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1.
Proc Natl Acad Sci U S A ; 115(20): E4623-E4632, 2018 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-29712850

RESUMO

Store-operated calcium entry (SOCE), an important mechanism of Ca2+ signaling in a wide range of cell types, is mediated by stromal interaction molecule (STIM), which senses the depletion of endoplasmic reticulum Ca2+ stores and binds and activates Orai channels in the plasma membrane. This inside-out mechanism of Ca2+ signaling raises an interesting question about the evolution of SOCE: How did these two proteins existing in different cellular compartments evolve to interact with each other? We investigated the gating mechanism of Caenorhabditis elegans Orai channels. Our analysis revealed a mechanism of Orai gating by STIM binding to the intracellular 2-3 loop of Orai in C. elegans that is radically different from Orai gating by STIM binding to the N and C termini of Orai in mammals. In addition, we found that the conserved hydrophobic amino acids in the 2-3 loop of Orai1 are important for the oligomerization and gating of channels and are regulated via an intramolecular interaction mechanism mediated by the N and C termini of Orai1. This study identifies a previously unknown SOCE mechanism in C. elegans and suggests that, while the STIM-Orai interaction is conserved between invertebrates and mammals, the gating mechanism for Orai channels differs considerably.


Assuntos
Caenorhabditis elegans/metabolismo , Canais de Cálcio/metabolismo , Cálcio/metabolismo , Ativação do Canal Iônico , Proteína ORAI1/metabolismo , Molécula 1 de Interação Estromal/metabolismo , Sequência de Aminoácidos , Animais , Caenorhabditis elegans/genética , Canais de Cálcio/química , Canais de Cálcio/genética , Sinalização do Cálcio , Membrana Celular/metabolismo , Retículo Endoplasmático/metabolismo , Evolução Molecular , Células HEK293 , Humanos , Proteína ORAI1/química , Proteína ORAI1/genética , Homologia de Sequência , Molécula 1 de Interação Estromal/química , Molécula 1 de Interação Estromal/genética
2.
Toxicol Appl Pharmacol ; 355: 68-79, 2018 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-29802913

RESUMO

Methamphetamine (MA), a psychostimulant abused worldwide, gives rise to neurotoxicity in the hippocampus, resulting in cognitive impairments and hippocampal volume reduction. The cellular and molecular mechanisms associated with hippocampal impairments due to MA remain unknown. The aim of this study was to investigate the effects of MA on structural alterations and gene expressions in the hippocampus. We analyzed the pattern of volumetric changes in the hippocampus using magnetic resonance imaging (MRI) after acute and chronic administration of MA to cynomolgus macaques. In addition, we performed large-scale transcriptome profiling in the hippocampus using RNA-Seq technology. The hippocampus in response to acute and chronic MA exhibited a significant volumetric atrophy compared with the hippocampus of controls. The genes associated with cytoskeleton organization and phagocytosis were downregulated in the acute MA-treated group compared to the control group. On the other hand, genes associated with synaptic transmission, regulation of neuron differentiation and regulation of neurogenesis were downregulated in the chronic MA-treated group. We confirmed that expression patterns for ADM, BMP4, CHRD, PDYN, UBA1, profilin 2 (PFN2), ENO2 and NSE mRNAs were similar to the results from RNA-Seq based on quantitative RT-PCR. In particular, PFN2 mRNA and protein expression levels, which play important roles in actin cytoskeleton dynamics, were decreased by acute and chronic MA administration. These results not only aid the understanding of cellular and molecular mechanisms regulated by MA in the hippocampus but also suggest basic information aiding biomarker and novel drug development for treating hippocampal impairment caused by MA abuse.


Assuntos
Estimulantes do Sistema Nervoso Central/toxicidade , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Metanfetamina/toxicidade , Transcriptoma/efeitos dos fármacos , Animais , Peso Corporal/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/ultraestrutura , Ingestão de Alimentos/efeitos dos fármacos , Feminino , Expressão Gênica/efeitos dos fármacos , Perfilação da Expressão Gênica , Hipocampo/diagnóstico por imagem , Macaca fascicularis , Imageamento por Ressonância Magnética , Neurogênese/efeitos dos fármacos , Fagocitose/efeitos dos fármacos , Transmissão Sináptica/efeitos dos fármacos
3.
Reprod Fertil Dev ; 29(7): 1306-1318, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27279419

