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1.
PLoS One ; 15(9): e0239284, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32941503

RESUMO

The Rho GTPase RAC1 is an important regulator of cytoskeletal dynamics, but the role of macrophage-specific RAC1 has not been explored during atherogenesis. We analyzed RAC1 expression in human carotid atherosclerotic plaques using immunofluorescence and found higher macrophage RAC1 expression in advanced plaques compared with intermediate human atherosclerotic plaques. We then produced mice with Rac1-deficient macrophages by breeding conditional floxed Rac1 mice (Rac1fl/fl) with mice expressing Cre from the macrophage-specific lysosome M promoter (LC). Atherosclerosis was studied in vivo by infecting Rac1fl/fl and Rac1fl/fl/LC mice with AdPCSK9 (adenoviral vector overexpressing proprotein convertase subtilisin/kexin type 9). Rac1fl/fl/LC macrophages secreted lower levels of IL-6 and TNF-α and exhibited reduced foam cell formation and lipid uptake. The deficiency of Rac1 in macrophages reduced the size of aortic atherosclerotic plaques in AdPCSK9-infected Rac1fl/fl/LC mice. Compare with controls, intima/media ratios, the size of necrotic cores, and numbers of CD68-positive macrophages in atherosclerotic plaques were reduced in Rac1-deficient mice. Moreover, we found that RAC1 interacts with actin-binding filamin A. Macrophages expressed increased RAC1 levels in advanced human atherosclerosis. Genetic inactivation of RAC1 impaired macrophage function and reduced atherosclerosis in mice, suggesting that drugs targeting RAC1 may be useful in the treatment of atherosclerosis.


Assuntos
Aterosclerose/metabolismo , Macrófagos/metabolismo , Neuropeptídeos/genética , Proteínas rac1 de Ligação ao GTP/genética , Animais , Antígenos CD/genética , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/genética , Antígenos de Diferenciação Mielomonocítica/metabolismo , Aterosclerose/genética , Aterosclerose/patologia , Células Cultivadas , Humanos , Interleucina-6/genética , Interleucina-6/metabolismo , Metabolismo dos Lipídeos , Camundongos , Camundongos Endogâmicos C57BL , Neuropeptídeos/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Proteínas rac1 de Ligação ao GTP/metabolismo
2.
PLoS Biol ; 18(8): e3000548, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32745077

RESUMO

Sleep is vital for survival. Yet under environmentally challenging conditions, such as starvation, animals suppress their need for sleep. Interestingly, starvation-induced sleep loss does not evoke a subsequent sleep rebound. Little is known about how starvation-induced sleep deprivation differs from other types of sleep loss, or why some sleep functions become dispensable during starvation. Here, we demonstrate that down-regulation of the secreted cytokine unpaired 2 (upd2) in Drosophila flies may mimic a starved-like state. We used a genetic knockdown strategy to investigate the consequences of upd2 on visual attention and sleep in otherwise well-fed flies, thereby sidestepping the negative side effects of undernourishment. We find that knockdown of upd2 in the fat body (FB) is sufficient to suppress sleep and promote feeding-related behaviors while also improving selective visual attention. Furthermore, we show that this peripheral signal is integrated in the fly brain via insulin-expressing cells. Together, these findings identify a role for peripheral tissue-to-brain interactions in the simultaneous regulation of sleep quality and attention, to potentially promote adaptive behaviors necessary for survival in hungry animals.


Assuntos
Atenção/fisiologia , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Comportamento Alimentar/fisiologia , Inanição/genética , Percepção Visual/fisiologia , Animais , Encéfalo/citologia , Encéfalo/metabolismo , Proteínas de Drosophila/deficiência , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Corpo Adiposo/metabolismo , Feminino , Regulação da Expressão Gênica , Técnicas de Silenciamento de Genes , Insulina/genética , Insulina/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Transdução de Sinais , Sono/fisiologia , Privação do Sono/genética , Privação do Sono/metabolismo , Inanição/metabolismo
3.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 40(7): 1001-1007, 2020 May 25.
Artigo em Chinês | MEDLINE | ID: mdl-32701233

