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We aimed to investigate the impact of heat stress (HS) on the expression of tight junction (TJ) proteins and the interaction between genes affecting intestinal barrier function using transcriptomics in the porcine jejunum. Twenty-four barrows (crossbred Yorkshire × Landrace × Duroc; average initial body weight, 56.71 ± 1.74 kg) were placed in different temperatures (normal temperature [NT]; HS) and reared for 56 days. At the end of the experiment, jejunal samples were collected from three pigs per treatment for transcriptome and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) analyses. We identified 43 differentially expressed genes, involving five Kyoto Encyclopedia of Genes and Genomes pathways, eight molecular functions, seven cellular components (CCs), and nine biological processes, using gene ontology enrichment analysis. Genes associated with the actin cytoskeleton, filament-binding pathways, and TJ proteins were selected and analyzed by RT-qPCR. Significant differences in relative mRNA expression showed that downregulated genes in the HS group included ZO1, CLDN1, OCLN, PCK1, and PCK2, whereas ACTG2, DES, MYL9, MYLK, TPM1, TPM2, CNN1, PDLIM3, and PCP4 were upregulated by HS (p < 0.05). These findings indicate that HS in growing-finishing pigs induces depression in gut integrity, which may be related to genes involved in the actin cytoskeleton and filaments of CC.
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BACKGROUND: The rising older adult population has led to an increase in the prevalence of chronic diseases and medical expenses. Women tend to have a longer healthy life expectancy than men and are more likely to be exposed to urological disorders around the age of 50, resulting in substantial healthcare expenses throughout their lifetime. Urological disorders often require continuous treatment owing to their high risk of recurrence, contributing to an increased financial burden from medical costs. This study aimed to identify factors influencing medical expense in female patients with urological disorders and propose strategies to alleviate the associated financial burden. METHODS: We used data from the Korea Health Panel Survey conducted from 2011 to 2016. The final sample comprised 2,932 patients who visited hospitals for urological disorders. To identify the factors influencing medical expense among female patients with urological disorders, we employed a generalized estimating equation model. RESULTS: The results indicated that younger people and patients with middle-income levels tended to incur higher medical expenses. Furthermore, patients receiving treatment at tertiary hospitals and those enrolled in National Health Insurance also incurred higher health expenses. CONCLUSIONS: This study suggests that effective management of medical expenses related to urological disorders in women requires improvements in healthcare accessibility to facilitate early detection and continuous disease management. In addition, the findings highlight the potential benefits of digital health and non-face-to-face treatments in addressing these needs.
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Circadian misalignment caused by differences in sleep duration between weekends and weekdays may be associated with adolescent mental health and sleep quality may be able to compensate for this problem. This study aimed to investigate the association between weekend catch-up sleep (CUS) ratio and sleep quality with depressive symptoms and suicidal ideation among South Korean adolescents. We used data from the Korea Youth Risk Behavior Web-based Survey 2015-2019 involving 270,619 adolescents. The weekend CUS ratio was calculated by dividing the average weekend sleep duration by the average weekday sleep duration (< 1.00, 1.00 ≤ CUS < 1.50, or ≥ 1.50). Subjective sleep quality was categorized as poor, moderate, or good. Multiple logistic regression analyses were performed. A weekend CUS ratio of < 1.00 and poor sleep quality was significantly associated with mental health. Absolutely short sleep duration (CUS < 1.00 and weekday sleep duration < 5 h) was most associated with depressive symptoms and suicidal ideation. Furthermore, adolescents with a CUS ratio of ≥ 1.50 showed increased odds of depressive symptoms despite having good sleep quality. Appropriate weekend CUS may benefit adolescents' mental health. When investigating the relationship between adolescents' sleep and mental health, a weekend CUS ratio should be considered in addition to sleep quality and duration.
