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1.
Nat Neurosci ; 23(3): 456-467, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32066983

RESUMO

Mammalian circadian behaviors are orchestrated by the suprachiasmatic nucleus (SCN) in the ventral hypothalamus, but the number of SCN cell types and their functional roles remain unclear. We have used single-cell RNA-sequencing to identify the basic cell types in the mouse SCN and to characterize their circadian and light-induced gene expression patterns. We identified eight major cell types, with each type displaying a specific pattern of circadian gene expression. Five SCN neuronal subtypes, each with specific combinations of markers, differ in their spatial distribution, circadian rhythmicity and light responsiveness. Through a complete three-dimensional reconstruction of the mouse SCN at single-cell resolution, we obtained a standardized SCN atlas containing the spatial distribution of these subtypes and gene expression. Furthermore, we observed heterogeneous circadian gene expression between SCN neuron subtypes. Such a spatiotemporal pattern of gene regulation within the SCN may have an important function in the circadian pacemaker.


Assuntos
Expressão Gênica/fisiologia , Neurônios/fisiologia , Análise de Célula Única , Núcleo Supraquiasmático/fisiologia , Animais , Atlas como Assunto , Ritmo Circadiano/fisiologia , Peptídeos e Proteínas de Sinalização do Ritmo Circadiano/genética , Expressão Gênica/efeitos da radiação , Regulação da Expressão Gênica/fisiologia , Genômica , Luz , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neurônios/classificação , Estimulação Luminosa , Núcleo Supraquiasmático/anatomia & histologia , Núcleo Supraquiasmático/citologia
2.
Genes Dev ; 34(5-6): 302-320, 2020 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-32029452

RESUMO

ADP-ribosylation (ADPRylation) is a posttranslational modification of proteins discovered nearly six decades ago, but many important questions remain regarding its molecular functions and biological roles, as well as the activity of the ADP-ribose (ADPR) transferase enzymes (PARP family members) that catalyze it. Growing evidence indicates that PARP-mediated ADPRylation events are key regulators of the protein biosynthetic pathway, leading from rDNA transcription and ribosome biogenesis to mRNA synthesis, processing, and translation. In this review we describe the role of PARP proteins and ADPRylation in all facets of this pathway. PARP-1 and its enzymatic activity are key regulators of rDNA transcription, which is a critical step in ribosome biogenesis. An emerging role of PARPs in alternative splicing of mRNAs, as well as direct ADPRylation of mRNAs, highlight the role of PARP members in RNA processing. Furthermore, PARP activity, stimulated by cellular stresses, such as viral infections and ER stress, leads to the regulation of mRNA stability and protein synthesis through posttranscriptional mechanisms. Dysregulation of PARP activity in these processes can promote disease states. Collectively, these results highlight the importance of PARP family members and ADPRylation in gene regulation, mRNA processing, and protein abundance. Future studies in these areas will yield new insights into the fundamental mechanisms and a broader utility for PARP-targeted therapeutic agents.


Assuntos
ADP-Ribosilação/fisiologia , Expressão Gênica/fisiologia , Poli(ADP-Ribose) Polimerases/metabolismo , Biossíntese de Proteínas/fisiologia , Proteostase/fisiologia , Animais , Humanos , Processamento de Proteína Pós-Traducional , RNA/metabolismo
3.
J Insect Sci ; 20(1)2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-32092133

RESUMO

Mating promotes reproductive activity, which may impact immune performance. Paradoxically, mating frequently challenges females' immunity (e.g., infections). Therefore, studies of postmating resource allocation between reproduction and survival are likely to shed new light on life-history trade-off and sexual selection. Here, we used RNAseq to test whether and how mating affected mRNA expression in genes related to reproduction and immunity in Spodoptera litura female moths. Results show a divergent change in the differentially expressed genes (DEGs) between reproduction and immunity: the immune response was largely downregulated shortly after mating (~6 h postmating), which has some recovery at 24 h postmating; reproductive response is trivial shortly after mating (~6 h postmating), but it largely upregulated at 24 h postmating (e.g., egg maturation related genes were highly upregulated). Considering the fact that most of the total DEGs downregulated from 0 to 6 h postmating (from 51/68 to 214/260) but most of the total DEGs upregulated at 24 h postmating (816/928), it is possible that trade-offs between reproduction and immunity occurred in mated females. For example, they may shut down immunity to favor sperm storage and save limited resources to support the increased energy required in reproduction (e.g., egg maturation and oviposition). Mating-induced infections should be trivial due to low polyandry in S. litura. A reduced immune defense may have no threat to S. litura survival but may benefit reproduction significantly. Furthermore, obvious expression changes were detected in genes related to hormone production, suggesting that endocrine changes could play important roles in postmating responses.


