RESUMEN
Methamphetamine (METH), an amphetamine derivate, may increase the risk of developing Parkinson's disease (PD). Human and animal studies have shown that METH produces persistent dopaminergic neurotoxicity in the nigrostriatal pathway, despite initial partial recovery. To determine the processes leading to early compensation, we studied the detailed morphology and distribution of tyrosine hydroxylase immunoreactive fibers (TH-ir) classified by their thickness (types I-IV) before and after METH. Applying three established neurotoxic regimens of METH: single high dose (1 × 30 mg/kg), multiple lower doses (3 × 5 mg/kg) or (3 × 10 mg/kg), we show that METH primarily damages type I fibers (the thinner ones), and to a much lesser extend types II-IV fibers including sterile axons. The striatal TH terminal partial recovery process, consisting of a progressive regrowth increases in types II, III, and IV fibers, demonstrated by co-localization of GAP-43, a sprouting marker, was observed 3 days post-METH treatment. In addition, we demonstrate the presence of growth-cone-like TH-ir structures, indicative of new terminal generation as well as improvement in motor functions after 3 days. A temporal relationship was observed between decreases in TH-expression and increases in silver staining, a marker of degeneration. Striatal regeneration was associated with an increase in astroglia and decrease in microglia expression, suggesting a possible role for the neuroimmune system in regenerative processes. Identification of regenerative compensatory mechanisms in response to neurotoxic agents could point to novel mechanisms in countering the neurotoxicity and/or enhancing the regenerative processes.
Asunto(s)
Estimulantes del Sistema Nervioso Central/toxicidad , Cuerpo Estriado/fisiopatología , Dopamina/metabolismo , Metanfetamina/toxicidad , Enfermedades Neurodegenerativas/inducido químicamente , Enfermedades Neurodegenerativas/patología , Animales , Proteínas de Unión al Calcio , Cuerpo Estriado/efectos de los fármacos , Cuerpo Estriado/ultraestructura , Modelos Animales de Enfermedad , Proteína GAP-43/metabolismo , Proteína Ácida Fibrilar de la Glía/metabolismo , Locomoción/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Proteínas de Microfilamentos , Terminales Presinápticos/efectos de los fármacos , Terminales Presinápticos/patología , Terminales Presinápticos/ultraestructura , Trastornos Psicomotores/etiología , Recuperación de la Función/efectos de los fármacos , Recuperación de la Función/fisiología , Tinción con Nitrato de Plata , Factores de Tiempo , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
Alzheimer's disease (AD) is multifactorial with unclear etiopathology. Due to the complexity of AD, many attempted single therapy treatments, like Aß immunization, have generally failed. Therefore, there is a need for drugs with multiple benefits. Naturally occurring phytochemicals with neuroprotective, anti-amyloidogenic, antioxidative, and anti-inflammatory properties could be a possible way out. In this study, the effect of Moringa oleifera (MO), a naturally occurring plant with high antioxidative, anti-inflammatory, and neuroprotective effects, was evaluated on hyperhomocysteinemia (HHcy) induced AD-like pathology in rats. Homocysteine (Hcy) injection for 14 days was used to induce AD-like pathology. Simultaneous MO extract gavage followed the injection as a preventive treatment or, after injection completion, MO gavage was performed for another 14 days as a curative treatment. MO was found to not only prevent but also rescue the oxidative stress and cognitive impairments induced by Hcy treatment. Moreover, MO recovered the decreased synaptic proteins PSD93, PSD95, Synapsin 1 and Synaptophysin, and improved neurodegeneration. Interestingly, MO decreased the Hyc-induced tau hyperphosphorylation at different sites including S-199, T-231, S-396, and S-404, and at the same time decreased Aß production through downregulation of BACE1. These effects in HHcy rats were accompanied by a decrease in calpain activity under MO treatment, supporting that calpain activation might be involved in AD pathogenesis in HHcy rats. Taken together, our data, for the first time, provided evidence that MO alleviates tau hyperphosphorylation and Aß pathology in a HHcy AD rat model. This and previous other studies support MO as a good candidate for, and could provide new insights into, the treatment of AD and other tauopathies.