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Calidad del Sueño , Ideación Suicida , Adolescente , Depresión/epidemiología , Humanos , República de Corea/epidemiología , SueñoRESUMEN
p53-binding protein 1 (53BP1) regulates the DNA double-strand break (DSB) repair pathway and maintains genomic integrity. Here we found that 53BP1 functions as a molecular scaffold for the nucleoside diphosphate kinase-mediated phosphorylation of ATP-citrate lyase (ACLY) which enhances the ACLY activity. This functional association is critical for promoting global histone acetylation and subsequent transcriptome-wide alterations in gene expression. Specifically, expression of a replication-dependent histone biogenesis factor, stem-loop binding protein (SLBP), is dependent upon 53BP1-ACLY-controlled acetylation at the SLBP promoter. This chain of regulation events carried out by 53BP1, ACLY, and SLBP is crucial for both quantitative and qualitative histone biogenesis as well as for the preservation of genomic integrity. Collectively, our findings reveal a previously unknown role for 53BP1 in coordinating replication-dependent histone biogenesis and highlight a DNA repair-independent function in the maintenance of genomic stability through a regulatory network that includes ACLY and SLBP.
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ATP Citrato (pro-S)-Liasa , Histonas , ATP Citrato (pro-S)-Liasa/genética , ATP Citrato (pro-S)-Liasa/metabolismo , Acetilación , Roturas del ADN de Doble Cadena , Reparación del ADN , Histonas/genética , Histonas/metabolismo , Proteína 1 de Unión al Supresor Tumoral P53/metabolismoRESUMEN
[This corrects the article DOI: 10.18632/oncotarget.10275.].
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The African swine fever virus (ASFV) was first detected in wild boar in the Demilitarized Zone, a bordered area between South and North Korea, on 2 October 2019. Phylogenetic analyses of ASFV genes encoding p72 and CD2v indicated that the causative strain belongs to genotype II and serogroup 8, respectively, and contained additional tandem repeat sequences between the I73R and the I329L protein genes.
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Fiebre Porcina Africana , Asfarviridae/genética , Fiebre Porcina Africana/diagnóstico , Fiebre Porcina Africana/epidemiología , Animales , Filogenia , República de Corea , Sus scrofa , PorcinosRESUMEN
There are widespread concerns that low-calorie sweeteners (LCSs) cause metabolic derangement. These concerns stem in part from prior studies linking LCS consumption to impaired glucose tolerance in humans and rodents. Here, we examined this linkage in mice. In experiment 1, we provided mice with chow, water, and an LCS-sweetened solution (saccharin, sucralose, or acesulfame K) for 28 days and measured glucose tolerance and body weight across the exposure period. Exposure to the LCS solutions did not impair glucose tolerance or alter weight gain. In experiment 2, we provided mice with chow, water, and a solution containing saccharin, glucose, or a mixture of both for 28 days, and tested for metabolic changes. Exposure to the saccharin solution increased the insulinemic response of mice to the glucose challenge, and exposure to the saccharin + glucose solution increased the rate of glucose uptake during the glucose challenge. However, neither of these test solutions altered glucose tolerance, insulin sensitivity, plasma triglycerides, or percent body fat. In contrast, exposure to the glucose solution increased glucose tolerance, early insulin response, insulin sensitivity, and percent body fat. We conclude that whereas the LCS-containing solutions induced a few metabolic changes, they were modest compared with those induced by the glucose solution.
