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Well-balanced and timed metabolism is essential for making a high-quality egg. However, the metabolic framework that supports oocyte development remains poorly understood. Here, we obtained the temporal metabolome profiles of mouse oocytes during in vivo maturation by isolating large number of cells at key stages. In parallel, quantitative proteomic analyses were conducted to bolster the metabolomic data, synergistically depicting the global metabolic patterns in oocytes. In particular, we discovered the metabolic features during meiotic maturation, such as the fall in polyunsaturated fatty acids (PUFAs) level and the active serine-glycine-one-carbon (SGOC) pathway. Using functional approaches, we further identified the key targets mediating the action of PUFA arachidonic acid (ARA) on meiotic maturation and demonstrated the control of epigenetic marks in maturing oocytes by SGOC network. Our data serve as a broad resource on the dynamics occurring in metabolome and proteome during oocyte maturation.
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Meiosis/fisiología , Oocitos/metabolismo , Animales , Epigénesis Genética/genética , Ácidos Grasos Insaturados/metabolismo , Femenino , Metaboloma/fisiología , Ratones , Ratones Endogámicos C57BL , Oogénesis/genética , Oogénesis/fisiología , Proteoma/metabolismo , ProteómicaRESUMEN
Meiotic maturation is an intricate and precisely regulated process orchestrated by various pathways and numerous proteins. However, little is known about the proteome landscape during oocytes maturation. Here, we obtained the temporal proteomic profiles of mouse oocytes during in vivo maturation. We successfully quantified 4694 proteins from 4500 oocytes in three key stages (germinal vesicle, germinal vesicle breakdown, and metaphase II). In particular, we discovered the novel proteomic features during oocyte maturation, such as the active Skp1-Cullin-Fbox pathway and an increase in mRNA decay-related proteins. Using functional approaches, we further identified the key factors controlling the histone acetylation state in oocytes and the vital proteins modulating meiotic cell cycle. Taken together, our data serve as a broad resource on the dynamics occurring in oocyte proteome and provide important knowledge to better understand the molecular mechanisms during germ cell development.
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Proteoma , Proteómica , Ratones , Animales , Proteoma/metabolismo , Oogénesis , Oocitos/metabolismo , Núcleo Celular/metabolismo , MeiosisRESUMEN
BACKGROUND: Pancreatic adenocarcinomas (PAADs) often exhibit a "cold" or immunosuppressive tumor milieu, which is associated with resistance to immune checkpoint blockade therapy; however, the underlying mechanisms are incompletely understood. Here, we aimed to improve our understanding of the molecular mechanisms occurring in the tumor microenvironment and to identify biomarkers, therapeutic targets, and potential drugs to improve PAAD treatment. METHODS: Patients were categorized according to immunologically hot or cold PAAD subtypes with distinct disease outcomes. Cox regression and weighted correlation network analysis were performed to construct a novel gene signature, referred to as 'Downregulated in hot tumors, Prognostic, and Immune-Related Genes' (DPIRGs), which was used to develop prognostic models for PAAD via machine learning (ML). The role of DPIRGs in PAAD was comprehensively analyzed, and biomarker genes able to distinguish PAAD immune subtypes and predict prognosis were identified by ML. The expression of biomarkers was verified using public single-cell transcriptomic and proteomic resources. Drug candidates for turning cold tumors hot and corresponding target proteins were identified via molecular docking studies. RESULTS: Using the DPIRG signature as input data, a combination of survival random forest and partial least squares regression Cox was selected from 137 ML combinations to construct an optimized PAAD prognostic model. The effects and molecular mechanisms of DPIRGs were investigated by analysis of genetic/epigenetic alterations, immune infiltration, pathway enrichment, and miRNA regulation. Biomarkers and potential therapeutic targets, including PLEC, TRPV1, and ITGB4, among others, were identified, and the cell type-specific expression of the biomarkers was validated. Drug candidates, including thalidomide, SB-431542, and bleomycin A2, were identified based on their ability to modulate DPIRG expression favorably. CONCLUSIONS: By combining multiple ML algorithms, we developed a novel prognostic model with excellent performance in PAAD cohorts. ML also proved to be powerful for identifying biomarkers and potential targets for improved PAAD patient stratification and immunotherapy.