RESUMO

Despite evidence of the presence of prostaglandin (PG) I2 in mammalian oviducts, its role in early development of in vitro-produced (IVP) embryos is largely unknown. Thus, in the present study we examined the effects of iloprost, a PGI2 analogue, on the in vitro developmental competence of early porcine embryos and the underlying mechanism(s). To examine the effects of iloprost on the development rate of IVF embryos, iloprost was added to the in vitro culture (IVC) medium and cultured for 6 days. Supplementation of the IVC medium with iloprost significantly improved developmental parameters, such as blastocyst formation rate, the trophectoderm:inner cell mass ratio and cell survival in IVF and parthenogenetically activated (PA) embryos. In addition, post-blastulation development into the expanded blastocyst stage was improved in iloprost-treated groups compared with controls. Interestingly, the phosphatidylinositol 3-kinase (PI3K)/AKT signalling pathway was significantly activated by iloprost supplementation in a concentration-dependent manner (10-1000nM), and the beneficial effects of iloprost on the early development of porcine IVF and PA embryos was completely ablated by treatment with 2.5µM wortmannin, a PI3K/AKT signalling inhibitor. Importantly, expression of the PI3K/AKT signalling pathway was significantly reduced in somatic cell nuclear transfer (SCNT) compared with IVF embryos, and iloprost supported the early development of SCNT embryos, as was the case for IVF and PA embryos, suggesting a consistent effect of iloprost on the IVC of IVP porcine embryos. Together, these results indicate that iloprost can be a useful IVC supplement for production of IVP early porcine embryos with high developmental competence.


Assuntos
Desenvolvimento Embrionário/efeitos dos fármacos , Iloprosta/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Sus scrofa/embriologia , Sus scrofa/metabolismo , Androstadienos/farmacologia , Animais , Meios de Cultura , Técnicas de Cultura Embrionária/métodos , Técnicas de Cultura Embrionária/veterinária , Desenvolvimento Embrionário/fisiologia , Epoprostenol/análogos & derivados , Epoprostenol/farmacologia , Feminino , Fertilização in vitro/métodos , Fertilização in vitro/veterinária , Modelos Biológicos , Técnicas de Transferência Nuclear/veterinária , Partenogênese , Transdução de Sinais/efeitos dos fármacos , Wortmanina
4.
J Reprod Dev ; 63(2): 149-155, 2017 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-28070055

RESUMO

The current study was performed to investigate the effect of oocyte donor status, including age and body weight, on metaphase II (MII) oocyte recovery using two superovulation methods in cynomolgus monkeys. The use of Method A [recombinant gonadotrophin (75 IU/kg, 3 ×, 3-day intervals) and human chorionic gonadotropin (hCG)] led to great increases in ovary size and the mean number of MII oocytes retrieved in age- and body-weight-dependent manner; in contrast, both the parameters were similar in Method B [recombinant gonadotrophin (60 IU, twice daily, 6 days), recombinant gonadotropin and recombinant human luteinizing hormone (rhLH) (60 IU, twice daily, 3 days), and hCG]. Importantly, Method A showed maximal MII oocyte recovery rate in > 60-month-old or 4.5-5.0-kg female monkeys, whereas Method B was equally effective regardless of the donor age and body weight. These results indicate that superovulatory responses depend on the interaction between oocyte donor status and the superovulation method used in cynomolgus monkeys.


Assuntos
Recuperação de Oócitos/veterinária , Oócitos/fisiologia , Indução da Ovulação/veterinária , Superovulação/fisiologia , Animais , Gonadotropina Coriônica/administração & dosagem , Células do Cúmulo/efeitos dos fármacos , Células do Cúmulo/fisiologia , Feminino , Hormônio Foliculoestimulante/administração & dosagem , Hormônio Luteinizante/administração & dosagem , Macaca fascicularis , Recuperação de Oócitos/métodos , Oócitos/efeitos dos fármacos , Indução da Ovulação/métodos , Superovulação/efeitos dos fármacos
5.
Int J Mol Sci ; 18(6)2017 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-28632163

RESUMO

Fragile X syndrome (FXS) is the most common monogenic form of autism spectrum disorder (ASD). FXS with ASD results from the loss of fragile X mental retardation (fmr) gene products, including fragile X mental retardation protein (FMRP), which triggers a variety of physiological and behavioral abnormalities. This disorder is also correlated with clock components underlying behavioral circadian rhythms and, thus, a mutation of the fmr gene can result in disturbed sleep patterns and altered circadian rhythms. As a result, FXS with ASD individuals may experience dysregulation of melatonin synthesis and alterations in melatonin-dependent signaling pathways that can impair vigilance, learning, and memory abilities, and may be linked to autistic behaviors such as abnormal anxiety responses. Although a wide variety of possible causes, symptoms, and clinical features of ASD have been studied, the correlation between altered circadian rhythms and FXS with ASD has yet to be extensively investigated. Recent studies have highlighted the impact of melatonin on the nervous, immune, and metabolic systems and, even though the utilization of melatonin for sleep dysfunctions in ASD has been considered in clinical research, future studies should investigate its neuroprotective role during the developmental period in individuals with ASD. Thus, the present review focuses on the regulatory circuits involved in the dysregulation of melatonin and disruptions in the circadian system in individuals with FXS with ASD. Additionally, the neuroprotective effects of melatonin intervention therapies, including improvements in neuroplasticity and physical capabilities, are discussed and the molecular mechanisms underlying this disorder are reviewed. The authors suggest that melatonin may be a useful treatment for FXS with ASD in terms of alleviating the adverse effects of variations in the circadian rhythm.


