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1.
Nat Commun ; 14(1): 455, 2023 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-36709207

RESUMO

Ectopic expression in fibroblasts of synapsin 1 and synaptophysin is sufficient to generate condensates of vesicles highly reminiscent of synaptic vesicle (SV) clusters and with liquid-like properties. Here we show that unlike synaptophysin, other major integral SV membrane proteins fail to form condensates with synapsin, but co-assemble into the clusters formed by synaptophysin and synapsin in this ectopic expression system. Another vesicle membrane protein, ATG9A, undergoes activity-dependent exo-endocytosis at synapses, raising questions about the relation of ATG9A traffic to the traffic of SVs. We find that both in fibroblasts and in nerve terminals ATG9A does not co-assemble into synaptophysin-positive vesicle condensates but localizes on a distinct class of vesicles that also assembles with synapsin but into a distinct phase. Our findings suggest that ATG9A undergoes differential sorting relative to SV proteins and also point to a dual role of synapsin in controlling clustering at synapses of SVs and ATG9A vesicles.


Assuntos
Sinapsinas , Vesículas Sinápticas , Vesículas Sinápticas/metabolismo , Sinapsinas/genética , Sinapsinas/metabolismo , Sinaptofisina/metabolismo , Sinapses/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo
2.
Sci Rep ; 12(1): 22028, 2022 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-36539544

RESUMO

The interrelationships between neuronal viability, synaptic integrity, and microglial responses remain in infancy. In dealing with the question, we induced a stretch injury to evaluate the mechanical effects of trauma on rat primary cortical neurons and BV2 microglial cells in a transwell culture system. The viability of primary neurons and BV2 cells was determined by MTT. Synaptic integrity was evaluated by determining the expression of beta-secretase 1 (BACE1), amyloid-beta (Aß), microtubule-associated protein 2 (MAP2), and synaptophysin (vehicle protein). Both CD16/32-positive (CD16/32+) and CD206-positive (CD206+) microglia cells were detected by immunofluorescence staining. The phagocytic ability of the BV2 cells was determined using pHrodo E. coli BioParticles conjugates and flow cytometry. We found that stretch injury BV2 cells caused reduced viability and synaptic abnormalities characterized by Aß accumulation and reductions of BACE1, MAP2, and synaptophysin in primary neurons. Intact BV2 cells exhibited normal phagocytic ability and were predominantly CD206+ microglia cells, whereas the injured BV2 cells exhibited reduced phagocytic ability and were predominantly CD16/32+ microglial cells. Like a stretch injury, the injured BV2 cells can cause both reduced viability and synaptic abnormalities in primary neurons; intact BV2 cells, when cocultured with primary neurons, can protect against the stretch-injured-induced reduced viability and synaptic abnormalities in primary neurons. We conclude that CD206+ and CD16/32+ BV-2 cells can produce neuroprotective and cytotoxic effects on primary cortical neurons.


Assuntos
Secretases da Proteína Precursora do Amiloide , Microglia , Ratos , Animais , Microglia/metabolismo , Sinaptofisina/metabolismo , Secretases da Proteína Precursora do Amiloide/metabolismo , Escherichia coli/metabolismo , Ácido Aspártico Endopeptidases/metabolismo , Neurônios/metabolismo , Peptídeos beta-Amiloides/metabolismo
3.
Wiad Lek ; 75(9 pt 2): 2256-2261, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36378705

RESUMO

OBJECTIVE: The aim: To study changes of the expression of synaptophysin (Syn) and vascular endothelial growth factor (VEGF) in neurons of the sensorimotor cortex (SMC) to reveal after unilateral ligation of the carotid artery, sensitization with brain antigen and their combination. PATIENTS AND METHODS: Materials and methods: Experimental animals - Wistar rats (260-290 g). Experimental models: mobilization of the left common carotid artery, ligation of the indicated artery, sensitization with cerebral antigen, combination of sensitization with cerebral antigen and ligation of the carotid artery. Methods: immunohistochemistry, quantitative densitometric assessment. RESULTS: Results: Dyscirculatory disorders of cerebral blood supply during unilateral mobilization or ligation of the common carotid artery, sensitization with cerebral antigen lead in rats to a transient decrease in synaptophysin expression and phase changes in VEGF expression in the SMC from the lesion side. These changes occur in the absence of morphological changes in the cerebral cortex. CONCLUSION: Conclusions: The absence of morphological changes in the SMC in the short term (10-30 days) after minor trauma to the common carotid artery (separation from the bed and n.vagus) or its ligation is accompanied by a transient decrease in Syn expression and some increase in VEGF, which may reflect a violation of synaptic function and the general metabolic activity of neurons. Sensitization with a brain antigen, leading to an increase in the level of anti-brain antibodies and immune complexes in the blood of rats, can act as an independent damaging factor for the brain, and also potentiates and prolongs changes caused by impaired blood circulation.


