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1.
Cell ; 184(15): 4016-4031.e22, 2021 07 22.
Artigo em Inglês | MEDLINE | ID: mdl-34081922

RESUMO

Cross-presentation of antigens from dead tumor cells by type 1 conventional dendritic cells (cDC1s) is thought to underlie priming of anti-cancer CD8+ T cells. cDC1 express high levels of DNGR-1 (a.k.a. CLEC9A), a receptor that binds to F-actin exposed by dead cell debris and promotes cross-presentation of associated antigens. Here, we show that secreted gelsolin (sGSN), an extracellular protein, decreases DNGR-1 binding to F-actin and cross-presentation of dead cell-associated antigens by cDC1s. Mice deficient in sGsn display increased DNGR-1-dependent resistance to transplantable tumors, especially ones expressing neoantigens associated with the actin cytoskeleton, and exhibit greater responsiveness to cancer immunotherapy. In human cancers, lower levels of intratumoral sGSN transcripts, as well as presence of mutations in proteins associated with the actin cytoskeleton, are associated with signatures of anti-cancer immunity and increased patient survival. Our results reveal a natural barrier to cross-presentation of cancer antigens that dampens anti-tumor CD8+ T cell responses.


Assuntos
Apresentação Cruzada/imunologia , Gelsolina/metabolismo , Imunidade , Lectinas Tipo C/metabolismo , Neoplasias/imunologia , Receptores Imunológicos/metabolismo , Receptores Mitogênicos/metabolismo , Actinas/metabolismo , Sequência de Aminoácidos , Animais , Antígenos de Neoplasias/metabolismo , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Apresentação Cruzada/efeitos dos fármacos , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/metabolismo , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Gelsolina/química , Gelsolina/deficiência , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Inibidores de Checkpoint Imunológico/farmacologia , Inibidores de Checkpoint Imunológico/uso terapêutico , Imunidade/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Mutação/genética , Neoplasias/tratamento farmacológico , Neoplasias/genética , Neoplasias/patologia , Ligação Proteica/efeitos dos fármacos , Análise de Sobrevida
2.
Cell ; 154(4): 843-58, 2013 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-23953115

RESUMO

Mononuclear phagocytes are classified as macrophages or dendritic cells (DCs) based on cell morphology, phenotype, or select functional properties. However, these attributes are not absolute and often overlap, leading to difficulties in cell-type identification. To circumvent this issue, we describe a mouse model to define DCs based on their ontogenetic descendence from a committed precursor. We show that precursors of mouse conventional DCs, but not other leukocytes, are marked by expression of DNGR-1. Genetic tracing of DNGR-1 expression history specifically marks cells traditionally ascribed to the DC lineage, and this restriction is maintained after inflammation. Notably, in some tissues, cells previously thought to be monocytes/macrophages are in fact descendants from DC precursors. These studies provide an in vivo model for fate mapping of DCs, distinguishing them from other leukocyte lineages, and thus help to unravel the functional complexity of the mononuclear phagocyte system.


Assuntos
Linhagem da Célula , Células Dendríticas/citologia , Lectinas Tipo C/metabolismo , Receptores Imunológicos/metabolismo , Animais , Células Dendríticas/metabolismo , Hematopoese , Inflamação/patologia , Rim/citologia , Lectinas Tipo C/genética , Células Progenitoras Linfoides/metabolismo , Macrófagos/citologia , Camundongos , Camundongos Endogâmicos C57BL , Fagócitos/citologia , Receptores de IgG/metabolismo , Receptores Imunológicos/genética
3.
Proc Natl Acad Sci U S A ; 115(2): 355-360, 2018 01 09.
Artigo em Inglês | MEDLINE | ID: mdl-29259119