Assuntos
Transtorno do Espectro Autista/complicações , Síndrome do Cromossomo X Frágil/complicações , Síndrome do Cromossomo X Frágil/tratamento farmacológico , Melatonina/uso terapêutico , Ansiedade , Relógios Circadianos/efeitos dos fármacos , Ritmo Circadiano/efeitos dos fármacos , Proteína do X Frágil da Deficiência Intelectual , Síndrome do Cromossomo X Frágil/genética , Humanos , Deficiência Intelectual , Aprendizagem , Melatonina/metabolismo , Melatonina/farmacologia , Plasticidade Neuronal/efeitos dos fármacos , Fatores de Risco , Convulsões/tratamento farmacológico , Transtornos do Sono-Vigília/complicações , Transtornos do Sono-Vigília/tratamento farmacológico
6.
Asian-Australas J Anim Sci ; 30(9): 1245-1252, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28231693

RESUMO

OBJECTIVE: Phellodendron amurense (P. amurense) and Humulus japonicus (H. japonicus) are closely involved in anti-oxidative response and increasing antioxidant enzymes activities. However, the effects of their extracts on development of preimplantation bovine embryos have not been investigated. Therefore, we investigated the effects of P. amurense and H. japonicus extracts on developmental competence and quality of preimplantation bovine embryos. METHODS: After in vitro fertilization, bovine embryos were cultured for 7 days in Charles Rosenkrans amino acid medium supplemented with P. amurense (0.01 µg/mL) and H. japonicus (0.01 µg/mL). The effect of this supplementation during in vitro culture on development competence and antioxidant was investigated. RESULTS: We observed that the blastocysts rate was significantly increased (p<0.05) in P. amurense (28.9%±2.9%), H. japonicus (30.9%±1.5%), and a mixture of P. amurense and H. japonicus (34.8%± 2.1%) treated groups compared with the control group (25.4%±1.6%). We next confirmed that the intracellular levels of reactive oxygen species (ROS) were significantly decreased (p<0.01) in P. amurense and/or H. japonicus extract treated groups when compared with the control group. Our results also showed that expression of cleaved caspase-3 and apoptotic cells of blastocysts were significantly decreased (p<0.05) in bovine blastocysts derived from both P. amurense and H. japonicus extract treated embryos. CONCLUSION: These results suggest that proper treatment with P. amurense and H. japonicus extracts in the development of preimplantation bovine embryos improves the quality of blastocysts, which may be related to the reduction of ROS level and apoptosis.

7.
J Phys Ther Sci ; 29(4): 760-762, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28533625

RESUMO

[Purpose] The purpose of the present review is to discuss recent published articles in the understanding of efficacy of interventional exercise on autistic Fragile X syndrome (FXS) with special emphasis on its significance in clinical application in patients. [Methods] This review article was identified scientifically and/or clinically relevant articles from PubMed that directly/indirectly met the inclusion criteria. [Results] Mutation of fragile X mental retardation 1 (fmr1) gene on the X chromosome is related with loss of fragile X mental retardation protein (FMRP) that affecting physiological and behavioral abnormalities. Autistic FXS individuals exhibit disturbed sleep and altered circadian behavior. Although the underlying molecular mechanisms are not been fully explored, interventional exercise in autistic FXS has been clinically used for the treatment of physiological and behavioral abnormalities as well as psychiatric disorder in autistic FXS. [Conclusion] This review describes beneficial efficacy of interventional exercise and its controversy in patients with autistic FXS. This review also provides interventional strategies for clinicians and scientists that the way of neurophysiological approaches according to the level of physical and behavioral abnormalities.

8.
Bioorg Med Chem Lett ; 26(1): 87-93, 2016 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-26602279

RESUMO

Protein tyrosine phosphatase sigma (PTPσ) is a potential target for the therapeutic treatment of neurological deficits associated with impaired neuronal recovery, as this protein is the receptor for chondroitin sulfate proteoglycan (CSPG), which is known to inhibit neuronal regeneration. Through a high-throughput screening approach started from 6400 representative compounds in the Korea Chemical Bank chemical library, we identified 11 novel PTPσ inhibitors that can be classified as flavonoid derivatives or analogs, with IC50 values ranging from 0.5 to 17.5µM. Biochemical assays and structure-based active site-docking simulation indicate that our inhibitors are accommodated at the catalytic active site of PTPσ as surrogates for the phosphotyrosine group. Treatments of these compounds on PC-12 neuronal cells led to the recovery of neurite extension attenuated by CSPG treatment, demonstrating their potential as antineurodegenerative agents.


