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1.
Nat Commun ; 11(1): 333, 2020 01 16.
Artigo em Inglês | MEDLINE | ID: mdl-31949145

RESUMO

Cutaneous malignant melanoma is an aggressive cancer of melanocytes with a strong propensity to metastasize. We posit that melanoma cells acquire metastatic capability by adopting an embryonic-like phenotype, and that a lineage approach would uncover metastatic melanoma biology. Using a genetically engineered mouse model to generate a rich melanoblast transcriptome dataset, we identify melanoblast-specific genes whose expression contribute to metastatic competence and derive a 43-gene signature that predicts patient survival. We identify a melanoblast gene, KDELR3, whose loss impairs experimental metastasis. In contrast, KDELR1 deficiency enhances metastasis, providing the first example of different disease etiologies within the KDELR-family of retrograde transporters. We show that KDELR3 regulates the metastasis suppressor, KAI1, and report an interaction with the E3 ubiquitin-protein ligase gp78, a regulator of KAI1 degradation. Our work demonstrates that the melanoblast transcriptome can be mined to uncover targetable pathways for melanoma therapy.


Assuntos
Perfilação da Expressão Gênica , Melanoma/genética , Melanoma/metabolismo , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/metabolismo , Transcriptoma , Animais , Linhagem Celular Tumoral , Retículo Endoplasmático , Feminino , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Proteína Kangai-1/genética , Proteína Kangai-1/metabolismo , Pulmão/patologia , Melanócitos/metabolismo , Melanoma/patologia , Camundongos , Camundongos Endogâmicos C57BL , Metástase Neoplásica/genética , Segunda Neoplasia Primária/patologia , Fenótipo , Receptores de Peptídeos/genética , Receptores de Peptídeos/metabolismo , Neoplasias Cutâneas/patologia , Ubiquitina-Proteína Ligases/metabolismo
2.
Proc Natl Acad Sci U S A ; 117(2): 1097-1106, 2020 01 14.
Artigo em Inglês | MEDLINE | ID: mdl-31843923

RESUMO

The molecular mechanisms by which animals integrate external stimuli with internal energy balance to regulate major developmental and reproductive events still remain enigmatic. We investigated this aspect in the marine bristleworm, Platynereis dumerilii, a species where sexual maturation is tightly regulated by both metabolic state and lunar cycle. Our specific focus was on ligands and receptors of the gonadotropin-releasing hormone (GnRH) superfamily. Members of this superfamily are key in triggering sexual maturation in vertebrates but also regulate reproductive processes and energy homeostasis in invertebrates. Here we show that 3 of the 4 gnrh-like (gnrhl) preprohormone genes are expressed in specific and distinct neuronal clusters in the Platynereis brain. Moreover, ligand-receptor interaction analyses reveal a single Platynereis corazonin receptor (CrzR) to be activated by CRZ1/GnRHL1, CRZ2/GnRHL2, and GnRHL3 (previously classified as AKH1), whereas 2 AKH-type hormone receptors (GnRHR1/AKHR1 and GnRHR2/AKHR2) respond only to a single ligand (GnRH2/GnRHL4). Crz1/gnrhl1 exhibits a particularly strong up-regulation in sexually mature animals, after feeding, and in specific lunar phases. Homozygous crz1/gnrhl1 knockout animals exhibit a significant delay in maturation, reduced growth, and attenuated regeneration. Through a combination of proteomics and gene expression analysis, we identify enzymes involved in carbohydrate metabolism as transcriptional targets of CRZ1/GnRHL1 signaling. Our data suggest that Platynereis CRZ1/GnRHL1 coordinates glycoprotein turnover and energy homeostasis with growth and sexual maturation, integrating both metabolic and developmental demands with the worm's monthly cycle.


Assuntos
Hormônio Liberador de Gonadotropina/metabolismo , Homeostase , Proteínas de Insetos/metabolismo , Lua , Neuropeptídeos/metabolismo , Poliquetos/fisiologia , Maturidade Sexual/fisiologia , Transdução de Sinais/fisiologia , Animais , Encéfalo , Proteínas de Ligação a DNA/genética , Técnicas de Silenciamento de Genes , Hormônio Liberador de Gonadotropina/genética , Hormônios de Inseto/genética , Hormônios de Inseto/metabolismo , Proteínas de Insetos/genética , Invertebrados/genética , Neuropeptídeos/genética , Filogenia , Poliquetos/genética , Poliquetos/crescimento & desenvolvimento , Receptores de Neuropeptídeos , Receptores de Peptídeos/genética , Transdução de Sinais/genética , Fatores de Transcrição
3.
BMC Evol Biol ; 19(1): 215, 2019 11 26.
Artigo em Inglês | MEDLINE | ID: mdl-31771521