RESUMO

Around the time of gastrulation in higher vertebrate embryos, inductive interactions direct cells to form central nervous system (neural plate) or sensory placodes. Grafts of different tissues into the periphery of a chicken embryo elicit different responses: Hensen's node induces a neural plate whereas the head mesoderm induces placodes. How different are these processes? Transcriptome analysis in time course reveals that both processes start by induction of a common set of genes, which later diverge. These genes are remarkably similar to those induced by an extraembryonic tissue, the hypoblast, and are normally expressed in the pregastrulation stage epiblast. Explants of this epiblast grown in the absence of further signals develop as neural plate border derivatives and eventually express lens markers. We designate this state as "preborder"; its transcriptome resembles embryonic stem cells. Finally, using sequential transplantation experiments, we show that the node, head mesoderm, and hypoblast are interchangeable to begin any of these inductions while the final outcome depends on the tissue emitting the later signals.


Assuntos
Sistema Nervoso Central/metabolismo , Indução Embrionária , Células-Tronco Embrionárias/metabolismo , Gástrula/metabolismo , Mesoderma/metabolismo , Placa Neural/metabolismo , Animais , Sistema Nervoso Central/embriologia , Embrião de Galinha , Gástrula/embriologia , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica no Desenvolvimento , Hibridização In Situ , Mesoderma/embriologia , Placa Neural/embriologia
4.
Development ; 139(5): 958-67, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22318627

RESUMO

The specification of the skeletal muscle lineage during craniofacial development is dependent on the activity of MYF5 and MYOD, two members of the myogenic regulatory factor family. In the absence of MYF5 or MYOD there is not an overt muscle phenotype, whereas in the double Myf5;MyoD knockout branchiomeric myogenic precursors fail to be specified and skeletal muscle is not formed. The transcriptional regulation of Myf5 is controlled by a multitude of regulatory elements acting at different times and anatomical locations, with at least five operating in the branchial arches. By contrast, only two enhancers have been implicated in the regulation of MyoD. In this work, we characterize an enhancer element that drives Myf5 expression in the branchial arches from 9.5 days post-coitum and show that its activity in the context of the entire locus is dependent on two highly conserved E-boxes. These binding sites are required in a subset of Myf5-expressing cells including both progenitors and those which have entered the myogenic pathway. The correct levels of expression of Myf5 and MyoD result from activation by musculin and TCF21 through direct binding to specific enhancers. Consistent with this, we show that in the absence of musculin the timing of activation of Myf5 and MyoD is not affected but the expression levels are significantly reduced. Importantly, normal levels of Myf5 expression are restored at later stages, which might explain the absence of particular muscles in the Msc;Tcf21 double-knockout mice.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Padronização Corporal/fisiologia , Região Branquial/embriologia , Regulação da Expressão Gênica no Desenvolvimento , Músculo Esquelético/fisiologia , Fator Regulador Miogênico 5/metabolismo , Fatores de Transcrição/metabolismo , Animais , Sequência de Bases , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Região Branquial/anatomia & histologia , Região Branquial/fisiologia , Embrião de Mamíferos/anatomia & histologia , Embrião de Mamíferos/fisiologia , Redes Reguladoras de Genes , Humanos , Camundongos , Camundongos Knockout , Dados de Sequência Molecular , Músculo Esquelético/anatomia & histologia , Mutação , Proteína MyoD/genética , Proteína MyoD/metabolismo , Fator Regulador Miogênico 5/genética , Sequências Reguladoras de Ácido Nucleico , Células-Tronco/citologia , Células-Tronco/fisiologia , Proteínas com Domínio T/genética , Proteínas com Domínio T/metabolismo , Fatores de Transcrição/genética
5.
J Hematol Oncol ; 17(1): 91, 2024 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-39380002