RESUMO

OBJECTIVE: To explore the effects of taurolithocholic acid (tLCA) and chenodeoxycholic acid (CDCA) on the expression of aorexigenic neuropeptide in mouse hypothalamus GT1-7 cells. METHODS: Mouse hypothalamic GT1-7 cells were treated with culture medium containing 10% FBS (control group, n=3) or with 10 nmol/L, 100 nmol/L, 1 µmol/L and 10 µmol/L tLCA (tLCA group, n=3) or CDCA (CDCA group, n=3) for 12, 24 or 48 h. Real-time PCR was performed to determine the expression levels of proopiomelanocortin (POMC) mRNA in the cells, and the production levels of α-melanocyte-stimulating hormone (α-MSH) were assessed using an ELISA kit. Signal transduction and activator of transcription 3 phosphorylation (p-STAT3), threonine kinase phosphorylation (p-AKT), suppressor of cytokine signaling 3 (SOCS3), G protein-coupled bile acid receptor-1 (TGR5) and farnesoid X receptor (FXR) protein were detected by Western blotting. RESULTS: Western blotting results showed that mouse hypothalamic GT1-7 cells expressed two bile acid receptors, TGR5 and FXR, whose expressions were regulated by bile acids. Real-time PCR showed that the expression of POMC mRNA was significantly increased in the cells after treatment with 10 µmol/L tLCA or CDCA for 24 h. POMC-derived anorexigenic peptide α-MSH increased significantly in GT1-7 cells after treatment with 10 µmol/L tLCA or CDCA for 24 h. Treatment of the cells with tLCA or CDCA significantly increased the expressions of intracellular signaling proteins including p-STAT3, p-AKT and SOCS3. CONCLUSIONS: Mouse hypothalamic GT1-7 cells express bile acid receptors TGR5 and FXR. Bile acids tLCA or CDCA can promote the expression of POMC mRNA and increase the production of the anorexigenic peptide α-MSH. The intracellular signaling proteins p-AKT, p-STAT3 and SOCS3 are likely involved in bile acid-induced anorexigenic peptide production.


Assuntos
Ácido Quenodesoxicólico , Regulação da Expressão Gênica , Pró-Opiomelanocortina , Transdução de Sinais , Ácido Taurolitocólico , alfa-MSH , Animais , Linhagem Celular , Ácido Quenodesoxicólico/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Hipotálamo/citologia , Camundongos , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Pró-Opiomelanocortina/genética , RNA Mensageiro/genética , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/efeitos dos fármacos , Proteína 3 Supressora da Sinalização de Citocinas/metabolismo , Ácido Taurolitocólico/farmacologia , alfa-MSH/genética
4.
Curr Opin Neurobiol ; 63: 176-188, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32679509

RESUMO

Neuropeptides, members of a large and evolutionarily ancient family of proteinaceous cell-cell signaling molecules, are widely recognized as extremely potent regulators of brain function and behavior. At the cellular level, neuropeptides are known to act mainly via modulation of ion channel and synapse function, but functional impacts emerging at the level of complex cortical synaptic networks have resisted mechanistic analysis. New findings from single-cell RNA-seq transcriptomics now illuminate intricate patterns of cortical neuropeptide signaling gene expression and new tools now offer powerful molecular access to cortical neuropeptide signaling. Here we highlight some of these new findings and tools, focusing especially on prospects for experimental and theoretical exploration of peptidergic and synaptic networks interactions underlying cortical function and plasticity.


Assuntos
Neuropeptídeos , Plasticidade Neuronal , Neuropeptídeos/genética , Transdução de Sinais , Sinapses
5.
Exp Anim ; 69(4): 448-460, 2020 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-32669479

RESUMO

The non-motor symptoms (NMS) of Parkinson's disease (PD) are found in more than 90% of patients with PD. Here, we explored the effects of electroacupuncture (EA) stimulation at Zhong wan (CV-12), Qihai (RN-7), Zusanli (ST-36) and Taichong (LR-3) on NMS and brain-gut peptides of PD. We found that EA intervention alleviated the motor deficit induced by 6-OHDA in rats indicated by the decreased abnormal involuntary movements (AIMs) scores and the net number of rotations and increased cylinder test grade. It also improved the spatial memory and attenuated anxiety-like and depression of PD model rats. EA treatment significantly inhibited neuronal apoptosis in PD model animals, as demonstrated by the increased number of TH positive cells and reduced number of apoptotic cells in the substantia nigra. The expression of cleaved caspase-3 and cleaved PARP in PD model rats was markedly suppressed by EA stimulation. Moreover, EA remarkably inhibited the inflammatory response in PD model rats, as revealed by the decreased levels of TNF-α, IL-1ß, and COX-2 mRNA expression. It also attenuated the oxidative stress in rats, as indicated by the increased levels of SOD and GSH and the decreased level of MDA. EA treatment contributed to alleviating PD by regulating brain-gut peptides in rats, such as NPY, CCK, SST, GAS, and PYY. In conclusion, EA stimulation at CV-12, RN-7, ST-36, and LR-3 effectively alleviates the NMS of PD partly through regulating the levels of brain-gut peptides.