Assuntos
Copulação , Spodoptera/genética , Spodoptera/imunologia , Animais , Feminino , Expressão Gênica/fisiologia , Masculino , RNA Mensageiro , Reprodução/genética , Spodoptera/metabolismo
4.
Cell Mol Life Sci ; 77(2): 243-251, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31407020

RESUMO

Transforming growth factor (TGF)-ß signalling pathways are intensively investigated because of their diverse association with physiological and pathophysiological states. Smad transcription factors are the key mediators of TGF-ß signalling. Smads can be directly phosphorylated in the carboxy terminal by the TGF-ß receptor or in the linker region via multiple intermediate serine/threonine kinases. Growth factors in addition to hormones and TGF-ß can activate many of the same kinases which can phosphorylate the Smad linker region. Historically, Smad linker region phosphorylation was shown to prevent nuclear translocation of Smads and inhibit TGF-ß signalling pathways; however, it was subsequently shown that Smad linker region phosphorylation can be a driver of gene expression. This review will cover the signalling pathways of Smad linker region phosphorylation that drive the expression of genes involved in pathology and pathophysiology. The role of Smad signalling in cell biology is expanding rapidly beyond its role in TGF-ß signalling and many signalling paradigms need to be re-evaluated in terms of Smad involvement.


Assuntos
Fosforilação/fisiologia , Transdução de Sinais/fisiologia , Proteínas Smad/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Animais , Expressão Gênica/fisiologia , Humanos
5.
Cell Prolif ; 53(2): e12737, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31821660

RESUMO

OBJECTIVE: Embryo implantation needs a reciprocal interaction between competent embryo and receptive endometrium. Adenosine triphosphate (ATP) produced by stressed or injured cells acts as an important signalling molecule. This study aims to investigate whether adenosine triphosphate (ATP) plays an important role in the dialogue of human blastocyst-endometrium. MATERIALS AND METHODS: The concentration of lactate was analysed in culture medium from human embryos collected from in vitro fertilization patients. Extracellular ATP was measured by ATP Bioluminescent Assay Kit. Ishikawa cells and T-HESCs were treated with ATP, ATP receptor antagonist, ATP hydrolysis enzyme or inhibitors of ATP metabolic enzymes. The levels of gene expression were evaluated by real-time PCR and immunoassay. RESULTS: We showed that injured human endometrial epithelial cells could rapidly release ATP into the extracellular environment as an important signalling molecule. In addition, blastocyst-derived lactate induces the release of non-lytic ATP from human endometrial receptive epithelial cells via connexins. Extracellular ATP stimulates the secretion of IL8 from epithelial cells to promote the process of in vitro decidualization. Extracellular ATP could also directly promote the decidualization of human endometrial stromal cells via P2Y-purinoceptors. More importantly, the supernatants of injured epithelial cells clearly induce the decidualization of stromal cells in time-dependent manner. CONCLUSION: Our results suggest that ATP should play an important role in human blastocyst-endometrium dialogue for the initiation of decidualization.


Assuntos
Trifosfato de Adenosina/metabolismo , Blastocisto/metabolismo , Blastocisto/fisiologia , Endométrio/metabolismo , Endométrio/fisiologia , Linhagem Celular , Técnicas de Cocultura/métodos , Implantação do Embrião/fisiologia , Células Epiteliais/metabolismo , Células Epiteliais/fisiologia , Feminino , Expressão Gênica/fisiologia , Humanos , Transdução de Sinais/fisiologia , Células Estromais/metabolismo , Células Estromais/fisiologia
6.
Nurs Res ; 69(1): 74-81, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31834118

RESUMO

BACKGROUND: Despite the emphasis on exercise to reduce pain and improve function among people with chronic low back pain (cLBP), little is known about the underlying mechanism of the impact of exercise on the neurophysiological and gene transcription alterations that characterize cLBP. OBJECTIVES: To present a study protocol to examine the feasibility, acceptability, and initial efficacy of Problem-Solving Pain to Enhance Living Well (PROPEL) with the support of nurse consultations and wearable activity-tracking technology on self-management (SM) knowledge, skills, physical activity, and pain and to examine the differential neurophysiological and gene expression profiles in cLBP participants from pre- to post-PROPEL. METHODS: A pretest and posttest study is employed on 40 adults ages 18-60 years with cLBP who do not have serious complications and/or comorbidities that affect sensorimotor function. Participants will receive video modules focused on SM and biweekly phone consultations to facilitate symptom monitoring and problem-solving while increasing physical activity frequency and duration. Participants will be assessed for outcomes including SM skills, physical activity, and pain every 2 weeks for 12 weeks. We will examine the participants' differential neurophysiological and gene expression profiles at 12 weeks postintervention and correlate these outcomes with the total duration of physical activity. RESULTS: The study began in September 2018. Of the 99 subjects that were screened, 23 were enrolled and 8 completed data collection. DISCUSSION: Comparing the neurophysiological and gene expression profiles of people with cLBP exposed to PROPEL could inform the development of interventions that offer personalized physical activity dosage along with general SM support. Web-based programs such as PROPEL have the potential to enhance accessibility of evidence-based interventions that improve functionality and quality of life among people living with cLBP.