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Metabolismo Energético/efectos de los fármacos , Edulcorantes/farmacología , Animales , Peso Corporal , Ingestión de Energía , Femenino , Prueba de Tolerancia a la Glucosa , Masculino , RatonesRESUMEN
BACKGROUND: A relatively high proportion of patients diagnosed with primary CNS lymphoma will experience recurrent disease, yet therapy options are limited in salvage therapy. This is the first study to evaluate a bendamustine-based combination regimen for the treatment of relapsed/refractory PCNSL and to characterize bendamustine pharmacokinetics in the human CSF. METHODS: Patients received bendamustine 75 mg/m2 for two days as part of R-B(O)AD administered intravenously every 4 weeks for up to 4 cycles. Response and adverse events of the regimen were assessed. A sparse sampling strategy and population based modeling approach was utilized for evaluation of plasma and CSF levels of bendamustine. RESULTS: Ten patients were enrolled into study of whom 70% were of refractory disease and with high IELSG prognostic risk scores. The ORR of R-BOAD was 50% (95% CI, 0.24 to 0.76) with one patient achieving CR and four PR. Primary toxicity of the regimen was reversible myelosuppression, mostly grade 3 or 4 neutropenia. The Cmax mean for plasma and CSF were 2669 ng/mL and 0.397 ng/mL, respectively, and patients with response at deep tumor sites displayed higher trends in peak exposure. Pharmacokinetic data was best described by a four-compartment model with first-order elimination of drug from central plasma and CSF compartments. CONCLUSIONS: R-BOAD is an effective salvage option for PCNSL, but with significant hematologic toxicity. Bendamustine CSF levels are minimal; however correspond to plasma exposure and response. TRIAL REGISTRATION: ClinicalTrials.gov NCT03392714 ; retrospectively registered January 8, 2018.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Clorhidrato de Bendamustina/administración & dosificación , Neoplasias del Sistema Nervioso Central/tratamiento farmacológico , Linfoma/tratamiento farmacológico , Terapia Recuperativa , Adulto , Anciano , Clorhidrato de Bendamustina/farmacocinética , Femenino , Humanos , Masculino , Persona de Mediana Edad , Proyectos PilotoRESUMEN
In previous studies, we observed that Zeta-chain-associated protein kinase 70 (Zap70) regulates spindle assembly and chromosome alignment in mouse oocyte and that Ran binding protein 2 (RanBP2) is a highly associated gene with Zap70 based on a microarray analysis. Because RanBP2 is related to nuclear envelope breakdown (NEBD) during mitosis, the aim of the present study was to elucidate the molecular mechanism of Zap70 with respect to RanBP2 in the germinal vesicle breakdown (GVBD) of oocytes. Results indicated that RanBP2 expression was regulated by Zap70 and that depletion of RanBP2 using RanBP2 RNAi manifested comparable phenotypes to those observed in Zap70 RNAi-treated oocytes, which presented faster processing of GVBD. Additionally, Zap70 RNAi-treated oocytes showed faster meiotic resumption with premature activation of maturation-promoting factor (MPF), premature division of chromosomes at approximately 6-8 h and more rapid degradation of securin. In conclusion, we report that Zap70 is a crucial factor for controlling the exact timing of meiotic progression in mouse oocytes.
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Meiosis , Chaperonas Moleculares/metabolismo , Proteínas de Complejo Poro Nuclear/metabolismo , Oocitos/citología , Oocitos/metabolismo , Proteína Tirosina Quinasa ZAP-70/metabolismo , Animales , Diferenciación Celular , Segregación Cromosómica , Cromosomas de los Mamíferos/metabolismo , Femenino , Mesotelina , Ratones Endogámicos ICR , Modelos Biológicos , Interferencia de ARN , Securina/metabolismo , Factores de TiempoRESUMEN
Inflammation is a potent inducer of tumorigenesis. Increased DNA damage or loss of genome integrity is thought to be one of the mechanisms linking inflammation and cancer development. It has been suggested that NF-κB-induced microRNA-146 (miR146a) may be a mediator of the inflammatory response. Based on our initial observation that miR146a overexpression strongly increases DNA damage, we investigated its potential role as a modulator of DNA repair. Here, we demonstrate that FANCM, a component in the Fanconi Anemia pathway, is a novel target of miR146a. miR146a suppressed FANCM expression by directly binding to the 3' untranslated region of the gene. miR146a-induced downregulation of FANCM was associated with inhibition of FANCD2 monoubiquitination, reduced DNA homologous recombination repair and checkpoint response, failed recovery from replication stress, and increased cellular sensitivity to cisplatin. These phenotypes were recapitulated when miR146a expression was induced by overexpressing the NF-κB subunit p65/RelA or Helicobacter pylori infection in a human gastric cell line; the phenotypes were effectively reversed with an anti-miR146a antagomir. These results suggest that undesired inflammation events caused by a pathogen or over-induction of miR146a can impair genome integrity via suppression of FANCM.
