RESUMO
Venom is a remarkable innovation found across the animal kingdom, yet the evolutionary origins of venom systems in various groups, including spiders, remain enigmatic. Here, we investigated the organogenesis of the venom apparatus in the common house spider, Parasteatoda tepidariorum. The venom apparatus consists of a pair of secretory glands, each connected to an opening at the fang tip by a duct that runs through the chelicerae. We performed bulk RNA-seq to identify venom gland-specific markers and assayed their expression using RNA in situ hybridisation experiments on whole-mount time-series. These revealed that the gland primordium emerges during embryonic stage 13 at the chelicera tip, progresses proximally by the end of embryonic development and extends into the prosoma post-eclosion. The initiation of expression of an important toxin component in late postembryos marks the activation of venom-secreting cells. Our selected markers also exhibited distinct expression patterns in adult venom glands: sage and the toxin marker were expressed in the secretory epithelium, forkhead and sum-1 in the surrounding muscle layer, while Distal-less was predominantly expressed at the gland extremities. Our study provides the first comprehensive analysis of venom gland morphogenesis in spiders, offering key insights into their evolution and development.
Assuntos
Organogênese , Venenos de Aranha , Aranhas , Animais , Aranhas/embriologia , Aranhas/metabolismo , Venenos de Aranha/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Glândulas Exócrinas/metabolismo , Glândulas Exócrinas/embriologiaRESUMO
Fluid secretion by exocrine glandular organs is essential to the survival of mammals. Each glandular unit within the body is uniquely organized to carry out its own specific functions, with failure to establish these specialized structures resulting in impaired organ function. Here, we review glandular organs in terms of shared and divergent architecture. We first describe the structural organization of the diverse glandular secretory units (the end-pieces) and their fluid transporting systems (the ducts) within the mammalian system, focusing on how tissue architecture corresponds to functional output. We then highlight how defects in development of end-piece and ductal architecture impacts secretory function. Finally, we discuss how knowledge of exocrine gland structure-function relationships can be applied to the development of new diagnostics, regenerative approaches and tissue regeneration.
Assuntos
Glândulas Exócrinas/anatomia & histologia , Morfogênese , Animais , Glândulas Exócrinas/embriologia , Glândulas Exócrinas/fisiologia , HumanosRESUMO
The mandibular gland is an important exocrine gland of worker bees, which mainly secretes fatty acids and pheromones. Lipids have important roles in energy storage, membrane structure stabilization, and signaling. However, molecular underpinnings of mandibular gland development and lipid remodeling at the different physiological stages of worker bees is still lacking. In this study, we used scanning and transmission electron microscopy to reveal the morphological changes in secretory cells, and liquid chromatography-mass spectrometry and RNA-seq to investigate the lipidome and gene transcripts during development. The morphology of secretory cells was flat in newly emerged workers, becoming vacuolated and turgid when they were activated in nurse bees and foragers. Transport vesicles became denser from newly emerged bees to 21-day worker bees. Concentrations of 10-HDA reached a maximum within 15d workers and changes in genes expression were consistent with 10-HDA content. Non-targeted lipidomics analysis of newly emerged, 6d, and 15d worker bees revealed that PC and TAG were the main lipids in mandibular gland, and lipids dramatically altered across developmental stages. TAG 54:4 was increased most strongly at 6d and 15d worker bees, meanwhile, the abundances of TAG 54:1 and TAG 54:2 were decreased sharply. Further, transcriptomics analysis showed that differentially expressed genes were significantly enriched in key nutrient metabolic pathways, particularly lipid metabolism, in 6d and 15d bees. This multi-omic perspective provides a unique resource and deeper insight into bee mandibular gland development and baseline data for further study of the mandibular gland in worker bees.