Assuntos
Inibidores Enzimáticos/farmacologia , Flavonoides/farmacologia , Neuritos/efeitos dos fármacos , Proteínas Tirosina Fosfatases Classe 2 Semelhantes a Receptores/antagonistas & inibidores , Animais , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Flavonoides/síntese química , Flavonoides/química , Modelos Moleculares , Estrutura Molecular , Neuritos/metabolismo , Células PC12 , Ratos , Proteínas Tirosina Fosfatases Classe 2 Semelhantes a Receptores/metabolismo , Bibliotecas de Moléculas Pequenas/síntese química , Bibliotecas de Moléculas Pequenas/química , Bibliotecas de Moléculas Pequenas/farmacologia , Relação Estrutura-Atividade
9.
J Reprod Dev ; 62(3): 249-55, 2016 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-26860251

RESUMO

Gangliosides are key lipid molecules required for the regulation of cellular processes such as proliferation, differentiation, and cell signaling, including signaling of epidermal growth factor receptor (EGFR). Epidermal growth factor (EGF) has long been considered a potential regulator of meiotic and cytoplasmic maturation in mammalian oocytes. However, there is no report on the direct effect of ganglioside GD1a in porcine oocyte maturation. In this study, we first investigated a functional link between GD1a and meiotic maturation during in vitro maturation (IVM) of porcine embryos. Moreover, we confirmed the effect of exogenous GD1a treatment on blastocyst development, quality, and fertilization rate in early embryonic development. First, we observed that the protein level of ST3GAL2, a GD1a synthesizing enzyme, significantly increased (P < 0.01) in cumulus-oocyte-complexes (COCs) during IVM progress. The proportion of arrested germinal vesicles (GV) increased in oocytes treated with EGF+GD1a (41.6 ± 1.5%) at the IVM I stage. Upon completion of meiotic maturation, the proportion of metaphase II (M II) was significantly higher (P < 0.05) in the EGF+GD1a (89.9 ± 3.6%) treated group. After IVF, the percentage of penetrated oocytes was significantly higher (P < 0.05) in the EGF+GD1a (89.1 ± 2.3%) treated group than in the control group. Furthermore, exogenous GD1a treatment improved the developmental competence and quality of blastocysts during preimplantation embryo development stage. These results suggest that ganglioside GD1a may play an important role in IVM mechanisms of porcine maturation capacity. Furthermore, our findings will be helpful for better promoting the embryo development and blastocyst quality in pigs.


Assuntos
Blastocisto/citologia , Gangliosídeo G(M1)/análogos & derivados , Oócitos/citologia , Animais , Apoptose , Núcleo Celular/metabolismo , Células Cultivadas , Fase de Clivagem do Zigoto , Células do Cúmulo/citologia , Desenvolvimento Embrionário , Fator de Crescimento Epidérmico/metabolismo , Receptores ErbB/metabolismo , Feminino , Fertilização , Gangliosídeo G(M1)/fisiologia , Meiose , Metáfase , Ovário/metabolismo , Sialiltransferases/metabolismo , Suínos , beta-Galactosídeo alfa-2,3-Sialiltransferase
10.
Anim Biotechnol ; 27(4): 223-30, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27565865

RESUMO

Transcription activator-like effector nucleases (TALENs) are a new type of engineered nuclease that is very effective for directed gene disruption in any genome sequence. We investigated the generation of mice with genetic knockout (KO) of the G protein-coupled receptor kinase (GRK) 5 gene by microinjection of TALEN mRNA. TALEN vectors were designed to target exons 1, 3, and 5 of the mouse GRK5 gene. Flow cytometry showed that the activity of the TALEN mRNAs targeted to exons 1, 3, and 5 was 8.7%, 9.7%, and 12.7%, respectively. The TALEN mRNA for exon 5 was injected into the cytoplasm of 180 one-cell embryos. Of the 53 newborns, three (5.6%) were mutant founders (F0) with mutations. Two clones from F028 showed a 45-bp deletion and F039 showed the same biallelic non-frame-shifting 3-bp deletions. Three clones from F041 were shown to possess a combination of frame-shifting 2-bp deletions. All of the mutations were transmitted through the germline but not to all progenies (37.5%, 37.5%, and 57.1% for the F028, F039, and F041 lines, respectively). The homozygote GRK5-KO mice for 28 and 41 lines created on F3 progenies and the homozygous genotype was confirmed by PCR, T7E1 assay and sequencing.


