas/crescimento & desenvolvimento , Feminino , Genes Homeobox , Genitália Feminina/embriologia , Genitália Feminina/crescimento & desenvolvimento , Proteínas de Homeodomínio , Humanos , Hibridização In Situ , Mesonefro/química , Mesonefro/crescimento & desenvolvimento , Camundongos , Ductos Paramesonéfricos/química , Ductos Paramesonéfricos/crescimento & desenvolvimento , Gravidez , Especificidade da Espécie , Útero/química , Útero/embriologia , Útero/crescimento & desenvolvimento , Vagina/química , Vagina/embriologia , Vagina/crescimento & desenvolvimentoRESUMO
Increasing evidence suggests that the renin-angiotensin system (RAS) is not only a potent regulator of blood pressure and fluid and electrolyte homeostasis, but that it also plays an important role in growth and differentiation in development as well as in pathological states. We, therefore, investigated the expression of all components of the RAS in the human embryo and fetus by in situ hybridization or immunohistochemistry. This study is the first to demonstrate the presence of all components of the RAS in very early human development (30-35 days of gestation). Angiotensinogen mRNA is expressed in very high amounts in the yolk sac, liver, and kidney, whereas renin mRNA and angiotensin-converting enzyme are expressed in the chorion, kidney, and heart, thus allowing fetal production of angiotensin II. This effector molecule of the RAS mediates its effects through binding to specific receptor types, AT1 and AT2. Both of these receptors are also expressed very early in development (24 days of gestation), suggesting a role for angiotensin II in organogenesis. Based on the expression pattern of these receptors, angiotensin II likely plays a role in the growth and differentiation of the kidney, adrenal gland, heart, and liver, all organs that are of major importance for the regulation of blood pressure later in life.
Assuntos
Desenvolvimento Embrionário e Fetal/fisiologia , Sistema Renina-Angiotensina/genética , Glândulas Suprarrenais/química , Glândulas Suprarrenais/embriologia , Glândulas Suprarrenais/fisiologia , Coração/embriologia , Coração/fisiologia , Humanos , Hibridização In Situ , Rim/química , Rim/embriologia , Rim/fisiologia , Fígado/química , Fígado/embriologia , Fígado/fisiologia , Mesonefro/química , Mesonefro/embriologia , Mesonefro/fisiologia , Distribuição TecidualRESUMO
The distribution of epidermal growth factor (EGF), transforming growth factor alpha (TGF alpha), and EGF/TGF alpha receptor were studied by means of immunohistochemical methods starting from the very early stages of human embryonic kidney development. Mesonephros and metanephros were examined in order to detect immunoreactive staining in serial sectioned embryos and fetal kidneys. Anti-EGF immunoprecipitates were found in the S-shaped mesonephric vesicles of 6-week old embryos as well as in the mesonephric duct albeit with a lower degree of reactivity. Intense reactivity was observed in the metanephros within the blastemic caps of the same gestational period; the reaction was weaker within the ureteric bud branches. Bowman's capsule, proximal tubules, and collecting ducts were also reactive in the fetal kidney to varying degrees. The distribution of TGF alpha reactivity in the mesonephros was similar to that observed for EGF but with a lower intensity. In contrast, there was no reactivity in the metanephros, at least during the embyronic periods examined. By the 11th week of gestation, an intense reactivity for TGF alpha polipeptide was shown in the fetal kidney at the level of the proximal tubules and Bowman's capsule; distal tubules as well as all urinary structures from the collecting ducts to the pelvis were less reactive. Finally, EGF/TGF alpha receptor reactivity was identified by the 6th week of development, being more intense in the mesonephros at the level of the mesonephric duct cells. In the metanephros, the ureteric bud-derived branches were reactive, whereas most of the blastemic tissue did not stain. By the 11th week, only the collecting ducts and the remaining urinary structures contained reaction products: Reactivity was distributed to the tissues originating from the ureteric bud branching. Taking into account recent advances in knowledge about the biology of growth factors, the hypothesis is proposed that the secretory components (vesicles, glomerulus, and tubules) of renal anlagen might release the growth factors while the cells of the urinary tract (i.e., collecting duct, pelvis, etc.) may be their targets.
Assuntos
Fator de Crescimento Epidérmico/análise , Receptores ErbB/análise , Proteínas Fetais/análise , Rim/química , Mesonefro/química , Fator de Crescimento Transformador alfa/análise , Idade Gestacional , Humanos , Técnicas Imunoenzimáticas , Rim/embriologia , Rim/ultraestrutura , Mesonefro/ultraestrutura , MorfogêneseRESUMO
Androgens are required for the development of male internal and external genitalia. Androgen action is mediated by an intracellular receptor which acts as a transcription factor following activation by ligand binding. The aim of the present study was to define the time of appearance of androgen receptor (AR) in the male fetal rat gonad using immunohistochemistry. Intact fetuses (days 13.5-16.5) or testicular tissue (days 16.5-20.5 and days 3-7 postnatal) were fixed in Bouins' solution and processed into paraffin wax. On day 16.5 nuclear AR were present in mesenchymal cells surrounding the Wolffian duct but those around the Mullerian duct were receptor negative. During the following day (17-18) the abundance of nuclear staining increased, becoming detectable in the epithelial cells of the Wolffian ducts. Within the testis some nuclear staining was apparent at day 17 but was confined to interstitial cells surrounding the seminiferous cords. As development of the testis proceeded the abundance of nuclear AR in peritubular and elongated mesenchymal cells increased. AR were not detected in fetal Leydig cells expressing 3 beta-hydroxysteroid dehydrogenase nor in the ovaries or associated ducts of female fetuses at the same ages. In conclusion, in the rat we have found AR expression detectable by immunohistochemistry in mesonephric mesenchyme to be confined to that underlying the Wolffian ducts and to be absent from the area around the degenerating Mullerian duct. On and after day 17 of gestation AR is present in Wolffian duct epithelial cell nuclei and within the testis it is confined to peritubular and interstitial cells which may have migrated from the mesonephros.(ABSTRACT TRUNCATED AT 250 WORDS)