Assuntos
Abelhas/embriologia , Glândulas Exócrinas/embriologia , Mandíbula/embriologia , Animais , Abelhas/metabolismo , Comportamento Animal/fisiologia , Glândulas Exócrinas/metabolismo , Perfilação da Expressão Gênica/métodos , Proteínas de Insetos/genética , Metabolismo dos Lipídeos/genética , Lipidômica/métodos , Mandíbula/metabolismo , Redes e Vias Metabólicas , Organogênese , Proteoma/metabolismo , Proteômica/métodos , Transcriptoma/genéticaRESUMO
We describe the morphology, histology, and histochemical characteristics of the uropygial gland (UG) of the monk parakeet Myiopsitta monachus. The UG has a heart-shape external appearance and adenomers extensively branched with a convoluted path, covered by a stratified epithelium formed by different cellular strata and divided into three zones (based on the epithelial height and lumen width), a cylindrical papilla with an internal structure of delicate type and two excretory pores surrounded by a feather tuft. Histochemical and lectin-histochemical techniques performed showed positivity against PAS, AB pH 2.5, AB-PAS, and some lectines, likely related to the granivorous feeding habits. Also, we describe the morphogenesis of the UG of the monk parakeet, which appears at embryological stage 34 as a pair of ectodermal invaginations. Heterochronic events in the onset development of the UG when compared with other birds could be recognized. Finally, to examine the phylogenetic occurrence of the UG within the Psittaciformes and infer its evolutionary history, we mapped its presence/absence over a molecular phylogeny. The reconstruction of the characters states at ancestral nodes revealed that the presence of the UG was the plesiomorphic feature for Psittaciformes and its loss evolved independently more than once.
Assuntos
Evolução Biológica , Glândulas Exócrinas/embriologia , Morfogênese , Periquitos/embriologia , Animais , Embrião não Mamífero/embriologia , Glândulas Exócrinas/anatomia & histologia , Periquitos/anatomia & histologia , FilogeniaRESUMO
The pancreas is derived from the foregut endoderm during embryonic development. After gastrulation and endoderm germ layer formation complex morphogenetic events coupled with cell differentiation programs pattern the gut tube and induce pancreas organogenesis. This results in formation of exocrine, ductal and hormone-producing endocrine cells. Among these, endocrine cells are responsible for blood glucose homeostasis and their malfunction leads to diabetes mellitus, which cannot be stopped or reversed by the current standard treatments. Thus, intense efforts to regenerate or replace the lost or dysfunctional insulin-producing ß-cells are on the way. This depends on identifying the factors that coordinate pancreas organogenesis. Here, we highlight the contribution of canonical and non-canonical Wnt signaling branches in orchestrating endoderm formation, pancreatic morphogenesis as well as endocrine cell formation and function.
Assuntos
Desenvolvimento Embrionário/fisiologia , Endoderma/embriologia , Pâncreas/embriologia , Via de Sinalização Wnt/fisiologia , Animais , Diferenciação Celular/fisiologia , Diabetes Mellitus/fisiopatologia , Glândulas Exócrinas/embriologia , Humanos , Insulina , OrganogêneseRESUMO
BACKGROUND: Aquatic species in several clades possess cement glands producing adhesive secretions of various strengths. In vertebrates, transient adhesive organs have been extensively studied in Xenopus laevis, other anurans, and in several fish species. However, the development of these structures is not fully understood. RESULTS: Here, we report on the development and functional morphology of the adhesive gland of a giant danio species, Devario malabaricus. We found that the gland is localized on the larval head, is composed of goblet-like secretory cells framed by basal, bordering, and intercalated apical epithelial cells, and is innervated by the trigeminal ganglion. The gland allows nonswimming larvae to adhere to various substrates. Its secretory cells differentiate by 12 hours postfertilization and begin to disappear in the second week of life. Exogenous retinoic acid disrupts the gland's patterning. More importantly, the single mature gland emerges from fusion of two differentiated secretory cells fields; this fusion is dependent on nonmuscle myosin II function. CONCLUSIONS: Taken together, our studies provide the first documentation of the embryonic development, structure, and function of the adhesive apparatus of a danioninae. To our knowledge, this is also the first report of a cement gland arising from convergence of two bilateral fields.