Assuntos
Encéfalo/metabolismo , Eletroacupuntura , Regulação da Expressão Gênica , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Doença de Parkinson/genética , Doença de Parkinson/terapia , Animais , Ansiedade , Modelos Animais de Doenças , Discinesias , Masculino , Estresse Oxidativo , Doença de Parkinson/metabolismo , Doença de Parkinson/psicologia , Ratos Sprague-Dawley , Memória Espacial
6.
PLoS One ; 15(6): e0233991, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32497060

RESUMO

Neuropeptides are secreted molecules that have conserved roles modulating many processes, including mood, reproduction, and feeding. Dysregulation of neuropeptide signaling is also implicated in neurological disorders such as epilepsy. However, much is unknown about the mechanisms regulating specific neuropeptides to mediate behavior. Here, we report that the expression levels of dozens of neuropeptides are up-regulated in response to circuit activity imbalance in C. elegans. acr-2 encodes a homolog of human nicotinic receptors, and functions in the cholinergic motoneurons. A hyperactive mutation, acr-2(gf), causes an activity imbalance in the motor circuit. We performed cell-type specific transcriptomic analysis and identified genes differentially expressed in acr-2(gf), compared to wild type. The most over-represented class of genes are neuropeptides, with insulin-like-peptides (ILPs) the most affected. Moreover, up-regulation of neuropeptides occurs in motoneurons, as well as sensory neurons. In particular, the induced expression of the ILP ins-29 occurs in the BAG neurons, which were previously shown to function in gas-sensing. We also show that this up-regulation of ins-29 in acr-2(gf) animals is activity-dependent. Our genetic and molecular analyses support cooperative effects for ILPs and other neuropeptides in promoting motor circuit activity in the acr-2(gf) background. Together, this data reveals that a major transcriptional response to motor circuit dysregulation is in up-regulation of multiple neuropeptides, and suggests that BAG sensory neurons can respond to intrinsic activity states to feedback on the motor circuit.


Assuntos
Proteínas de Caenorhabditis elegans/genética , Caenorhabditis elegans/genética , Neuropeptídeos/genética , Receptores Nicotínicos/genética , Transcriptoma , Animais , Caenorhabditis elegans/fisiologia , Perfilação da Expressão Gênica , Neurônios Motores/metabolismo , Mutação , Células Receptoras Sensoriais/metabolismo
7.
Elife ; 92020 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-32538354

RESUMO

Experiments in sea cucumbers reveal how the physiological responses regulated by a neuropeptide called kisspeptin have evolved.


Assuntos
Neuropeptídeos , Pepinos-do-Mar , Animais , Evolução Biológica , Kisspeptinas/genética , Kisspeptinas/metabolismo , Neuropeptídeos/genética , Pepinos-do-Mar/metabolismo , Transdução de Sinais
8.
Am J Physiol Lung Cell Mol Physiol ; 319(1): L48-L60, 2020 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-32460521

RESUMO

Chronic obstructive pulmonary disease (COPD) is associated with features of accelerated aging, including cellular senescence, DNA damage, oxidative stress, and extracellular matrix (ECM) changes. We propose that these features are particularly apparent in patients with severe, early-onset (SEO)-COPD. Whether fibroblasts from COPD patients display features of accelerated aging and whether this is also present in relatively young SEO-COPD patients is unknown. Therefore, we aimed to determine markers of aging in (SEO)-COPD-derived lung fibroblasts and investigate the impact on ECM. Aging hallmarks and ECM markers were analyzed in lung fibroblasts from SEO-COPD and older COPD patients and compared with fibroblasts from matched non-COPD groups (n = 9-11 per group), both at normal culture conditions and upon Paraquat-induced senescence. COPD-related differences in senescence and ECM expression were validated in lung tissue. Higher levels of cellular senescence, including senescence-associated ß-galactosidase (SA-ß-gal)-positive cells (19% for COPD vs. 13% for control) and p16 expression, DNA damage (γ-H2A.X-positive nuclei), and oxidative stress (MGST1) were detected in COPD compared with control-derived fibroblasts. Most effects were also different in SEO-COPD, with SA-ß-gal-positive cells only being significant in SEO-COPD vs. matched controls. Lower decorin expression in COPD-derived fibroblasts correlated with higher p16 expression, and this association was confirmed in lung tissue. Paraquat treatment induced cellular senescence along with clear changes in ECM expression, including decorin. Fibroblasts from COPD patients, including SEO-COPD, display higher levels of cellular senescence, DNA damage, and oxidative stress. The association between cellular senescence and ECM expression changes may suggest a link between accelerated aging and ECM dysregulation in COPD.