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Adenocarcinoma , Biomarcadores de Tumor , Regulación Neoplásica de la Expresión Génica , Aprendizaje Automático , Neoplasias Pancreáticas , Humanos , Neoplasias Pancreáticas/tratamiento farmacológico , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/patología , Neoplasias Pancreáticas/inmunología , Biomarcadores de Tumor/metabolismo , Adenocarcinoma/tratamiento farmacológico , Adenocarcinoma/genética , Adenocarcinoma/inmunología , Adenocarcinoma/metabolismo , Pronóstico , Simulación del Acoplamiento Molecular , Microambiente Tumoral , Antineoplásicos/uso terapéutico , Antineoplásicos/farmacología , Masculino , Transcriptoma/genética , FemeninoRESUMEN
BACKGROUND: Obesity is a crucial risk factor for asthma. Observational studies have examined the association between abdominal obesity and asthma symptoms. This study aimed to investigate the causal relationship between visceral adipose tissue (VAT) and asthma and its potential as an independent indicator. METHODS: This study utilized data from the National Health and Nutrition Examination Survey spanning 2011-8. Multivariable logistic regression and stratified variable selection were employed to identify associations between asthma and VAT. Moreover, a two-sample Mendelian randomization analysis, using 221 genetic variants as instrumental variables, was conducted to assess this relationship further. RESULTS: Our findings indicated that individuals with higher VAT levels were more likely to develop asthma. Visceral obesity remained a significant risk factor for asthma after adjusting for demographic characteristics. Genetic predictions suggest a positive association between VAT and an elevated risk of asthma (odds ratio [OR] = 1.393, 95% confidence interval [CI]: 1.266-1.534, and P = 1.43E-11). No significant polymorphisms were detected using the Mendelian randomization-Egger intercept test. CONCLUSIONS: This study presents potential evidence supporting the causal role of VAT in asthma development. Furthermore, the findings from the Mendelian randomization analysis further reinforce the relationship between VAT and asthma risk.
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Objective: To assess the effects of the clinical nursing pathway (CNP) on blood glucose and urine ketones in patients with diabetic ketoacidosis (DKA). Methods: A total of 60 patients with DKA (20 type I and 40 type II) treated in the Department of Endocrinology at Anhui Second People's Hospital from January 2018 to May 2022 were recruited and randomly assigned to receive routine nursing (control group) or CNP plus routine nursing (observation group), with 30 patients in each group. The observation group received the clinical nursing pathway (CNP) along with routine nursing care. As part of the CNP, low-flow oxygen therapy was administered to the patients. Low-flow oxygen therapy involves the delivery of oxygen at a lower flow rate compared to high-flow oxygen therapy. In this study, a flow rate of 2 L/min was used. The low-flow oxygen was administered to the patients through a nasal cannula or a similar device. Outcome measures included symptom relief and length of hospital stay. Results: The observation group showed a significantly higher decline rate of blood glucose in patients than in the control group. Patients in the observation group had a more rapid disappearance of urine ketones versus those in the control group. CNP plus routine nursing resulted in a significantly shorter length of hospital stay versus routine nursing (RR:0.79, 95% CI (1.078, 4.511), P < .05). Conclusion: CNP plus continuous low-flow oxygen breathing facilitates the decline of blood glucose, removes ketone bodies, mitigates DKA symptoms, and shortens the length of hospital stay.
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BACKGROUND: Photodynamic therapy (PDT) features high biocompatibility and high spatiotemporal selectivity, showing a great potential in glioblastoma (GBM) treatment. However, its application was restricted by the poor therapeutic efficacy and side effect. RESULTS: In this study, a therapeutic nanoplatform (UCNPs@Ce6/3HBQ@CM) with combination of PDT and CO therapy was constructed, in which a photoCORM and a photosensitizer were loaded onto the surface of upconversion nanoparticles (UCNPs) functioning as photon transducer. Benefitting from NIR excitation and multicolor emission of UCNPs, the penetration depth of excitation light is enhanced and meanwhile simultaneous generation of CO and ROS in tumor site can be achieved. The as-prepared nanocomposite possessed an elevated therapeutic efficiency with the assistance of CO through influencing mitochondrial respiration and depleting ATP, accompanying with the reduced inflammatory responses. By wrapping a homologous cell membrane, the nanocomposite can target GBM and accumulate in the tumor site, affording a powerful tool for precise and efficient treatment of GBM. CONCLUSION: This therapeutic nanoplatform UCNPs@Ce6/3HBQ@CM, which combines PDT and CO therapy enables precise and efficient treatment of refractory glioblastoma.