Assuntos
Doença Crônica/terapia , Terapia por Exercício/métodos , Exercício Físico/fisiologia , Exercício Físico/psicologia , Expressão Gênica/fisiologia , Dor Lombar/terapia , Neurofisiologia/métodos , Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
7.
Lipids Health Dis ; 18(1): 215, 2019 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-31823799

RESUMO

BACKGROUND: Macrophage are specialized cells that contributes to the removal of detrimental contents via phagocytosis. Lipid accumulation in macrophages, whether from phagocytosis of dying cells or from circulating oxidized low-density lipoproteins, alters macrophage biology and functionality. It is known that carnitine palmitoyl transferase 1-a (CPT1a) gene encodes an enzyme involved in fatty acid oxidation and, therefore, lipid content. However, the potential of CPT1a to activate macrophage phagocytic function have not been elucidated. METHODS: Using a murine macrophage cell line, RAW264.7, we determine if intracellular accumulation of 7-ketocholesterol (7-KC) modulates macrophage phagocytic function through CPT1a gene expression. In addition, the effects of CPT1a genetic modification on macrophage phenotype and phagocytosis has been studied. RESULTS: Our results revealed that CPT1a gene expression decreased by the accumulation of 7-KC at the higher dose of 7-KC. This was concomitant with an impair ability to phagocytize bioparticles and an inflammatory phenotype. GW3965 treatment, which have shown to facilitate the efflux of cholesterol, eliminated the intracellular lipid droplets of 7-KC-laden macrophages, increased the gene expression of CPT1a, diminished the gene expression of the inflammatory marker iNOS and restored macrophage phagocytosis. Furthermore, CPT1a Knockdown per se was detrimental for macrophage phagocytosis whereas transcriptional activation of CPT1a heightened the uptake of bioparticles. CONCLUSIONS: Altogether, our findings indicate that downregulation of CPT1a by lipid content modulates macrophage phagocytosis and inflammatory phenotype.


Assuntos
Carnitina O-Palmitoiltransferase/genética , Expressão Gênica/fisiologia , Inflamação , Cetocolesteróis/fisiologia , Macrófagos/fisiologia , Fagocitose/fisiologia , Animais , Carnitina O-Palmitoiltransferase/fisiologia , Regulação para Baixo , Técnicas de Silenciamento de Genes , Cetocolesteróis/farmacologia , Ativação de Macrófagos/fisiologia , Camundongos , Células RAW 264.7 , Transfecção
8.
PLoS Pathog ; 15(12): e1008221, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31881074

RESUMO

Kaposi's sarcoma (KS) is an AIDS-defining cancer caused by the KS-associated herpesvirus (KSHV). Unanswered questions regarding KS are its cellular ontology and the conditions conducive to viral oncogenesis. We identify PDGFRA(+)/SCA-1(+) bone marrow-derived mesenchymal stem cells (Pα(+)S MSCs) as KS spindle-cell progenitors and found that pro-angiogenic environmental conditions typical of KS are critical for KSHV sarcomagenesis. This is because growth in KS-like conditions generates a de-repressed KSHV epigenome allowing oncogenic KSHV gene expression in infected Pα(+)S MSCs. Furthermore, these growth conditions allow KSHV-infected Pα(+)S MSCs to overcome KSHV-driven oncogene-induced senescence and cell cycle arrest via a PDGFRA-signaling mechanism; thus identifying PDGFRA not only as a phenotypic determinant for KS-progenitors but also as a critical enabler for viral oncogenesis.