Assuntos
Senescência Celular , Matriz Extracelular/metabolismo , Doença Pulmonar Obstrutiva Crônica/patologia , Adulto , Idade de Início , Biomarcadores/metabolismo , Células Cultivadas , Dano ao DNA , Feminino , Fibroblastos/patologia , Regulação da Expressão Gênica , Humanos , Pulmão/patologia , Pulmão/fisiopatologia , Masculino , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Pessoa de Meia-Idade , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Estresse Oxidativo , Paraquat/toxicidade , Doença Pulmonar Obstrutiva Crônica/genética , Doença Pulmonar Obstrutiva Crônica/fisiopatologia
9.
Tunis Med ; 98(2): 161-163, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32395807

RESUMO

We report the case of a 23-year-old woman with a not yet described (to the best of our knowledge) association of left ventricle non-compaction with both atrial and ventricular defects. Family genetic survey concluded to, a probably sporadic, E101K gene mutation.


Assuntos
Cardiopatias Congênitas/diagnóstico , Comunicação Interatrial/diagnóstico , Ventrículos do Coração/anormalidades , Substituição de Aminoácidos/genética , Proteínas Associadas à Distrofina/genética , Feminino , Ácido Glutâmico/genética , Cardiopatias Congênitas/genética , Comunicação Interatrial/complicações , Comunicação Interatrial/genética , Humanos , Lisina/genética , Mutação de Sentido Incorreto , Neuropeptídeos/genética , Adulto Jovem
10.
Biochim Biophys Acta Mol Cell Res ; 1867(8): 118718, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32289337

RESUMO

Alternative splicing enables G protein-coupled receptor (GPCR) genes to greatly increase the number of structurally and functionally distinct receptor isoforms. However, the functional role and relevance of the individual GPCR splice variants in regulating physiological processes are still to be assessed. A naturally occurring alternative splice variant of Bombyx CAPA-PVK receptor, BomCAPA-PVK-R1-Δ341, has been shown to act as a dominant-negative protein to regulate cell surface expression and function of the canonical CAPA-PVK receptor. Herein, using functional assays, we identify the splice variant Δ341 as a specific receptor for neuropeptide CAPA-PK, and upon activation, Δ341 signals to ERK1/2 pathway. Further characterization demonstrates that Δ341 couples to Gαi/o, distinct from the Gαq-coupled canonical CAPA-PVK receptor, triggering ERK1/2 phosphorylation through Gßγ-PI3K-PKCζ signaling cascade. Moreover, our ELISA data show that the ligand-dependent internalization of the splice variant Δ341 is significantly impaired due to lack of GRKs-mediated phosphorylation sites. Our findings highlight the potential of this knowledge for molecular, pharmacological and physiological studies on GPCR splice variants in the future.


Assuntos
Bombyx/genética , Bombyx/metabolismo , Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/genética , Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/metabolismo , Processamento de RNA/fisiologia , Receptores Acoplados a Proteínas-G/genética , Receptores Acoplados a Proteínas-G/metabolismo , Processamento Alternativo , Animais , Clonagem Molecular , Proteínas de Ligação ao GTP/genética , Proteínas de Ligação ao GTP/metabolismo , Células HEK293 , Humanos , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Larva , Sistema de Sinalização das MAP Quinases , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Fosforilação , Isoformas de Proteínas/genética , Transdução de Sinais , Transcriptoma
11.
J Neurosci ; 40(21): 4219-4229, 2020 05 20.
Artigo em Inglês | MEDLINE | ID: mdl-32303647

RESUMO

In Drosophila, the mushroom bodies (MB) constitute the central brain structure for olfactory associative memory. As in mammals, the cAMP/PKA pathway plays a key role in memory formation. In the MB, Rutabaga (Rut) adenylate cyclase acts as a coincidence detector during associative conditioning to integrate calcium influx resulting from acetylcholine stimulation and G-protein activation resulting from dopaminergic stimulation. Amnesiac encodes a secreted neuropeptide required in the MB for two phases of aversive olfactory memory. Previous sequence analysis has revealed strong homology with the mammalian pituitary adenylate cyclase-activating peptide (PACAP). Here, we examined whether amnesiac is involved in cAMP/PKA dynamics in response to dopamine and acetylcholine co-stimulation in living flies. Experiments were conducted with both sexes, or with either sex. Our data show that amnesiac is necessary for the PKA activation process that results from coincidence detection in the MB. Since PACAP peptide is cleaved by the human membrane neprilysin hNEP, we searched for an interaction between Amnesiac and Neprilysin 1 (Nep1), a fly neprilysin involved in memory. We show that when Nep1 expression is acutely knocked down in adult MB, memory deficits displayed by amn hypomorphic mutants are rescued. Consistently, Nep1 inhibition also restores normal PKA activation in amn mutant flies. Taken together, the results suggest that Nep1 targets Amnesiac degradation to terminate its signaling function. Our work thus highlights a key role for Amnesiac in establishing within the MB the PKA dynamics that sustain middle-term memory (MTM) formation, a function modulated by Nep1.SIGNIFICANCE STATEMENT The Drosophila amnesiac gene encodes a secreted neuropeptide whose expression is required for specific memory phases in the mushroom bodies (MB), the olfactory memory center. Here, we show that Amnesiac is required for PKA activation resulting from coincidence detection, a mechanism by which the MB integrate two spatially distinct stimuli to encode associative memory. Furthermore, our results uncover a functional relationship between Amnesiac and Neprilysin 1 (Nep1), a membrane peptidase involved in memory and expressed in the MB. These results suggest that Nep1 modulates Amnesiac levels. We propose that on conditioning, Amnesiac release from the MB allows, via an autocrine process, the sustaining of PKA activation-mediating memory, which subsequently is inactivated by Nep1 degradation.