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Glioblastoma , Nanopartículas , Fotoquimioterapia , Humanos , Glioblastoma/tratamiento farmacológico , Línea Celular Tumoral , Fármacos Fotosensibilizantes/farmacología , Fármacos Fotosensibilizantes/uso terapéutico , Rayos Infrarrojos , Nanopartículas/uso terapéuticoRESUMEN
Oncogene HER2 is amplified in 20%-25% of human breast cancers and 6.1%-23.0% of gastric cancers, and HER2-directed therapy significantly improves the outcome for patients with HER2-positive cancers. However, drug resistance is still a clinical challenge due to primary or acquired mutations and drug-induced negative regulatory feedback. In this study, we discovered a potent irreversible HER2 kinase inhibitor, CHMFL-26, which covalently targeted cysteine 805 of HER2 and effectively overcame the drug resistance caused by HER2 V777L, HER2 L755S, HER2 exon 20 insertions, and p95-HER2 truncation mutations. CHMFL-26 displayed potent antiproliferation efficacy against HER2-amplified and mutant cells through constant HER2-mediated signaling pathway inhibition and apoptosis induction. In addition, CHMFL-26 suppressed tumor growth in a dose-dependent manner in xenograft mouse models. Together, these results suggest that CHMFL-26 may be a potential novel anti-HER2 agent for overcoming drug resistance in HER2-positive cancer therapy.
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Neoplasias de la Mama , Receptor ErbB-2 , Animales , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Línea Celular Tumoral , Cisteína , Resistencia a Antineoplásicos , Femenino , Humanos , Ratones , Mutación , Inhibidores de Proteínas Quinasas/farmacología , Inhibidores de Proteínas Quinasas/uso terapéutico , Receptor ErbB-2/genética , Receptor ErbB-2/metabolismo , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
BACKGROUND: To compare MMA-related gene mutations in MMA children and the population in Qingdao, discuss the blood propionyl carnitine (C3), free carnitine (C0) methionine (MET), the mutual ratio and division difference in normal group, carrier group, and MMA group to analyze the relationship between some hotspot mutations and biochemical indicators. METHODS: In total 3,700 newborns testing negative in tandem mass spectrometry (MS/MS) were selected at random and submitted for testing 8 pathogenic sites in MMACHC and 10 in MMUT. The gene mutations in 84 cases with detected mutation genes and 42 diagnosed children were compared. The levels and concentration distribution of C3, C0, MET, C3/C2, C3/C0, C3/MET in the blood samples of three groups were analyzed as well as the difference of biochemical indicators in newborns with hotspot mutations (c.609A>G, c.482G>A, and c.658-660delAAG). RESULTS: All 8 pathogenic mutations in MMACHC in the population were detected and were basically consistent with the mutation types and frequency order in MMA group. The first three were c.609G>A, c.482G>A, and c.658_660delAAG. There were more types of mutation sites detected in MMA group than carrier group. Five out of 10 MMUT gene mutations were detected in the population, and 9 MMUT gene mutation sites were detected in MMA group. The findings in the two groups and the preset sites were not completely consistent. C3, C0, C3/C2, C3/C0, C3/MET in MMA group were higher than carrier and normal groups, and the difference was statistically significant; the MET in MMA group was lower than carrier and normal groups, and the difference was statistical¬ly significant. Based on the three sets of data distribution graphs, C3, C3/C2, C3/C0, and C3/MET were well distinguished. There were differences in the average C3 and C0 levels between carrier and normal groups, but with an obvious cross distribution in the graphs, and no difference in other indicators. In contrast to non-carrier group, C0, C3, C3/C0, C3/C2, and C3/MET concentration levels were higher in 609A>G mutation group, while MET level was lower, with statistical significance; in c.482G>A mutation group, C3, C3/C0, C3/C2, and C3/MET concentration levels were lower than non-carrier group, while MET level was higher, with statistical significance; in c.658-660delAAG mutation group, C0, C3, C3/C0, C3/C2, MET, and C3/MET concentration levels were not statistically different in contrast to other groups. CONCLUSIONS: The top three mutations in MMA children in Qingdao area are c.609A>G, c.482G>A, c.658-660del AAG mutations in MMAHC; C3, C3/C2, C3/C0 can be used as specific prompt indicators for MMA screening; C3, C3/C2, C3/C0, C3/MET can be used as specific prompt indicators for combined MMA screening; abnormalities in biochemical indicators in hotspot mutation group intuitively explains c.609A>G mutation and early-onset MMA. c.482G>A mutation links with late-onset MMA.