Assuntos
Quinase 5 de Receptor Acoplado a Proteína G/genética , Marcação de Genes/métodos , Camundongos Knockout/genética , Nucleases dos Efetores Semelhantes a Ativadores de Transcrição/genética , Animais , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mutação
11.
J Neurochem ; 132(6): 687-702, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25359615

RESUMO

Mitochondrial dysfunction is implicated in age-related degenerative disorders such as Alzheimer's disease (AD). Maintenance of mitochondrial dynamics is essential for regulating mitochondrial function. Aß oligomers (AßOs), the typical cause of AD, lead to mitochondrial dysfunction and neuronal loss. AßOs have been shown to induce mitochondrial fragmentation, and their inhibition suppresses mitochondrial dysfunction and neuronal cell death. Oxidative stress is one of the earliest hallmarks of AD. Cyclin-dependent kinase 5 (Cdk5) may cause oxidative stress by disrupting the antioxidant system, including Prx2. Cdk5 is also regarded as a modulator of mitochondrial fission; however, a precise mechanistic link between Cdk5 and mitochondrial dynamics is lacking. We estimated mitochondrial morphology and alterations in mitochondrial morphology-related proteins in Neuro-2a (N2a) cells stably expressing the Swedish mutation of amyloid precursor protein (APP), which is known to increase AßO production. We demonstrated that mitochondrial fragmentation by AßOs accompanies reduced mitofusin 1 and 2 (Mfn1/2) levels. Interestingly, the Cdk5 pathway, including phosphorylation of the Prx2-related oxidative stress, has been shown to regulate Mfn1 and Mfn2 levels. Furthermore, Mfn2, but not Mfn1, over-expression significantly inhibits the AßO-mediated cell death pathway. Therefore, these results indicate that AßO-mediated oxidative stress triggers mitochondrial fragmentation via decreased Mfn2 expression by activating Cdk5-induced Prx2 phosphorylation. Mitochondrial fragmentation induced by amyloid-beta oligomer (AßOs) which is generated from the Swedish mutation of amyloid precursor protein (APP) accompanies reduced Mfn1/2 levels. Interestingly, the Cdk5 pathway, including phosphorylation of the Prx2-related oxidative stress, has been shown to regulate Mfn1/2. Furthermore, Mfn2 over-expression significantly inhibits the AßO-mediated neuronal cells death pathway, but not Mfn1 over-expression. Therefore, these results indicate that AßO-mediated oxidative stress triggers mitochondrial fragmentation via decreased Mfn2 expression by activating Cdk5-induced Prx2 phosphorylation. ATP, adenosine triphosphate; Bax, Bcl-2-associated X protein; Bcl-2, B-cell lymphoma 2; Cdk5, Cyclin-dependent kinase; Cyt C, cytochrome C; Mfn2, mitofusin 2; Prx2, peroxiredoxin 2; ROS, reactive oxygen species.


Assuntos
Peptídeos beta-Amiloides/toxicidade , Quinase 5 Dependente de Ciclina/metabolismo , GTP Fosfo-Hidrolases/biossíntese , Mitocôndrias/metabolismo , Neurônios/metabolismo , Estresse Oxidativo/fisiologia , Peptídeos beta-Amiloides/metabolismo , Animais , Linhagem Celular Tumoral , GTP Fosfo-Hidrolases/deficiência , Camundongos , Mitocôndrias/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos
12.
Stem Cells ; 32(4): 998-1011, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24715692

RESUMO

Redox balance has been suggested as an important determinant of "stemness" in embryonic stem cells (ESCs). In this study, we demonstrate that peroxiredoxin (Prx) plays a pivotal role in maintenance of ESC stemness during neurogenesis through suppression of reactive oxygen species (ROS)-sensitive signaling. During neurogenesis, Prx I and Oct4 are expressed in a mutually dependent manner and their expression is abruptly downregulated by an excess of ROS. Thus, in Prx I(-/-) or Prx II(-/-) ESCs, rapid loss of stemness can occur due to spontaneous ROS overload, leading to their active commitment into neurons; however, stemness is restored by the addition of an antioxidant or an inhibitor of c-Jun N-terminal kinase (JNK). In addition, Prx I and Prx II appear to have a tight association with the mechanism underlying the protection of ESC stemness in developing teratomas. These results suggest that Prx functions as a protector of ESC stemness by opposing ROS/JNK cascades during neurogenesis. Therefore, our findings have important implications for understanding of maintenance of ESC stemness through involvement of antioxidant enzymes and may lead to development of an alternative stem cell-based therapeutic strategy for production of high-quality neurons in large quantity.