Assuntos
Aprendizagem da Esquiva/fisiologia , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Proteínas de Drosophila/genética , Memória/fisiologia , Corpos Pedunculados/metabolismo , Neprilisina/metabolismo , Neuropeptídeos/genética , Animais , Animais Geneticamente Modificados , Proteínas de Drosophila/metabolismo , Drosophila melanogaster , Neuropeptídeos/metabolismo , Olfato/fisiologia
12.
Poult Sci ; 99(3): 1409-1420, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32115028

RESUMO

Lysine is the second most limiting amino acid after methionine and is considered the most limiting amino acid for growth in poultry. Lysine requirement for broiler chickens has changed over the years. Leptin and adiponectin represent 2 adipokines that mediate metabolism by eliciting satiety effects whereas ghrelin peptide hormone influences appetite. We hypothesize that this affects growth performance of chicks. This study evaluates the effect of varying dietary lysine homeostasis on performance of broiler chickens through satiety- and appetite-mediating hormones. In 3 replications, 270 one-day-old chicks were reared for 8 wk feeding on diets comprising 0.85, 1.14, and 1.42% lysine during the starter period and 0.75, 1.00, and 1.25% lysine during the grower period. These concentrations of lysine represent 75% (low lysine), 100% (control), and 125% (high lysine) of National Research Council recommendation for broiler chickens. Feed and water were provided for ad libitum consumption. At 8 wk of age, liver, pancreas, brain, and hypothalamus tissues were collected from 18 birds randomly selected from each treatment, snap frozen in liquid nitrogen, and stored at -80°C until use. Total RNA was extracted, and cDNA was synthesized for quantitative real-time PCR assays. Low lysine concentration caused slow growth and high mortality. There was significant upregulation of ghrelin in the hypothalamus and pancreas, and leptin and adiponectin in the hypothalamus and liver, and downregulation of ghrelin in the intestines. At low lysine concentrations, adiponectin was not expressed in both pancreas and intestines. High lysine concentration exhibited increased growth, upregulation of ghrelin in the liver, and downregulation of ghrelin in the intestines, and both adiponectin and leptin in the liver. The expression of ghrelin was negatively correlated with the expression of adiponectin and leptin (P < 0.05) in the liver, hypothalamus, and pancreas. Expression of leptin was positively correlated with adiponectin in the hypothalamus and liver (P < 0.05), exhibiting satiety effects when the concentrations of lysine were low.


Assuntos
Apetite/genética , Galinhas/fisiologia , Lisina/metabolismo , Neuropeptídeos/genética , Hormônios Peptídicos/genética , Saciação , Adiponectina/genética , Adiponectina/metabolismo , Ração Animal/análise , Animais , Galinhas/genética , Galinhas/crescimento & desenvolvimento , Dieta/veterinária , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga , Regulação para Baixo , Perfilação da Expressão Gênica/veterinária , Grelina/genética , Grelina/metabolismo , Homeostase , Leptina/genética , Leptina/metabolismo , Lisina/administração & dosagem , Neuropeptídeos/metabolismo , Hormônios Peptídicos/metabolismo , Distribuição Aleatória , Regulação para Cima
13.
Exp Eye Res ; 193: 107993, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32147400

RESUMO

Bombina variegata 8 (Bv8), also known as prokineticin-2 (PK-2), is a potent pro-angiogenic factor. However, its role in retinal neovascularization (RNV) remains unknown. In this study, we explored the role of Bv8 in the pathogenesis of RNV. We found that the expression of Bv8 was significantly increased in two different models of retinal neovascularization: the oxygen-induced retinopathy (OIR) mouse model and the rhodopsin promoter (rho)/VEGF transgenic mouse model. Neutralization of Bv8 by intravitreal injections of its antibody, not only inhibited retinal and subretinal neovascularization but also decreased the mRNA and protein levels of several pro-angiogenic factors. Our in vitro assay showed that recombinant human Bv8 (RhBv8) protein promoted human retinal microvascular endothelial cells (HRECs) tube-formation, cell proliferation and vascular endothelial growth factor receptor 1 (VEGFR1) and receptor 2 (VEGFR2) expression. Our findings suggest that Bv8 could be used as a novel target for the treatment of RNV-related ocular diseases.