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Errores Innatos del Metabolismo de los Aminoácidos , Espectrometría de Masas en Tándem , Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Errores Innatos del Metabolismo de los Aminoácidos/genética , Niño , Humanos , Recién Nacido , Ácido Metilmalónico , Biología Molecular , Mutación , OxidorreductasasRESUMEN
Background and objectives: The COVID-19 pandemic continues worldwide, and there is no effective treatment to treat it. Chinese medicine is considered the recommended treatment for COVID-19 in China. This study aimed to examine the effectiveness of tetrandrine in treating COVID-19, which is originally derived from Chinese medicine. Materials and Methods: A total of 60 patients, categorized into three types (mild, moderate, severe), from Daye Hospital of Chinese Medicine with a diagnosis of COVID-19 were included in this study. Demographics, medical history, treatment, and results were collected. We defined two main groups according to the clinical outcome between improvement and recovery. All underlying factors including clinical outcomes were assessed in the total number of COVID-19 patients and moderate-type patients. Results: In a total of 60 patients, there were significant differences in the clinical outcome underlying treatment with antibiotics, tetrandrine, and arbidol (p < 0.05). When the comparison was limited to the moderate type, treatment with tetrandrine further increased recovery rate (p = 0.007). However, the difference disappeared, and no association was indicated between the clinical outcome and the treatment with and without antibiotic (p = 0.224) and arbidol (p = 0.318) in the moderate-type patients. In all-type and moderate-type patients, tetrandrine improved the rate of improvement in cough and fatigue on day 7 (p < 0.05). Conclusions: Tetrandrine may improve clinical outcome in COVID-19 patientsand could be a promising potential natural antiviral agent for the prevention and treatment of COVID-19.
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Tratamiento Farmacológico de COVID-19 , Antibacterianos/uso terapéutico , Antivirales/uso terapéutico , Bencilisoquinolinas , Humanos , Pandemias , SARS-CoV-2 , Resultado del TratamientoRESUMEN
When fabricating ratiometric optical probes using lanthanide-doped upconversion nanoparticles (UCNPs), which are promising luminescent materials that have widely been utilized in biosensing and bioimaging as energy donors, it is still a challenge to obtain the emission signal of energy acceptors with reasons unclear so far. Herein, we reveal that the energy-transfer efficiency and brightness of UCNPs as well as the aggregation-caused quenching (ACQ) of energy accepting dyes are the main factors restricting the emission of energy acceptors, and we have circumvented this problem by modulating the structure of UCNPs and the assembly manner of the energy donor-acceptor pair. On this basis, a proof-of-concept ratiometric upconversion nanoprobe was constructed for hydrogen sulfide (H2S) detection with an elaborate dye Fl-1 as an energy acceptor. As the H2S concentration increased, the emission intensity of Fl-1 at 525 nm increased gradually, accompanied by a decrease of upconversion luminescence at 480 nm, thus providing a ratiometric signal of F480/F525 dependent on the H2S concentration. This probe was able to track H2S in living cells and zebrafish and visualize the H2S level of mice in physiological processes.
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Elementos de la Serie de los Lantanoides , Nanopartículas , Animales , Concentración de Iones de Hidrógeno , Luminiscencia , Ratones , Pez CebraRESUMEN
Ribes meyeri leaves are used as traditional Kazakh medicine in China. However, no study on the characterization of the phenolic compounds in R. meyeri leaves has been reported, resulting in the lack of quality control measures and poor standardization. This study was conducted to identify the phenolic compounds in R. meyeri leaves and evaluate their antioxidant and antidiabetic activities. A total of 77 phenolics were tentatively identified by liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry. Ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry was applied to simultaneously quantify 12 phenolics in R. meyeri leaves. Rutin, epigallocatechin, isoquercitrin, epicatechin, protocatechuic acid, and kaempferol-3-O-rutinoside were abundant in the R. meyeri leaves. The methanol extract and four different extracts enhanced the glucose uptake in 3T3-L1 adipocytes. The ethyl acetate extracts showed a total phenolic content of 966.89 ± 3.59 mg gallic acid equivalents/g, a total flavonoid content of 263.58 ± 17.09 mg catechin equivalents/g, and good protein-tyrosine phosphatase-1B inhibitory activities (IC50 : 0.60 ± 0.03 µg/mL). To our knowledge, this work is the first to identify and quantify the major phenolics in R. meyeri leaves.