RESUMO

BACKGROUND: The epigenetic factors KAT6A (MOZ/MYST3) and KMT2A (MLL/MLL1) interact in normal hematopoiesis to regulate progenitors' self-renewal. Both proteins are recurrently translocated in AML, leading to impairment of critical differentiation pathways in these malignant cells. We evaluated the potential of different KAT6A therapeutic targeting strategies to alter the growth of KAT6A and KMT2A rearranged AMLs. METHODS: We investigated the action and potential mechanisms of the first-in-class KAT6A inhibitor, WM-1119 in KAT6A and KMT2A rearranged (KAT6Ar and KMT2Ar) AML using cellular (flow cytometry, colony assays, cell growth) and molecular (shRNA knock-down, CRISPR knock-out, bulk and single-cell RNA-seq, ChIP-seq) assays. We also used two novel genetic murine KAT6A models combined with the most common KMT2Ar AML, KMT2A::MLLT3 AML. In these murine models, the catalytic activity of KAT6A, or the whole protein, can be conditionally abrogated or deleted. These models allowed us to compare the effects of specific KAT6A KAT activity inhibition with the complete deletion of the whole protein. Finally, we also tested these therapeutic approaches on human AML cell lines and primary patient AMLs. RESULTS: We found that WM-1119 completely abrogated the proliferative and clonogenic potential of KAT6Ar cells in vitro. WM-1119 treatment was associated with a dramatic increase in myeloid differentiation program. The treatment also decreased stemness and leukemia pathways at the transcriptome level and led to loss of binding of the fusion protein at critical regulators of these pathways. In contrast, our pharmacologic and genetic results indicate that the catalytic activity of KAT6A plays a more limited role in KMT2Ar leukemogenicity, while targeting the whole KAT6A protein dramatically affects leukemic potential in murine KMT2A::MLLT3 AML. CONCLUSION: Our study indicates that inhibiting KAT6A KAT activity holds compelling promise for KAT6Ar AML patients. In contrast, targeted degradation of KAT6A, and not just its catalytic activity, may represent a more appropriate therapeutic approach for KMT2Ar AMLs.


Assuntos
Histona Acetiltransferases , Histona-Lisina N-Metiltransferase , Leucemia Mieloide Aguda , Proteína de Leucina Linfoide-Mieloide , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/tratamento farmacológico , Animais , Proteína de Leucina Linfoide-Mieloide/genética , Humanos , Camundongos , Histona-Lisina N-Metiltransferase/genética , Histona Acetiltransferases/genética , Histona Acetiltransferases/antagonistas & inibidores , Rearranjo Gênico , Linhagem Celular Tumoral
6.
Elife ; 122023 03 03.
Artigo em Inglês | MEDLINE | ID: mdl-36867045

RESUMO

During early vertebrate development, signals from a special region of the embryo, the organizer, can redirect the fate of non-neural ectoderm cells to form a complete, patterned nervous system. This is called neural induction and has generally been imagined as a single signalling event, causing a switch of fate. Here, we undertake a comprehensive analysis, in very fine time course, of the events following exposure of competent ectoderm of the chick to the organizer (the tip of the primitive streak, Hensen's node). Using transcriptomics and epigenomics we generate a gene regulatory network comprising 175 transcriptional regulators and 5614 predicted interactions between them, with fine temporal dynamics from initial exposure to the signals to expression of mature neural plate markers. Using in situ hybridization, single-cell RNA-sequencing, and reporter assays, we show that the gene regulatory hierarchy of responses to a grafted organizer closely resembles the events of normal neural plate development. The study is accompanied by an extensive resource, including information about conservation of the predicted enhancers in other vertebrates.


Assuntos
Redes Reguladoras de Genes , Sistema Nervoso , Animais , Sistema Nervoso/metabolismo , Galinhas , Desenvolvimento Embrionário , Organizadores Embrionários , Vertebrados
7.
Dev Biol ; 355(2): 372-80, 2011 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-21527258