Assuntos
Córtex Sensório-Motor , Fator A de Crescimento do Endotélio Vascular , Animais , Ratos , Sinaptofisina/metabolismo , Ratos Wistar , Artérias Carótidas/metabolismo , Artérias Carótidas/patologia , Artéria Carótida Primitiva/metabolismo , Fatores de Crescimento do Endotélio Vascular/metabolismo , Ligadura
4.
Exp Gerontol ; 169: 111981, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36270545

RESUMO

The incidence of aging-related cognitive decline is increasing with population aging. It is urgent to explore ways to ameliorate aging-related cognitive decline. Cognitive-exercise dual-task intervention has shown beneficial effects on improving cognition in aging cohorts, but the mechanisms of the effects remain unclear. In this study, 18-month-old Sprague Dawley rats served as a model of natural aging. First, the performance in the Morris water maze test and the change in synaptophysin content in the hippocampus were used to investigate the cognitive decline of 18-month-old rats. Then, a batch of 18-month-old rats was treated with cognitive, exercise, or cognitive-exercise dual-task intervention for 12 weeks. The novel object recognition test was used to assess cognitive ability. Enzyme-linked immunosorbent assay and Western blotting were used to detect the levels of oxidative stress molecules and synaptic plasticity-related proteins. We found that cognitive-exercise dual-task intervention improved the discrimination index of natural aging rats. After dual-task intervention, the expression levels of synaptophysin, brain-derived neurotrophic factor, superoxide dismutase, and glutathione peroxidase were increased, and the expression level of lipid peroxide malondialdehyde was decreased. Furthermore, the effect of dual-task intervention on synaptic plasticity-related proteins and oxidative stress indicators was greater than that of single cognitive or exercise intervention. In conclusion, cognitive-exercise dual-task intervention can significantly ameliorate aging-related cognitive decline, and the improvement might be related to the reduction of oxidative stress and the enhancement of synaptic plasticity. The effect of cognitive-exercise dual-task intervention may be better than that of single cognitive or exercise intervention.


Assuntos
Disfunção Cognitiva , Plasticidade Neuronal , Ratos , Animais , Sinaptofisina/metabolismo , Aprendizagem em Labirinto , Ratos Sprague-Dawley , Envelhecimento/psicologia , Disfunção Cognitiva/prevenção & controle , Disfunção Cognitiva/metabolismo , Hipocampo/metabolismo , Estresse Oxidativo , Cognição
5.
Anticancer Res ; 42(11): 5475-5478, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36288880

RESUMO

BACKGROUND/AIM: Cholangioblastic variant of intrahepatic cholangiocarcinoma (CVICC) is an exceedingly rare primary biliary tract tumor and typically occurs in young patients with a median age of 24.5-year-old. It can mimic metastatic well-differentiated neuroendocrine tumors in the liver with its similar histologic and immunophenotypic features. CASE REPORT: We hereby report a CVICC in a 68-year-old female patient with distinctive biphasic cytologic features. The patient was diagnosed and treated as a metastatic well differentiated neuroendocrine tumor. The recurrent liver tumor was resected and the tumor cells were strongly positive for Inhibin A and cytokeratin 19 (CK19), focally and weakly positive for synaptophysin and chromogranin, and negative for Insulinoma associated protein 1 (INSM1). Ribonucleic acid (RNA) sequencing showed that the tumor bared a characteristic Nipped-B-like protein (NIPBL)-Nucleus accumbens-associated protein 1 (NACC1) gene fusion. CONCLUSION: To the best of our knowledge, this is the first documented case in an elder patient of this entity with NIPPL-NACC1 gene fusion. Acknowledgment of the biphasic cytology, screening with Inhibin A in suspicious cases, and coupled with a molecular study may facilitate accurate classification of this aggressive tumor and lead to proper clinical management.


Assuntos
Neoplasias dos Ductos Biliares , Colangiocarcinoma , Neoplasias Hepáticas , Feminino , Humanos , Idoso , Adulto Jovem , Adulto , Sinaptofisina/metabolismo , Queratina-19/metabolismo , Cromograninas/metabolismo , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Colangiocarcinoma/genética , Colangiocarcinoma/cirurgia , Colangiocarcinoma/diagnóstico , Neoplasias Hepáticas/patologia , Neoplasias dos Ductos Biliares/genética , Neoplasias dos Ductos Biliares/cirurgia , Neoplasias dos Ductos Biliares/metabolismo , Ductos Biliares Intra-Hepáticos/patologia , RNA/metabolismo , Proteínas Repressoras/metabolismo , Proteínas de Ciclo Celular/metabolismo , Proteínas de Neoplasias/metabolismo
6.
Sci Rep ; 12(1): 17084, 2022 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-36224264

RESUMO

Slowing down age-related neurocognitive impairment has been a challenge. We evaluated the therapeutic effects of metformin in D-galactose-induced aging. Additionally, we studied the potential molecular mechanisms that could be responsible for metformin's anti-aging effects. Thirty male rats were equally divided into: 1-control group, which received saline solution, 2-D-galactose (D-gal) group, which received D-galactose (100 mg/kg/day) by gastric lavage for eight weeks, and 3-D-galactose + Metformin (D-gal + Met) treated group, which received D-galactose + metformin (200 mg/kg/day) by gastric lavage for eight weeks. Neurocognitive assessment was done. Measurement of inflammatory, oxidative stress, and BDNF biomarkers was performed. AMPK and PI3K genes expression were assessed. Hippocampal tissues were dissected for histopathological and immunohistochemical studies. D-gal resulted in neurocognitive impairments, elevation of inflammatory biomarkers, altered oxidative stress markers, decreased BDNF, decreased expression of synaptophysin and Bcl2 with increased expression of Caspase-3, and down-regulation of AMPK and PI3K genes. Neurodegenerative changes were present in the hippocampus. Metformin restored significantly D-gal induced neurodegenerative changes. We concluded that metformin could alleviate age-induced neurocognitive deficit via amelioration of neuroinflammation, attenuation of oxidative stress, reduction of apoptosis, as well as promotion of synaptic plasticity. These mechanisms could be mediated via the activation of the AMPK/BDNF/PI3K pathway.