Asunto(s)
Enfermedad de Alzheimer/tratamiento farmacológico , Enfermedad de Alzheimer/patología , Trastornos del Conocimiento , Homocisteína/toxicidad , Moringa oleifera , Extractos Vegetales/uso terapéutico , Enfermedad de Alzheimer/inducido químicamente , Secretasas de la Proteína Precursora del Amiloide/metabolismo , Péptidos beta-Amiloides/metabolismo , Precursor de Proteína beta-Amiloide/metabolismo , Animales , Ácido Aspártico Endopeptidasas/metabolismo , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Trastornos del Conocimiento/inducido químicamente , Trastornos del Conocimiento/tratamiento farmacológico , Modelos Animales de Enfermedad , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Proteínas del Tejido Nervioso/metabolismo , Neuronas/patología , Neuronas/ultraestructura , Estrés Oxidativo/efectos de los fármacos , Fragmentos de Péptidos/metabolismo , Fosforilación/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Tinción con Nitrato de Plata , Proteínas tau/metabolismoRESUMEN
BACKGROUND: Thalamic dysfunction has been implicated in overall chronic neurological dysfunction after traumatic brain injury (TBI), however little is known about the underlying histopathology. In experimental diffuse TBI (dTBI), we hypothesize that persisting histopathological changes in the ventral posteromedial (VPM) nucleus of the thalamus is indicative of progressive circuit reorganization. Since circuit reorganization in the VPM impacts the whisker sensory system, the histopathology could explain the development of hypersensitivity to whisker stimulation by 28days post-injury; similar to light and sound hypersensitivity in human TBI survivors. METHODS: Adult, male Sprague-Dawley rats underwent craniotomy and midline fluid percussion injury (FPI) (moderate severity; 1.8-2.0atm) or sham surgery. At 1d, 7d, and 28days post-FPI (d FPI) separate experiments confirmed the cytoarchitecture (Giemsa stain) and evaluated neuropathology (silver stain), activated astrocytes (GFAP), neuron morphology (Golgi stain) and microglial morphology (Iba-1) in the VPM. RESULTS: Cytoarchitecture was unchanged throughout the time course, similar to previously published data; however, neuropathology and astrocyte activation were significantly increased at 7d and 28d and activated microglia were present at all time points. Neuron morphology was dynamic over the time course with decreased dendritic complexity (fewer branch points; decreased length of processes) at 7d FPI and return to sham values by 28d FPI. CONCLUSIONS: These data indicate that dTBI results in persisting thalamic histopathology out to a chronic time point. While these changes can be indicative of either adaptive (recovery) or maladaptive (neurological dysfunction) circuit reorganization, they also provide a potential mechanism by which maladaptive circuit reorganization could contribute to the development of chronic neurological dysfunction. Understanding the processes that mediate circuit reorganization is critical to the development of future therapies for TBI patients.
Asunto(s)
Lesiones Traumáticas del Encéfalo/patología , Tálamo/lesiones , Tálamo/patología , Enfermedad Aguda , Animales , Astrocitos/patología , Astrocitos/fisiología , Lesiones Traumáticas del Encéfalo/fisiopatología , Enfermedad Crónica , Modelos Animales de Enfermedad , Imagenología Tridimensional , Inmunohistoquímica , Masculino , Microglía/patología , Microglía/fisiología , Plasticidad Neuronal , Neuronas/patología , Neuronas/fisiología , Ratas Sprague-Dawley , Tinción con Nitrato de Plata , Tálamo/fisiopatología , Factores de TiempoRESUMEN
Objetivou-se com este estudo avaliar o processo de cicatrização de feridas de coelhos tratadas com extrato de barbatimão (S. adstringens) associado a células mononucleares autólogas da medula óssea (CMMO). Utilizaram-se 20 coelhos, distribuídos em quatro grupos: B, extrato de barbatimão; CB, CMMO com extrato de barbatimão; CS, CMMO com solução fisiológica; S, solução fisiológica. Foi avaliada a presença de crosta, hiperemia, secreção, hemorragia, reepitelização, área da ferida e tempo de cicatrização. No terceiro, sétimo, 14º e 21º dias pós-operatório, realizou-se a biópsia das feridas e avaliaram-se os indicadores dos processos de inflamação e de reparo, com destaque para o colágeno, na coloração picrosírius, bem como de proliferação celular, na coloração AgNOR. Houve maior deposição de fibras colágenas nos grupos B e CB (P=0,00003) e formação de crostas mais espessas no sétimo dia, com fibras colágenas mais organizadas no 21º dia. Conclui-se que o barbatimão estimula a produção de fibras colágenas e promove a formação de crostas mais espessas sobre a ferida na fase inicial da cicatrização e, na fase de remodelação, favorece a orientação das fibras colágenas. Além disso, a associação desse fitoterápico com CMMO não estimula a cicatrização de feridas.(AU)
This study aimed to evaluate the healing process of wounds of rabbits in response to treatment with barbatiman extract (S. adstringens) associated with autologous mononuclear bone marrow cells (BM-MNC). We used 20 rabbits, divided into four groups: B, 10% barbatiman extract with 9.48% of total tannins; CB, BM-MNC with barbatiman extract; CS, BM-MNC with NaCl 0.9% solution; S, NaCl 0.9% solution. Clinical evaluation was performed by observing the presence of crust, redness, discharge, bleeding, re-epithelialization, the wound area and healing time in days. In the third, seventh, 14th and 21st postoperative days wounds were biopsied for microscopic evaluation of inflammation and repair process indicators, especially collagen, in picrosirius staining, and cell proliferation, in AgNOR staining. There was a greater deposition of collagen fibers in groups B and CB (p=0.00003) on the seventh day and formation of thicker crusts, and more organized collagen fibers on the 21st day in these groups. In conclusion, in the initial phase of healing, barbatiman extract stimulates the production of collagen fibers and promotes the formation of more exuberant crusts on the wounds and remodeling phase favors the orientation of collagen fibers, but when combined with BMMC does not stimulate wound healing in rabbits.(AU)
Asunto(s)
Animales , Conejos , Tratamiento Basado en Trasplante de Células y Tejidos/veterinaria , Colágeno/análisis , Fabaceae , Cicatrización de Heridas , Tinción con Nitrato de Plata/veterinariaRESUMEN
OBJECTIVES: We investigated whether Ginkgo biloba extract (EGb761) can provide neuroprotective effects and enhance the efficacy of bone marrow-derived mesenchymal stem cells (BMSCs) in a rat model of experimental autoimmune encephalomyelitis (EAE). METHODS: We examined the synergistic action of BMSCs combined with EGb761 treatment in EAE rats. The immunized rats received an intravenous injection of BMSCs or intraperitoneal administration of EGb761 or both on the day of the onset of clinical symptoms and for the following 21 days. Clinical severity scores were recorded daily and histopathological examination of the spinal cord and cytokine concentrations in the serum were studied on days 14 and 31 postimmunization. RESULTS: Our results showed that combined treatment with BMSCs and EGb761 further decreased the disease severity, maximal clinical score and number of infiltrated mononuclear cells, especially CD3-positive T cells. We observed that the demyelination score and the density of axonal loss in the spinal cord were significantly reduced in mice receiving the combination therapy. The serum concentrations of the phosphorylated neurofilament heavy chain, tumor necrosis factor-α and interferon-γ were reduced in the combination-treatment group. CONCLUSION: Our results suggest that combined treatment with BMSCs and EGb761 have a synergistic effect in rats with EAE by inhibiting the secretion of proinflammatory cytokines, demyelination and protecting axons and neurons.