Assuntos
Cyprinidae/embriologia , Embrião não Mamífero/embriologia , Glândulas Exócrinas/embriologia , Células Caliciformes/metabolismo , Organogênese/fisiologia , Animais , Embrião não Mamífero/citologia , Glândulas Exócrinas/citologia , Células Caliciformes/citologia , Organogênese/efeitos dos fármacos , Tretinoína/farmacologiaRESUMO
Mucus secretion and mucociliary clearance are crucial processes required to maintain pulmonary homeostasis. In the trachea and nasal passages, mucus is secreted by submucosal glands (SMGs) that line the airway, with an additional contribution from goblet cells of the surface airway epithelium. The SMG mucus is rich in mucins and antimicrobial enzymes. Defective tracheal SMGs contribute to hyper-secretory respiratory diseases, such as cystic fibrosis, asthma, and chronic obstructive pulmonary disease, however little is known about the signals that regulate their morphogenesis and patterning. Here, we show that Fgf10 is essential for the normal development of murine tracheal SMGs, with gland development arresting at the early bud stage in the absence of FGF10 signalling. As Fgf10 knockout mice are lethal at birth, inducible knockdown of Fgf10 at late embryonic stages was used to follow postnatal gland formation, confirming the essential role of FGF10 in SMG development. In heterozygous Fgf10 mice the tracheal glands formed but with altered morphology and restricted distribution. The reduction in SMG branching in Fgf10 heterozygous mice was not rescued with time and resulted in a reduction in overall tracheal mucus secretion. Fgf10 is therefore a key signal in SMG development, influencing both the number of glands and extent of branching morphogenesis, and is likely, therefore, to play a role in aspects of SMG-dependent respiratory health.
Assuntos
Glândulas Exócrinas/embriologia , Fator 10 de Crescimento de Fibroblastos/metabolismo , Mucosa Respiratória/embriologia , Traqueia/embriologia , Animais , Cruzamentos Genéticos , Feminino , Fator 10 de Crescimento de Fibroblastos/deficiência , Fator 10 de Crescimento de Fibroblastos/genética , Masculino , Camundongos , Morfogênese , Muco/metabolismo , Traqueia/metabolismoAssuntos
Glândulas Exócrinas/metabolismo , Fator XIIIa/metabolismo , Células Secretoras de Insulina/metabolismo , Ilhotas Pancreáticas/metabolismo , Peptídeo Hidrolases/metabolismo , Glândulas Endócrinas/citologia , Glândulas Endócrinas/embriologia , Glândulas Endócrinas/metabolismo , Glândulas Exócrinas/citologia , Glândulas Exócrinas/embriologia , Células Secretoras de Glucagon/metabolismo , Humanos , Imuno-Histoquímica , Ilhotas Pancreáticas/citologia , Ilhotas Pancreáticas/embriologia , Células Secretoras de Somatostatina/metabolismoRESUMO
An avian uropygial gland is located on the mid-dorsum of the tail, and is the only external gland found in birds. Most studies have focused on the function, gross anatomy and chemical nature of this gland, with little research on its ontogeny. The purpose of this study was to examine the development of this gland in a series of Laysan Albatross (Phoebastria immutabilis) embryos. Specimens were examined anatomically and histologically. It was found that grooves preceded glandular development by many stages. The embryogenesis of the uropygial gland was divided into 6 phases: preinception, groove inception, mesodermal separation, migrating mesodermal cells, oval shaped "depressions", constriction and finally glandular inception. No other gland is known to develop similarly, though there may be parallels with femoral gland development. In comparison to other bird species, the length of the development period in the Albatross, as well as other compounding factors, make it difficult to determine the significance of these observations. The development of a mesodermal band, soon to be a connective tissue capsule, is more complex than originally described in ducks. Thus, the unique nature of this gland is established, but the significance of the observations required further studies into uropygial gland development. Anat Rec, 300:1420-1428, 2017. © 2017 Wiley Periodicals, Inc.
Assuntos
Aves/anatomia & histologia , Aves/embriologia , Desenvolvimento Embrionário/fisiologia , Glândulas Exócrinas/anatomia & histologia , Glândulas Exócrinas/embriologia , AnimaisRESUMO
Exocrine glands of blood-feeding parasitic copepods are believed to be important in host immune response modulation and inhibition of host blood coagulation, but also in the production of substances for integument lubrication and antifouling. In this study, we aimed to characterize the distribution of different types of salmon louse (Lepeophtheirus salmonis) exocrine glands and their site of secretion. The developmental appearance of each gland type was mapped and genes specifically expressed by glands were identified. Three types of tegumental (teg 1-3) glands and one labial gland type were found. The first glands to appear during development were teg 1 and teg 2 glands. They have ducts extending both dorsally and ventrally suggested to be important in lubricating the integument. Teg 1 glands were found to express two astacin metallopeptidases and a gene with fibronectin II domains, while teg 2 glands express a heme peroxidase. The labial glands were first identified in planktonic copepodids, with reservoirs that allows for storage of glandular products. The last gland type to appear during development was named teg 3 and was not seen before the preadult I stage when the lice become more virulent. Teg 3 glands have ducts ending ventrally at the host-parasite contact area, and may secrete substances important for the salmon lice virulence. Salmon lice teg 3 and labial glands are thus likely to be especially important in the host-parasite interaction. Proteins secreted from the salmon louse glands to its salmonid host skin or blood represents a potential interface where the host immune system can meet and elicit effective responses to sea lice antigens. The present study thus represents a fundamental basis for further functional studies and identification of possible vaccine candidates. J. Morphol. 277:1616-1630, 2016. © 2016 Wiley Periodicals, Inc.