RESUMO

The transcriptional regulation of the Mrf4/Myf5 locus depends on a multitude of enhancers that, in equilibria with transcription balancing sequences and the promoters, regulate the expression of the two genes throughout embryonic development and in the adult. Transcription in a particular set of muscle progenitors can be driven by the combined outputs of several enhancers that are not able to recapitulate the entire expression pattern in isolation, or by the action of a single enhancer the activity of which in isolation is equivalent to that within the context of the locus. We identified a new enhancer element of this second class, ECR111, which is highly conserved in all vertebrate species and is necessary and sufficient to drive Myf5 expression in ventro-caudal and ventro-rostral somitic compartments in the mouse embryo. EMSA analyses and data obtained from binding-site mutations in transgenic embryos show that a binding site for a TEA Domain (TEAD) transcription factor is essential for the function of this new enhancer, while ChIP assays show that at least two members of the family of transcription factors bind to it in vivo.


Assuntos
Proteínas de Ligação a DNA/metabolismo , Elementos Facilitadores Genéticos/genética , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Desenvolvimento Muscular/fisiologia , Fator Regulador Miogênico 5/metabolismo , Somitos/embriologia , Fatores de Transcrição/metabolismo , Animais , Sítios de Ligação/genética , Imunoprecipitação da Cromatina , Ensaio de Desvio de Mobilidade Eletroforética , Camundongos , Somitos/metabolismo , Fatores de Transcrição de Domínio TEA
8.
Elife ; 102021 10 12.
Artigo em Inglês | MEDLINE | ID: mdl-34636321

RESUMO

Lung squamous cell carcinoma (LSCC) is a considerable global health burden, with an incidence of over 600,000 cases per year. Treatment options are limited, and patient's 5-year survival rate is less than 5%. The ubiquitin-specific protease 28 (USP28) has been implicated in tumourigenesis through its stabilization of the oncoproteins c-MYC, c-JUN, and Δp63. Here, we show that genetic inactivation of Usp28-induced regression of established murine LSCC lung tumours. We developed a small molecule that inhibits USP28 activity in the low nanomole range. While displaying cross-reactivity against the closest homologue USP25, this inhibitor showed a high degree of selectivity over other deubiquitinases. USP28 inhibitor treatment resulted in a dramatic decrease in c-MYC, c-JUN, and Δp63 proteins levels and consequently induced substantial regression of autochthonous murine LSCC tumours and human LSCC xenografts, thereby phenocopying the effect observed by genetic deletion. Thus, USP28 may represent a promising therapeutic target for the treatment of squamous cell lung carcinoma.


Assuntos
Proteínas de Ligação a DNA/genética , Deleção de Genes , Neoplasias Pulmonares/genética , Neoplasias de Células Escamosas/genética , Fatores de Transcrição/genética , Ubiquitina Tiolesterase/genética , Animais , Proteínas de Ligação a DNA/metabolismo , Modelos Animais de Doenças , Humanos , Camundongos , Fatores de Transcrição/metabolismo , Ubiquitina Tiolesterase/metabolismo
11.
J Mol Endocrinol ; 34(3): 767-79, 2005 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15956346

RESUMO

Gonadotrophin-releasing hormone (GnRH) is the main neurohormone controlling gonadotrophin release in all vertebrates, and in teleost fish also of growth hormone and possibly of other adenohypophyseal hormones. Over 20 GnRHs have been identified in vertebrates and protochoordates and shown to bind cognate G-protein couple receptors (GnRHR). We have searched the puffer fish, Fugu rubripes, genome sequencing database, identified five GnRHR genes and proceeded to isolate the corresponding complementary DNAs in European sea bass, Dicentrachus labrax. Phylogenetic analysis clusters the European sea bass, puffer fish and all other vertebrate receptors into two main lineages corresponding to the mammalian type I and II receptors. The fish receptors could be subdivided in two GnRHR1 (A and B) and three GnRHR2 (A, B and C) subtypes. Amino acid sequence identity within receptor subtypes varies between 70 and 90% but only 50-55% among the two main lineages in fish. All European sea bass receptor mRNAs are expressed in the anterior and mid brain, and all but one are expressed in the pituitary gland. There is differential expression of the receptors in peripheral tissues related to reproduction (gonads), chemical senses (eye and olfactory epithelium) and osmoregulation (kidney and gill). This is the first report showing five GnRH receptors in a vertebrate species and the gene expression patterns support the concept that GnRH and GnRHRs play highly diverse functional roles in the regulation of cellular functions, besides the "classical" role of pituitary function regulation.