Asunto(s)
Encefalomielitis Autoinmune Experimental/terapia , Trasplante de Células Madre Mesenquimatosas/métodos , Células Madre Mesenquimatosas/fisiología , Fármacos Neuroprotectores/uso terapéutico , Extractos Vegetales/uso terapéutico , Animales , Complejo CD3/metabolismo , Movimiento Celular/efectos de los fármacos , Citocinas/antagonistas & inhibidores , Citocinas/metabolismo , Modelos Animales de Enfermedad , Encefalomielitis Autoinmune Experimental/patología , Femenino , Ginkgo biloba , Células Madre Mesenquimatosas/efectos de los fármacos , Ratas , Ratas Wistar , Índice de Severidad de la Enfermedad , Tinción con Nitrato de Plata , Médula Espinal/efectos de los fármacos , Médula Espinal/patología , Médula Espinal/ultraestructura , Estadísticas no ParamétricasRESUMEN
Dietary n-3 polyunsaturated fatty acids (PUFAs) are critical components of inflammatory response and memory impairment. However, the mechanisms underlying the sensitizing effects of low n-3 PUFAs in the brain for the development of memory impairment following inflammation are still poorly understood. In this study, we examined how a 2-month n-3 PUFAs deficiency from pre-puberty to adulthood could increase vulnerability to the effect of inflammatory event on spatial memory in mice. Mice were given diets balanced or deficient in n-3 PUFAs for a 2-month period starting at post-natal day 21, followed by a peripheral administration of lipopolysaccharide (LPS), a bacterial endotoxin, at adulthood. We first showed that spatial memory performance was altered after LPS challenge only in n-3 PUFA-deficient mice that displayed lower n-3/n-6 PUFA ratio in the hippocampus. Importantly, long-term depression (LTD), but not long-term potentiation (LTP) was impaired in the hippocampus of LPS-treated n-3 PUFA-deficient mice. Proinflammatory cytokine levels were increased in the plasma of both n-3 PUFA-deficient and n-3 PUFA-balanced mice. However, only n-3 PUFA-balanced mice showed an increase in cytokine expression in the hippocampus in response to LPS. In addition, n-3 PUFA-deficient mice displayed higher glucocorticoid levels in response to LPS as compared with n-3 PUFA-balanced mice. These results indicate a role for n-3 PUFA imbalance in the sensitization of the hippocampal synaptic plasticity to inflammatory stimuli, which is likely to contribute to spatial memory impairment.
Asunto(s)
Ácidos Grasos Omega-3/metabolismo , Inflamación/complicaciones , Trastornos de la Memoria/etiología , Animales , Animales Recién Nacidos , Corticosterona/sangre , Citocinas/sangre , Citocinas/genética , Espinas Dendríticas/efectos de los fármacos , Espinas Dendríticas/patología , Espinas Dendríticas/ultraestructura , Modelos Animales de Enfermedad , Ácidos Grasos Omega-3/administración & dosificación , Regulación de la Expresión Génica/efectos de los fármacos , Hipocampo/metabolismo , Hipocampo/patología , Hipocampo/fisiopatología , Inflamación/sangre , Inflamación/inducido químicamente , Lipopolisacáridos/toxicidad , Depresión Sináptica a Largo Plazo/efectos de los fármacos , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Aprendizaje por Laberinto/fisiología , Trastornos de la Memoria/patología , Ratones , Ratones Endogámicos C57BL , Neuronas/patología , Neuronas/fisiología , Neuronas/ultraestructura , Técnicas de Placa-Clamp , Tinción con Nitrato de PlataRESUMEN
Alzheimer's disease (AD) is a progressive neurodegenerative disorder, characterized by irreversible decline of mental faculties, emotional and behavioral changes, loss of motor skills, and dysfunction of autonomic nervous system and disruption of circadian rhythms (CRs). We attempted to describe the morphological findings of the hypothalamus in early cases of AD, focusing our study mostly on the suprachiasmatic nucleus (SCN), the supraoptic nucleus (SON), and the paraventricular nucleus (PVN). Samples were processed for electron microscopy and silver impregnation techniques. The hypothalamic nuclei demonstrated a substantial decrease in the neuronal population, which was particularly prominent in the SCN. Marked abbreviation of dendritic arborization, in association with spinal pathology, was also seen. The SON and PVN demonstrated a substantial number of dystrophic axons and abnormal spines. Alzheimer's pathology, such as deposits of amyloid-ß peptide and neurofibrillary degeneration, was minimal. Electron microscopy revealed mitochondrial alterations in the cell body and the dendritic branches. The morphological alterations of the hypothalamic nuclei in early cases of AD may be related to the gradual alteration of CRs and the instability of autonomic regulation.