RESUMO

BACKGROUND: In mammals, the natriuretic system contains three natriuretic peptides, NPPA, NPPB and NPPC, that bind to three transmembrane receptors, NPR1, NPR2 and NPR3. The natriuretic peptides are known only in vertebrates. In contrast, the receptors have orthologs in all the animal taxa and in plants. However, in non-vertebrates, these receptors do not have natriuretic properties, and most of their ligands are unknown. How was the interaction of the NP receptors and the NP established in vertebrates? Do natriuretic peptides have orthologs in non-vertebrates? If so, what was the function of the interaction? How did that function change? If not, are the NP homologous to ancestral NPR ligands? Or did the receptor's binding pocket completely change during evolution? METHODS: In the present study, we tried to determine if the pairs of natriuretic receptors and their ligands come from an ancestral pair, or if the interaction only appeared in vertebrates. Alignments, modeling, docking, research of positive selection, and motif research were performed in order to answer this question. RESULTS: We discovered that the binding pocket of the natriuretic peptide receptors was completely remodeled in mammals. We found several peptides in non vertebrates that could be related to human natriuretic peptides, but a set of clues, as well as modeling and docking analysis, suggest that the natriuretic peptides undoubtedly appeared later than their receptors during animal evolution. We suggest here that natriuretic peptide receptors in non vertebrates bind to other ligands. CONCLUSIONS: The present study further support that vertebrate natriuretic peptides appeared after their receptors in the tree of life. We suggest the existence of peptides that resemble natriuretic peptides in non-vertebrate species, that might be the result of convergent evolution.


Assuntos
Peptídeos Natriuréticos/genética , Vertebrados/genética , Sequência de Aminoácidos , Animais , Humanos , Ligantes , Modelos Moleculares , Peptídeos Natriuréticos/química , Peptídeos Natriuréticos/metabolismo , Filogenia , Ligação Proteica , Receptores de Peptídeos/genética , Seleção Genética , Vertebrados/metabolismo
4.
Genes (Basel) ; 10(9)2019 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-31491999

RESUMO

Equine neuroaxonal dystrophy/equine degenerative myeloencephalopathy (eNAD/EDM) is a neurologic disease that has been reported in young horses from a wide range of breeds. The disease is inherited and associated with vitamin E deficiency during the first two years of life, resulting in bilateral symmetric ataxia. A missense mutation (chr3:71,917,591 C > T) within adhesion G protein-coupled receptor L3 (ADGRL3) was recently associated with risk for EDM in the Caspian breed. In order to confirm these findings, genotyping of this missense mutation, along with the three other associated single nucleotide polymorphisms (SNPs) in the genomic region, was carried out on 31 postmortem-confirmed eNAD/EDM cases and 43 clinically phenotyped controls from various breeds. No significant association was found between eNAD/EDM confirmed cases and genotype at any of the four identified SNPs (P > 0.05), including the nonsynonymous variant (EquCab2.0 chr3:71,917,591; allelic P = 0.85). These findings suggest that the four SNPs, including the missense variant in the ADGRL3 region, are not associated with risk for eNAD/EDM across multiple breeds of horses.


Assuntos
Doenças dos Cavalos/genética , Cavalos/genética , Distrofias Neuroaxonais/genética , Polimorfismo de Nucleotídeo Único , Receptores Acoplados a Proteínas-G/genética , Receptores de Peptídeos/genética , Animais , Mutação de Sentido Incorreto , Distrofias Neuroaxonais/veterinária
5.
Genetics ; 213(2): 529-553, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31399485

RESUMO

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


Assuntos
Hormônio Antimülleriano/genética , Genitália Feminina/crescimento & desenvolvimento , Processos de Determinação Sexual , Peixe-Zebra/genética , Animais , Feminino , Gônadas/crescimento & desenvolvimento , Ductos Paramesonéfricos/crescimento & desenvolvimento , Folículo Ovariano/crescimento & desenvolvimento , Ovário/crescimento & desenvolvimento , Receptores de Peptídeos/genética , Receptores de Fatores de Crescimento Transformadores beta/genética , Peixe-Zebra/crescimento & desenvolvimento
6.
Cells ; 8(8)2019 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-31426340