Assuntos
Filogenia , Receptores LHRH/isolamento & purificação , Sequência de Aminoácidos , Animais , Sequência de Bases , Bass , Southern Blotting , Clonagem Molecular , Primers do DNA , DNA Complementar , Dados de Sequência Molecular , RNA Mensageiro/genética , Receptores LHRH/química , Receptores LHRH/classificação , Receptores LHRH/genética , Receptores LHRH/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de Aminoácidos , Takifugu
12.
Aquat Toxicol ; 63(2): 127-37, 2003 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-12657488

RESUMO

During the last decade, special attention has been focused on the consequences of exposure to environmental estrogens on reproduction in wild fish species. For this reason, characterization of biomarkers of such exposures could result in useful tools for these studies. The detection of vitellogenin (Vtg), a precursor of yolk proteins, is being intensely studied since its synthesis in the liver is regulated by the estradiol-17beta and is influenced by other estrogenic compounds. The aim of this work was to assess the presence of Vtg in the surface mucus of males of Cichlasoma dimerus (Teleostei, Perciformes), a typical South American freshwater cichlid, after hormonal treatment with estradiol-17beta (intraperitoneal injections of 10 microg E(2)/g fish). Plasma and mucus from females and treated males analyzed by Western blot revealed that different heterologous antisera against Vtg bind to putative protein of 180 & 120 kDa and 120 & 110 kDa, respectively, whereas no reaction was found in samples of untreated males. The same profile was observed in mucus samples using Dot blot, a very easy and direct technique. Using immunocytochemistry techniques, immunoreactive Vtg (ir-Vtg) producing cells in the liver of females and treated males were detected. Testes and liver tissues were also assessed by histological techniques. Marked alterations in both organs were observed, such as lower sperm production, presence of immature germ cells in the lobular lumen of testes, and some morphology changes in the hepatocytes due to the accumulation of Vtg. This is the first report about the effects of an estrogen in the Vtg synthesis and their consequences on liver and gonads of a South American fresh water cichlid. These results also support the possibility of using Vtg from surface mucus as a potential biomarker for estrogenic compounds through a noninvasive, useful and easy assay to monitor the presence of endocrine disrupting chemicals in the environment.


Assuntos
Ciclídeos/fisiologia , Estradiol/efeitos adversos , Vitelogeninas/biossíntese , Animais , Biomarcadores/análise , Exposição Ambiental , Feminino , Gônadas/química , Imuno-Histoquímica , Fígado/química , Masculino , Muco/química , Vitelogeninas/análise
13.
FEBS J ; 280(17): 3980-90, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23751110

RESUMO

The transcriptional regulatory network that controls the determination and differentiation of skeletal muscle cells in the embryo has at its core the four myogenic regulatory factors (MRFs) Myf5, MyoD, Mrf4 and MyoG. These basic helix-loop-helix transcription factors act by binding, as obligate heterodimers with the ubiquitously expressed E proteins, to the E-box sequence CANNTG. While all skeletal muscle cells have the same underlying function their progenitors arise at many sites in the embryo and it has become apparent that the upstream activators of the cascade differ in these various populations so that it can be switched on by a variety of inductive signals, some of which act by initiating transcription, some by maintaining it. The application of genome-wide approaches has provided important new information as to how the MRFs function to activate the terminal differentiation programme and some of these data provide significant mechanistic insights into questions which have exercised the field for many years. We also consider the emerging roles played by micro-RNAs in the regulation of both upstream activators and terminal differentiation genes.