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ADN Helicasas/biosíntesis , Regulación de la Expresión Génica/genética , MicroARNs/genética , Línea Celular , Línea Celular Tumoral , Transformación Celular Neoplásica/genética , Daño del ADN/fisiología , ADN Helicasas/genética , Reparación del ADN/fisiología , Humanos , Inflamación/genética , Inflamación/metabolismo , Inflamación/patologíaRESUMEN
Mouse oocytes begin to mature in vitro once liberated from ovarian follicles. Previously, we showed that oocyte-specific homeobox 4 (Obox4) is critical for maintaining the intact nuclear membrane of the germinal vesicle (GV) in oocytes and for completing meiosis at the metaphase I-II (MI-MII) transition. This study further examines the molecular mechanisms of OBOX4 in regulating GV nuclear membrane breakdown. Maturation-promoting factor (MPF) and MAPK are normally inactive in GV stage oocytes but were activated prematurely in arrested GV stage oocytes by 3-isobutyl-1-metyl-xanthine (IBMX) in vitro after Obox4 RNA interference (RNAi). Furthermore, signal transducer and activator of transcription 3 (STAT3) was significantly activated by Obox4 RNAi. We confirmed that this Obox4 RNAi-induced premature STAT3 and MPF/MAPK activation at the GV stage provoked subsequent GV breakdown (GVBD) despite the opposing force of high cAMP in the IBMX-supplemented medium to maintain intact GV. When cumulus-oocyte complexes were exposed to interferon α (IFNA), a STAT3 activator, oocytes matured and cumulus cells expanded to resume nuclear maturation in IBMX-supplemented medium, suggesting that STAT3 activation is sufficient for stimulating the continuation of meiosis. Using Stattic, a specific STAT3 inhibitor, we confirmed that GVBD involves STAT3 activation in Obox4-silenced oocytes. Based on these findings, we concluded that i) Obox4 is an important upstream regulator of MPF/MAPK and STAT3 signaling, and ii) Obox4 is a key regulator of the GV arrest mechanism in oocytes.
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Proteínas Ligadas a GPI/metabolismo , Silenciador del Gen , Proteínas de Homeodominio/antagonistas & inhibidores , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Membrana Nuclear/metabolismo , Oocitos/metabolismo , Factor de Transcripción STAT3/metabolismo , Animales , Western Blotting , Núcleo Celular/metabolismo , Células Cultivadas , Femenino , Técnica del Anticuerpo Fluorescente , Proteínas Ligadas a GPI/genética , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Técnicas para Inmunoenzimas , Meiosis/fisiología , Mesotelina , Ratones , Proteínas Quinasas Activadas por Mitógenos/genética , Oocitos/citología , Folículo Ovárico/citología , Folículo Ovárico/metabolismo , Fosforilación , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Transcripción STAT3/genéticaRESUMEN
MDC1 is critical component of the DNA damage response (DDR) machinery and orchestrates the ensuring assembly of the DDR protein at the DNA damage sites, and therefore loss of MDC1 results in genomic instability and tumorigenicity. However, the molecular mechanisms controlling MDC1 expression are currently unknown. Here, we show that miR-22 inhibits MDC1 translation via direct binding to its 3' untranslated region, leading to impaired DNA damage repair and genomic instability. We demonstrated that activated Akt1 and senescence hinder DDR function of MDC1 by upregulating endogenous miR-22. After overexpression of constitutively active Akt1, homologous recombination was inhibited by miR-22-mediated MDC1 repression. In addition, during replicative senescence and stress-induced premature senescence, MDC1 was downregulated by upregulating miR-22 and thereby accumulating DNA damage. Our results demonstrate a central role of miR-22 in the physiologic regulation of MDC1-dependent DDR and suggest a molecular mechanism for how aberrant Akt1 activation and senescence lead to increased genomic instability, fostering an environment that promotes tumorigenesis.