Assuntos
Galactose , Metformina , Proteínas Quinases Ativadas por AMP/metabolismo , Envelhecimento/metabolismo , Animais , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Caspase 3/metabolismo , Galactose/farmacologia , Masculino , Metformina/farmacologia , Estresse Oxidativo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos , Solução Salina/farmacologia , Sinaptofisina/metabolismo
7.
Differentiation ; 128: 43-56, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36306693

RESUMO

Major histocompatibility complex class I (MHC-I) has been implicated in several types of neuroplasticity phenomena. Interferon beta-1b (IFN-ß) increases MHC-I expression by motoneurons after sciatic nerve crush in mice, improving axonal growth and functional recovery. Additionally, IFN-ß induces glial hypertrophy associated with upregulation of glial fibrillary acidic protein (GFAP) and MHC-I in murine astrocytes in vitro. As knowledge about MHC-I and its role in synaptic plasticity in human astrocytes (HAs) is scarce, we investigated these aspects in mature HAs obtained from the neocortex of patients undergoing surgery due to hippocampal sclerosis. Cells were exposed to media in the absence (0 IU/ml) or presence of IFN-ß for 5 days (500 IU/ml). Beta-2 microglobulin (ß2m), a component of the MHC-I, GFAP and vimentin proteins, was quantified by flow cytometry (FC) and increased by 100%, 60% and 46%, respectively, after IFN-ß exposure. We also performed qRT-PCR gene expression analyses for ß2m, GFAP, vimentin, and pro- and anti-inflammatory cytokines. Our data showed that IFN-ß-treated astrocytes displayed ß2m and GFAP gene upregulation. Additionally, they presented a proinflammatory profile with increase in the IL-6 and IL-1ß genes and a tendency to upregulate TNF-α. Moreover, we evaluated the effect of HAs conditioned medium (CM) on the formation/maintenance of neurites/synapses by the PC12 lineage. Synaptophysin protein expression was quantified by FC. The CM of IFN-ß-activated astrocytes was not harmful to PC12 neurites, and there was no change in synaptophysin protein expression. Therefore, IFN-ß activated HAs by increasing GFAP, vimentin and MHC-I protein expression. Like MHC-I modulation and astrocyte activation may be protective after peripheral nerve damage and in some neurodegenerative conditions, this study opens perspectives on the pathophysiological roles of astroglial MHC-I in the human CNS.


Assuntos
Astrócitos , Interferon beta , Humanos , Animais , Camundongos , Astrócitos/metabolismo , Sinaptofisina/genética , Sinaptofisina/metabolismo , Sinaptofisina/farmacologia , Vimentina/genética , Vimentina/metabolismo , Vimentina/farmacologia , Interferon beta/genética , Interferon beta/metabolismo , Interferon beta/farmacologia , Antígenos de Histocompatibilidade Classe I/genética , Antígenos de Histocompatibilidade Classe I/metabolismo , Complexo Principal de Histocompatibilidade , Fenótipo
8.
Int J Med Sci ; 19(10): 1586-1595, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36185335

RESUMO

Background: Resveratrol, a natural antioxidant polyphenol, has the functions of anti-inflammation, anti-cancer, liver protection and cardioprotection. Microorganism biotransformation-produced resveratrol (MBR) product shows higher purity than the natural source of resveratrol and costs less than the chemically synthesized resveratrol. The aim of the present study was to investigate the protective effects of MBR in hamsters treated with a high-fat diet (HFD). Methods: MBR was obtained by the fermentative process of piceid. Hamsters were randomly divided into four groups: HFD plus oral administration of MBR 0 (C), 5 (L), 20 (M) or 50 mg/kg (H), respectively. After six-week of treatment, hamsters were sacrificed, and tissues were collected for further analysis. Results: MBR at these three dosages did not influence the appetite or growth of the hamsters. Liver enzymes, blood glucose, total cholesterol, triglyceride, and liver weight were significantly reduced in the MBR groups than in the control group. Additionally, high-density lipoprotein-cholesterol (HDL-C) was also elevated in all MBR groups. On the other hand, serum low-density lipoprotein-cholesterol (LDL-C) was decreased in the MBR groups. Triglyceride (TG) in liver tissue and fatty liver level were lower in group H. Memory-associated proteins, phosphorylation of calmodulin-dependent protein kinase II (p-CaMK II) and synaptophysin (SYP), were increased in the brains of MBR groups. Conclusion: The high yield- and short procedure-produced MBR has the potential to protect animals fed with HFD from hyperlipidemia, hepatic steatosis, hyperglycemia, and synaptic impairment, which might be beneficial for patients with these types of diseases.