Assuntos
Proteínas de Anfíbios/genética , Regulação da Expressão Gênica , Neuropeptídeos/genética , Neovascularização Retiniana/tratamento farmacológico , Rodopsina/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Proteínas de Anfíbios/metabolismo , Animais , Animais Recém-Nascidos , Proliferação de Células , Modelos Animais de Doenças , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Neuropeptídeos/metabolismo , Oxigênio/toxicidade , Regiões Promotoras Genéticas , RNA/genética , Neovascularização Retiniana/metabolismo , Neovascularização Retiniana/patologia , Vasos Retinianos/metabolismo
14.
BMC Mol Cell Biol ; 21(1): 8, 2020 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-32111164

RESUMO

BACKGROUND: TgDCX is a doublecortin-domain protein associated with the conoid fibers, a set of strongly curved non-tubular tubulin-polymers in Toxoplasma. TgDCX deletion impairs conoid structure and parasite invasion. TgDCX contains two tubulin-binding domains: a partial P25α and the DCX/doublecortin domain. Orthologues are found in apicomplexans and their free-living relatives Chromera and Vitrella. RESULTS: We report that isolated TgDCX-containing conoid fibers retain their pronounced curvature, but loss of TgDCX destabilizes the fibers. We crystallized and determined the 3D-structure of the DCX-domain, which is similar to those of human doublecortin and well-conserved among TgDCX orthologues. However, the orthologues vary widely in targeting to the conoid in Toxoplasma and in modulating microtubule organization in Xenopus cells. Several orthologues bind to microtubules in Xenopus cells, but only TgDCX generates short, strongly curved microtubule arcs. EM analysis shows microtubules decorated with TgDCX bundled into rafts, often bordered on one edge by a "C"-shaped incomplete tube. A Chromera orthologue closely mimics TgDCX targeting in Toxoplasma and binds to microtubules in Xenopus cells, but does not generate arcs or "C"-shaped tubes, and fails to rescue the defects of the TgDCX-knockout parasite. CONCLUSIONS: These observations suggest that species-specific features of TgDCX enable it to generate strongly curved tubulin-polymers to support efficient host-cell invasion.


Assuntos
Proteínas Associadas aos Microtúbulos/química , Neuropeptídeos/química , Toxoplasma/metabolismo , Tubulina (Proteína)/metabolismo , Animais , Células Cultivadas , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/metabolismo , Técnicas de Inativação de Genes , Interações Hospedeiro-Parasita/genética , Microscopia Eletrônica , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Proteínas Associadas aos Microtúbulos/ultraestrutura , Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Microtúbulos/ultraestrutura , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Polímeros/metabolismo , Regiões Promotoras Genéticas , Ligação Proteica , Domínios Proteicos/genética , Proteínas Recombinantes , Toxoplasma/química , Toxoplasma/efeitos dos fármacos , Toxoplasma/ultraestrutura , Tubulina (Proteína)/química , Xenopus
15.
Proc Natl Acad Sci U S A ; 117(13): 7401-7408, 2020 03 31.
Artigo em Inglês | MEDLINE | ID: mdl-32179671

RESUMO

The intake of macronutrients is crucial for the fitness of any animal and is mainly regulated by peripheral signals to the brain. How the brain receives and translates these peripheral signals or how these interactions lead to changes in feeding behavior is not well-understood. We discovered that 2 crustacean cardioactive peptide (CCAP)-expressing neurons in Drosophila adults regulate feeding behavior and metabolism. Notably, loss of CCAP, or knocking down the CCAP receptor (CCAP-R) in 2 dorsal median neurons, inhibits the release of neuropeptide F (NPF), which regulates feeding behavior. Furthermore, under starvation conditions, flies normally have an increased sensitivity to sugar; however, loss of CCAP, or CCAP-R in 2 dorsal median NPF neurons, inhibited sugar sensitivity in satiated and starved flies. Separate from its regulation of NPF signaling, the CCAP peptide also regulates triglyceride levels. Additionally, genetic and optogenetic studies demonstrate that CCAP signaling is necessary and sufficient to stimulate a reflexive feeding behavior, the proboscis extension reflex (PER), elicited when external food cues are interpreted as palatable. Dopaminergic signaling was also sufficient to induce a PER. On the other hand, although necessary, NPF neurons were not able to induce a PER. These data illustrate that the CCAP peptide is a central regulator of feeding behavior and metabolism in adult flies, and that NPF neurons have an important regulatory role within this system.