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Reparación del ADN , Inestabilidad Genómica , MicroARNs/fisiología , Proteínas Nucleares/genética , Transactivadores/genética , Proteínas Adaptadoras Transductoras de Señales , Adolescente , Anciano , Animales , Proteínas de Ciclo Celular , Senescencia Celular , Daño del ADN , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Células HCT116 , Células HEK293 , Humanos , Ratones , Persona de Mediana Edad , Proteínas Nucleares/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Interferencia de ARN , Transactivadores/metabolismo , Adulto JovenRESUMEN
Perilla (Perilla frutescens L.) leaves have shown therapeutic efficacy in the treatment of inflammatory disorders, allergies, bronchial asthma, and systemic damage due to free radicals. In the present study we analyzed the active constituents in perilla leaves using high-performance liquid chromatography (HPLC) and isolated luteolin, a polyphenolic flavonoid. We investigated the anti-inflammatory and antipruritic properties of luteolin. Luteolin inhibited the secretion of inflammatory cytokines such as interleukin-1ß (IL-1 ß) and tumor necrosis factor-α (TNF-α) from human mast cells (HMC-1) stimulated with phorbol myristate acetate plus calcium ionophore A23187 in a dose-dependent manner. Luteolin also significantly reduced the histamine release from rat peritoneal mast cells stimulated by compound 48/80, a potent histamine liberator. Furthermore, the administration of luteolin markedly inhibited the scratching behavior and vascular permeability induced by pruritogens, such as compound 48/80 or serotonin, in ICR mice. These results suggested that luteolin has potential as a therapeutic agent against inflammation and itch-related skin diseases.
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Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Antipruriginosos/farmacología , Antipruriginosos/uso terapéutico , Luteolina/uso terapéutico , Perilla/química , Animales , Calcimicina/farmacología , Humanos , Interleucina-1beta/efectos de los fármacos , Interleucina-1beta/metabolismo , Luteolina/farmacología , Masculino , Mastocitos/efectos de los fármacos , Mastocitos/metabolismo , Ratones , Ratones Endogámicos ICR , Prurito/tratamiento farmacológico , Ratas , Ratas Sprague-Dawley , Acetato de Tetradecanoilforbol/farmacología , Factor de Necrosis Tumoral alfa/metabolismo , p-Metoxi-N-metilfenetilamina/farmacologíaRESUMEN
Thioredoxin-interacting protein (Txnip) regulates intracellular redox state and prompts oxidative stress by binding to and inhibiting Thioredoxin (Trx). In addition, via a Trx-independent mechanism, Txnip regulates glucose metabolism and thus maintains intracellular glucose levels. Previously, we found Txnip mRNA highly expressed in immature germinal vesicle (GV) oocytes, but currently there is no report describing the role of Txnip in oocytes. Therefore, we conducted the present study to determine the function of Txnip in mouse oocytes' maturation and meiosis by using RNA interference (RNAi) method. Upon specific depletion of Txnip, 79.5% of oocytes were arrested at metaphase I (MI) stage. Time-lapse video microscopy analysis revealed that the formation of granules in the oocyte cytoplasm increased concurrent with retarded cytoplasmic streaming after Txnip RNAi treatment. Txnip RNAi-treated oocytes had upregulated glucose uptake and lactate production. To confirm the supposition that mechanism responsible for these observed phenomena involves increased lactate in oocytes, we cultured oocytes in high lactate medium and observed the same increased granule formation and retarded cytoplasmic streaming as found by Txnip RNAi. The MI-arrested oocytes exhibited scattered microtubules and aggregated chromosomes indicating that actin networking was disturbed by Txnip RNAi. Therefore, we conclude that Txnip is a critical regulator of glucose metabolism in oocytes and is involved in maintaining cytoplasmic streaming in mouse oocytes.