Asunto(s)
Enfermedad de Alzheimer/patología , Neuronas/ultraestructura , Núcleo Hipotalámico Paraventricular/ultraestructura , Núcleo Supraquiasmático/ultraestructura , Núcleo Supraóptico/ultraestructura , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Espinas Dendríticas/ultraestructura , Femenino , Aparato de Golgi/ultraestructura , Humanos , Hipotálamo/ultraestructura , Masculino , Microscopía Electrónica , Persona de Mediana Edad , Mitocondrias/ultraestructura , Tinción con Nitrato de PlataRESUMEN
Alzheimer's disease (AD) has multiple etiopathogenic factors, yet the definitive cause remains unclear and the therapeutic strategies have been elusive. Combination therapy, as one of the promising treatments, has been studied for years and may exert synergistic beneficial effects on AD through polytherapeutic targets. In this study, we tested the effects of a synthesized juxtaposition (named SCR1693) composed of an acetylcholinesterase inhibitor (AChEI) and a calcium channel blocker (CCB) on the hyperhomocysteinemia (HHcy)-induced AD rat model, and found that SCR1693 remarkably improved the HHcy-induced memory deficits and preserved dendrite morphologies as well as spine density by upregulating synapse-associated proteins PSD95 and synapsin-1. In addition, SCR1693 attenuated HHcy-induced tau hyperphosphorylation at multiple AD-associated sites by regulating the activity of protein phosphatase-2A and glycogen synthase kinase-3ß. Furthermore, SCR1693 was more effective than individual administration of both donepezil and nilvadipine which were used as AChEI and CCB, respectively, in the clinical practice. In conclusion, our data suggest that the polytherapeutic targeting juxtaposition SCR1693 (AChEI-CCB) is a promising therapeutic candidate for AD.
Asunto(s)
Encefalopatías , Bloqueadores de los Canales de Calcio/uso terapéutico , Inhibidores de la Colinesterasa/uso terapéutico , Hiperhomocisteinemia/complicaciones , Trastornos de la Memoria/tratamiento farmacológico , Trastornos de la Memoria/etiología , Animales , Encefalopatías/tratamiento farmacológico , Encefalopatías/etiología , Encefalopatías/patología , Modelos Animales de Enfermedad , Donepezilo , Quimioterapia Combinada , Hipocampo/efectos de los fármacos , Hipocampo/patología , Hipocampo/ultraestructura , Homocisteína/toxicidad , Hiperhomocisteinemia/inducido químicamente , Indanos/uso terapéutico , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Proteínas del Tejido Nervioso/metabolismo , Nifedipino/análogos & derivados , Nifedipino/uso terapéutico , Piperidinas/uso terapéutico , Corteza Prefrontal/efectos de los fármacos , Corteza Prefrontal/patología , Corteza Prefrontal/ultraestructura , Ratas , Ratas Sprague-Dawley , Tinción con Nitrato de Plata , Tacrina/análogos & derivados , Tacrina/farmacología , Tacrina/uso terapéuticoRESUMEN
The present study examined the potential application of Juglans mandshurica Maxim extracts (HT) for cancer therapy by assessing their antiproliferative activity, reduction of telomerase activity, induction of apoptosis and cell cycle arrest in S phase in HeLa cells. From the perspective of using HT as a herbal medicine, photomicroscopy and florescent microscopy techniques were utilized to characterize the effect of the extracts on telomerase activity and cell morphology. Flow cytometry was employed to study apoptosis and cell cycle of HeLa cells, and DNA laddering was performed. The results showed that HT inhibited cell proliferation and telomerase activity, induced apoptosis and caused S phase arrest of HeLa cells in vitro. HT inhibited HeLa cell proliferation significantly, and the highest inhibition rate was 83.7%. A trapsilver staining assay showed that HT was capable of markedly decreasing telomerase activity of HeLa cells and this inhibition was enhanced in a time and dosedependent manner. Results of a Hoechst 33258 staining assay showed that HeLa cells treated by HT induced cell death. Through DNA agarose gel electrophoresis, DNA ladders of HeLa cells treated with HT were observed, indicating apoptosis. In conclusion, the present study demonstrated that HT exhibited antitumor effects comprising the inhibition of growth and telomerase activity as well as apoptosis and cell cycle arrest in HeLa cells.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Juglans/química , Extractos Vegetales/farmacología , Antineoplásicos Fitogénicos/química , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , ADN de Neoplasias/metabolismo , Electroforesis en Gel de Agar , Fluorescencia , Células HeLa , Humanos , Naftoquinonas/química , Naftoquinonas/farmacología , Necrosis , Tinción con Nitrato de Plata , Coloración y Etiquetado , Telomerasa/metabolismoRESUMEN
Epigallocatechin gallate (EGCG), the major flavonoid in green tea, is consumed via tea products and dietary supplements, and has been tested in clinical trials. However, EGCG can cause hepatotoxicity in humans and animals by unknown mechanisms. Here EGCG effects on rat liver mitochondria were examined. EGCG showed negligible effects on oxidative phosphorylation at 7.5-100µM in normal mitochondria. However, respiratory chain complexes (RCCs) were profoundly inhibited by EGCG in mitochondria undergoing Ca(2+) overload-induced mitochondrial permeability transition (MPT). As RCCs are located in mitochondrial inner membranes (IM) and matrix, it was reasoned that EGCG could not readily pass through IM to affect RCCs in normal mitochondria but may do so when IM integrity is compromised. This speculation was substantiated in three ways. (1) Purified EGCG-bound proteins were barely detectable in normal mitochondria and contained no RCCs as determined by Western blotting, but swelling mitochondria contained about 1.5-fold more EGCG-bound proteins which included four RCC subunits together with cyclophilin D that locates in mitochondrial matrix. (2) Swelling mitochondria consumed more EGCG than normal ones. (3) The MPT blocker cyclosporine A diminished the above-mentioned difference. Among four subunits of RCC II, only SDHA and SDHB which locate in mitochondrial matrix, but not SDHC or SDHD which insert into the IM, were found to be EGCG targets. Interestingly, EGCG promoted Ca(2+) overload-induced MPT only when moderate MPT already commenced. This study identified hepatic RCCs as targets for EGCG in swelling but not normal mitochondria, suggesting EGCG may trigger hepatotoxicity by worsening pre-existing mitochondria abnormalities.