Assuntos
Células-Tronco Mesenquimais/virologia , Neovascularização Patológica/virologia , Receptor alfa de Fator de Crescimento Derivado de Plaquetas/metabolismo , Sarcoma de Kaposi/virologia , Animais , Carcinogênese/metabolismo , Expressão Gênica/fisiologia , Herpesvirus Humano 8/genética , Células-Tronco Mesenquimais/citologia , Camundongos , Transdução de Sinais/fisiologia
10.
Int J Mol Sci ; 20(22)2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31731578

RESUMO

Pancreatic cancer is one of the leading causes of cancer-related mortality worldwide. The molecular basis for the pathogenesis of this disease remains elusive. In this study, we have investigated the role of wild-type Carboxypeptidase E (CPE-WT) and a 40 kDa N-terminal truncated isoform, CPE-ΔN in promoting proliferation and invasion of Panc-1 cells, a pancreatic cancer cell line. Both CPE-WT and CPE-ΔN were expressed in Panc-1 and BXPC-3 pancreatic cancer cells. Immunocytochemical studies revealed that in CPE transfected Panc-1 cells, CPE-ΔN was found primarily in the nucleus, whereas CPE-WT was present exclusively in the cytoplasm as puncta, characteristic of secretory vesicles. Endogenous CPE-WT was secreted into the media. Overexpression of CPE-ΔN in Panc-1 cells resulted in enhancement of proliferation and invasion of these cells, as determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cell proliferation assay and Matrigel invasion assay, respectively. In contrast, the expression of CPE-WT protein at comparable levels to CPE-ΔN in Panc-1 cells resulted in promotion of proliferation but not invasion. Importantly, there was an upregulation of the expression of CXCR2 mRNA and protein in Panc-1 cells overexpressing CPE-ΔN, and these cells exhibited significant increase in proliferation in a CXCR2-dependent manner. Thus, CPE-ΔN may play an important role in promoting pancreatic cancer growth and malignancy through upregulating the expression of the metastasis-related gene, CXCR2.


Assuntos
Carboxipeptidase H/metabolismo , Proliferação de Células/fisiologia , Neoplasias Pancreáticas/metabolismo , Carboxipeptidase H/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Expressão Gênica/genética , Expressão Gênica/fisiologia , Regulação Neoplásica da Expressão Gênica/genética , Regulação Neoplásica da Expressão Gênica/fisiologia , Humanos , Imuno-Histoquímica , Imunoprecipitação , Neoplasias Pancreáticas/genética , Plasmídeos/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa
11.
J Plant Physiol ; 243: 153054, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31648109

RESUMO

Adhesion of the barley husk to the underlying caryopsis requires the development of a cuticular cementing layer on the caryopsis surface. Differences in adhesion quality among genotypes have previously been correlated with cementing layer composition, which is thought to influence caryopsis cuticle permeability, the hypothesised mechanism of adhesion mediation. It is not yet known whether differences in adhesion quality among genotypes are determined by changes in caryopsis cuticle permeability. We examined changes in candidate cementing layer biosynthetic and regulatory genes to investigate the genetic mechanisms behind husk adhesion quality. We used both commercially relevant UK malting cultivars and older European lines to ensure phenotypic diversity in adhesion quality. An ethylene responsive transcription factor (NUD) is required for the development of the cementing layer. To examine correlations between gene expression, cementing layer permeability and husk adhesion quality we also treated cultivars with ethephon (2-chloroethylphosphonic acid) which breaks down to ethylene, and silver thiosulphate which inhibits ethylene reception, and measured caryopsis cuticle permeability. Differential adhesion qualities among genotypes are not determined by NUD expression during development of the cementing material alone, but could result from differences in biosynthetic gene expression during cementing layer development in response to longer-term NUD expression patterns. Altered caryopsis cuticle permeability does result in altered adhesion quality, but the correlation is not consistently positive or negative. Cuticle permeability is therefore not the mechanism that determines husk adhesion quality, but is likely a consequence of the required cuticular compositional changes that determine adhesion.


Assuntos
Etilenos/metabolismo , Hordeum/fisiologia , Compostos Organofosforados/farmacologia , Reguladores de Crescimento de Planta/farmacologia , Sementes/fisiologia , Tiossulfatos/farmacologia , Adesividade , Etilenos/antagonistas & inibidores , Expressão Gênica/fisiologia , Hordeum/genética , Permeabilidade , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Sementes/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
12.
Psychiatry Res ; 282: 112621, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31648143