Assuntos
Copépodes/anatomia & histologia , Copépodes/embriologia , Glândulas Exócrinas/embriologia , Glândulas Exócrinas/metabolismo , Animais , Glândulas Exócrinas/anatomia & histologia , Feminino , Interações Hospedeiro-Parasita , Hibridização In Situ , Pele/metabolismoRESUMO
Hypertrophy, hyperplasia and altered mucus secretion from the respiratory submucosal glands (SMG) are characteristics of airway diseases such as cystic fibrosis, asthma and chronic bronchitis. More commonly, hyper-secretion of the nasal SMGs contributes to allergic rhinitis and upper airway infection. Considering the role of these glands in disease states, there is a significant dearth in understanding the molecular signals that regulate SMG development and patterning. Due to the imperative role of FGF signalling during the development of other branched structures, we investigated the role of Fgf10 during initiation and branching morphogenesis of murine nasal SMGs. Fgf10 is expressed in the mesenchyme around developing SMGs while expression of its receptor Fgfr2 is seen within glandular epithelial cells. In the Fgf10 null embryo, Steno's gland and the maxillary sinus gland were completely absent while other neighbouring nasal glands showed normal duct elongation but defective branching. Interestingly, the medial nasal glands were present in Fgf10 homozygotes but missing in Fgfr2b mutants, with expression of Fgf7 specifically expressed around these developing glands, indicating that Fgf7 might compensate for loss of Fgf10 in this group of glands. Intriguingly the lateral nasal glands were only mildly affected by loss of FGF signalling, while these glands were missing in Eda mutant mice, where the Steno's and maxillary sinus gland developed as normal. This analysis reveals that regulation of nasal gland development is complex with different subsets of glands being regulated by different signalling pathways. This analysis helps shed light on the nasal gland defects observed in patients with hypohidrotic ectodermal dysplasia (HED) (defect EDA pathway) and LADD syndrome (defect FGFR2b pathway).
Assuntos
Ectodisplasinas/fisiologia , Glândulas Exócrinas/embriologia , Fator 10 de Crescimento de Fibroblastos/fisiologia , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/fisiologia , Transdução de Sinais/fisiologia , Animais , Ectodisplasinas/deficiência , Ectodisplasinas/genética , Ressecção Endoscópica de Mucosa , Glândulas Exócrinas/metabolismo , Glândulas Exócrinas/ultraestrutura , Feminino , Fator 10 de Crescimento de Fibroblastos/deficiência , Fator 10 de Crescimento de Fibroblastos/genética , Fator 7 de Crescimento de Fibroblastos/fisiologia , Masculino , Seio Maxilar/embriologia , Seio Maxilar/ultraestrutura , Mesoderma/metabolismo , Camundongos , Morfogênese , Mucosa Nasal/embriologia , Mucosa Nasal/ultraestrutura , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/deficiência , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/genéticaRESUMO
Salivary gland secretion is altered in Drosophila embryos with loss of function of the sage gene. Saliva has a reduced volume and an increased electron density according to transmission electron microscopy, resulting in regions of tube dilation and constriction with intermittent tube closure. However, the precise functions of Bmsage in silkworm (Bombyx mori) are unknown, although its sequence had been deposited in SilkDB. From this, Bmsage is inferred to be a transcription factor that regulates the synthesis of silk fibroin and interacts with another silk gland-specific transcription factor, namely, silk gland factor-1. In this study, we introduced a germline mutation of Bmsage using the Cas9/sgRNA system, a genome-editing technology, resulting in deletion of Bmsage from the genome of B. mori. Of the 15 tested samples, seven displayed alterations at the target site. The mutagenesis efficiency was about 46.7% and there were no obvious off-target effects. In the screened homozygous mutants, silk glands developed poorly and the middle and posterior silk glands (MSG and PSG) were absent, which was significantly different from the wild type. The offspring of G0 mosaic silkworms had indel mutations causing 2- or 9-bp deletions at the target site, but exhibited the same abnormal silk gland structure. Mutant larvae containing different open-reading frames of Bmsage had the same silk gland phenotype. This illustrated that the mutant phenotype was due to Bmsage knockout. We conclude that Bmsage participates in embryonic development of the silk gland.