Assuntos
Comportamento Alimentar/fisiologia , Neuropeptídeos/metabolismo , Animais , Encéfalo/metabolismo , Ritmo Circadiano/fisiologia , Dopamina/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Neurônios/metabolismo , Neuropeptídeos/genética , Transdução de Sinais , Inanição/metabolismo
16.
Sci Rep ; 10(1): 4140, 2020 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-32139752

RESUMO

An animal's behavioral and physiological response to stressors includes changes to its responses to stimuli. How such changes occur is not well understood. Here we describe a Caenorhabditis elegans quiescent behavior, post-response quiescence (PRQ), which is modulated by the C. elegans response to cellular stressors. Following an aversive mechanical or blue light stimulus, worms respond first by briefly moving, and then become more quiescent for a period lasting tens of seconds. PRQ occurs at low frequency in unstressed animals, but is more frequent in animals that have experienced cellular stress due to ultraviolet light exposure as well as in animals following overexpression of epidermal growth factor (EGF). PRQ requires the function of the carboxypeptidase EGL-21 and the calcium-activated protein for secretion (CAPS) UNC-31, suggesting it has a neuropeptidergic mechanism. Although PRQ requires the sleep-promoting neurons RIS and ALA, it is not accompanied by decreased arousability, and does not appear to be homeostatically regulated, suggesting that it is not a sleep state. PRQ represents a simple, tractable model for studying how neuromodulatory states like stress alter behavioral responses to stimuli.


Assuntos
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/metabolismo , Animais , Caenorhabditis elegans/genética , Caenorhabditis elegans/efeitos da radiação , Proteínas de Caenorhabditis elegans/genética , Fator de Crescimento Epidérmico/genética , Fator de Crescimento Epidérmico/metabolismo , Mecanotransdução Celular/fisiologia , Neurônios/metabolismo , Neurônios/efeitos da radiação , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Raios Ultravioleta
17.
Gene ; 743: 144605, 2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32199950

RESUMO

Atrijuglans hetaohei Yang (Lepidoptera: Gelechioidea), is one of the major pests that can seriously damage the walnut fruits. Neuropeptides and their receptors regulate most physiological functions in insects and represent new targets for the development of control agents. To identify the neuropeptides and their receptors from A. hetaohei, we sequenced and analyzed its head transcriptomic data, identified 32 neuropeptides and 39 neuropeptide receptor genes. Sequence comparisons and phylogenetic analyses suggest that A. hetaohei neuropeptides and receptor genes have high homology with those in Bombyx mori, Chilo suppressalis, Plutella xylostella and Helicoverpa armigera. Moreover, gene expression patterns revealed that neuropeptide genes such as AKH1, CP, MS and PTTH were expressed specifically in male head, while CAP3, DH, NPLP1, PBAN and SIF showed higher expression in the female head. Bur showed abdomen biased expression in both male and female. Neuropeptide receptor genes such as A8, A11, A15 and LGR were highly expressed in male head, whereas A24 and LGR2 were preferentially expressed in female head. This is the first sequencing, identification and expression analyses of neuropeptides and neuropeptide receptor genes from A. hetaohei. Our results could provide a powerful background that will facilitate the further investigations using transcriptomics to determine neuropeptides and their receptors presence, functions, and indicates potential targets in A. hetaohei for a novel pest management strategy.


Assuntos
Proteínas de Insetos/genética , Mariposas/genética , Neuropeptídeos/genética , Receptores de Neuropeptídeos/genética , Transcriptoma/genética , Animais , Clonagem Molecular , Feminino , Perfilação da Expressão Gênica , Genes de Insetos , Juglans/parasitologia , Masculino , Controle de Pragas/métodos , Filogenia , Fatores Sexuais
18.
Exp Cell Res ; 389(2): 111911, 2020 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-32061832

RESUMO

Phospholipid Phosphatase-Related Protein Type 1 (PLPPR1) is a six-transmembrane protein that belongs to the family of plasticity-related gene proteins, which is a novel brain-specific subclass of the lipid phosphate phosphatase superfamily. PLPPR1-5 have prominent roles in synapse formation and axonal pathfinding. We found that PLPPR1 overexpression in the mouse neuroblastoma cell line (Neuro2a) results in increase in cell adhesion and reduced cell migration. During migration, these cells leave behind long fibrous looking extensions of the plasma membrane causing a peculiar phenotype. Cells expressing PLPPR1 showed decreased actin turnover and decreased disassembly of focal adhesions. PLPPR1 also reduced active Rac1, and expressing dominant negative Rac1 produced a similar phenotype to overexpression of PLPPR1. The PLPPR1-induced phenotype of long fibers was reversed by introducing constitutively active Rac1. In summary, we show that PLPPR1 decreases active Rac1 levels that leads to cascade of events which increases cell adhesion.