Assuntos
Fígado Gorduroso , Hiperlipidemias , Animais , Antioxidantes/farmacologia , Biotransformação , Glicemia/metabolismo , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/farmacologia , HDL-Colesterol , LDL-Colesterol , Cricetinae , Dieta Hiperlipídica/efeitos adversos , Fígado Gorduroso/tratamento farmacológico , Fígado Gorduroso/etiologia , Fígado Gorduroso/metabolismo , Hiperlipidemias/tratamento farmacológico , Hiperlipidemias/etiologia , Fígado , Polifenóis/metabolismo , Polifenóis/farmacologia , Resveratrol/farmacologia , Sinaptofisina/metabolismo , Triglicerídeos
9.
Eur J Neurosci ; 56(11): 6003-6021, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36226387

RESUMO

Type 2 diabetes mellitus (T2DM) increases the risk of developing Alzheimer's disease (AD), which has been proposed to be driven by an abnormal neuroinflammatory response affecting cognitive function. However, the impact of T2DM on hippocampal function and synaptic integrity during aging has not been investigated. Here, we investigated the effects of aging in T2DM on AD-like pathology using the leptin receptor-deficient db/db mouse model of T2DM. Our results indicate that adult T2DM mice exhibited impaired spatial acquisition in the Morris water maze (MWM). Morphological analysis showed an age-dependent neuronal loss in the dentate gyrus. We found that astrocyte density was significantly decreased in all regions of the hippocampus in T2DM mice. Our analysis showed that microglial activation was increased in the CA3 and the dentate gyrus of the hippocampus in an age-dependent manner in T2DM mice. However, the expression of presynaptic marker protein (synaptophysin) and the postsynaptic marker protein [postsynaptic density protein 95 (PSD95)] was unchanged in the hippocampus of adult T2DM mice. Interestingly, synaptophysin and PSD95 expression significantly decreased in the hippocampus of aged T2DM mice, suggesting an impaired hippocampal synaptic integrity. Cytokine profiling analysis displayed a robust pro-inflammatory cytokine profile in the hippocampus of aged T2DM mice compared with the younger cohort, outlining the role of aging in exacerbating the neuroinflammatory profile in the diabetic state. Our results suggest that T2DM impairs cognitive function by promoting neuronal loss in the dentate gyrus and triggering an age-dependent deterioration in hippocampal synaptic integrity, associated with an aberrant neuroinflammatory response.


Assuntos
Doença de Alzheimer , Diabetes Mellitus Tipo 2 , Navegação Espacial , Camundongos , Animais , Sinaptofisina/metabolismo , Hipocampo/metabolismo , Doença de Alzheimer/metabolismo , Inflamação/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças
10.
PLoS One ; 17(9): e0274478, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36178902

RESUMO

The neuroendocrine tumours paraganglioma and pheochromocytoma (PPGLs) are commonly associated with succinate dehydrogenase (SDH) gene variants, but no human SDH-related PPGL-derived cell line has been developed to date. The aim of this study was to systematically explore practical issues related to the classical 2D-culture of SDH-related human paragangliomas and pheochromocytomas, with the ultimate goal of identifying a viable tumour-derived cell line. PPGL tumour tissue/cells (chromaffin cells) were cultured in a variety of media formulations and supplements. Tumour explants and dissociated primary tumour cells were cultured and stained with a range of antibodies to identify markers suitable for use in human PPGL culture. We cultured 62 PPGLs, including tumours with confirmed SDHB, SDHC and SDHD variants, as well as several metastatic tumours. Testing a wide range of basic cell culture media and supplements, we noted a marked decline in chromaffin cell numbers over a 4-8 week period but the persistence of small numbers of synaptophysin/tyrosine hydroxylase-positive chromaffin cells for up to 99 weeks. In cell culture, immunohistochemical staining for chromogranin A and neuron-specific enolase was generally negative in chromaffin cells, while staining for synaptophysin and tyrosine hydroxylase was generally positive. GFAP showed the most consistent staining of type II sustentacular cells. Of the media tested, low serum or serum-free media best sustained relative chromaffin cell numbers, while lactate enhanced the survival of synaptophysin-positive cells. Synaptophysin-positive PPGL tumour cells persist in culture for long periods but show little evidence of proliferation. Synaptophysin was the most consistent cell marker for chromaffin cells and GFAP the best marker for sustentacular cells in human PPGL cultures.


Assuntos
Neoplasias das Glândulas Suprarrenais , Paraganglioma , Feocromocitoma , Neoplasias das Glândulas Suprarrenais/patologia , Cromogranina A/metabolismo , Meios de Cultura Livres de Soro , Mutação em Linhagem Germinativa , Humanos , Lactatos , Paraganglioma/genética , Paraganglioma/patologia , Feocromocitoma/patologia , Fosfopiruvato Hidratase/metabolismo , Succinato Desidrogenase/genética , Succinato Desidrogenase/metabolismo , Sinaptofisina/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismo
11.
Neuroreport ; 33(14): 597-603, 2022 10 05.
Artigo em Inglês | MEDLINE | ID: mdl-36062510