Assuntos
Bombyx/fisiologia , Glândulas Exócrinas/embriologia , Proteínas de Insetos/metabolismo , Fatores de Transcrição/metabolismo , Animais , Bombyx/embriologia , Bombyx/genética , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Embrião não Mamífero , Glândulas Exócrinas/fisiologia , Feminino , Proteínas de Insetos/genética , Larva/genética , Larva/crescimento & desenvolvimento , Mutação , Fatores de Transcrição/genéticaRESUMO
Silk production in spiders is considered a key innovation, and to have been vital for the diversification of the clade. The evolutionary origin of the organs involved in spider silk production, however, and in particular of the silk glands, is poorly understood. Homologies have been proposed between these and other glands found in arachnids, but lacking knowledge of the embryonic development of spider silk glands hampers an evaluation of hypotheses. This study focuses on the embryonic origin of the largest silk glands of the spider Cupiennius salei, the major and minor ampullate glands. We show how the ampullate glands originate from ectodermal invaginations on the embryonic spinneret limb buds, in relation to morphogenesis of these buds. Moreover, we visualize the subsequent growth of the ampullate glands in sections of the early postembryonic stages. The invaginations are shown to correlate with expression of the proneural gene CsASH2, which is remarkable since it has been proposed that spider silk glands and their nozzles originate from sensory bristles. Hence, by confirming the ectodermal origin of spider silk glands, and by describing the (post-)embryonic morphogenesis of the ampullate glands, this work provides a starting point for further investigating into the genetic program that underlies their development.
Assuntos
Aranhas/embriologia , Aranhas/crescimento & desenvolvimento , Animais , Glândulas Exócrinas/embriologia , Glândulas Exócrinas/crescimento & desenvolvimento , Glândulas Exócrinas/ultraestrutura , Feminino , Masculino , Microscopia Eletrônica de Varredura , Seda , Aranhas/ultraestruturaRESUMO
BACKGROUND: The submucosal glands (SMGs) of the respiratory system are specialized structures essential for maintaining airway homeostasis. The significance of SMGs is highlighted by their involvement in respiratory diseases such as cystic fibrosis, asthma and chronic bronchitis, where their phenotype and function are severely altered. Uncovering the normal development of the airway SMGs is essential to elucidate their role in these disorders, however, very little is known about the cellular mechanisms and intracellular signals involved in their morphogenesis. RESULTS: This review describes in detail the embryonic developmental journey of the nasal SMGs and the postnatal development of the tracheal SMGs in the mouse. Current knowledge of the genes and signalling molecules involved in SMG organogenesis is also explored. CONCLUSION: Here we review the temporal localisation and development of the murine respiratory glands in the hope of stimulating further research into the mechanisms required for successful SMG patterning and function.