Asunto(s)
Catequina/análogos & derivados , Proteínas del Complejo de Cadena de Transporte de Electrón/antagonistas & inhibidores , Mitocondrias Hepáticas/efectos de los fármacos , Mitocondrias Hepáticas/metabolismo , Dilatación Mitocondrial/efectos de los fármacos , Té/química , Animales , Western Blotting , Catequina/farmacología , Proteínas del Complejo de Cadena de Transporte de Electrón/metabolismo , Electroforesis en Gel de Poliacrilamida , Masculino , Proteínas de Transporte de Membrana Mitocondrial/metabolismo , Poro de Transición de la Permeabilidad Mitocondrial , Fosforilación Oxidativa/efectos de los fármacos , Unión Proteica/efectos de los fármacos , Subunidades de Proteína/metabolismo , Ratas , Ratas Sprague-Dawley , Tinción con Nitrato de PlataRESUMEN
Endothelial cells are highly sensitive to high doses of ionizing radiation and the cellular response leads to acute damage of the endothelium. This chapter describes how to measure the effects of ionizing radiation on the proteome of endothelial cells, here showing analysis at 4 and 24 h after exposure. Two complementary proteomic strategies, namely "stable isotope labeling by amino acids in cell culture" (SILAC) and 2D-DIGE analysis are used. In the example given, the exposure triggers considerable alterations in the endothelial protein expression with deregulated proteins categorized into four key pathways: (1) glycolysis/gluconeogenesis, (2) oxidative phosphorylation, (3) Rho-mediated cell motility, and (4) non-homologous end joining (NHEJ). After exposure to high-dose radiation, an immediate down-regulation is seen in the Ku70/Ku80 heterodimer and proliferating cell nuclear antigen (PCNA) proteins belonging to the NHEJ DNA repair pathway. Later time points show significant decrease in the expression levels of proteins of the oxidative phosphorylation (OXPHOS) pathway along with a significant expression increase in the enzymes of the glycolytic pathway. The methods to reproduce our analysis are presented here.
Asunto(s)
Proteómica , Aminoácidos/metabolismo , Animales , Técnicas de Cultivo de Célula , Línea Celular , Medios de Cultivo , Humanos , Marcaje Isotópico , Tinción con Nitrato de Plata , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Espectrometría de Masas en Tándem , Electroforesis Bidimensional Diferencial en GelRESUMEN
This study addresses the relationship between cochlear oxidative damage and auditory cortical injury in a rat model of repeated noise exposure. To test the effect of increased antioxidant defenses, a water-soluble coenzyme Q10 analog (Qter) was used. We analyzed auditory function, cochlear oxidative stress, morphological alterations in auditory cortices and cochlear structures, and levels of coenzymes Q9 and Q10 (CoQ9 and CoQ10, respectively) as indicators of endogenous antioxidant capability. We report three main results. First, hearing loss and damage in hair cells and spiral ganglion was determined by noise-induced oxidative stress. Second, the acoustic trauma altered dendritic morphology and decreased spine number of II-III and V-VI layer pyramidal neurons of auditory cortices. Third, the systemic administration of the water-soluble CoQ10 analog reduced oxidative-induced cochlear damage, hearing loss, and cortical dendritic injury. Furthermore, cochlear levels of CoQ9 and CoQ10 content increased. These findings indicate that antioxidant treatment restores auditory cortical neuronal morphology and hearing function by reducing the noise-induced redox imbalance in the cochlea and the deafferentation effects upstream the acoustic pathway.
