RESUMEN
BACKGROUND: MR-guided focused ultrasound (MRgFUS) thalamotomy is a novel and effective treatment for medication-refractory tremor in essential tremor (ET), but how the brain responds to this deliberate lesion is not clear. OBJECTIVE: The current study aimed to evaluate the immediate and longitudinal alterations of functional networks after MRgFUS thalamotomy. METHODS: We retrospectively obtained preoperative and postoperative 30-day, 90-day, and 180-day data of 31 ET patients subjected with MRgFUS thalamotomy from 2018 to 2020. Their archived resting-state functional MRI data were used for functional network comparison as well as graph-theory metrics analysis. Both partial least squares (PLS) regression and linear regression were conducted to associate functional features to tremor symptoms. RESULTS: MRgFUS thalamotomy dramatically abolished tremors, while global functional network only sustained immediate fluctuation within one week after the surgery. Network-based statistics have identified a long-term enhanced corticostriatal subnetwork by comparison between 180-day and preoperative data (P = 0.019). Within this subnetwork, network degree, global efficiency and transitivity were significantly recovered in ET patients right after MRgFUS thalamotomy compared to the pre-operative timepoint (P < 0.05), as well as hemisphere lateralization (P < 0.001). The PLS main component significantly accounted for 33.68 % and 34.16 % of the total variances of hand tremor score and clinical rating scale for tremor (CRST)-total score (P = 0.037 and 0.027). Network transitivity of this subnetwork could serve as a reliable biomarker for hand tremor score control prediction at 180-day after the surgery (ß = 2.94, P = 0.03). CONCLUSION: MRgFUS thalamotomy promoted corticostriatal connectivity activation correlated with tremor improvement in ET patient after MRgFUS thalamotomy.
Asunto(s)
Temblor Esencial , Imagen por Resonancia Magnética , Tálamo , Humanos , Tálamo/diagnóstico por imagen , Tálamo/cirugía , Tálamo/fisiopatología , Femenino , Masculino , Temblor Esencial/cirugía , Temblor Esencial/diagnóstico por imagen , Temblor Esencial/fisiopatología , Anciano , Persona de Mediana Edad , Imagen por Resonancia Magnética/métodos , Estudios Retrospectivos , Red Nerviosa/diagnóstico por imagen , Red Nerviosa/fisiopatología , Cuerpo Estriado/diagnóstico por imagen , Cuerpo Estriado/cirugía , Cuerpo Estriado/fisiopatología , Corteza Cerebral/diagnóstico por imagen , Corteza Cerebral/fisiopatología , Corteza Cerebral/cirugía , Vías Nerviosas/fisiopatología , Vías Nerviosas/diagnóstico por imagenRESUMEN
Glioblastoma (GBM) is the most aggressive malignant glioma, with a very poor prognosis; as such, efforts to explore new treatments and GBM's etiology are a priority. We previously described human GBM cells (R2J-GS) as exhibiting the properties of cancer stem cells (growing in serum-free medium and proliferating into nude mice when orthotopically grafted). Sodium selenite (SS)-an in vitro attractive agent for cancer therapy against GBM-was evaluated in R2J-GS cells. To go further, we launched a preclinical study: SS was given orally, in an escalation-dose study (2.25 to 10.125 mg/kg/day, 5 days on, 2 days off, and 5 days on), to evaluate (1) the absorption of selenium in plasma and organs (brain, kidney, liver, and lung) and (2) the SS toxicity. A 6.75 mg/kg SS dose was chosen to perform a tumor regression assay, followed by MRI, in R2J-GS cells orthotopically implanted in nude mice, as this dose was nontoxic and increased brain selenium concentration. A group receiving TMZ (5 mg/kg) was led in parallel. Although not reaching statistical significance, the group of mice treated with SS showed a slower tumor growth vs. the control group (p = 0.08). No difference was observed between the TMZ and control groups. We provide new insights of the mechanisms of SS and its possible use in chemotherapy.