Assuntos
Células-Tronco Embrionárias/enzimologia , MAP Quinase Quinase 4/metabolismo , Neurogênese/fisiologia , Peroxirredoxinas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Animais , Células-Tronco Embrionárias/citologia , MAP Quinase Quinase 4/genética , Camundongos , Camundongos Knockout , Peroxirredoxinas/genética
13.
Reprod Fertil Dev ; 27(3): 481-90, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24411581

RESUMO

γ-tocotrienol (GTT), an isomer of vitamin E, has been the subject of increasing interest due to its strong anti-oxidant effects. Therefore, in this study, the effects of GTT on blastocyst development, expression levels of reactive oxygen species (ROS) and apoptotic index were investigated in preimplantation porcine embryos. After in vitro maturation and fertilisation, porcine embryos were cultured for 6 days in porcine zygote medium 3 supplemented with or without GTT (200µM) under oxidative stress conditions (200µM hydrogen peroxide (H2O2)). Blastocyst development was significantly improved in the GTT-treated group when compared with the H2O2-treated group (P<0.05). Subsequent evaluation of the intracellular levels of ROS and numbers of apoptotic nuclei in GTT-treated blastocysts revealed that ROS levels of GTT-treated porcine blastocysts were decreased (P<0.05) and the numbers of apoptotic nuclei were reduced by GTT treatment in porcine embryos. Moreover, the total cell numbers of blastocysts were significantly increased in the GTT-treated group relative to the untreated group under H2O2-induced oxidative stress (P<0.05). The expression levels of apoptosis-related genes (BCL-XL, BAX) in GTT-treated blastocysts were then investigated using real-time reverse transcription polymerase chain reaction. Expression of the anti-apoptotic BCL-XL gene was shown to be increased in the GTT-treated blastocyst group, whereas expression of the pro-apoptotic BAX gene was decreased. Taken together, these results suggest that GTT (200µM) under H2O2-induced oxidative stress, thereby improving the developmental competence of porcine embryos via modulation of intracellular levels of ROS and the apoptotic index during the preimplantation stage.


Assuntos
Antioxidantes/farmacologia , Blastocisto/efeitos dos fármacos , Cromanos/farmacologia , Desenvolvimento Embrionário/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Vitamina E/análogos & derivados , Animais , Apoptose/efeitos dos fármacos , Blastocisto/metabolismo , Técnicas de Cultura Embrionária , Glutationa/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Suínos , Vitamina E/farmacologia
14.
J Reprod Dev ; 61(2): 81-9, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25501014

RESUMO

Mitochondria are highly dynamic organelles that undergo constant fusion/fission as well as activities orchestrated by large dynamin-related GTPases. These dynamic mitochondrial processes influence mitochondrial morphology, size and function. Therefore, this study was conducted to evaluate the effects of mitochondrial fission inhibitor, mdivi-1, on developmental competence and mitochondrial function of porcine embryos and primary cells. Presumptive porcine embryos were cultured in PZM-3 medium supplemented with mdivi-1 (0, 10 and 50 µM) for 6 days. Porcine fibroblast cells were cultured in growth medium with mdivi-1 (0 and 50 µM) for 2 days. Our results showed that the rate of blastocyst production and cell growth in the mdivi-1 (50 µM) treated group was lower than that of the control group (P < 0.05). Moreover, loss of mitochondrial membrane potential in the mdivi-1 (50 µM) treated group was increased relative to the control group (P < 0.05). Subsequent evaluation revealed that the intracellular levels of reactive oxygen species (ROS) and the apoptotic index were increased by mdivi-1 (50 µM) treatment (P < 0.05). Finally, the expression of mitochondrial fission-related protein (Drp 1) was lower in the embryos and cells in the mdivi-1-treated group than the control group. Taken together, these results indicate that mdivi-1 treatment may inhibit developmental competence and mitochondrial function in porcine embryos and primary cells.


Assuntos
Apoptose/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Quinazolinonas/farmacologia , Animais , Apoptose/fisiologia , Proteínas Quinases Associadas com Morte Celular/metabolismo , Técnicas de Cultura Embrionária , Desenvolvimento Embrionário/fisiologia , Potencial da Membrana Mitocondrial/fisiologia , Mitocôndrias/metabolismo , Dinâmica Mitocondrial/efeitos dos fármacos , Dinâmica Mitocondrial/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Suínos
15.
Int J Mol Sci ; 16(8): 19657-70, 2015 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-26295390

RESUMO

Neural diseases including injury by endogenous factors, traumatic brain injury, and degenerative neural injury are eventually due to reactive oxygen species (ROS). Thus ROS generation in neural tissues is a hallmark feature of numerous forms of neural diseases. Neural degeneration and the neural damage process is complex, involving a vast array of tissue structure, transcriptional/translational, electrochemical, metabolic, and functional events within the intact neighbors surrounding injured neural tissues. During aging, multiple changes involving physical, chemical, and biochemical processes occur from the molecular to the morphological levels in neural tissues. Among many recommended therapeutic candidates, melatonin also plays a role in protecting the nervous system from anti-inflammation and efficiently safeguards neuronal cells via antioxidants and other endogenous/exogenous beneficial factors. Therefore, given the wide range of mechanisms responsible for neuronal damage, multi-action drugs or therapies for the treatment of neural injury that make use of two or more agents and target several pathways may have greater efficacy in promoting functional recovery than a single therapy alone.