Assuntos
Adesão Celular , Adesões Focais , Regulação Neoplásica da Expressão Gênica , Neuroblastoma/patologia , Neuropeptídeos/metabolismo , Monoéster Fosfórico Hidrolases/metabolismo , Proteínas rac1 de Ligação ao GTP/metabolismo , Actinas/metabolismo , Animais , Movimento Celular , Camundongos , Neuroblastoma/metabolismo , Neuropeptídeos/genética , Monoéster Fosfórico Hidrolases/genética , Transdução de Sinais , Células Tumorais Cultivadas , Proteínas rac1 de Ligação ao GTP/genética
19.
Sci Rep ; 10(1): 3007, 2020 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-32080271

RESUMO

In commercial flocks of laying hens, keel bone fractures (KBFs) are prevalent and associated with behavioural indicators of pain. However, whether their impact is severe enough to induce a depressive-like state of chronic stress is unknown. As chronic stress downregulates adult hippocampal neurogenesis (AHN) in mammals and birds, we employ this measure as a neural biomarker of subjective welfare state. Radiographs obtained longitudinally from Lohmann Brown laying hens housed in a commercial multi-tier aviary were used to score the severity of naturally-occurring KBFs between the ages of 21-62 weeks. Individual birds' transitions between aviary zones were also recorded. Focal hens with severe KBFs at 3-4 weeks prior to sampling (n = 15) had lower densities of immature doublecortin-positive (DCX+) multipolar and bipolar neurons in the hippocampal formation than focal hens with minimal fractures (n = 9). KBF severity scores at this time also negatively predicted DCX+ cell numbers on an individual level, while hens that acquired fractures earlier in their lives had fewer DCX+ neurons in the caudal hippocampal formation. Activity levels 3-4 weeks prior to sampling were not associated with AHN. KBFs thus lead to a negative affective state lasting at least 3-4 weeks, and management steps to reduce their occurrence are likely to have significant welfare benefits.


Assuntos
Bem-Estar do Animal/ética , Fraturas Ósseas/complicações , Hipocampo/fisiopatologia , Doenças das Aves Domésticas/psicologia , Esterno/lesões , Estresse Psicológico/etiologia , Criação de Animais Domésticos/ética , Animais , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Galinhas , Feminino , Fraturas Ósseas/patologia , Fraturas Ósseas/psicologia , Expressão Gênica , Hipocampo/metabolismo , Abrigo para Animais/ética , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Neurogênese/fisiologia , Neurônios/metabolismo , Neurônios/patologia , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Doenças das Aves Domésticas/patologia , Reprodução/genética , Estresse Psicológico/patologia , Estresse Psicológico/psicologia , Índices de Gravidade do Trauma
20.
Gen Comp Endocrinol ; 292: 113443, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32097662

RESUMO

We de novo assembled a transcriptome for early life-stages of the Aotearoa-New Zealand crayfish, Paranephrops zealandicus, establishing the first genetic resource for this under-developed aquaculture species and for the Paranephrops genus. Mining of this transcriptome for neuropeptides and their putative cognate G protein-coupled receptors (GPCRs) yielded a comprehensive catalogue of neuropeptides, but few putative neuropeptide GPCRs. Of the neuropeptides commonly identified from decapod transcriptomes, only crustacean female sex hormone and insulin-like peptide were absent from our trinity de novo transcriptome assembly, and also RNA-sequence reads. We identified 63 putative neuropeptide precursors from 43 families, predicted to yield 122 active peptides. Transcripts encoding 26 putative neuropeptide GPCRs were identified but were often incomplete. Putative GPCRs for 15 of the neuropeptides identified here were absent from our transcriptome and RNAseq reads. These data highlight the diverse neuropeptide systems already present at the early development life stages sampled here for P. zealandicus.


Assuntos
Astacoidea/embriologia , Astacoidea/genética , Neuropeptídeos/genética , Receptores Acoplados a Proteínas-G/genética , Transcriptoma/genética , Sequência de Aminoácidos , Animais , Embrião não Mamífero/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Neuropeptídeos/química , Neuropeptídeos/metabolismo , Nova Zelândia , Receptores Acoplados a Proteínas-G/metabolismo , Transdução de Sinais
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