RESUMO

Alterations in GABAergic interneurons and glutamic acid decarboxylase (GAD) are observed in the brains of people with schizophrenia. Studies also show increased density of interstitial white matter neurons (IWMN), including those containing GAD and somatostatin (SST) in the brain in schizophrenia. Maternal immune activation can be modelled in rodents to investigate the relationship between prenatal exposure to infections and increased risk of developing schizophrenia. We reported that maternal immune activation induced an increase in density of somatostatin-positive IWMN in the adult rat offspring. Here we hypothesised that maternal immune activation induced in pregnant rats by polyinosinic:polycytidylic acid would alter SST and GAD gene expression as well as increase the density of GAD-positive IWMNs in the adult offspring. SST gene expression was significantly reduced in the cingulate cortex of adult offspring exposed to late gestation maternal immune activation. There was no change in cortical GAD gene expression nor GAD-positive IWMN density in adults rats exposed to maternal immune activation at either early or late gestation. This suggests that our model of maternal immune activation induced by prenatal exposure of rats to polyinosinic:polycytidylic acid during late gestation is able to recapitulate changes in SST but not other GABAergic neuropathologies observed in schizophrenia.


Assuntos
Neurônios GABAérgicos , Expressão Gênica/fisiologia , Glutamato Descarboxilase/metabolismo , Giro do Cíngulo , Efeitos Tardios da Exposição Pré-Natal , Esquizofrenia , Somatostatina/metabolismo , Substância Branca , Animais , Modelos Animais de Doenças , Feminino , Neurônios GABAérgicos/imunologia , Neurônios GABAérgicos/metabolismo , Glutamato Descarboxilase/genética , Giro do Cíngulo/imunologia , Giro do Cíngulo/metabolismo , Masculino , Gravidez , Efeitos Tardios da Exposição Pré-Natal/imunologia , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Ratos , Ratos Wistar , Esquizofrenia/genética , Esquizofrenia/imunologia , Esquizofrenia/metabolismo , Somatostatina/genética , Substância Branca/imunologia , Substância Branca/metabolismo
13.
J Plant Physiol ; 243: 153021, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31639534

RESUMO

Kiwifruit (Actinidia chinensis var. deliciosa (A. Chev) A. Chev.) is a widely cultivated crop due to the nutritional value of its fruits. Its commercialization is related to the fruit size, which is directly linked with the number of seeds and, consequently, with pollination. In this dioecious species pollination is dependent on a short effective pollination period which is related to a Programmed Cell Death (PCD) process. At the same time, this PCD process allows the growth of many pollen tubes. Several studies suggest that ethylene can play an important role in PCD in a number of systems. In this report, we determined the full sequence of the AcACS gene, encoding the enzyme that catalyses a rate-limiting step of the ethylene synthesis. Next, we monitored the expression pattern of this gene as well as of other genes involved in ethylene synthesis (ACO2-5) and signalling (AdERS1a, AdERS1b, AdETR1, AdETR2, AdETR3, AdCTR1, AdCTR2, AdEIL1) in pollinated and non-pollinated stigmatic arms of kiwifruit female flowers. The relative expression patterns observed for AcACS, ACOs and ethylene perception and signalling genes (AdERS1, AdETR1, AdCTR1 and AdEIL1) showed that they are expressed before anthesis. After anthesis, expression of the studied genes was detected earlier in pollinated than in non-pollinated stigmatic arms, as it was previously determined for PCD hallmarks. In addition, the expression pattern of the studied genes showed a clear relationship with the PCD hallmarks described in a previous report in the secretory tissue both in non-pollinated stigmatic arms (related to the short EPP in this species) and in pollinated ones (related to the growth of many pollen tubes during progamic phase). Overall, these results suggest an involvement of ethylene with PCD contributing to the high reproductive success of this species.


Assuntos
Actinidia/fisiologia , Apoptose/genética , Etilenos/biossíntese , Expressão Gênica/fisiologia , Genes de Plantas/fisiologia , Transdução de Sinais/genética , Actinidia/genética , Perfilação da Expressão Gênica
14.
Exp Eye Res ; 188: 107787, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31479653