RESUMO

BACKGROUND: Stress is not scarce in peoples' daily life that may result in mental diseases and cognitive impairments. Chronic restraint stress (CRS) is a well-validated animal model used to investigate the mechanism of stress-associated depression and cognitive impairments. Dl-3-n-butylphthalide (NBP) possesses anti-oxidant, anti-inflammatory and anti-apoptotic, promoting neurogenesis and neuroplasticity that exerts neuroprotective effects. However, the effects of NBP on CRS-induced depression and cognitive impairments remain unclear. METHODS: C57BL/6 male mice were randomly divided into the control group, stress group and stress+NBP group. Mice were exposed to CRS for three consecutive weeks and mice in the NBP treatment group were administered with NBP before the CRS procedure. After that, depression and cognition behaviors were evaluated followed by phosphorylation of Ca2+/calmodulin-dependent protein kinase II (p-CaMKII), phosphorylation of cAMP-response element-binding protein (p-CREB), brain-derived neurotrophic factor (BDNF) proteins expression, immunohistochemistry of hippocampal postsynaptic density 95 (PSD95) and synaptophysin, and hippocampal morphology. RESULTS: Our results showed that mice exhibited depression-like behaviors and cognitive deficits after 3 weeks exposure to CRS. Additionally, CRS downregulated CaMKII/CREB/BDNF signaling pathway, reduced PSD95 and synaptophysin expression and induced hippocampal CA1 and dentate gyrus ment significantly reversed the hippocampal pathological and molecular changes induced by CRS. CONCLUSION: In conclusion, these results reveal that NBP exerts a neuroprotective effect on depression and cognitive deficit through activating CaMKII/CREB/BDNF pathway, enhancing PSD95 and synaptophysin expression and protecting hippocampal morphology.


Assuntos
Disfunção Cognitiva , Fármacos Neuroprotetores , Animais , Benzofuranos , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Disfunção Cognitiva/tratamento farmacológico , Disfunção Cognitiva/etiologia , Disfunção Cognitiva/prevenção & controle , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Depressão/etiologia , Depressão/metabolismo , Depressão/prevenção & controle , Hipocampo/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fármacos Neuroprotetores/farmacologia , Transdução de Sinais , Sinaptofisina/metabolismo
12.
J Microbiol Biotechnol ; 32(9): 1154-1167, 2022 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-36039041

RESUMO

In this study, we investigated the anti-amnesic effect of Korean red pine (Pinus densiflora) bark extract (KRPBE) against amyloid beta1-42 (Aß1-42)-induced neurotoxicity. We found that treatment with KRPBE improved the behavioral function in Aß-induced mice, and also boosted the antioxidant system in mice by decreasing malondialdehyde (MDA) content, increasing superoxide dismutase (SOD) activities, and reducing glutathione (GSH) levels. In addition, KRPBE improved the cholinergic system by suppressing reduced acetylcholine (ACh) content while also activating acetylcholinesterase (AChE), regulating the expression of choline acetyltransferase (ChAT), postsynaptic density protein-95 (PSD-95), and synaptophysin. KRPBE also showed an ameliorating effect on cerebral mitochondrial deficit by regulating reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and ATP levels. Moreover, KRPBE modulated the expression levels of neurotoxicity indicators Aß and phosphorylated tau (p-tau) and inflammatory cytokines TNF-α, p-IκB-α, and IL-1ß. Furthermore, we found that KRPBE improved the expression levels of neuronal apoptosis-related markers BAX and BCl-2 and increased the expression levels of BDNF and p-CREB. Therefore, this study suggests that KRPBE treatment has an anti-amnestic effect by modulating cholinergic system dysfunction and neuroinflammation in Aß1-42-induced cognitive impairment in mice.


Assuntos
Disfunção Cognitiva , Fármacos Neuroprotetores , Pinus , Acetilcolina/metabolismo , Acetilcolina/farmacologia , Acetilcolinesterase/metabolismo , Trifosfato de Adenosina/metabolismo , Peptídeos beta-Amiloides , Animais , Antioxidantes/farmacologia , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Colina O-Acetiltransferase/metabolismo , Colina O-Acetiltransferase/farmacologia , Colinérgicos/farmacologia , Disfunção Cognitiva/induzido quimicamente , Disfunção Cognitiva/tratamento farmacológico , Glutationa/metabolismo , Malondialdeído/metabolismo , Malondialdeído/farmacologia , Camundongos , Inibidor de NF-kappaB alfa/metabolismo , Doenças Neuroinflamatórias , Fármacos Neuroprotetores/farmacologia , Estresse Oxidativo , Casca de Planta , Extratos Vegetais/farmacologia , Espécies Reativas de Oxigênio/metabolismo , República da Coreia , Superóxido Dismutase/metabolismo , Sinaptofisina/metabolismo , Sinaptofisina/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Proteína X Associada a bcl-2/metabolismo
13.
J Small Anim Pract ; 63(11): 809-815, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-35986507

RESUMO

OBJECTIVES: Diabetes mellitus is a common condition that requires intensive treatment and markedly impacts the welfare of affected cats. The aim of this study was to identify diabetes mellitus-associated perturbations in the feline pancreatic islet microenvironment. The utility of "clear, unobstructed brain/body imaging cocktails and computational analysis" (CUBIC) for three-dimensional pancreatic analysis was investigated. METHODS: Formalin-fixed paraffin-embedded tissues from cats with diabetes mellitus, or control cats without pancreatic pathology, were retrospectively identified. Immunohistochemistry for synaptophysin and ionised calcium binding adaptor molecule 1, and immunofluorescence for insulin and synaptophysin, were used to assess changes in islets. An image analysis pipeline was developed to analyse images acquired from two-dimensional immunofluorescence. CUBIC was used to optically clear selected pancreas samples before immunofluorescence and deep three-dimensional confocal microscopy. RESULTS: Diabetic cats have a significant reduction in synaptophysin-positive islet area. Whilst islets from diabetic patients have similar numbers of ß cells to islets from control cats, significantly lower intensity of insulin expression can be observed in the former. CUBIC facilitates clear visualisation of pancreatic islets in three dimensions. CLINICAL SIGNIFICANCE: The data presented support the theory that there is a decrease in function of ß cells before their destruction, suggesting a potentially significant step in the pathogenesis of feline diabetes mellitus. In parallel, we demonstrate CUBIC as a valuable new tool to visualise the shape of feline pancreatic islets and to interrogate pathology occurring in the islets of diabetic pets.