Assuntos
Glândulas Exócrinas/embriologia , Sistema Respiratório/embriologia , Adipócitos/metabolismo , Animais , Animais Geneticamente Modificados , Membrana Basal/metabolismo , Colágeno Tipo IV/metabolismo , Drosophila melanogaster , Matriz Extracelular/metabolismo , Hemócitos/metabolismo , Camundongos , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Mutação , Necrose , Osteonectina/metabolismo , Fenótipo , TemperaturaRESUMO
Epithelial gland systems play an important role in marine molluscs in fabricating lubricants, repellents, fragrances, adhesives or enzymes. In cephalopods the typically single layered epithelium provides a highly dynamic variability and affords a rapid rebuilding of gland cells. While the digestive hatching gland (also named Hoyle organ) is obligatory for most cephalopods, only four genera (Nautilus, Sepia, Euprymna and Idiosepius) produce adhesive secretions by means of glandular cells in an adhesive area on the mantle or tentacles. In Idiosepius this adhesive organ is restricted to the posterior part of the fin region on the dorsal mantle side and well developed in the adult stage. Two gland cell types could be distinguished, which produce different contents of the adhesive. During the embryonic development the same body area is occupied by the temporary hatching gland. The question arises, in which way the hatching gland degrades and is replaced by the adhesive gland. Ultrastructural analyses as well as computer tomography scans were performed to monitor the successive post hatching transformation in the mantle epithelium from hatching gland degradation to the formation of the adhesive organ. According to our investigations the hatching gland cells degrade within about 1 day after hatching by a type of programmed cell death and leave behind a temporary cellular gap in this area. First glandular cells of the adhesive gland arise 7 days after hatching and proceed evenly over the posterior mantle epithelium. In contrast, the accompanying reduction of a part of the dorsal mantle musculature is already established before hatching. The results demonstrate a distinct independence between the two gland systems and illustrate the early development of the adhesive organ as well as the corresponding modifications within the mantle.
Assuntos
Decapodiformes/citologia , Células Epiteliais/fisiologia , Epitélio/embriologia , Animais , Apoptose , Decapodiformes/embriologia , Glândulas Exócrinas/citologia , Glândulas Exócrinas/embriologia , Feminino , Masculino , OrganogêneseRESUMO
The cement gland in batrachians is a temporal ectodermic organ which is necessary for an embryo's attachment to the substrate. In this review, some notions about the origin of the cement gland of Xenopus laevis frogs, its functioning, genes being expressed in it, and regulation of its formation and development are provided. The role of some homologies of agrgenes of the cement gland in Xenopus laevis is noted at different conditions of other animals and man.
Assuntos
Glândulas Exócrinas/embriologia , Proteínas de Xenopus/genética , Xenopus laevis/embriologia , Animais , Embrião não Mamífero , Glândulas Exócrinas/anatomia & histologia , Regulação da Expressão Gênica no Desenvolvimento , Fatores de Transcrição Otx/genética , Xenopus laevis/genéticaRESUMO
The clear cells of Toker are a mysterious population of intra-epidermal glandular cells. They were originally described in nipples, but were recently observed in the vulva as well. It was hypothesized that intra-epidermal embryonic remnants or underlying glands were a potential source. The embryological aspects were investigated by studying specimens of the anogenital region of 18 male and 15 female fetuses between 12 and 39 weeks gestation. The search for Toker cells was enhanced by cytokeratin (CK) 7 immunohistochemistry. The investigation showed that Toker cell elements are a normal, though highly variable constituent of the developing anogenital region. The study revealed the following: (1) single intra-epidermal glandular vesicles near follicular anlages in interlabial sulcuses of female fetuses of 15 and 16.5 weeks gestation; (2) CK7+ solitary cells, clusters, and vesicles which were related to developing intra-epidermal follicular canal tracks and tended to disperse inside the epidermis in fetuses of approximately 18 weeks gestation; (3) dispersed CK7+ cells in fetuses of 19-23 weeks gestation; (4) characteristic CK7+ Toker cell proliferations in fetuses more than 23 weeks gestation. These observations indicate that in the anogenital region, primordial follicular cells programmed to participate in the formation of apocrine and mammary-like glands, become displaced into the epidermis where they disperse, and proliferate into Toker cell populations. However, the proximity of Toker cells to CK7+ cells in excretory ducts of late fetal apocrine and mammary-like glands suggested a possible additional source. Consequences for Toker cells of the breast and primary Paget disease are discussed.