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Neoplasias Encefálicas/tratamiento farmacológico , Cuerpo Estriado/cirugía , Glioblastoma/tratamiento farmacológico , Células Madre Neoplásicas/trasplante , Selenito de Sodio/efectos adversos , Oligoelementos/efectos adversos , Carga Tumoral/efectos de los fármacos , Ensayos Antitumor por Modelo de Xenoinjerto/métodos , Animales , Apoptosis/efectos de los fármacos , Neoplasias Encefálicas/patología , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cuerpo Estriado/metabolismo , Glioblastoma/patología , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Selenio/metabolismo , Selenito de Sodio/administración & dosificación , Temozolomida/administración & dosificación , Oligoelementos/administración & dosificación , Resultado del TratamientoRESUMEN
OBJECTIVE: It has been proposed that Tourette syndrome is associated with dysfunction in widespread cortical areas and globus pallidus externus hyperactivity secondary to dopaminergic hyperactivity and serotonergic/dynorphinergic hypoactivity. The main objective of this study was to test this hypothesis by developing an animal model of Tourette syndrome via striatotomy, followed by administration of drugs that mimic the neurotransmitter environment, so as to induce globus pallidus externus hyperactivity. METHODS: Rats were assigned to 3 groups: stereotactic striatotomy (STT) and striatal sham -lesion (SHAM) groups, treated with anterior and posterior striatum procedures in both hemispheres, and a group of nonoperated animals (NAIVE). Postoperatively, all rodents were blindly administered 3 drug protocols: levodopa/benserazide; levodopa/benserazide/ergotamine/naloxone (MIX); and saline. The animals were filmed at the peak action of these drugs. The videos were evaluated by a single blinded researcher. RESULTS: Six types of involuntary movements (IMs) were observed: cephalic, trunk jerks, oromandibular, forepaw jerks, dystonic, and locomotive. The number of animals with IM and the mean number of IM after both levodopa/benserazide and MIX was significantly higher in the STT compared with the SHAM and NAIVE groups. In the SHAM and NAIVE, MIX was superior to levodopa/benserazide in the induction of IM. In the STT, MIX was superior to levodopa/benserazide in the induction of trunk jerks. Appendicular IM were more common after posterior than after anterior striatotomy. CONCLUSIONS: These results show that striatotomy, followed by administration of levodopa/benserazide alone or associated with ergotamine and naloxone, is efficacious in inducing IM, supporting the hypothesis that led to this study.
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Cuerpo Estriado/patología , Cuerpo Estriado/cirugía , Dopaminérgicos/administración & dosificación , Técnicas Estereotáxicas/efectos adversos , Síndrome de Tourette/tratamiento farmacológico , Síndrome de Tourette/patología , Analgésicos no Narcóticos/administración & dosificación , Animales , Benserazida/administración & dosificación , Cuerpo Estriado/efectos de los fármacos , Método Doble Ciego , Combinación de Medicamentos , Ergotamina/administración & dosificación , Femenino , Globo Pálido/efectos de los fármacos , Globo Pálido/patología , Globo Pálido/cirugía , Levodopa/administración & dosificación , Naloxona/administración & dosificación , Estudios Prospectivos , Ratas , Ratas WistarRESUMEN
An SU-8 probe with an array of nine, individually addressable gold microband electrodes (100 µm long, 4 µm wide, separated by 4-µm gaps) was photolithographically fabricated and characterized for detection of low concentrations of chemicals in confined spaces and in vivo studies of biological tissues. The probe's shank (6 mm long, 100 µm wide, 100 µm thick) is flexible, but exhibits sufficient sharpness and rigidity to be inserted into soft tissue. Laser micromachining was used to define probe geometry by spatially revealing the underlying sacrificial aluminum layer, which was then etched to free the probes from a silicon wafer. Perfusion with fluorescent nanobeads showed that, like a carbon fiber electrode, the probe produced no noticeable damage when inserted into rat brain, in contrast to damage from an inserted microdialysis probe. The individual addressability of the electrodes allows single and multiple electrode activation. Redox cycling is possible, where adjacent electrodes serve as generators (that oxidize or reduce molecules) and collectors (that do the opposite) to amplify signals of small concentrations without background subtraction. Information about electrochemical mechanisms and kinetics may also be obtained. Detection limits for potassium ferricyanide in potassium chloride electrolyte of 2.19, 1.25, and 2.08 µM and for dopamine in artificial cerebral spinal fluid of 1.94, 1.08, and 5.66 µM for generators alone and for generators and collectors during redox cycling, respectively, were obtained.