Assuntos
Doenças do Gato , Diabetes Mellitus , Ilhotas Pancreáticas , Gatos , Animais , Insulina , Sinaptofisina/metabolismo , Estudos Retrospectivos , Ilhotas Pancreáticas/metabolismo , Ilhotas Pancreáticas/patologia , Diabetes Mellitus/veterinária , Diabetes Mellitus/metabolismo , Diabetes Mellitus/patologia , Doenças do Gato/patologia
14.
J Psychiatr Res ; 154: 224-232, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35961178

RESUMO

BACKGROUND: Single Prolonged Stress (SPS) is a valid animal model that reflects the core of post-traumatic stress disorder (PTSD) phenotypes. Although SPS has been a pivotal tool, it can bring ethics approval difficulties due to the use of ether as a stressor. The present study evaluated if changing a chemical (ether) with a social stressor resembles the PTSD hallmark symptoms. METHODS: Female and male young adult rats were distributed in Sham and Social-SPS groups. Rats in Social-SPS groups were subjected to stress, whereas those in Sham groups remained undisturbed. One set of animals performed the behavioral tests, elevated plus-maze (EPM) and Y-maze. Plasma corticosterone levels and cortical and hippocampal molecular protein contents were analyzed. Another set of animals performed the dexamethasone suppression test. RESULTS: A significant decrease in the percentage of time spent and the number of entries in open arms and an increase in anxiety index in the EPM were observed in rats of the social-SPS groups. In the Social-SPS groups, rats reduced the spontaneous alternations in Y-maze. The Social-SPS exposure enhanced the HPA-axis feedback and increased glucocorticoid receptor contents in the cerebral cortex and hippocampus of rats. A decrease in the content of synaptic integrity-related proteins, synaptophysin, and PSD-95, were found in the cortex and hippocampus of rats subjected to social-SPS. There were no statistical differences between males and females in any parameter analyzed. LIMITATIONS: The absence of a task to recap criterion E 'arousal' and predictive validity experiments. CONCLUSIONS: This study reveals that social-SPS recapitulated the main clusters required for a candidate animal model of PTSD.


Assuntos
Transtornos de Estresse Pós-Traumáticos , Animais , Feminino , Masculino , Ratos , Corticosterona , Dexametasona , Modelos Animais de Doenças , Éter/metabolismo , Hipocampo/metabolismo , Receptores de Glucocorticoides/metabolismo , Transtornos de Estresse Pós-Traumáticos/metabolismo , Sinaptofisina/metabolismo
15.
Neurochem Res ; 47(11): 3402-3413, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36028734

RESUMO

Vascular dementia (VaD) is the second most common form of dementia globally, yet there are no efficient treatments. Naringenin, a natural flavonoid, exerts antioxidative, anti-inflammatory, and neuroprotective properties; however, its potential effect on VaD remain unclear. Herein, the purpose of present study was to elucidate whether naringenin attenuates cognitive dysfunction in VaD via inhibiting hippocampal oxidative stress and inflammatory response, and promoting N-methyl-D-aspartate receptors (NMDARs) signaling pathway. A rat model of VaD was established by permanent bilateral common carotid artery occlusion [2-vessel occlusion (2VO)]. Behavioral performance analyses results revealed that administration of naringenin improves cognitive impairment in rats with VaD according to the new object recognition test and the Morris water maze test. In addition, naringenin attenuated hippocampal oxidative stress by reducing reactive oxygen species generation, decreasing malondialdehyde content and recombinant reactive oxygen species modulator 1 (Romo-1) expression, and increasing superoxide dismutase and glutathione peroxidase activities in the hippocampus of VaD rats. Moreover, naringenin decreased the proinflammatory cytokines (IL-1ß, IL-6, and TNF-α) levels and increased the anti-inflammatory cytokines (IL-10 and IL-4) levels in the hippocampus of 2VO surgery-treated rats, attenuating hippocampal inflammatory response during VaD. Furthermore, naringenin promoted synaptophysin (SYP), postsynaptic density protein 95 (PSD95), N-methyl-Daspartic acid receptor 1 (NR1) and N-methyl-D-aspartate receptor subunit 2B (NR2B) expressions levels in hippocampus of VaD rats. Collectively, these findings indicated that naringenin mitigates cognitive impairment in VaD rats partly via inhibiting hippocampal oxidative stress and inflammatory response and restoring NMDARs signaling pathway.