Assuntos
Diferenciação Celular , Desenvolvimento Muscular/fisiologia , Músculos/citologia , Fatores de Regulação Miogênica/metabolismo , Animais , Humanos , Músculos/metabolismo
14.
Dev Cell ; 18(4): 655-61, 2010 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-20412779

RESUMO

Hox genes are essential for the patterning of the axial skeleton. Hox group 10 has been shown to specify the lumbar domain by setting a rib-inhibiting program in the presomitic mesoderm (PSM). We have now produced mice with ribs in every vertebra by ectopically expressing Hox group 6 in the PSM, indicating that Hox genes are also able to specify the thoracic domain. We show that the information provided by Hox genes to specify rib-containing and rib-less areas is first interpreted in the myotome through the regional-specific control of Myf5 and Myf6 expression. This information is then transmitted to the sclerotome by a system that includes FGF and PDGF signaling to produce vertebrae with or without ribs at different axial levels. Our findings offer a new perspective of how Hox genes produce global patterns in the axial skeleton and support a redundant nonmyogenic role of Myf5 and Myf6 in rib formation.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/metabolismo , Fator Regulador Miogênico 5/fisiologia , Fatores de Regulação Miogênica/fisiologia , Animais , Desenvolvimento Ósseo , Fatores de Crescimento de Fibroblastos/metabolismo , Proteínas Homeobox A10 , Camundongos , Camundongos Transgênicos , Modelos Biológicos , Modelos Genéticos , Desenvolvimento Muscular , Fenótipo , Fator de Crescimento Derivado de Plaquetas/metabolismo
15.
Proc Natl Acad Sci U S A ; 104(43): 16952-7, 2007 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-17940039

RESUMO

Homeodomain-containing transcription factors are important regulators of stem cell behavior. HOXB4 mediates expansion of adult and embryo-derived hematopoietic stem cells (HSCs) when expressed ectopically. To define the underlying molecular mechanisms, we performed gene expression profiling in combination with subsequent functional analysis with enriched adult HSCs and embryonic derivatives expressing inducible HOXB4. Thereby, we identified a set of overlapping genes that likely represent "universal" targets of HOXB4. A substantial number of loci are involved in signaling pathways important for controlling self-renewal, maintenance, and differentiation of stem cells. Functional assays performed on selected pathways confirmed the biological coherence of the array results. HOXB4 activity protected adult HSCs from the detrimental effects mediated by the proinflammatory cytokine TNF-alpha. This protection likely contributes to the competitive repopulation advantage of HOXB4-expressing HSCs observed in vivo. The concept of TNF-alpha inhibition may also prove beneficial for patients undergoing bone marrow transplantation. Furthermore, we demonstrate that HOXB4 activity and FGF signaling are intertwined. HOXB4-mediated expansion of adult and ES cell-derived HSCs was enhanced by specific and complete inhibition of FGF receptors. In contrast, the expanding activity of HOXB4 on hematopoietic progenitors in day 4-6 embryoid bodies was blunted in the presence of basic FGF (FGF2), indicating a dominant negative effect of FGF signaling on the earliest hematopoietic cells. In summary, our results strongly suggest that HOXB4 modulates the response of HSCs to multiple extrinsic signals in a concerted manner, thereby shifting the balance toward stem cell self-renewal.


Assuntos
Células-Tronco Embrionárias/citologia , Células-Tronco Hematopoéticas/citologia , Proteínas de Homeodomínio/metabolismo , Fatores de Transcrição/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células-Tronco Embrionárias/efeitos dos fármacos , Células-Tronco Embrionárias/metabolismo , Fatores de Crescimento de Fibroblastos/farmacologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Redes Reguladoras de Genes , Células-Tronco Hematopoéticas/efeitos dos fármacos , Células-Tronco Hematopoéticas/metabolismo , Proteínas de Homeodomínio/genética , Camundongos , Camundongos Endogâmicos C57BL , Análise de Sequência com Séries de Oligonucleotídeos , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Fatores de Tempo , Fatores de Transcrição/genética , Fator de Necrose Tumoral alfa/farmacologia
16.
Artigo em Inglês | MEDLINE | ID: mdl-16716622