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Antioxidantes/farmacología , Inhibidores Enzimáticos/farmacología , Fenoles/farmacología , Ribes/química , Células 3T3-L1 , Animales , Antioxidantes/análisis , Benzotiazoles/antagonistas & inhibidores , Compuestos de Bifenilo/antagonistas & inhibidores , Cromatografía Líquida de Alta Presión , Inhibidores Enzimáticos/análisis , Glicósido Hidrolasas/antagonistas & inhibidores , Glicósido Hidrolasas/metabolismo , Humanos , Ratones , Fenoles/análisis , Picratos/antagonistas & inhibidores , Proteína Tirosina Fosfatasa no Receptora Tipo 1/antagonistas & inhibidores , Proteína Tirosina Fosfatasa no Receptora Tipo 1/metabolismo , Ácidos Sulfónicos/antagonistas & inhibidores , Espectrometría de Masas en TándemRESUMEN
Maternal obesity has been reported to impair oocyte quality in mice, however, the underlying mechanisms remain unclear. In the present study, by conducting a comparative proteomic analysis, we identified a reduced expression of TIGAR (TP53-induced glycolysis and apoptosis regulator) protein in ovulated oocytes from high-fat diet (HFD)-fed mice. Specific depletion of TIGAR in mouse oocytes results in the marked elevation of reactive oxygen species (ROS) levels and the failure of meiotic apparatus assembly. Importantly, forced expression of TIGAR in HFD oocytes not only attenuates ROS production, but also partly prevents spindle disorganization and chromosome misalignment during meiosis. Meantime, we noted that TIGAR knockdown in oocytes induces a strong activation of autophagy, whereas overexpression of TIGAR significantly reduces the LC3 accumulation in HFD oocytes. By anti-oxidant treatment, we further demonstrated that such an autophagic response is dependent on the TIGAR-controlled ROS production. In summary, our data indicate a role for TIGAR in modulating redox homeostasis during oocyte maturation, and uncover that loss of TIGAR is a critical pathway mediating the effects of maternal obesity on oocyte quality.
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Meiosis , Oocitos/metabolismo , Oocitos/patología , Estrés Oxidativo , Proteínas/metabolismo , Animales , Proteínas Reguladoras de la Apoptosis , Autofagia , Cromosomas de los Mamíferos/metabolismo , Dieta Alta en Grasa , Femenino , Técnicas de Silenciamiento del Gen , Ratones Endogámicos ICR , Ratones Obesos , Ovulación , Monoéster Fosfórico Hidrolasas , Proteómica , Especies Reactivas de Oxígeno , Huso Acromático/metabolismoRESUMEN
Diabetic nephropathy (DN) is a chronic complication of diabetes, and thus the present investigation evaluates the nephroprotective effect of anemonin against streptozotocin (STZ)-induced DN rats. Diabetes was induced by intraperitoneal administration of STZ (50 mg/kg) on day 2 and 3 postnatal, and rats were kept as such for the duration of 12 weeks. Thereafter, rats were treated with anemonin 75 and 150 mg/kg per oral for the period 4 week which means between the period of 12-16 weeks. Effect of anemonin was estimated by determining the blood glucose, markers of nephropathy, and mediators of inflammation in the serum and activity of tumor necrosis factor-α (TNF-α)converting enzyme (TACE) in the kidney tissue of DN rats. Moreover, reverse transcriptase polymerase chain reaction and western blot assay were determined in the kidney tissue homogenate of DN rats. Histopathology study was done by Periodic acid-Schiff's and masson staining for the pathological changes and apoptosis of podocytes in the kidney tissue of DN rats. Moreover, production of reactive oxygen species (ROS) was estimated in the kidney tissue by 2',7'-dichlorofluorescein staining. Data of study reveal that anemonin significantly reduces (p < 0.01) the blood glucose and markers of renal injury in the serum and urine of DN rats. There was a reduction in the level of cytokines in the serum, and production of ROS and activity of TACE were reduced in the kidney tissue of the anemonin-treated group than in the DN group. Expression of iRhom-2, TACE, TNF-α, and inducible nitric oxide synthase protein and histopathology of kidney tissue were attenuated in the anemonin-treated group in DN rats. In conclusion, data of study reveal that treatment with anemonin ameliorates progression of renal injury by regulating TACE/iRhom-2 signaling pathway.