RESUMO

Attention Deficit Hyperactivity Disorder (ADHD) is a highly heritable and prevalent neurodevelopmental disorder that frequently persists into adulthood. Strong evidence from genetic studies indicates that single nucleotide polymorphisms (SNPs) harboured in the ADGRL3 (LPHN3), SNAP25, FGF1, DRD4, and SLC6A2 genes are associated with ADHD. We genotyped 26 SNPs harboured in genes previously reported to be associated with ADHD and evaluated their potential association in 386 individuals belonging to 113 nuclear families from a Caribbean community in Barranquilla, Colombia, using family-based association tests. SNPs rs362990-SNAP25 (T allele; p = 2.46 × 10-4), rs2282794-FGF1 (A allele; p = 1.33 × 10-2), rs2122642-ADGRL3 (C allele, p = 3.5 × 10-2), and ADGRL3 haplotype CCC (markers rs1565902-rs10001410-rs2122642, OR = 1.74, Ppermuted = 0.021) were significantly associated with ADHD. Our results confirm the susceptibility to ADHD conferred by SNAP25, FGF1, and ADGRL3 variants in a community with a significant African American component, and provide evidence supporting the existence of specific patterns of genetic stratification underpinning the susceptibility to ADHD. Knowledge of population genetics is crucial to define risk and predict susceptibility to disease.


Assuntos
Transtorno do Deficit de Atenção com Hiperatividade/genética , Haplótipos/genética , Polimorfismo de Nucleotídeo Único/genética , Adolescente , Adulto , Afro-Americanos/genética , Estudos de Casos e Controles , Criança , Colômbia , Feminino , Fator 1 de Crescimento de Fibroblastos/genética , Predisposição Genética para Doença , Humanos , Masculino , Receptores Acoplados a Proteínas-G/genética , Receptores de Peptídeos/genética , Proteína 25 Associada a Sinaptossoma/genética
7.
J Assist Reprod Genet ; 36(6): 1281-1289, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31089932

RESUMO

OBJECTIVE: Our study aimed to investigate the relationship between polymorphisms (Apa1, Bsm1, Fok1, and Cdx2) in the VDR gene as well as AMH and AMHR2 genes and their influence on AMH and 25(OH)D levels in PCOS women. STUDY DESIGN: Seventy-five patients with PCOS and 23 control women were included. Serum AMH and 25(OH)D levels in patients and controls were measured by enzyme-linked immunosorbent assay (ELISA). Polymorphisms in VDR gene Fok1 C/T (rs2228587), Bsm1 A/G (rs1544410), Apa1 A/C (rs7975232), and Cdx2 A/G (rs11568820) polymorphisms as well as AMH G/T (rs10407022) and AMHR2 A/G (rs2002555) were analyzed using real-time PCR. RESULTS: Analysis of the VDR Cdx2 polymorphism showed a significantly higher frequency of the homozygous GG (mutant) genotype in the PCOS group as compared with the control group (p < 0.05). The analysis revealed a statistically significant correlation between the presence of FokI and ApaI polymorphisms and AMH levels in PCOS women (p < 0.05). The presence of mutant genotypes (CT, TT) in the Fok1 and (CA, CC) in the Apa1 polymorphisms were associated with higher AMH level in PCOS women (p < 0.05). No statistically significant correlations between AMH and AMHR2 polymorphisms and AMH level were found. Moreover, there was no correlation between AMH and 25(OH)D levels in the PCOS or in the control group. CONCLUSION: It seems that the elevated AMH level is associated with VDR Fokl and Apal polymorphisms, but not with 25(OH)D levels in PCOS women. Further research is needed to determine the role of VDR polymorphism in AMH level in PCOS.


Assuntos
Hormônio Antimülleriano/sangue , Síndrome do Ovário Policístico/sangue , Receptores de Calcitriol/sangue , Receptores de Peptídeos/sangue , Receptores de Fatores de Crescimento Transformadores beta/sangue , Adulto , Hormônio Antimülleriano/genética , Feminino , Genótipo , Humanos , Ovulação/genética , Síndrome do Ovário Policístico/genética , Síndrome do Ovário Policístico/fisiopatologia , Polimorfismo de Nucleotídeo Único/genética , Receptores de Calcitriol/genética , Receptores de Peptídeos/genética , Receptores de Fatores de Crescimento Transformadores beta/genética , Vitamina D/sangue
8.
Gen Comp Endocrinol ; 280: 185-191, 2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31054903

RESUMO

Myosuppressin is one of essential peptides controlling biological processes including feeding behavior. Here we identified and characterized the cDNAs that encode myosuppressin precursor and its receptor in the two-spotted cricket Gryllus bimaculatus. The presence of the mature peptide (Grybi-MS) was confirmed by direct measurement of adult brain. RT-PCR revealed the tissue distribution of these transcripts; myosuppressin is expressed predominantly in the brain and central nervous system, whereas its receptor is ubiquitously expressed in the cricket body. To address the function of Grybi-MS, we performed several bioassays to test concerning feeding behavior and digestive function upon exposure to Grybi-MS. Administration of synthetic Grybi-MS resulted in increased feeding motivation, accompanied by an increase in food intake. Meanwhile, the hemolymph lipid and carbohydrate titers were both elevated after Grybi-MS injection. As the intestinal contraction is significantly inhibited by the exposure to Grybi-MS, the upregulating feeding index might be complicated in the cricket body. The current data indicate that Grybi-MS modulates feeding behavior to control the physiological processes in the cricket.