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Proteínas Portadoras/metabolismo , Glucosa/metabolismo , Oocitos/metabolismo , Tiorredoxinas/metabolismo , Citoesqueleto de Actina/ultraestructura , Animales , Proteínas Portadoras/antagonistas & inhibidores , Proteínas Portadoras/genética , Cromosomas de los Mamíferos/ultraestructura , Citoplasma/metabolismo , Citoplasma/ultraestructura , Femenino , Expresión Génica , Ácido Láctico/biosíntesis , Meiosis , Metafase , Ratones , Ratones Endogámicos ICR , Microtúbulos/ultraestructura , Oocitos/citología , Oocitos/ultraestructura , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Tiorredoxinas/antagonistas & inhibidores , Tiorredoxinas/genética , Imagen de Lapso de Tiempo , Grabación en VideoRESUMEN
Resveratrol has been clinically shown to possess a number of human health benefits. As a result, many attempts have been made to engineer resveratrol production in major cereal grains but have been largely unsuccessful. In this study, we report the creation of a transgenic rice plant that accumulates 1.9 µg resveratrol/g in its grain, surpassing the previously reported anti-metabolic syndrome activity of resveratrol through a synergistic interaction between the transgenic resveratrol and the endogenous properties of the rice. Consumption of our transgenic resveratrol-enriched rice significantly improved all aspects of metabolic syndrome and related diseases in animals fed a high-fat diet. Compared with the control animals, the resveratrol-enriched rice reduced body weight, blood glucose, triglycerides, total cholesterol, and LDL-cholesterol by 24.7%, 22%, 37.4%, 27%, and 59.6%, respectively. The resveratrol-enriched rice from our study may thus provide a safe and convenient means of preventing metabolic syndrome and related diseases without major lifestyle changes or the need for daily medications. These results also suggest that future transgenic plants could be improved if the synergistic interactions of the transgene with endogenous traits of the plant are considered in the experimental design.
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Alimentos Fortificados , Síndrome Metabólico/tratamiento farmacológico , Oryza/genética , Estilbenos/uso terapéutico , Aciltransferasas/genética , Aciltransferasas/metabolismo , Tejido Adiposo , Animales , Glucemia/metabolismo , Peso Corporal , Cromatografía Líquida de Alta Presión , Femenino , Glucósidos/metabolismo , Glucosiltransferasas/metabolismo , Humanos , Lípidos/sangre , Síndrome Metabólico/sangre , Ratones , Ratones Endogámicos C57BL , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/enzimología , Plantas Modificadas Genéticamente , Resveratrol , Semillas/efectos de los fármacos , Semillas/metabolismo , Sirtuina 1/metabolismo , Estilbenos/metabolismo , Estilbenos/farmacologíaRESUMEN
A specific inhibitor of RNA polymerase II, α-amanitin is broadly used to block transcriptional activities in cells. Previous studies showed that α-amanitin affects in vitro maturation of cumulus-oocyte-complex (COC). In this study, we evaluated the target of α-amanitin, and whether it affects oocytes or cumulus cells (CCs), or both. We treated α-amanitin with different time period during in vitro culture of denuded oocytes (DOs) or COCs in comparison, and observed the changes in morphology and maturation status. Although DOs did not show any change in morphology and maturation rates with α-amanitin treatment, oocytes from COCs were arrested at metaphase I (MI) stage and CCs were more scattered than control groups. To discover causes of meiotic arrest and scattering of CCs, we focused on changes of cumulus expansion, gap junctions, and cellular metabolism which to be the important factors for the successful in vitro maturation of COCs. Expression of genes for cumulus expansion markers (Ptx3, Has2, and Tnfaip6) and gap junctional proteins (Gja1, Gja4, and Gjc1) decreased in α-amanitin-treated CCs. However, these changes were not observed in oocytes. In addition, expression of genes related to metabolism (Prps1, Rpe, Rpia, Taldo1, and Tkt) decreased in α -amanitin-treated CCs but not in oocytes. Therefore, we concluded that the transcriptional activities of CCs for supporting suitable transcripts, especially for its metabolic activities and formation of gap junctions among CCs as well as with oocytes, are important for oocytes maturation in COCs.