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Proteína ADAM17/metabolismo , Diabetes Mellitus Experimental/tratamiento farmacológico , Nefropatías Diabéticas/tratamiento farmacológico , Furanos/uso terapéutico , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Sustancias Protectoras/uso terapéutico , Proteína ADAM17/genética , Animales , Animales Recién Nacidos , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patología , Nefropatías Diabéticas/genética , Nefropatías Diabéticas/metabolismo , Nefropatías Diabéticas/patología , Furanos/farmacología , Péptidos y Proteínas de Señalización Intracelular/genética , Riñón/efectos de los fármacos , Riñón/metabolismo , Estrés Oxidativo/efectos de los fármacos , Sustancias Protectoras/farmacología , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
SET-domain-containing 2 (SETD2), a member of the histone lysine methyltransferase family, has been reported to be involved in multiple biological processes. However, the function of SETD2 during oocyte maturation has not been addressed. In this study, we find that mouse oocytes are incapable of progressing through meiosis completely once SETD2 is specifically depleted. These oocytes present an abnormal spindle morphology and deficient chromosome movement, with disrupted kinetochore-microtubule attachments, consequently producing aneuploidy eggs. In line with this, the BubR1 signal is markedly elevated in metaphase kinetochores of oocytes with SETD2 depletion, indicative of the activation of spindle assembly checkpoint. In addition, we note that loss of SETD2 results in a drastic decrease in the trimethylation level of H3K36 in oocytes. Collectively, our data demonstrate that SETD2 is required for oocyte maturation and indicate a novel mechanism controlling the meiotic apparatus.
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Proteínas de Ciclo Celular/genética , N-Metiltransferasa de Histona-Lisina/genética , Meiosis/genética , Oocitos/crecimiento & desarrollo , Proteínas Serina-Treonina Quinasas/genética , Aneuploidia , Animales , Segregación Cromosómica/genética , Cinetocoros/metabolismo , Ratones , Oocitos/metabolismo , Huso Acromático/genéticaRESUMEN
Intersectins (ITSNs), an evolutionarily conserved adaptor protein family, have been implicated in multiple biologic processes; however, their functions in mammalian oocytes have not been addressed. Here, we report delayed meiotic resumption and defective cytokinesis upon specific depletion of ITSN2 in mouse oocytes. In particular, abnormal spindle, misaligned chromosomes, and loss of cortical actin cap are readily observed in ITSN2-depleted oocytes. Similarly, a small molecule that targets the Cdc42-ITSN interaction also disrupts oocyte maturation and actin polymerization. Moreover, we find that ITSN2 depletion reduces the activity of Cdc42 in oocytes and, of note, that forced expression of the dominant-positive mutant of Cdc42, in part, prevents the effects of ITSN2 knockdown on actin cap formation. In addition, the localization of WASP and Arp2, the downstream effector proteins of Cdc42, is altered in ITSN2-depleted oocytes accordingly. In summary, our data support a model in which ITSN2 depletion induces the inactivation of Cdc42, which, in turn, influences the distribution and function of Arp2/3 and WASP, consequently disrupting oocyte polarity establishment and meiotic division.-Zhang, J., Ma, R., Li, L., Wang, L., Hou, X., Han, L., Ge, J., Li, M., Wang, Q. Intersectin 2 controls actin cap formation and meiotic division in mouse oocytes through the Cdc42 pathway.
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Actinas/metabolismo , Proteínas Adaptadoras del Transporte Vesicular/metabolismo , Meiosis/fisiología , Oocitos/metabolismo , Huso Acromático/metabolismo , Proteína de Unión al GTP cdc42/metabolismo , Citoesqueleto de Actina/metabolismo , Animales , Polaridad Celular/fisiología , Citocinesis , Femenino , Ratones , Proteína de Unión al GTP cdc42/genéticaRESUMEN
Pyruvate dehydrogenase kinases (PDKs) modulate energy homeostasis in multiple tissues and cell types, under various nutrient conditions, through phosphorylation of the α subunit (PDHE1α, also known as PDHA1) of the pyruvate dehydrogenase (PDH) complex. However, the roles of PDKs in meiotic maturation are currently unknown. Here, by undertaking knockdown and overexpression analysis of PDK paralogs (PDK1-PDK4) in mouse oocytes, we established the site-specificity of PDKs towards the phosphorylation of three serine residues (Ser232, Ser293 and Ser300) on PDHE1α. We found that PDK3-mediated phosphorylation of Ser293-PDHE1α results in disruption of meiotic spindle morphology and chromosome alignment and decreased total ATP levels, probably through inhibition of PDH activity. Unexpectedly, we discovered that PDK1 and PDK2 promote meiotic maturation, as their knockdown disturbs the assembly of the meiotic apparatus, without significantly altering ATP content. Moreover, phosphorylation of Ser232-PDHE1α was demonstrated to mediate PDK1 and PDK2 action in meiotic maturation, possibly through a mechanism that is distinct from PDH inactivation. These findings reveal that there are divergent roles of PDKs during oocyte maturation and indicate a new mechanism controlling meiotic structure.