Assuntos
Digestão/fisiologia , Gryllidae/fisiologia , Neuropeptídeos/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Metabolismo dos Carboidratos , DNA Complementar/genética , Sistema Digestório/metabolismo , Comportamento Alimentar , Gryllidae/metabolismo , Metabolismo dos Lipídeos , Masculino , Contração Muscular , Neuropeptídeos/química , Neuropeptídeos/genética , Receptores de Peptídeos/química , Receptores de Peptídeos/genética , Receptores de Peptídeos/metabolismo , Distribuição Tecidual
9.
Cell Tissue Res ; 377(3): 293-308, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31079207

RESUMO

The digestive system is responsible for nutrient intake and defense against pathogenic microbes. Thus, identification of regulatory factors for digestive functions and immune systems is a key step to the verification of the life cycle, homeostasis, survival strategy and evolutionary aspects of an organism. Over the past decade, there have been increasing reports on neuropeptides, their receptors, variable region-containing chitin-binding proteins (VCBPs) and Toll-like receptors (TLRs) in the ascidian, Ciona intestinalis. Mass spectrometry-based peptidomes and genome database-searching detected not only Ciona orthologs or prototypes of vertebrate peptides and their receptors, including cholecystokinin, gonadotropin-releasing hormones, tachykinin, calcitonin and vasopressin but also Ciona-specific neuropeptides including Ci-LFs and Ci-YFVs. The species-specific regulation of GnRHergic signaling including unique signaling control via heterodimerization among multiple GnRH receptors has also been revealed. These findings shed light on the remarkable significance of ascidians in investigations of the evolution and diversification of the peptidergic systems in chordates. In the defensive systems of C. intestinalis, VCBPs and TLRs have been shown to play major roles in the recognition of exogenous microbes in the innate immune system. These findings indicate both common and species-specific functions of the innate immunity-related molecules between C. intestinalis and vertebrates. In this review article, we present recent advances in molecular and functional features and evolutionary aspects of major neuropeptides, their receptors, VCBPs and TLRs in C. intestinalis.


Assuntos
Ciona intestinalis , Sistema Digestório , Neuropeptídeos , Receptores de Peptídeos , Receptores Toll-Like , Animais , Ciona intestinalis/imunologia , Ciona intestinalis/metabolismo , Sistema Digestório/imunologia , Sistema Digestório/metabolismo , Neuropeptídeos/química , Neuropeptídeos/genética , Filogenia , Receptores de Peptídeos/química , Receptores de Peptídeos/genética , Especificidade da Espécie , Receptores Toll-Like/química , Receptores Toll-Like/genética
10.
Cell Physiol Biochem ; 52(4): 850-868, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30958660

RESUMO

BACKGROUND/AIMS: Endoplasmic reticulum (ER)-resident proteins with a C-terminal KDEL ERretention sequence are captured in the Golgi apparatus by KDEL receptors (KDELRs). The binding of such proteins to these receptors induces their retrograde transport. Nevertheless, some KDEL proteins, such as Protein Disulfide Isomerases (PDIs), are found at the cell surface. PDIs target disulfide bridges in the extracellular domains of proteins, such as integrins or A Disintegrin And Metalloprotease 17 (ADAM17) leading to changes in the structure and function of these molecules. Integrins become activated and ADAM17 inactivated upon disulfide isomerization. The way that PDIs escape from retrograde transport and reach the plasma membrane remains far from clear. Various mechanisms might exist, depending on whether a local cell surface association or a more global secretion is required. METHODS: To get a more detailed insight in the transport of PDIs to the cell surface, methods such as cell surface biotinylation, flow cytometric analysis, immunoprecipitation, fluorescence microscopy as well as labeling of cells with fluorescence labled recombinant PDIA6 was performed. RESULTS: Here, we show that the C-terminal KDEL ER retention sequence is sufficient to prevent secretion of PDIA6 into the extracellular space but is mandatory for its association with the cell surface. The cell surface trafficking of PDIA1, PDIA3, and PDIA6 is dependent on KDELR1, which travels in a dynamic manner to the cell surface. This transport is assumed to result in PDI cell surface association, which differs from PDI inducible secretion into the extracellular space. Distinct PDIs differ in their trafficking properties. Endogenous KDELR1, detectable at the cell surface, might be involved not only in the transport of cell-surface-associated PDIs, but also in their retrieval and internalization from the extracellular space. CONCLUSION: Beside their ER retention motive PDIs travel to the cell surface. Here they target different proteins to render their function. To escape the ER PDIs travel via various pathways. One of them depends on the KDELR1, which can transport its target to the cell surface, where it is to be expected to release its cargo in close vicinity to its target molecules. Hence, the KDEL sequence is needed for cell surface association of PDIs, such as PDIA6.