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The COP9 (constitutive photomorphogenic) signalosome (CSN), composed of eight subunits, is a highly conserved protein complex that regulates processes such as cell cycle progression and kinase signalling. Previously, we found the expression of the COP9 constitutive photomorphogenic homolog subunit 3 (CSN3) and subunit 5 (CSN5) changes as oocytes mature for the first time, and there is no report regarding roles of COP9 in the mammalian oocytes. Therefore, in the present study, we examined the effects of RNA interference (RNAi)-mediated transient knockdown of each subunit on the meiotic cell cycle in mice oocytes. Following knockdown of either CSN3 or CSN5, oocytes failed to complete meiosis I. These arrested oocytes exhibited a disrupted meiotic spindle and misarranged chromosomes. Moreover, down-regulation of each subunit disrupted the activity of maturation-promoting factor (MPF) and concurrently reduced degradation of the anaphase-promoting complex/cyclosome (APC/C) substrates Cyclin B1 and Securin. Our data suggest that the CSN3 and CSN5 are involved in oocyte meiosis by regulating degradation of Cyclin B1 and Securin via APC/C.
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Proteínas Portadoras/metabolismo , Factor Promotor de Maduración/metabolismo , Meiosis , Complejos Multiproteicos/metabolismo , Oocitos/citología , Oocitos/metabolismo , Péptido Hidrolasas/metabolismo , Proteolisis , Animales , Complejo del Señalosoma COP9 , Puntos de Control del Ciclo Celular , Diferenciación Celular/genética , Cromosomas de los Mamíferos/metabolismo , Ciclina B1/metabolismo , Regulación hacia Abajo , Femenino , Técnica del Anticuerpo Fluorescente , Técnicas de Silenciamiento del Gen , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Mesotelina , Ratones , Ratones Endogámicos ICR , Modelos Biológicos , Proteínas Nucleares/metabolismo , Proteínas Quinasas/metabolismo , Proteínas Proto-Oncogénicas , Interferencia de ARN , SecurinaRESUMEN
Previously, we found that the growth arrest-specific gene 6 (Gas6) is more highly expressed in germinal vesicle (GV) oocytes than in metaphase II (MII) oocytes using annealing control primer (ACP)-PCR technology. The current study was undertaken to investigate the role of Gas6 in oocyte maturation and fertilization using RNA interference (RNAi). Interestingly, despite the specific and marked decrease in Gas6 mRNA and protein expression in GVs after Gas6 RNAi, nuclear maturation including spindle structures and chromosome segregation was not affected. The only discernible effect induced by Gas6 RNAi was a change in maturation promoting factor (MPF) activity. After parthenogenetic activation, Gas6 RNAi-treated oocytes at the MII stage had not developed further and arrested at MII (90.0%). After stimulation with Sr(2+), Gas6-silenced MII oocytes had markedly reduced Ca(2+) oscillation and exhibited no exocytosis of cortical granules. In these oocytes, sperm penetration occurred during fertilization but not pronucleus (PN) formation. By roscovitine and colcemid treatment, we found that the Gas6 knockdown affected cytoplasmic maturation directly, independent to the changed MPF activity. These results strongly suggest that 1) the Gas6 signaling itself is important to the cytoplasmic maturation, but not nuclear maturation, and 2) the decreased Gas6 expression and decreased MPF activity separately or mutually influence sperm head decondensation and PN formation.
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Núcleo Celular/metabolismo , Citoplasma/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Factor Promotor de Maduración/metabolismo , Animales , Western Blotting , Células Cultivadas , Cromosomas/metabolismo , Embrión de Mamíferos/metabolismo , Femenino , Fertilización/genética , Fertilización/fisiología , Péptidos y Proteínas de Señalización Intercelular/genética , Masculino , Factor Promotor de Maduración/genética , Mesotelina , Ratones , Oocitos/metabolismo , Embarazo , Interferencia de ARN , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Intracranial aneurysm (IA) is characterized by an abnormal bulging of one of the arteries in the brain and is heavily affected by genetic factors. Although IA is a very serious disease because of its severity and prevalence in the general public, the gene causing IA has not yet been identified due mainly to the lack of definitive genetic loci for the disease. Following a model-based family collection that recruited families from a geographically limited area that inherited IA as an autosomal dominant trait, we conducted a genome-wide linkage analysis. Significant evidence of linkage to IA was found on chromosome 8p22.2 with a maximum two-point logarithm of the odds ratio score of 3.61 under an autosomal dominant model of inheritance. The methods described in this study could be applied to localize disease-causing genes of other complex diseases through either a genome-wide linkage analysis or a genome-wide association study.