Asunto(s)
Cóclea/patología , Pérdida Auditiva Provocada por Ruido , Estrés Oxidativo/fisiología , Ubiquinona/uso terapéutico , Corteza Visual/patología , Fascículo Atrioventricular Accesorio , Estimulación Acústica , Aldehídos/metabolismo , Análisis de Varianza , Animales , Antioxidantes/metabolismo , Antioxidantes/farmacología , Antioxidantes/uso terapéutico , Vías Auditivas/efectos de los fármacos , Vías Auditivas/patología , Vías Auditivas/ultraestructura , Lesiones Encefálicas/tratamiento farmacológico , Lesiones Encefálicas/etiología , Lesiones Encefálicas/patología , Cóclea/fisiopatología , Modelos Animales de Enfermedad , Etidio/análogos & derivados , Etidio/metabolismo , Potenciales Evocados Auditivos del Tronco Encefálico/efectos de los fármacos , Potenciales Evocados Auditivos del Tronco Encefálico/fisiología , Células Ciliadas Auditivas/patología , Células Ciliadas Auditivas/ultraestructura , Pérdida Auditiva Provocada por Ruido/complicaciones , Pérdida Auditiva Provocada por Ruido/tratamiento farmacológico , Pérdida Auditiva Provocada por Ruido/patología , Masculino , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Wistar , Tinción con Nitrato de Plata , Ubiquinona/análogos & derivados , Ubiquinona/metabolismo , Ubiquinona/farmacología , Corteza Visual/efectos de los fármacosRESUMEN
Green tea (GT) displays strong anti-oxidant and anti-inflammatory properties mostly attributed to (-)-epigallocatechin-3-gallate (EGCG), while experiments focusing on other catechins are scarce. With the present work we intended to analyze the neuroprotective effects of prolonged consumption of a GT extract (GTE) rich in catechins but poor in EGCG and other GT bioactive components that could also afford benefit. The endpoints evaluated were aging-induced biochemical and morphological changes in the rat hippocampal formation (HF) and behavioral alterations. Male Wistar rats aged 12 months were treated with GTE until 19 months of age. This group of animals was compared with control groups aged 19 (C-19M) or 12 months (C-12M). We found that aging increased oxidative markers but GTE consumption protected proteins and lipids against oxidation. The age-associated increase in lipofuscin content and lysosomal volume was also prevented by treatment with GTE. The dendritic arborizations of dentate granule cells of GTE-treated animals presented plastic changes accompanied by an improved spatial learning evaluated with the Morris water maze. Altogether our results demonstrate that the consumption of an extract rich in catechins other than EGCG protected the HF from aging-related declines contributing to improve the redox status and preventing the structural damage observed in old animals, with repercussions on behavioral performance.
Asunto(s)
Catequina/uso terapéutico , Hipocampo/efectos de los fármacos , Trastornos de la Memoria/prevención & control , Fármacos Neuroprotectores/uso terapéutico , Percepción Espacial/efectos de los fármacos , Factores de Edad , Animales , Catequina/análogos & derivados , Dendritas/efectos de los fármacos , Dendritas/patología , Dendritas/ultraestructura , Modelos Animales de Enfermedad , Hipocampo/patología , Hipocampo/ultraestructura , Lisosomas/efectos de los fármacos , Lisosomas/patología , Masculino , Malondialdehído/metabolismo , Aprendizaje por Laberinto/efectos de los fármacos , Trastornos de la Memoria/patología , Microscopía Electrónica de Transmisión , Mitocondrias/efectos de los fármacos , Mitocondrias/ultraestructura , Carbonilación Proteica/efectos de los fármacos , Células Piramidales/efectos de los fármacos , Células Piramidales/patología , Células Piramidales/ultraestructura , Distribución Aleatoria , Ratas , Ratas Wistar , Tinción con Nitrato de PlataRESUMEN
Cognitive impairment or its recovery has been associated with the absence or reestablishment of estrogenic actions in the central nervous system of female experimental animals or women. It has been proposed that these cognitive phenomena are related to estrogen-mediated modulatory activity of synaptic transmission in brain structures involved in cognitive functions. In the present work a morphological study was conducted in adult female ovariectomized rats to evaluate estradiol-dependent dendritic spine sprouting in hippocampal pyramidal neurons, and changes in the presynaptic marker synaptophysin. Three or ten days after estradiol treatment (10 µg/day, twice) in the ovariectomized rats, a significant increase of synaptophysin was observed, which was coincident with a significant higher numerical density of thin (22%), stubby (36%), mushroom (47%) and double spines (125%), at day 3, without significant changes of spine density at day 10, after treatment. These results may be interpreted as evidence of pre- and postsynaptic plastic events that may be involved in the modulation of cognitive-related behavioral performance after estrogen replacement therapy.
Asunto(s)
Región CA1 Hipocampal/citología , Espinas Dendríticas/efectos de los fármacos , Estradiol/farmacología , Estrógenos/farmacología , Células Piramidales/ultraestructura , Análisis de Varianza , Animales , Región CA1 Hipocampal/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Ovariectomía , Células Piramidales/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Tinción con Nitrato de Plata , Sinaptofisina/metabolismoRESUMEN
Reliable evidence supports the role of sleep in learning and memory processes. In rodents, sleep deprivation (SD) negatively affects consolidation of hippocampus-dependent memories. As memory is integral to post-traumatic stress symptoms, the effects of post-exposure SD on various aspect of the response to stress in a controlled, prospective animal model of post-traumatic stress disorder (PTSD) were evaluated. Rats were deprived of sleep for 6 h throughout the first resting phase after predator scent stress exposure. Behaviors in the elevated plus-maze and acoustic startle response tests were assessed 7 days later, and served for classification into behavioral response groups. Freezing response to a trauma reminder was assessed on day 8. Urine samples were collected daily for corticosterone levels, and heart rate (HR) was also measured. Finally, the impact of manipulating the hypothalamus-pituitary-adrenal axis and adrenergic activity before SD was assessed. Mifepristone (MIFE) and epinephrine (EPI) were administered systemically 10-min post-stress exposure and behavioral responses and response to trauma reminder were measured on days 7-8. Hippocampal expression of glucocorticoid receptors (GRs) and morphological assessment of arborization and dendritic spines were subsequently evaluated. Post-exposure SD effectively ameliorated long-term, stress-induced, PTSD-like behavioral disruptions, reduced trauma reminder freezing responses, and decreased hippocampal expression of GR compared with exposed-untreated controls. Although urine corticosterone levels were significantly elevated 1 h after SD and the HR was attenuated, antagonizing GRs with MIFE or stimulation of adrenergic activity with EPI effectively abolished the effect of SD. MIFE- and EPI-treated animals clearly demonstrated significantly lower total dendritic length, fewer branches and lower spine density along dentate gyrus dendrites with increased levels of GR expression 8 days after exposure, as compared with exposed-SD animals. Intentional prevention of sleep in the early aftermath of stress exposure may well be beneficial in attenuating traumatic stress-related sequelae. Post-exposure SD may disrupt the consolidation of aversive or fearful memories by facilitating correctly timed interactions between glucocorticoid and adrenergic systems.