RESUMO

Gonadotropin-releasing hormone (GnRH) is the final common signaling molecule used by the brain to regulate reproduction in all vertebrates. Until now, a total of 24 GnRH structural variants have been characterized from vertebrate, protochordate and invertebrate nervous tissue. Almost all vertebrates already investigated have at least two GnRH forms coexisting in the central nervous system. Furthermore, it is now well accepted that three GnRH forms are present both in early and late evolved teleostean fishes. The number and taxonomic distribution of the different GnRH variants also raise questions about the phylogenetic relationships between them. Most of the GnRH phylogenetic analyses are in agreement with the widely accepted idea that the GnRH family can be divided into three main groups. However, the examination of the gnathostome GnRH phylogenetic relationships clearly shows the existence of two main paralogous GnRH lineages: the ''midbrain GnRH" group and the "forebrain GnRH" group. The first one, represented by chicken GnRH-II forms, and the second one composed of two paralogous lineages, the salmon GnRH cluster (only represented in teleostean fish species) and the hypophysotropic GnRH cluster, also present in tetrapods. This analysis suggests that the two forebrain clades share a common precursor and reinforces the idea that the salmon GnRH branch has originated from a duplication of the hypophysotropic lineage. GnRH ligands exert their activity through G protein-coupled receptors of the rhodopsin-like family. As with the ligands, multiple GnRHRs are expressed in individual vertebrate species and phylogenetic analyses have revealed that all vertebrate GnRHRs cluster into three main receptor types. However, new data and a new phylogenetic analysis propose a two GnRHR type model, in which different rounds of gene duplications may have occurred in different groups within each lineage.


Assuntos
Evolução Molecular , Hormônio Liberador de Gonadotropina/fisiologia , Filogenia , Receptores LHRH/fisiologia , Vertebrados/fisiologia , Animais , Hormônio Liberador de Gonadotropina/química , Hormônio Liberador de Gonadotropina/classificação , Ligantes , Receptores LHRH/química , Vertebrados/classificação
17.
Gen Comp Endocrinol ; 143(1): 21-32, 2005 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-15993101

RESUMO

The brain-type aromatase (CYP19A2) cDNA from pejerrey Odontesthes bonariensis was characterized. Its sequence differs from the ovarian-derived aromatase (CYP19A1) previously reported for the same species. The cDNA is 2305bp in length and the deduced protein comprises 501 amino-acids. The percentage of identity was higher when compared to other brain-derived aromatase proteins (85-63%) and lower with ovarian-derived aromatases (64-57%). Pejerrey aromatases share 61% of identity. The tissue expression analysis showed that CYP19A2 was expressed in the kidney, brain, and pituitary gland of both sexes and also in the ovary, but not in the eye, spleen, liver, gill, and testis. Semi-quantitative RT-PCR analysis of different brain areas revealed that CYP19A2 was expressed significantly higher in anterior male brain areas than in the corresponding female areas, and also when compared to posterior brain areas from both sexes. An immunological analysis using a polyclonal anti-teleost aromatase showed immunoreactive aromatase cells bordering the telencephalic ventricle and a strong signal in the ependymal cells of the preoptic area and the hypothalamus. In the optic tectum immunoreactive aromatase cells were labeled in the ventral wall and in the ependymal layer of the third and fourth ventricle with lateral projections. In the pituitary gland immunoreactive aromatase cells could be found in the rostral and proximal pars distalis. In this gland, aromatase fibers were also detected in different areas; many of them concentrated around blood vessels.


Assuntos
Aromatase/metabolismo , Encéfalo/enzimologia , DNA Complementar , Peixes , Regulação Enzimológica da Expressão Gênica , Proteínas de Peixe-Zebra/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA Complementar/análise , Feminino , Imunoquímica , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA
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