RESUMO

Cataract-associated gene discovery in human and animal models have informed on key aspects of human lens development, homeostasis and pathology. Additionally, in vitro models such as the culture of permanent human lens epithelium-derived cell lines (LECs) have also been utilized to understand the molecular biology of lens cells. However, these resources remain uncharacterized, specifically regarding their global gene expression and suitability to model lens cell biology. Therefore, we sought to molecularly characterize gene expression in the human LEC, SRA01/04, which is commonly used in lens studies. We first performed short tandem repeat (STR) analysis and validated SRA01/04 LEC for its human origin, as recommended by the eye research community. Next, we used Illumina HumanHT-12 v3.0 Expression BeadChip arrays to gain insights into the global gene expression profile of SRA01/04. Comparative analysis of SRA01/04 microarray data was performed using other resources such as the lens expression database iSyTE (integrated Systems Tool for Eye gene discovery), the cataract gene database Cat-Map and the published lens literature. This analysis showed that SRA01/04 significantly expresses >40% of the top iSyTE lens-enriched genes (313 out of 749) across different developmental stages. Further, SRA01/04 also significantly expresses ~53% (168 out of 318) of cataract-associated genes in Cat-Map. We also performed comparative gene expression analysis between SRA01/04 cells and the previously validated mouse LEC 21EM15. To gain insight into whether SRA01/04 reflects epithelial or fiber cell characteristics, we compared its gene expression profile to previously reported differentially expressed genes in isolated mouse lens epithelial and fiber cells. This analysis suggests that SRA01/04 has reduced expression of several fiber cell-enriched genes. In agreement with these findings, cell culture analysis demonstrates that SRA01/04 has reduced potential to initiate spontaneous lentoid body formation compared to 21EM15 cells. Next, to independently validate SRA01/04 microarray gene expression, we subjected several candidate genes to RT-PCR and RT-qPCR assays. This analysis demonstrates that SRA01/04 supports expression of many key genes associated with lens development and cataract, including CRYAB, CRYBB2, CRYGS, DKK3, EPHA2, ETV5, GJA1, HSPB1, INPPL1, ITGB1, PAX6, PVRL3, SFRP1, SPARC, TDRD7, and VIM, among others, and therefore can be relevant for understanding the mechanistic basis of these factors. At the same time, SRA01/04 cells do not exhibit robust expression of several genes known to be important to lens biology and cataract such as ALDH1A1, COL4A6, CP, CRYBA4, FOXE3, HMX1, HSF4, MAF, MEIS1, PITX3, PRX, SIX3, and TRPM3, among many others. Therefore, the present study offers a rich transcript-level resource for case-by-case evaluation of the potential advantages and limitations of SRA01/04 cells prior to their use in downstream investigations. In sum, these data show that the human LEC, SRA01/04, exhibits lens epithelial cell-like character reflected in the expression of several lens-enriched and cataract-associated genes, and therefore can be considered as a useful in vitro resource when combined with in vivo studies to gain insight into specific aspects of human lens epithelial cells.


Assuntos
Biomarcadores , Células Epiteliais/citologia , Proteínas do Olho/genética , Cristalino/citologia , Linhagem Celular , Efrina-A2/genética , Células Epiteliais/metabolismo , Expressão Gênica/fisiologia , Humanos , Cristalino/metabolismo , Biologia Molecular , Análise de Sequência com Séries de Oligonucleotídeos , Osteonectina/genética , Fator de Transcrição PAX6/genética , Reação em Cadeia da Polimerase em Tempo Real , Ribonucleoproteínas/genética , Cadeia B de alfa-Cristalina/genética , Cadeia B de beta-Cristalina/genética
15.
Biotech Histochem ; 94(7): 540-545, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31537133

RESUMO

Interleukin-23 (IL-23, IL-23p19) is a proinflammatory cytokine in the IL-12-related family. Although inflammatory cells in herniated discs have been shown to contain IL-23, little is known about the presence and role of IL-23 in human disc cells. We analyzed disc specimens for IL-23 localization using immunohistochemistry in control, herniated and non-herniated discs from which annulus fibrosus (annulus) cells were isolated and cultured to identify IL-23 gene expression and production. Microarray analysis was used to assess the expression of IL-23 in disc tissue and in cells exposed to two proinflammatory cytokines, IL-1ß and TNF-α. IL-23 was present in annulus cells at the protein level and its expression was up-regulated significantly in herniated compared to control disc tissue. Direct measurement of medium components confirmed production of IL-23 and its receptor, IL-23R, by annulus cells in vitro. Annulus cells in three-dimensional culture exposed to TNF-α, but not IL-1ß, resulted in significant up-regulation of IL-23 expression compared to control cells. Our findings are evidence for the constitutive presence of IL-23 in the human disc and that its expression in vitro is modified by exposure to TNF-α.