Assuntos
Proteína ADAM17/metabolismo , Membrana Celular/metabolismo , Retículo Endoplasmático/metabolismo , Isomerases de Dissulfetos de Proteínas/metabolismo , Receptores de Peptídeos/metabolismo , Proteína ADAM17/genética , Membrana Celular/genética , Retículo Endoplasmático/genética , Células HEK293 , Humanos , Isomerases de Dissulfetos de Proteínas/genética , Transporte Proteico/fisiologia , Receptores de Peptídeos/genética
11.
Res Vet Sci ; 124: 223-227, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30928654

RESUMO

Gastrointestinal hormone based therapies are being investigated for treating diabetes in cats; however, the tissue distribution of these hormones and their cognate receptors remain largely understudied. We determined the distribution of transcripts for the gut hormones proglucagon (Gcg), glucose-dependent insulinotropic peptide (Gip), peptide YY (Pyy), and their receptors (Glp1r, Gipr, Npy2r), in feline peripheral tissues. The Gcg, Gip and Pyy mRNA were expressed in the gut, with higher Gcg and Pyy abundance in the lower gut. Interestingly, Glp1r and Npy2r mRNA were expressed in multiple peripheral tissues including the gut, pancreas and liver, whereas, Gipr mRNA was restricted to the stomach and adipose tissues. The localized mRNA expression of Gcg and Pyy in the gut, but the extensive distribution of Glp1r and Npy2r in several peripheral tissues suggests that these hormones may have pleiotropic physiological functions in cats.


Assuntos
Gatos/genética , Polipeptídeo Inibidor Gástrico/genética , Peptídeo YY/genética , Proglucagon/genética , Receptores dos Hormônios Gastrointestinais/genética , Receptores de Peptídeos/genética , Animais , Gatos/metabolismo , Polipeptídeo Inibidor Gástrico/metabolismo , Perfilação da Expressão Gênica , Peptídeo YY/metabolismo , Proglucagon/metabolismo , Receptores dos Hormônios Gastrointestinais/metabolismo , Receptores de Peptídeos/metabolismo , Distribuição Tecidual , Transcrição Genética
12.
Int J Dev Biol ; 63(1-2): 29-35, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30919913

RESUMO

Latrophilin2 (Lphn2) is an adhesion-class of G protein-coupled receptor with an unknown function in development. Here, we show that Xenopus laevis lphn2 (Xlphn2) is involved in the migration and differentiation of neural crest (NC) cells and placode patterning in Xenopus laevis embryos. Although Xlphn2 mRNA was detected throughout embryogenesis, it was expressed more abundantly in the placode region. Morpholino antisense oligonucleotide-mediated knockdown of Xlphn2 caused abnormal migration of NC cells, irregular epibranchial placode segmentation, and defective cartilage formation. Transplantation of fluorescently-labeled NC regions of wild-type embryos into Xlphn2 morpholino-injected embryos reproduced the defective NC cell migration, indicating that Xlphn2 regulates the migration of NC cells in a non-cell autonomous manner. Our results suggest that Xlphn2 is essential for placode patterning and as a guidance molecule for NC cells.


Assuntos
Padronização Corporal , Movimento Celular , Ectoderma/fisiologia , Crista Neural/fisiologia , Receptores de Peptídeos/metabolismo , Proteínas de Xenopus/metabolismo , Xenopus laevis/fisiologia , Animais , Diferenciação Celular , Células Cultivadas , Ectoderma/embriologia , Embrião não Mamífero/citologia , Embrião não Mamífero/metabolismo , Desenvolvimento Embrionário , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Crista Neural/embriologia , Organogênese , Receptores de Peptídeos/genética , Proteínas de Xenopus/genética , Xenopus laevis/embriologia
14.
Artigo em Inglês | MEDLINE | ID: mdl-30857124