Asunto(s)
Agonistas Adrenérgicos/uso terapéutico , Epinefrina/uso terapéutico , Antagonistas de Hormonas/uso terapéutico , Mifepristona/uso terapéutico , Privación de Sueño/fisiopatología , Trastornos por Estrés Postraumático/prevención & control , Estimulación Acústica/efectos adversos , Agonistas Adrenérgicos/farmacología , Análisis de Varianza , Animales , Corticosterona/orina , Dendritas/efectos de los fármacos , Dendritas/metabolismo , Dendritas/ultraestructura , Modelos Animales de Enfermedad , Electrocardiografía , Reacción Cataléptica de Congelación/efectos de los fármacos , Reacción Cataléptica de Congelación/fisiología , Regulación de la Expresión Génica/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Hipocampo/ultraestructura , Antagonistas de Hormonas/farmacología , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Mifepristona/farmacología , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Neuronas/ultraestructura , Ratas , Ratas Sprague-Dawley , Receptores de Glucocorticoides/metabolismo , Reflejo de Sobresalto/efectos de los fármacos , Reflejo de Sobresalto/fisiología , Tinción con Nitrato de Plata , Trastornos por Estrés Postraumático/fisiopatología , Telemetría , Factores de TiempoRESUMEN
Photoperiodism is a biological phenomenon in which environmental day length is monitored to ascertain time of year to engage in seasonally appropriate adaptations. This trait is common among organisms living outside of the tropics. White-footed mice (Peromyscus leucopus) are small photoperiodic rodents which display a suite of adaptive responses to short day lengths, including reduced hippocampal volume, impairments in hippocampal-mediated memory, and enhanced hypothalamic-pituitary-adrenal axis reactivity. Because these photoperiodic changes in brain and behavior mirror some of the etiology of post-traumatic stress disorder (PTSD), we hypothesized that photoperiod may also alter fear memory and neuronal morphology within the hippocampus-basolateral amygdala-prefrontal cortex fear circuit. Ten weeks of exposure to short days increased fear memory in an auditory-cued fear conditioning test. Short days also increased dendritic spine density of the neurons of the basolateral amygdala, without affecting morphology of pyramidal neurons within the infralimbic region of the medial prefrontal cortex. Taken together, photoperiodic phenotypic changes in brain morphology and physiology induced by a single environmental factor, exposure to short day lengths, affect responses to fearful stimuli in white-footed mice. These results have potential implications for understanding seasonal changes in fear responsiveness, as well as for expanding translational animal models for studying gene-environment interactions underlying psychiatric diseases, such as PTSD.
Asunto(s)
Amígdala del Cerebelo/citología , Espinas Dendríticas/fisiología , Miedo , Neuronas/ultraestructura , Fotoperiodo , Estimulación Acústica/efectos adversos , Amígdala del Cerebelo/fisiología , Análisis de Varianza , Animales , Condicionamiento Psicológico , Corticosterona/sangre , Reacción Cataléptica de Congelación , Masculino , Peromyscus , Radioinmunoensayo , Tinción con Nitrato de PlataRESUMEN
Changes in neuron morphology, stemming from experiences in early life or adulthood, may be the basis for changes in behavior and their underlying functional mechanisms. For example, reproductive experience has been shown to significantly alter neuron morphology in the hippocampus and prefrontal cortex. In contrast to the effects of reproductive experience, a form of enrichment, on neuron morphology, our understanding of the effects of early social isolation on adult neuron morphology is limited. Therefore, the present study examined changes in neuron morphology in the dorsal (caudate nucleus) and ventral (nucleus accumbens, shell region) striatum and the medial preoptic area of adult virgin and postpartum females exposed to either artificial or maternal rearing during development. Primary results show that regardless of early social isolation, neurons in the caudate nucleus of postpartum females have decreased dendritic complexity compared to virgin females. Maternal experience also increased dendritic complexity in neurons of the nucleus accumbens shell. However, both early social isolation and maternal experience in adulthood influenced dendritic complexity in the medial preoptic area. Together these findings suggest that hypothalamic and striatal neurons show experience-dependent dendritic plasticity and the type and timing of these experiences differentially affect the location and degree of these morphological changes.