Assuntos
Subunidade p19 da Interleucina-23/metabolismo , Degeneração do Disco Intervertebral/metabolismo , Deslocamento do Disco Intervertebral/metabolismo , Disco Intervertebral/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Anel Fibroso/metabolismo , Células Cultivadas , Citocinas/metabolismo , Expressão Gênica/fisiologia , Humanos , Degeneração do Disco Intervertebral/diagnóstico , Deslocamento do Disco Intervertebral/diagnóstico , Regulação para Cima
16.
Biomed Res Int ; 2019: 3726721, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31531351

RESUMO

Identification of protein complex is very important for revealing the underlying mechanism of biological processes. Many computational methods have been developed to identify protein complexes from static protein-protein interaction (PPI) networks. Recently, researchers are considering the dynamics of protein-protein interactions. Dynamic PPI networks are closer to reality in the cell system. It is expected that more protein complexes can be accurately identified from dynamic PPI networks. In this paper, we use the undulating degree above the base level of gene expression instead of the gene expression level to construct dynamic temporal PPI networks. Further we convert dynamic temporal PPI networks into dynamic Temporal Interval Protein Interaction Networks (TI-PINs) and propose a novel method to accurately identify more protein complexes from the constructed TI-PINs. Owing to preserving continuous interactions within temporal interval, the constructed TI-PINs contain more dynamical information for accurately identifying more protein complexes. Our proposed identification method uses multisource biological data to judge whether the joint colocalization condition, the joint coexpression condition, and the expanding cluster condition are satisfied; this is to ensure that the identified protein complexes have the features of colocalization, coexpression, and functional homogeneity. The experimental results on yeast data sets demonstrated that using the constructed TI-PINs can obtain better identification of protein complexes than five existing dynamic PPI networks, and our proposed identification method can find more protein complexes accurately than four other methods.


Assuntos
Mapas de Interação de Proteínas/fisiologia , Proteínas/metabolismo , Algoritmos , Análise por Conglomerados , Biologia Computacional/métodos , Expressão Gênica/fisiologia
17.
ACS Chem Biol ; 14(10): 2243-2251, 2019 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-31513382

RESUMO

Ralstonia solanacearum strains are devastating plant pathogens with global distribution, a wide host range, and genetic diversity, and they are now also referred to as the R. solanacearum species complex (RSSC). RSSC strains employ the quorum sensing (QS) system composed of the phcBSR operon to regulate their virulence on plants. The RSSC strains previously examined produce either (R)-methyl 3-hydroxymyristate (3-OH MAME) or (R)-methyl 3-hydroxypalmitate (3-OH PAME) as their QS signals. Analogously, the phylogenetic analyses of the signal synthase PhcB and the signal receptor PhcS from 15 RSSC strains revealed that these proteins have two clades dependent on their QS signal types. However, the biochemical mechanism underlying this selectivity of QS signal production remains to be elucidated. We demonstrated that the PhcB methyltransferases synthesize QS signals from the cognate fatty acids (R)-3-hydroxymyristic acid or (R)-3-hydroxypalmitic acid. The RSSC strains used here produced both fatty acids, and thus the selectivity of QS signal production depends on the activity of PhcB enzymes. On the other hand, the enantioselective supply of the precursors functioned in the production of enantiopure QS signals. The opposite QS signals weakly induced the production of virulence factors in the RSSC strains. Furthermore, the complementation of the phcB gene encoding the 3-OH PAME-type synthase to the phcB-deletion mutant of the 3-OH MAME-producing strain did not rescue its virulence on tomato plants. Taken together, we propose that the specific production of 3-OH MAME/3-OH PAME ensures full virulence of the RSSC strains.


Assuntos
Proteínas de Bactérias/metabolismo , Metiltransferases/metabolismo , Percepção de Quorum/fisiologia , Ralstonia solanacearum/fisiologia , Fatores de Virulência/biossíntese , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Escherichia coli/genética , Expressão Gênica/fisiologia , Metiltransferases/química , Metiltransferases/genética , Miristatos/metabolismo , Ácidos Mirísticos/química , Ácidos Mirísticos/metabolismo , Ácidos Palmíticos/química , Ácidos Palmíticos/metabolismo , Ralstonia solanacearum/patogenicidade , Estereoisomerismo , Especificidade por Substrato , Transcriptoma/fisiologia
18.
Plant Physiol Biochem ; 143: 257-264, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31525603

RESUMO

Wild ginseng (Panax ginseng) can survive in their natural habitat for hundreds of years, reflecting a remarkable plasticity. Plant stem cells (SCs) play a key role in the regenerative capacity and lifelong activity of these plants. WUSCHEL-RELATED HOMEOBOX (WOX) genes are master regulators of plant SC pluripotency, but their functions in medicinal plants have not been previously reported. To investigate whether these genes define different SC niches in ginseng, we cloned and analysed five WOX genes in ginseng (PgWOXs) and found that they might regulate root reconstruction. Then, the whole-mount RNA in situ hybridization was used to characterize the 3D gene expression pattern of PgWOXs in ginseng seedlings and cultured adventitious roots. PgWOX4 was expressed in vascular cambium SCs; PgWOX5 and PgWOX11 were mainly expressed in the tips of seedling and adventitious roots, which are the energetic centre of the meristem; and PgWOX13a and PgWOX13b were detected in the parenchyma cells of the main root of seedlings and cultured adventitious roots, suggesting that they are important for maintaining the balance between SC differentiation and self-renewal in the phloem and xylem. This is the first report of SC regulation in medical herbs; we expect that P. ginseng can serve as a model herb for investigating the relationship between SCs and their herbal morphological features, which would be a new research direction to improve the yield and quality of the medicinal materials by regulating the herbal SCs.