RESUMO

The aim of this study was to examine the effects of single-nucleotide polymorphisms (SNPs) in the anti-Müllerian hormone (AMH) and AMH type II receptor (AMHRII) genes on in vitro fertilization (IVF) outcomes. In this prospective cohort study, we genotyped the AMH 146 T > G, AMHRII -482 A > G and AMHRII IVS1 +149 T > A variants in 635 women undergoing their first cycle of controlled ovarian stimulation for IVF. DNA was extracted from the peripheral blood of all participants, and the SNPs were genotyped by real-time polymerase chain reaction. The distributions, frequencies of genes, and correlation with clinical pregnancy of IVF were analyzed. The AMH 146 T > G G/G genotype in women was associated with a lower clinical pregnancy rate (T/T: 55.0%, T/G: 51.8%, G/G: 40.0%; p < 0.05). Women with the AMH 146 T > G GG genotype were half as likely to have a clinical pregnancy compared with women with TT genotypes (OR = 0.55, 95% CI: 0.34⁻0.88, p = 0.014). With multivariate analysis, the AMH 146 T > G GG genotype remains as a significant independent factor to predict clinical pregnancy (p = 0.014). No significant difference was found between AMHRII polymorphisms and clinical pregnancy outcomes of IVF. In conclusion, our results show that AMH 146 T > G seems to be a susceptibility biomarker capable of predicting IVF pregnancy outcomes. Further studies should focus on the mechanism of these associations and the inclusion of other ethnic populations to confirm the findings of this study.


Assuntos
Hormônio Antimülleriano/genética , Fertilização In Vitro , Polimorfismo de Nucleotídeo Único , Taxa de Gravidez , Receptores de Peptídeos/genética , Receptores de Fatores de Crescimento Transformadores beta/genética , Adulto , Hormônio Antimülleriano/sangue , Grupos Étnicos , Feminino , Genótipo , Humanos , Reação em Cadeia da Polimerase , Gravidez , Estudos Prospectivos , Taiwan
15.
Horm Mol Biol Clin Investig ; 38(1)2019 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-30904901

RESUMO

Background Infertile women may have underlying genetic abnormalities. There is, at present, a significant number of studies on the relation between the follicle stimulating hormone receptor (FSHR) or anti-Müllerian hormone type II receptor (AMHRII) polymorphisms and response to in-vitro fertilisation (IVF) treatment. However, it is not yet clear which genotype or combination of genotypes is favourable towards a better ovarian stimulation and pregnancy outcome. Materials and methods In this study we assessed the distribution of the genotypes of FSHR Ser680Asn and of AMHRII -482A>G gene polymorphisms in a group of 126 infertile women and a control group of 100 fertile women by using real-time polymerase chain reaction (RT-PCR). Results Statistical analysis showed that the frequency of the genotypes is similar in both control and IVF/ intracytoplasmic sperm injection (ICSI) groups. Further investigation of the frequency of the nine possible combinations of these polymorphisms in the groups revealed no correlation between infertility and combination of the polymorphisms. Women with one polymorphism have on average 5.5 units higher levels of AMH compared to women carrying no polymorphism. In women with no polymorphisms, for each unit of FSH increase, the average concentration of blood AMH is expected to be 72% lower. Conclusion The distribution of the FSHR Ser680Asn and of the AMHRII -482A>G gene polymorphisms, in the Greek population is similar in fertile and infertile women. The study showed that FSH and AMH correlated levels in certain cases could be used to estimate a patient's ovarian reserve.


Assuntos
Infertilidade Feminina/genética , Polimorfismo de Nucleotídeo Único , Receptores do FSH/genética , Receptores de Peptídeos/genética , Receptores de Fatores de Crescimento Transformadores beta/genética , Adulto , Feminino , Humanos , Infertilidade Feminina/terapia , Reserva Ovariana , Receptores do FSH/sangue , Receptores de Peptídeos/sangue , Receptores de Fatores de Crescimento Transformadores beta/sangue , Injeções de Esperma Intracitoplásmicas/estatística & dados numéricos
16.
Bull Entomol Res ; 109(4): 534-543, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30789108

RESUMO

Latrophilin (LPH) is known as an adhesion G-protein-coupled receptor which involved in multiple physiological processes in organisms. Previous studies showed that lph not only involved the susceptibility to anticholinesterase insecticides but also affected fecundity in Tribolium castaneum. However, its regulatory mechanisms in these biological processes are still not clear. Here, we identified two potential downstream carboxylesterase (cce) genes of Tclph, esterase4 and esterase6, and further characterized their interactions with Tclph. After treatment of T. castaneum larvae with carbofuran or dichlorvos insecticides, the transcript levels of Tcest4 and Tcest6 were significantly induced from 12 to 72 h. RNAi against Tcest4 or Tcest6 led to the higher mortality compared with the controls after the insecticides treatment, suggesting that these two genes play a vital role in detoxification of insecticides in T. castaneum. Furthermore, with insecticides exposure to Tclph knockdown beetles, the expression of Tcest4 was upregulated but Tcest6 was downregulated, indicating that beetles existed a compensatory response against the insecticides. Additionally, RNAi of Tcest6 resulted in 43% reductions in female egg laying and completely inhibited egg hatching, which showed the similar phenotype as that of Tclph knockdown. These results indicated that Tclph affected fecundity by positively regulating Tcest6 expression. Our findings will provide a new insight into the molecular mechanisms of Tclph involved in physiological functions in T. castaneum.