Assuntos
Canal Anal/embriologia , Glândulas Exócrinas/embriologia , Períneo/embriologia , Escroto/embriologia , Vulva/embriologia , Canal Anal/química , Biomarcadores/análise , Proliferação de Células , Glândulas Exócrinas/química , Feminino , Idade Gestacional , Humanos , Imuno-Histoquímica , Queratina-7/análise , Masculino , Escroto/química , Vulva/químicaRESUMO
Phoneutria (Ctenidae) is among the most dangerous venomous spiders in Brazil. Its venom is composed of a mixture of pharmacologically active components, some of which have been quite extensively studied due to their potentiality as models for new pharmaceutical drugs. Nevertheless, literature data on the venom-producing glands are very limited. In the present study, we follow the biological development of intra-cocoon stages of Phoneutria nigriventer spiders, mainly regarding the formation of the venomous apparatus and venom production. The results showed that the venom glands of Phoneutria are already present in the early 1st pre-larva stage. The venomous apparatus is completely formed in the larva, a stage that precedes the spider eclosion from the cocoon. At embryo stages, transcripts of a vertebrate-active neurotoxin (PhTx1) were shown to be present, as well as, unidentified venom proteins that were immunolabeled by anti-venom antibodies. It seems that venom toxins play roles in the protection and survival of those early developmental stages of Phoneutria spiders.
Assuntos
Embrião não Mamífero/embriologia , Desenvolvimento Embrionário/fisiologia , Venenos de Aranha/metabolismo , Aranhas/embriologia , Animais , Embrião não Mamífero/fisiologia , Embrião não Mamífero/ultraestrutura , Glândulas Exócrinas/embriologia , Glândulas Exócrinas/metabolismo , Feminino , Expressão Gênica , Larva/crescimento & desenvolvimento , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , RNA Mensageiro/metabolismo , Aranhas/fisiologiaRESUMO
The basic helix-loop-helix transcription factor Neurog3 (Neurogenin3 or Ngn3) actively drives endodermal progenitor cells towards endocrine islet cell differentiation during embryogenesis. Here, we manipulate Neurog3 expression levels in endocrine progenitor cells without altering its expression pattern using heterozygosity and a hypomorph. Lowered Neurog3 gene dosage in the developing pancreatic epithelium reduces the overall production of endocrine islet cells without significantly affecting the proportions of various islet cell types that do form. A reduced Neurog3 production level in the endocrine-directed pancreatic progenitor population activates the expression of Neurog3 in an increased number of epithelial progenitors. Yet a significant number of these Neurog3+ cells detected in heterozygous and hypomorphic pancreata, possibly those that express low levels of Neurog3, move on to adopt pancreatic ductal or acinar fates. These data directly demonstrate that achieving high levels of Neurog3 expression is a critical step for endocrine commitment from multipotent pancreatic progenitors. These findings also suggest that a high level of Neurog3 expression could mediate lateral inhibition or other unknown feedback mechanisms to regulate the number of cells that initiate Neurog3 transcription and protein production. The control of Neurog3+ cell number and the Neurog3 threshold-dependent endocrine differentiation mechanism combine to select a specific proportion of pancreatic progenitor cells to adopt the islet cell fate.
Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Glândulas Endócrinas/embriologia , Glândulas Exócrinas/embriologia , Dosagem de Genes , Proteínas do Tecido Nervoso/genética , Pâncreas/embriologia , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/fisiologia , Linhagem da Célula , Glândulas Endócrinas/citologia , Glândulas Exócrinas/citologia , Camundongos , Camundongos Transgênicos , Proteínas do Tecido Nervoso/fisiologia , Pâncreas/citologia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Regulation of the lymphoid enhancer factor 1 (Lef-1) transcription factor is important for the inductive formation of many epithelial-derived appendages including airway submucosal glands (SMGs). Although Wnts have been linked to developmental processes involving transcriptional activation of the Lef-1 protein, there is little in vivo information directly linking Wnts with the transcriptional regulation of the Lef-1 promoter. In the present study, we hypothesized that Wnt3a directly regulates Lef-1 gene expression required for SMG morphogenesis in mice. In support of this hypothesis, TOPGAL reporter mice demonstrated activation of beta-catenin/Tcf complexes during early phases of SMG development and immunolocalization studies confirmed abundant expression of Tcf4, but not Tcf1 or Tcf3, at this stage. ChIP analysis in primary airway epithelial cells revealed that Tcf4 associates with a known Wnt Responsive Region in the Lef-1 promoter and transfection of Cos-1 cells with dominant active beta-catenin and Tcf4 synergistically activated the Lef-1 promoter. Using Wnt3a deficient and Lef-1 promoter-GFP reporter mice, we also demonstrate that Wnt3a induces Lef-1 gene expression in newly forming SMG buds of mice and is required for the maintenance of gland bud growth. These findings provide the first in vivo evidence that Wnt3a can transcriptionally regulate the Lef-1 gene.