Assuntos
Disfunção Cognitiva , Demência Vascular , Flavanonas , Animais , Anti-Inflamatórios/farmacologia , Ácido Aspártico/metabolismo , Disfunção Cognitiva/metabolismo , Demência Vascular/tratamento farmacológico , Modelos Animais de Doenças , Proteína 4 Homóloga a Disks-Large/metabolismo , Flavanonas/farmacologia , Flavanonas/uso terapêutico , Glutationa Peroxidase/metabolismo , Hipocampo/metabolismo , Interleucina-10/metabolismo , Interleucina-4/metabolismo , Interleucina-4/farmacologia , Interleucina-4/uso terapêutico , Interleucina-6/metabolismo , Malondialdeído/metabolismo , Aprendizagem em Labirinto , Estresse Oxidativo , Ratos , Espécies Reativas de Oxigênio/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Transdução de Sinais , Superóxido Dismutase/metabolismo , Sinaptofisina/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
16.
Tissue Eng Regen Med ; 19(5): 1063-1075, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35857260

RESUMO

BACKGROUND: Mesenchymal stem cells (MSCs) are considered a potential tool for regenerating damaged tissues due to their great multipotency into various cell types. Here, we attempted to find the appropriate conditions for neuronal differentiation of tonsil-derived MSCs (TMSCs) and expand the potential application of TMSCs for treating neurological diseases. METHODS: The TMSCs were differentiated in DMEM/F-12 (Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12) supplemented with various neurotrophic factors for 7-28 days to determine the optimal neuronal differentiation condition for the TMSCs. The morphologies as well as the levels of the neural markers and neurotransmitters were assessed to determine neuronal differentiation potentials and the neuronal lineages of the differentiated TMSCs. RESULTS: Our initial study demonstrated that DMEM/F12 supplemented with 50 ng/mL basic fibroblast growth factor with 10 µM forskolin was the optimal condition for neuronal differentiation for the TMSCs. TMSCs had higher protein expression of neuronal markers, including neuron-specific enolase (NSE), GAP43, postsynaptic density protein 95 (PSD95), and synaptosomal-associated protein of 25 kDa (SNAP25) compared to the undifferentiated TMSCs. Immunofluorescence staining also validated the increased mature neuron markers, NeuN and synaptophysin, in the differentiated TMSCs. The expression of glial fibrillar acidic protein and ionized calcium-binding adaptor molecule 1 the markers of astrocytes and microglia, were also slightly increased. Additionally, the differentiated TMSCs released a significantly higher level of acetylcholine, the cholinergic neurotransmitter, as analyzed by the liquid chromatography-tandem mass spectrometry and showed an enhanced choline acetyltransferase immunoreactivity compared to the undifferentiated cells. CONCLUSION: Our study suggests that the optimized condition favors the TMSCs to differentiate into cholinergic neuron-like phenotype, which could be used as a possible therapeutic tool in treating certain neurological disorders such as Alzheimer's disease.


Assuntos
Células-Tronco Mesenquimais , Tonsila Palatina , Acetilcolina/metabolismo , Cálcio/metabolismo , Colina O-Acetiltransferase/metabolismo , Colinérgicos/metabolismo , Colforsina/metabolismo , Proteína 4 Homóloga a Disks-Large/metabolismo , Fator 2 de Crescimento de Fibroblastos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Fatores de Crescimento Neural/metabolismo , Fosfopiruvato Hidratase/metabolismo , Sinaptofisina/metabolismo
17.
STAR Protoc ; 3(3): 101495, 2022 09 16.
Artigo em Inglês | MEDLINE | ID: mdl-35776639

RESUMO

Following the release of neurotransmitters at synaptic vesicles via exocytosis, endocytosis is initiated to retrieve vesicles that have fused with the plasma membrane of nerve terminals and recycle them, thus sustaining synaptic transmission. Here, we describe imaging-based protocols for quantitative measurements of endocytosis at cultured synapses. These protocols include (1) primary culture of mouse hippocampal neurons, (2) studying endocytosis at neurons transfected with a pH-sensitive synaptophysin-pHluorin2× using fluorescent microscopy, and (3) imaging endocytosis at fixed neurons with electron microscopy. For complete details on the use and execution of this protocol, please refer to Wu et al. (2016) and Wu et al. (2021).


Assuntos
Elétrons , Vesículas Sinápticas , Animais , Endocitose/fisiologia , Hipocampo , Camundongos , Microscopia Eletrônica , Neurotransmissores/metabolismo , Vesículas Sinápticas/metabolismo , Sinaptofisina/metabolismo
18.
Hum Pathol ; 127: 102-111, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35690220

RESUMO

INSM1, ASCL1, and POU2F3 are novel transcription factors involved in neuroendocrine (NE) differentiation of neoplasms in several organs, but data on their expression in breast carcinomas (BCs) are limited. We retrospectively evaluated the expression of these markers in a series of 97 BCs (58 with NE morphology and 39 with otherwise uncommon morphology) tested prospectively using immunohistochemistry (IHC). Nuclear staining in >50% of the cells was used as the positive cut-off. Thirty-two of the 97 BCs (33%) were INSM1-positive. INSM1-positivity correlated significantly with histologic type and presence of stromal mucin. INSM1 also correlated with synaptophysin and chromogranin, established markers of NE differentiation (P < .0001 and P = .0023, respectively). In BC with NE morphology, the expression of INSM1 supported NE differentiation, and INSM1 was more specific than synaptophysin and more sensitive and specific than chromogranin. INSM1 was the most expressed NE marker in 17 BCs. INSM1-positive BCs included 56% of solid papillary BCs, 88% of BCs with solid papillary features, and 75% of high-grade NE carcinomas. Of 35 BCs tested for POU2F3 and ASCL1, only 1 and 4 cases were positive, respectively. Our results show that INSM1 is a sensitive marker of NE differentiation in BC and should be included with synaptophysin and chromogranin in the IHC panel used to evaluate NE differentiation in BC with NE morphology. ASCL1 and POU2F3 are uncommon in BC and their routine assessment is not warranted.