Assuntos
Hidrolases de Éster Carboxílico/genética , Proteínas de Insetos/genética , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Receptores de Peptídeos/genética , Tribolium/efeitos dos fármacos , Animais , Carbofurano/farmacologia , Hidrolases de Éster Carboxílico/metabolismo , Diclorvós/farmacologia , Fertilidade/genética , Proteínas de Insetos/metabolismo , Receptores de Peptídeos/metabolismo , Tribolium/genética
17.
Theranostics ; 9(3): 620-632, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30809297

RESUMO

Rationale: Endometriosis is a highly prevalent gynecological disease in women of reproductive age that markedly reduces life quality and fertility. Unfortunately, there is no cure for this disease, which highlights that more efforts are needed to investigate the underlying mechanism for designing novel therapeutic regimens. This study aims to investigate druggable membrane receptors distinctively expressed in endometriotic cells. Methods: Bioinformatic analysis of public databases was employed to identify potential druggable candidates. Normal endometrial tissues and ectopic endometriotic lesions were obtained for the determination of target genes. Primary endometrial and endometriotic stromal cells as well as two different mouse models of endometriosis were used to characterize molecular mechanisms and therapeutic outcomes of endometriosis, respectively. Results: Anthrax toxin receptor 2 (ANTXR2) mRNA and protein are upregulated in the endometriotic specimens. Elevation of ANTXR2 promotes endometriotic cell adhesion, proliferation, and angiogenesis. Furthermore, hypoxia is the driving force for ANTXR2 upregulation via altering histone modification of ANTXR2 promoter by reducing the repressive mark, histone H3 lysine 27 (H3K27) trimethylation, and increasing the active mark, H3K4 trimethylation. Activation of ANTXR2 signaling leads to increased Yes-associated protein 1 (YAP1) nuclear translocation and transcriptional activity, which contributes to numerous pathological processes of endometriosis. Pharmacological blocking of ANTXR2 signaling not only prevents endometriotic lesion development but also causes the regression of established lesion. Conclusion: Taken together, we have identified a novel target that contributes to the disease pathogenesis of endometriosis and provided a potential therapeutic regimen to treat it.


Assuntos
Endometriose/patologia , Endometriose/terapia , Receptores de Peptídeos/antagonistas & inibidores , Receptores de Peptídeos/análise , Animais , Células Cultivadas , Modelos Animais de Doenças , Feminino , Perfilação da Expressão Gênica , Humanos , Hipóxia , Camundongos Endogâmicos C57BL , Receptores de Peptídeos/genética
18.
Mol Cell Endocrinol ; 487: 2-11, 2019 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-30703485

RESUMO

The goal of this paper is to establish Japanese medaka (Oryzias latipes) as a model for relaxin family peptide research, particularly for studying the functions of RLN3 and INSL5, hormones playing roles in neuroendocrine regulation. Medaka, like other teleosts, retained duplicate copies of rln3, insl5 and their rxfp3/4-type receptors following fish-specific whole genome duplication (WGD) and paralogous copies of these genes may have sub-functionalised providing an intuitive model for teasing apart the pleiotropic roles of the corresponding genes in mammals. To this end, we provide experimental evidence for the expression of the relaxin family genes in medaka that had previously only been identified in-silico, confirm the gene structure of five of the ligand genes, characterise gene expression across multiple tissues and during embryonic development, perform in situ hybridization with anti-sense insl5a on embryos and in adult brain and intestinal samples, and compare these results to the data available in zebrafish. We find broad similarities but also some differences in the expression of relaxin family genes in zebrafish versus medaka, and find support for the hypothesis that the rln3a/rln3b and insl5a/insl5b paralogues have been subfunctionalized. Given that medaka has a suite of relaxin family genes more similar to other teleosts, and has retained the gene for rxfp4 (which is lost in zebrafish), our results suggest that O. latipes may be a good model for delineating the ancestral function of the relaxin family genes involved in neuroendocrine regulation.