Assuntos
Panax/metabolismo , Proteínas de Plantas/metabolismo , Diferenciação Celular/genética , Diferenciação Celular/fisiologia , Expressão Gênica/genética , Expressão Gênica/fisiologia , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Panax/genética , Proteínas de Plantas/genética
19.
Philos Trans R Soc Lond B Biol Sci ; 374(1785): 20190287, 2019 11 11.
Artigo em Inglês | MEDLINE | ID: mdl-31544607

RESUMO

Nerve injury leads to devastating and often untreatable neuropathic pain. While acute noxious sensation (nociception) is a crucial survival mechanism and is conserved across phyla, chronic neuropathic pain is considered a maladaptive response owing to its devastating impact on a patient's quality of life. We have recently shown that a neuropathic pain-like response occurs in adult Drosophila. However, the mechanisms underlying this phenomenon are largely unknown. Previous studies have shown that the α2δ peripheral calcium channel subunit straightjacket (stj) is a conserved factor required for thermal pain perception. We demonstrate here that stj is required in peripheral ppk+ sensory neurons for acute thermal responses and that it mediates nociceptive hypersensitivity in an adult Drosophila model of neuropathic pain-like disease. Given that calcium channels are the main targets of gabapentinoids (pregabalin and gabapentin), we assessed if these drugs can alleviate nociceptive hypersensitivity. Our findings suggest that gabapentinoids may prevent nociceptive hypersensitivity by preserving central inhibition after nerve injury. Together, our data further highlight the similarity of some mechanisms for pain-like conditions across phyla and validates the scientific use of Drosophila neuropathic sensitization models for analgesic drug discovery. This article is part of the Theo Murphy meeting issue 'Evolution of mechanisms and behaviour important for pain'.


Assuntos
Canais de Cálcio/genética , Proteínas de Drosophila/genética , Drosophila melanogaster/fisiologia , Neuralgia/genética , Animais , Canais de Cálcio/metabolismo , Modelos Animais de Doenças , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Expressão Gênica/fisiologia , Larva/genética , Larva/fisiologia , Neuralgia/fisiopatologia
20.
Fish Shellfish Immunol ; 94: 628-633, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31541776

RESUMO

The present study evaluated the effects of cyclical short-term food deprivation and refeeding on growth performance, body composition, and gene expression of SOD, GPX and HSP70 in Schizothorax wangchiachii.The experimental design included four feeding protocols for eight weeks: feeding every day of the week (control), starvation for one day and refeeding for six days per week (S1F6 treatment), starvation for two days and refeeding for five days per week (S2F5 treatment), and starvation for three days and refeeding for four days per week (S3F4 treatment). The results showed that no significant difference in final body weight, specific growth rate and feed conversion efficiency were observed among the treatments (P > 0.05).The feeding rate significantly increased with the duration of food deprivation per week compared to the control (P < 0.05). The expression levels of HSP70 showed no significant differences in the gill, liver and spleen of S.wangchiachii subjected to different feed restriction regimes(P > 0.05), but in the kidney, the expression levels of HSP70 were significantly downregulated in S1F6 and S2F5 compared to the control(P < 0.05). The expression levels of SOD and GPX in the examined tissues were not affected by the different feed restriction regimes(P > 0.05). In conclusion, full compensatory growth was observed in S.wangchiachii under eight cycles of food deprivation and refeeding. Hyperphagia was the main mechanism of compensatory growth of S.wangchiachii.


Assuntos
Cyprinidae/genética , Proteínas de Peixes/genética , Privação de Alimentos/fisiologia , Expressão Gênica/fisiologia , Ração Animal/análise , Animais , Peso Corporal , Cyprinidae/crescimento & desenvolvimento , Cyprinidae/imunologia , Dieta/veterinária , Proteínas de Peixes/metabolismo , Glutationa Peroxidase/genética , Glutationa Peroxidase/metabolismo , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Tamanho do Órgão , Distribuição Aleatória , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo
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