Assuntos
Quimioterapia Combinada/métodos , Degeneração Neural/prevenção & controle , Fármacos Neuroprotetores/farmacologia , Traumatismos do Sistema Nervoso/prevenção & controle , Envelhecimento/fisiologia , Animais , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Humanos , Melatonina/farmacologia , Melatonina/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo
16.
Int J Mol Sci ; 16(11): 26880-913, 2015 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-26569225

RESUMO

Ionizing radiation (IR), such as X-rays and gamma (γ)-rays, mediates various forms of cancer cell death such as apoptosis, necrosis, autophagy, mitotic catastrophe, and senescence. Among them, apoptosis and mitotic catastrophe are the main mechanisms of IR action. DNA damage and genomic instability contribute to IR-induced cancer cell death. Although IR therapy may be curative in a number of cancer types, the resistance of cancer cells to radiation remains a major therapeutic problem. In this review, we describe the morphological and molecular aspects of various IR-induced types of cell death. We also discuss cytogenetic variations representative of IR-induced DNA damage and genomic instability. Most importantly, we focus on several pathways and their associated marker proteins responsible for cancer resistance and its therapeutic implications in terms of cancer cell death of various types and characteristics. Finally, we propose radiation-sensitization strategies, such as the modification of fractionation, inflammation, and hypoxia and the combined treatment, that can counteract the resistance of tumors to IR.


Assuntos
Aberrações Cromossômicas , Terapia Combinada/métodos , Raios gama/uso terapêutico , Neoplasias/terapia , Tolerância a Radiação/efeitos dos fármacos , Radiossensibilizantes/uso terapêutico , Apoptose/genética , Apoptose/efeitos da radiação , Autofagia/genética , Autofagia/efeitos da radiação , Análise Citogenética , Dano ao DNA/efeitos da radiação , Instabilidade Genômica , Humanos , Mitose/efeitos da radiação , Necrose/genética , Necrose/patologia , Neoplasias/genética , Neoplasias/patologia , Terapia por Raios X
17.
Int J Mol Sci ; 16(2): 2386-402, 2015 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-25622254

RESUMO

The accumulation and aggregation of misfolded proteins in the brain, such as amyloid-ß (Aß) and hyperphosphorylated tau, is a neuropathological hallmark of Alzheimer's disease (AD). Previously, we developed and validated a novel non-human primate model for sporadic AD (sAD) research using intracerebroventricular administration of streptozotocin (icv STZ). To date, no characterization of AD-related genes in different brain regions has been performed. Therefore, in the current study, the expression of seven amyloid precursor protein (APP) pathway-related and five tau phosphorylation-related genes was investigated by quantitative real-time PCR experiments, using two matched-pair brain samples from control and icv STZ-treated cynomolgus monkeys. The genes showed similar expression patterns within the control and icv STZ-treated groups; however, marked differences in gene expression patterns were observed between the control and icv STZ-treated groups. Remarkably, other than ß-secretase (BACE1) and cyclin-dependent kinase 5 (CDK5), all the genes tested showed similar expression patterns in AD models compared to controls, with increased levels in the precuneus and occipital cortex. However, significant changes in gene expression patterns were not detected in the frontal cortex, hippocampus, or posterior cingulate. Based on these results, we conclude that APP may be cleaved via the general metabolic mechanisms of increased α- and γ-secretase levels, and that hyperphosphorylation of tau could be mediated by elevated levels of tau protein kinase, specifically in the precuneus and occipital cortex.