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Oocitos/citología , Oocitos/enzimología , Proteínas Serina-Treonina Quinasas/metabolismo , Animales , Diferenciación Celular , Cromosomas de los Mamíferos/metabolismo , Regulación Enzimológica de la Expresión Génica , Técnicas de Silenciamiento del Gen , Meiosis , Ratones Endogámicos ICR , Modelos Biológicos , Fosforilación , Fosfoserina/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Piruvato Deshidrogenasa Quinasa Acetil-Transferidora , Huso Acromático/metabolismoRESUMEN
Maternal obesity in humans is associated with poor outcomes across the reproductive spectrum. Emerging evidence indicates that these defects are likely attributed to factors within the oocyte. Although various molecules and pathways may contribute to impaired oocyte quality, prevention of fertility issues associated with maternal obesity is a challenge. Using mice fed a high-fat diet (HFD) as an obesity model, we document spindle disorganization, chromosome misalignment, and elevated reactive oxygen species (ROS) levels in oocytes from obese mice. Oral administration of melatonin to HFD mice not only reduces ROS generation, but also prevents spindle/chromosome anomalies in oocytes, consequently promoting the developmental potential of early embryos. Consistent with this finding, we find that melatonin supplement during in vitro maturation also markedly attenuates oxidative stress and meiotic defects in HFD oocytes. Finally, by performing morpholino knockdown and acetylation-mimetic mutant overexpression assays, we reveal that melatonin ameliorates maternal obesity-induced defective phenotypes in oocytes through the SIRT3-SOD2-dependent mechanism. In sum, our data uncover the marked beneficial effects of melatonin on oocyte quality from obese females; this opens a new area for optimizing culture system as well as fertility management.
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Antioxidantes/uso terapéutico , Melatonina/uso terapéutico , Obesidad/complicaciones , Oocitos/efectos de los fármacos , Complicaciones del Embarazo/prevención & control , Acetilación/efectos de los fármacos , Animales , Antioxidantes/farmacología , Dieta Alta en Grasa , Evaluación Preclínica de Medicamentos , Desarrollo Embrionario/efectos de los fármacos , Femenino , Meiosis/efectos de los fármacos , Melatonina/farmacología , Ratones Endogámicos ICR , Oocitos/metabolismo , Estrés Oxidativo/efectos de los fármacos , Embarazo , Complicaciones del Embarazo/etiología , Especies Reactivas de Oxígeno/metabolismo , Sirtuina 3/metabolismo , Superóxido Dismutasa/metabolismoRESUMEN
Multiwalled carbon nanotubes (MWCNTs) have been explored in pharmaceutical applications such as tumor targeting and delivery of drugs, in which MWCNTs are given through intravenous injection. However, the biosafety of MWCNTs is of concern for such application. Therefore, in the current study, we used a fatty liver model to investigate the possible toxicity of MWCNTs to the liver, as MWCNTs were retained mainly in the liver of mice after intravenous injection. Male Sprague Dawley rats were used to generate the fatty liver model, and the effects of intravenous administration of MWCNTs on fatty liver were studied. Hematoxylin and eosin staining for hepatocellular anatomy and Masson trichrome staining for hepatic fibrosis were conducted. Histologically, MWCNTs aggravated steatohepatitis with higher nonalcoholic fatty liver disease scores. Analysis of liver injury markers indicated that MWCNTs administration resulted in chronic hepatitis, along with increased liver fat and altered liver oxidation, including the increase of P6 protein and the depletion of glutathione. In conclusion, our results suggest that MWCNTs can aggravate nonalcoholic steatohepatitis in Sprague Dawley rats, and oxidative injury may be involved in this process.