Assuntos
Família Multigênica , Sistemas Neurossecretores/metabolismo , Oryzias/genética , Receptores Acoplados a Proteínas-G/genética , Receptores de Peptídeos/genética , Relaxina/genética , Homologia de Sequência de Aminoácidos , Animais , Cromossomos/metabolismo , Embrião não Mamífero/metabolismo , Regulação da Expressão Gênica , Oryzias/embriologia , Receptores Acoplados a Proteínas-G/metabolismo , Receptores de Peptídeos/metabolismo , Relaxina/metabolismo , Especificidade da Espécie
19.
J Biol Chem ; 294(13): 5008-5022, 2019 03 29.
Artigo em Inglês | MEDLINE | ID: mdl-30709904

RESUMO

The hormone relaxin is considered a potential therapy for idiopathic pulmonary fibrosis (IPF). We have previously shown that a potential limitation to relaxin-based IPF therapy is decreased expression of a relaxin receptor, relaxin/insulin-like family peptide receptor 1 (RXFP1), in IPF fibroblasts. The mechanism that down-regulates RXFP1 in IPF remains unclear. To determine whether microRNAs (miRs) regulate RXFP1 gene expression, here we employed a bioinformatics approach to identify miRs predicted to target RXFP1 and identified a putative miR-144-3p target site in the RXFP1 mRNA. In situ hybridization of IPF lung biopsies revealed that miR-144-3p is expressed in fibroblastic foci. Furthermore, we found that miR-144-3p is up-regulated in IPF fibroblasts compared with lung fibroblasts from healthy donors. Transforming growth factor ß increased miR-144-3p expression in both healthy and IPF lung fibroblasts in a SMAD family 2/3 (SMAD2/3)-dependent manner, and Jun proto-oncogene AP-1 transcription factor subunit (AP-1) was required for constitutive miR-144-3p expression. Overexpression of an miR-144-3p mimic significantly reduced RXFP1 mRNA and protein levels and increased expression of the myofibroblast marker α-smooth muscle actin (α-SMA) in healthy lung fibroblasts. IPF lung fibroblasts transfected with anti-miR-144-3p had increased RXFP1 expression and reduced α-SMA expression. Of note, a lentiviral luciferase reporter carrying the WT 3' UTR of RXFP1 was significantly repressed in IPF lung fibroblasts, whereas a reporter carrying a mutated miR-144-3p-binding site exhibited less sensitivity toward endogenous miR-144-3p expression, indicating that miR-144-3p down-regulates RXFP1 in IPF lung fibroblasts by targeting its 3' UTR. We conclude that miR-144-3p directly represses RXFP1 mRNA and protein expression.


Assuntos
Fibroblastos/patologia , Fibrose Pulmonar Idiopática/genética , Pulmão/patologia , MicroRNAs/genética , Receptores Acoplados a Proteínas-G/genética , Receptores de Peptídeos/genética , Regiões 3' não Traduzidas , Células Cultivadas , Fibroblastos/metabolismo , Regulação da Expressão Gênica , Humanos , Fibrose Pulmonar Idiopática/epidemiologia , Fibrose Pulmonar Idiopática/patologia , Pulmão/metabolismo , RNA Mensageiro/genética
20.
Science ; 363(6429)2019 02 22.
Artigo em Inglês | MEDLINE | ID: mdl-30792275

RESUMO

Bidirectional signaling by cell adhesion molecules is thought to mediate synapse formation, but the mechanisms involved remain elusive. We found that the adhesion G protein-coupled receptors latrophilin-2 and latrophilin-3 selectively direct formation of perforant-path and Schaffer-collateral synapses, respectively, to hippocampal CA1-region neurons. Latrophilin-3 binds to two transcellular ligands: fibronectin leucine-rich repeat transmembrane proteins (FLRTs) and teneurins. In transgenic mice in vivo, both binding activities were required for input-specific synapse formation, which suggests that coincident binding of both ligands is necessary for synapse formation. In cultured neurons in vitro, teneurin or FLRT alone did not induce excitatory synapse formation, whereas together they potently did so. Thus, postsynaptic latrophilins promote excitatory synapse formation by simultaneous binding of two unrelated presynaptic ligands, which is required for formation of synaptic inputs at specific dendritic localizations.


Assuntos
Região CA1 Hipocampal/fisiologia , Proteínas de Membrana/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neurônios/fisiologia , Receptores Acoplados a Proteínas-G/fisiologia , Receptores de Peptídeos/metabolismo , Sinapses/fisiologia , Animais , Região CA1 Hipocampal/citologia , Células HEK293 , Humanos , Camundongos , Camundongos Knockout , Ligação Proteica , Receptores Acoplados a Proteínas-G/genética , Receptores Acoplados a Proteínas-G/metabolismo , Receptores de Peptídeos/genética , Sinapses/genética
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