Assuntos
Neoplasias da Mama , Carcinoma Neuroendócrino , Fatores de Transcrição Hélice-Alça-Hélice Básicos , Biomarcadores Tumorais/metabolismo , Carcinoma Neuroendócrino/patologia , Cromograninas , Feminino , Humanos , Mucinas , Fatores de Transcrição de Octâmero , Proteínas Repressoras/metabolismo , Estudos Retrospectivos , Sinaptofisina/metabolismo , Fatores de Transcrição
19.
Am J Dermatopathol ; 44(10): 718-727, 2022 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-35642978

RESUMO

ABSTRACT: This study sought to reveal the clinicopathologic characteristics of large cell neuroendocrine carcinoma (LCNEC) of the skin/conjunctiva. The retrieved patients included 3 men and 3 women with a median age of 85 (63-95) years. All lesions occurred on the face, including the ears, with a median tumor size of 11.5 (7-65) mm. Lymph node metastasis was observed in 5 (83%) of 6 cases, and distant metastasis was noted in 2 (33%). One patient (17%) who had a 13-mm-sized tumor died of the tumor 13 months after excision. All tumors were mainly located in the dermis, and one of them also exhibited intraepithelial spreading. The cytology resembled that of an LCNEC in other organs. No adnexal differentiation was observed. Five cases were of the pure type, but one had a component of squamous cell carcinoma. Immunoreactivities for CAM5.2, CK7, CK19, BerEP4, epithelial membrane antigen, neuron-specific enolase, synaptophysin, c-KIT, GATA3, and bcl-2 were frequently present, but CK20, neurofilament, Merkel cell polyomavirus large T antigen, mammaglobin, estrogen receptor, HER2, and TTF1 were completely negative in all cases. Mutant-pattern immunostaining of p53, PTEN, and Rb was frequently observed. The Ki67 rate exceeded 70% in all cases. LCNEC of the skin/conjunctiva is a morphologically-defined group of primary cutaneous/conjunctival neuroendocrine neoplasm, although it may be heterogeneous similar to other-site LCNEC or Merkel cell carcinoma. This study highlighted the predominant location for the face, high metastatic and lethal potential, possible combination with other tumor components, and frequent mutant-type immunoexpressions of p53, PTEN, and Rb in this tumor group.


Assuntos
Carcinoma de Célula de Merkel , Carcinoma Neuroendócrino , Carcinoma de Células Escamosas , Neoplasias Cutâneas , Idoso de 80 Anos ou mais , Antígenos Virais de Tumores , Biomarcadores Tumorais/metabolismo , Carcinoma de Célula de Merkel/patologia , Carcinoma Neuroendócrino/patologia , Túnica Conjuntiva/metabolismo , Túnica Conjuntiva/patologia , Feminino , Humanos , Imuno-Histoquímica , Antígeno Ki-67/metabolismo , Masculino , Mucina-1/metabolismo , Fosfopiruvato Hidratase/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Receptores de Estrogênio , Neoplasias Cutâneas/patologia , Sinaptofisina/metabolismo , Proteína Supressora de Tumor p53
20.
Mol Neurobiol ; 59(7): 4488-4500, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35575872

RESUMO

The pathological characteristics of Alzheimer's disease (AD) include formation of senile plaques resulting from amyloid-ß (Aß) deposition and neurofibrillary tangles caused by tau hyperphosphorylation. Reducing tau hyperphosphorylation is crucial for treatment of AD. Network pharmacology analysis showed that CTS may reduce tau hyperphosphorylation by regulating the phosphatidylinositol 3 kinases/protein kinase B/ glycogen synthase kinase-3ß (PI3K/Akt/GSK3ß) pathway. We investigated the ability of cryptotanshinone (CTS) to reduce Aß-induced tau hyperphosphorylation and characterized the underlying mechanisms. Amyloid-ß42 oligomers (AßO) were used to establish an AD model in HT22 cells. The expression levels of tau and related proteins in PI3K/Akt/GSK3ß pathway were measured using western blot and immunofluorescence staining. The above-mentioned proteins were then evaluated in an okadaic acid (OKA)-induced AD cell model to verify the results. Synapse-associated proteins including post-synaptic density protein-95 (PSD95) and synaptophysin were also evaluated. We found that CTS significantly reduced tau hyperphosphorylation at Ser202, Ser404, Thr181, and Thr231 in AßO- and OKA-induced cell models. Moreover, we also found that CTS reversed AßO-induced reductions in the levels of PSD95 and synaptophysin. We used LY294002 to block PI3K and the results showed that CTS exerted neuroprotective effects through regulation of the PI3K/Akt/GSK3ß signaling pathway. In summary, we showed for the first time that CTS inhibited AD-related tau hyperphosphorylation and reduced the effects of AßO on the expression levels of PSD95 and synaptophysin via the PI3K/Akt/GSK3ß pathway in HT22 cells.


Assuntos
Doença de Alzheimer , Fosfatidilinositol 3-Quinases , Doença de Alzheimer/tratamento farmacológico , Doença de Alzheimer/patologia , Peptídeos beta-Amiloides/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , Humanos , Ácido Okadáico/farmacologia , Fenantrenos , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Sinaptofisina/metabolismo , Proteínas tau/metabolismo
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