Asunto(s)
Hígado/efectos de los fármacos , Nanotubos de Carbono/toxicidad , Enfermedad del Hígado Graso no Alcohólico , Animales , Aspartato Aminotransferasas/sangre , Dieta Alta en Grasa , Ácidos Grasos no Esterificados/metabolismo , Glutatión/metabolismo , Glutatión Peroxidasa/sangre , Inyecciones Intravenosas , Hígado/metabolismo , Hígado/patología , Masculino , Malondialdehído/metabolismo , Enfermedad del Hígado Graso no Alcohólico/sangre , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Enfermedad del Hígado Graso no Alcohólico/patología , Ratas Sprague-Dawley , Superóxido Dismutasa/metabolismo , Triglicéridos/metabolismo , gamma-Glutamiltransferasa/sangreRESUMEN
BACKGROUND: Homer is a family of post synaptic density proteins functionally and physically attached to target proteins at proline-rich sequences. Reducing Homer1b/c expression has been shown in previous studies to be protective against excitotoxic insults, implicating Homer1b/c in the physiological regulation of aberrant neuronal excitability. METHODS: To test the efficacy of a Homer1b/c reducing therapy for disorders with a detrimental hyperexcitability profile in mice, we used small interfere RNA (siRNA) to decrease endogenous Homer1b/c expression in mouse hippocampus. The baseline motor and cognitive behavior was measured by sensorimotor tests, Morris water maze and elevated plus maze tasks. The anti-epileptic effects of Homer1b/c knockdown were determined in two chemically induced seizure models induced by Picrotoxin (PTX) or pentylenetetrazole (PTZ) administration. RESULTS: The results of sensorimotor tests, Morris water maze and elevated plus maze tasks showed that Homer1b/c reduction had no effect on baseline motor or cognitive behavior. In two chemically induced seizure models, mice with reduced Homerb/c protein had less severe seizures than control mice. Total Homer1b/c protein levels and seizure severity were highly correlated, such that those mice with the most severe seizures also had the highest levels of Homer1b/c. In addition, the phosphorylation of mammalian target of rapamycin (mTOR) and its target protein S6 was significantly inhibited in Homer1b/c down-regulated mice. Homer1b/c knockdown-induced inhibition of mTOR pathway was partially ablated by the metabotropic glutamate receptor 5 (mGluR5) agonist CHPG. CONCLUSION: Our results demonstrate that endogenous Homer1b/c is integral for regulating neuronal hyperexcitability in adult animals and suggest that reduction of Homer1b/c could protect against chemically induced seizures through inhibition mTOR pathway.
Asunto(s)
Proteínas Portadoras/metabolismo , Serina-Treonina Quinasas TOR/metabolismo , Animales , Conducta Animal , Proteínas Portadoras/antagonistas & inhibidores , Proteínas Portadoras/genética , Modelos Animales de Enfermedad , Regulación hacia Abajo/efectos de los fármacos , Glicina/análogos & derivados , Glicina/farmacología , Hipocampo/metabolismo , Proteínas de Andamiaje Homer , Aprendizaje por Laberinto , Ratones , Células PC12 , Pentilenotetrazol/toxicidad , Fenilacetatos/farmacología , Fosforilación , Picrotoxina/toxicidad , Interferencia de ARN , Ratas , Receptor del Glutamato Metabotropico 5/agonistas , Receptor del Glutamato Metabotropico 5/metabolismo , Proteínas Quinasas S6 Ribosómicas/metabolismo , Convulsiones/inducido químicamente , Convulsiones/metabolismo , Convulsiones/patología , Transducción de Señal/efectos de los fármacos , Serina-Treonina Quinasas TOR/antagonistas & inhibidoresRESUMEN
Various lanthanide oxides (Sm2 O3 and Gd2 O3 ) nanostructures were synthesized by a facile hydrothermal method. The loss of surfactants on the nanocrystals surface, followed by the resultant assembly is responsible for the formation of ultrathin nanosheets. Owing to strong surface effects, the different morphologies of the Sm2 O3 :5 % Eu and Gd2 O3 :5 % Eu nanocrystals present unique photoluminescence properties. As a proof-of-concept application, the as-obtained Sm2 O3 and Gd2 O3 ultrathin nanosheets exhibit promising pH-controlled anticancer drug-delivery behavior.