Assuntos
Doença de Alzheimer/patologia , Precursor de Proteína beta-Amiloide/metabolismo , Proteínas tau/metabolismo , Doença de Alzheimer/induzido quimicamente , Doença de Alzheimer/metabolismo , Secretases da Proteína Precursora do Amiloide/genética , Secretases da Proteína Precursora do Amiloide/metabolismo , Animais , Encéfalo/metabolismo , Calpaína/genética , Calpaína/metabolismo , Quinase 5 Dependente de Ciclina/genética , Quinase 5 Dependente de Ciclina/metabolismo , Modelos Animais de Doenças , Feminino , Quinase 3 da Glicogênio Sintase/genética , Quinase 3 da Glicogênio Sintase/metabolismo , Glicogênio Sintase Quinase 3 beta , Macaca fascicularis , Fosforilação , RNA Mensageiro/metabolismo , Estreptozocina/toxicidade
18.
J Phys Ther Sci ; 27(6): 1743-5, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26180311

RESUMO

[Purpose] The purpose of this study was to determine whether muscle activity and pressure-induced pain in the upper extremities are affected by smartphone use, and to compare the effects of phone handling with one hand and with both hands. [Subjects] The study subjects were asymptomatic women 20-22 years of age. [Methods] The subjects sat in a chair with their feet on the floor and the elbow flexed, holding a smartphone positioned on the thigh. Subsequently, the subjects typed the Korean anthem for 3 min, one-handed or with both hands. Each subject repeated the task three times, with a 5-min rest period between tasks to minimize fatigue. Electromyography (EMG) was used to record the muscle activity of the upper trapezius (UT), extensor pollicis longus (EPL), and abductor pollicis (AP) during phone operation. We also used a dolorimeter to measure the pressure-induced pain threshold in the UT. [Results] We observed higher muscle activity in the UT, AP, and EPL in one-handed smartphone use than in its two-handed use. The pressure-induced pain threshold of the UT was lower after use of the smartphone, especially after one-handed use. [Conclusion] Our results show that smartphone operation with one hand caused greater UT pain and induced increased upper extremity muscle activity.

19.
J Phys Ther Sci ; 27(12): 3933-5, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26834383

RESUMO

[Purpose] The intensity of therapeutic physical exercise is complex and sometimes controversial in patients with neural injuries. This review assessed whether therapeutic physical exercise is beneficial according to the intensity of the physical exercise. [Methods] The authors identified clinically or scientifically relevant articles from PubMed that met the inclusion criteria. [Results] Exercise training can improve body strength and lead to the physiological adaptation of skeletal muscles and the nervous system after neural injuries. Furthermore, neurophysiological and neuropathological studies show differences in the beneficial effects of forced therapeutic exercise in patients with severe or mild neural injuries. Forced exercise alters the distribution of muscle fiber types in patients with neural injuries. Based on several animal studies, forced exercise may promote functional recovery following cerebral ischemia via signaling molecules in ischemic brain regions. [Conclusions] This review describes several types of therapeutic forced exercise and the controversy regarding the therapeutic effects in experimental animals versus humans with neural injuries. This review also provides a therapeutic strategy for physical therapists that grades the intensity of forced exercise according to the level of neural injury.

20.
Biol Reprod ; 90(5): 104, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24695629

RESUMO

The stress produced by the coupling of reactive oxygen species (ROS) and endoplasmic reticulum (ER) has been explored extensively, but little is known regarding their roles in the early development of mammalian embryos. Here, we demonstrated that the early development of in vitro-produced (IVP) bovine embryos was governed by the cooperative action between ROS and ER stress. Compared with the tension produced by 5% O2, 20% O2 significantly decreased the blastocyst formation rate and cell survival, which was accompanied by increases in ROS and in levels of sXBP-1 transcript, which is an ER stress indicator. In addition, treatment with glutathione (GSH), a ROS scavenger, decreased ROS levels, which resulted in increased blastocyst formation and cell survival rates. Importantly, levels of sXBP-1 and ER stress-associated transcripts were reduced by GSH treatment in developing bovine embryos. Consistent with this observation, tauroursodeoxycholate (TUDCA), an ER stress inhibitor, improved blastocyst developmental rate, trophectoderm proportion, and cell survival. Moreover, ROS and sXBP-1 transcript levels were markedly decreased by supplementation with TUDCA, suggesting a possible mechanism governing the mutual regulation between ROS and ER stress. Interestingly, knockdown of XBP-1 transcripts resulted in both elevation of ROS and decrease of antioxidant transcripts, which ultimately reduced in vitro developmental competence of bovine embryos. Based on these results, in vitro developmental competence of IVP bovine embryos was highly dependent on the coupled response between oxidative and ER stresses. These results increase our understanding of the mechanism(s) governing early embryonic development and may improve strategies for the generation of IVP embryos with high developmental competence.


Assuntos
Apoptose/fisiologia , Bovinos/embriologia , Embrião de Mamíferos/fisiologia , Desenvolvimento Embrionário/fisiologia , Estresse do Retículo Endoplasmático/fisiologia , Animais , Western Blotting/veterinária , Feminino , Glutationa/farmacologia , Marcação In Situ das Extremidades Cortadas/veterinária , Microscopia de Fluorescência/veterinária , Gravidez , RNA Interferente Pequeno/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Ácido Taurodesoxicólico/farmacologia
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