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Cuerpo Estriado/citología , Dendritas/fisiología , Hipotálamo/citología , Conducta Materna , Neuronas/citología , Aislamiento Social/psicología , Factores de Edad , Análisis de Varianza , Animales , Animales Recién Nacidos , Peso Corporal , Dendritas/ultraestructura , Femenino , Modelos Lineales , Tamaño de los Órganos , Ratas , Ratas Sprague-Dawley , Reproducción/fisiología , Tinción con Nitrato de PlataRESUMEN
OBJECTIVE: To evaluate the surface treatment and hydrophilicity of the bonding agent on the composite repair strength (RS), silver nitrate uptake (SNU), surface roughness (SR) and estimation of surface area (SA). METHODS: Fifty resin blocks (Opallis, FGM) were polished and divided in 5 groups: no treatment (NT); roughening with a fine (FDB); medium (MDB); coarse-grit (CDB) and 50-µm aluminium oxide sandblasting (AO). A hydrophobic (Adhesive, Scotchbond Multi-Purpose Plus, 3M ESPE) or hydrophilic (Adper Single Bond 2, 3M ESPE) adhesive was applied, followed by composite placement (Opallis, FGM). Composite-composite bonded sticks (0.9 mm2) were tested in tension (1.0mm/min) immediately [IM] or after 6 months [6M] of water storage. Two sticks from each tooth, at each time, were immersed in a 50% silver nitrate solution and evaluated by SEM. A profilometer was used to measure SR. SEM images were used to estimate the SA using the Image J software. Data were analysed by ANOVA and Tukey's tests. RESULTS: The AO showed the highest CR strength for both adhesives, whilst the NT group showed the lowest, irrespective of the storage period. The diamond burs had intermediate performance. Silver nitrate deposits were seen after 6M for the hydrophilic adhesive. The ranking of SR values (Ra) from the lowest to the highest was: NTAsunto(s)
Resinas Compuestas/química
, Materiales Dentales/química
, Reparación de Restauración Dental
, Óxido de Aluminio/química
, Bisfenol A Glicidil Metacrilato/química
, Cementos Dentales/química
, Grabado Dental/métodos
, Diamante/química
, Humanos
, Interacciones Hidrofóbicas e Hidrofílicas
, Ensayo de Materiales
, Metacrilatos/química
, Microscopía Electrónica de Rastreo
, Polietilenglicoles/química
, Ácidos Polimetacrílicos/química
, Poliuretanos/química
, Cementos de Resina/química
, Tinción con Nitrato de Plata
, Estrés Mecánico
, Propiedades de Superficie
, Factores de Tiempo
, Agua/química
RESUMEN
VIDEO ABSTRACT: The precise connectivity of inputs and outputs is critical for cerebral cortex function; however, the cellular mechanisms that establish these connections are poorly understood. Here, we show that the secreted molecule Sonic Hedgehog (Shh) is involved in synapse formation of a specific cortical circuit. Shh is expressed in layer V corticofugal projection neurons and the Shh receptor, Brother of CDO (Boc), is expressed in local and callosal projection neurons of layer II/III that synapse onto the subcortical projection neurons. Layer V neurons of mice lacking functional Shh exhibit decreased synapses. Conversely, the loss of functional Boc leads to a reduction in the strength of synaptic connections onto layer Vb, but not layer II/III, pyramidal neurons. These results demonstrate that Shh is expressed in postsynaptic target cells while Boc is expressed in a complementary population of presynaptic input neurons, and they function to guide the formation of cortical microcircuitry.
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Corteza Cerebral/citología , Regulación del Desarrollo de la Expresión Génica/fisiología , Proteínas Hedgehog/metabolismo , Red Nerviosa/metabolismo , Neuronas/metabolismo , Tractos Piramidales/fisiología , Factores de Edad , Animales , Animales Recién Nacidos , Corteza Cerebral/crecimiento & desarrollo , Channelrhodopsins , Cuerpo Calloso/citología , Cuerpo Calloso/crecimiento & desarrollo , Proteínas de Unión al ADN/metabolismo , Espinas Dendríticas/metabolismo , Espinas Dendríticas/fisiología , Estimulación Eléctrica , Electroporación/métodos , Fluorobencenos/metabolismo , Lateralidad Funcional/genética , Furanos/metabolismo , Regulación del Desarrollo de la Expresión Génica/genética , Proteínas Hedgehog/genética , Inmunoglobulina G/genética , Inmunoglobulina G/metabolismo , Técnicas In Vitro , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Proteínas de Unión a la Región de Fijación a la Matriz/metabolismo , Potenciales de la Membrana/genética , Ratones , Ratones Transgénicos , Mutación/genética , Red Nerviosa/citología , Neuronas/ultraestructura , Proteínas Nucleares/metabolismo , Técnicas de Placa-Clamp , Fosfopiruvato Hidratasa/metabolismo , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Proteínas Represoras/metabolismo , Tinción con Nitrato de Plata/métodos , Estilbamidinas/metabolismo , Sinapsis/metabolismo , Sinapsis/ultraestructura , Sinaptofisina/genética , Sinaptofisina/metabolismo , Factores de Transcripción/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Ubiquitina-Proteína Ligasas , Ácido gamma-Aminobutírico/metabolismoRESUMEN
PURPOSE: To analyze the cytoarchitectural characteristics of the remaining pyramidal neurons in the hippocampal CA1 subfield of rats, four months after global cerebral ischemia (GCI) and progesterone treatment. METHODS: Dendritic arborization, and density and shape of the dendritic spines of CA1 pyramidal neurons in brains of intact rats, or rats submitted 120 days earlier to GCI and treatment with progesterone (8 mg/kg) or its vehicle, at 15 min, and 2, 6, 24, 48, and 72 h after the onset of reperfusion, were analyzed in samples processed by a modified Golgi method. RESULTS: Few impregnated CA1 pyramidal neurons were identified in the ischemic vehicle-treated rats, with a short apical dendrite devoid of bifurcations and dendritic spines. In contrast, the remaining CA1 pyramidal neurons sampled from ischemic progesterone-treated rats showed sinuously branched dendrites with similar number of bifurcations and whole density of spines, and higher proportional density of mushroom spines than those in the intact group. CONCLUSIONS: These cytoarchitectural characteristics may underlie the long-term preservation of place learning and memory functions seen after ischemia and progesterone neuroprotective treatment, possibly compensating for the severe reduction in neuronal population.