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Dopamina/líquido cefalorraquídeo , Técnicas Electroquímicas/instrumentación , Microelectrodos , Animales , Calibración , Cuerpo Estriado/cirugía , Técnicas Electroquímicas/métodos , Electrólitos/química , Ferricianuros/análisis , Ferricianuros/química , Oro , Rayos Láser , Masculino , Microelectrodos/efectos adversos , Microtecnología , Oxidación-Reducción , Polímeros/química , Cloruro de Potasio/química , Ratas Sprague-DawleyRESUMEN
Cellular transplant therapy is one of the most common therapeutic strategies used to mitigate symptoms of neurodegenerative diseases such as Huntington's disease (HD). Briefly, the main goal of the present study was to investigate HD's motor deficits through the olfactory ecto-mesenchymals stem cells (OE-MSC) secretome. OE-MSCs were characterized immunophenotypically by the positive expression of CD73, CD90 and CD105. Also, three specific markers of OE-MSCs were obtained from the nasal cavity of human volunteers. The main features of OE-MSCs are their high proliferation, ease of harvesting and growth factor secretion. All animals were randomly assigned to three groups: control, 3-NP + vehicle treated and 3-NP + Cell groups. In both experimental groups, the subjects received intraperitoneal 3-NP (30 mg/kg) injections once a day for five consecutive days, followed by the bilateral intra-striatal implantation of OE-MSCs in the 3-NP + Cell group. Muscular function was assessed by electromyography and rotarod test, and the locomotor function was evaluated using the open field test. According to our findings, striatal transplants of OE-MSCs reduced microglial inflammatory factor, the tumor necrosis factor (TNFα) in the 3-NP + Cell group, with a significant reduction in RIP3, the markers of necroptosis in striatum. In addition to the remarkable recovery of the striatal volume after engraftment, the motor activities were enhanced in the 3-NP + cell group compared to the 3-NP + vehicle group. Taken together, our results demonstrated the in vivo advantages of OE-MSCs treatment in an HD rat model with numerous positive paracrine effects including behavioral and anatomical recovery.
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Cuerpo Estriado/cirugía , Enfermedad de Huntington/terapia , Trasplante de Células Madre Mesenquimatosas/métodos , Actividad Motora/fisiología , Necroptosis/fisiología , Animales , Conducta Animal/fisiología , Cuerpo Estriado/patología , Modelos Animales de Enfermedad , Humanos , Enfermedad de Huntington/patología , Enfermedad de Huntington/fisiopatología , Ratas , Prueba de Desempeño de Rotación con Aceleración Constante , Resultado del TratamientoRESUMEN
The dorsal striatum (dS) has been implicated in storing procedural memories and controlling movement kinematics. Since procedural memories are expressed through movements, the exact nature of the dS function has proven difficult to delineate. Here, we challenged rats in complementary locomotion-based tasks designed to alleviate this confound. Surprisingly, dS lesions did not impair the rats' ability to remember the procedure for the successful completion of motor routines. However, the speed and initiation of the reward-oriented phase of the routines were irreversibly altered by the dS lesion. Further behavioral analyses, combined with modeling in the optimal control framework, indicated that these kinematic alterations were well explained by an increased sensitivity to effort. Our work provides evidence supporting a primary role of the dS in modulating the kinematics of reward-oriented actions, a function that may be related to the optimization of the energetic costs of moving.
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Cuerpo Estriado/fisiología , Carrera/fisiología , Animales , Cuerpo Estriado/cirugía , Metabolismo Energético/fisiología , Masculino , Modelos Animales , Ratas , Ratas Long-Evans , Recompensa , Técnicas EstereotáxicasRESUMEN
Human pluripotent stem cells (hPSCs) are a powerful tool for modelling human development. In recent years, hPSCs have become central in cell-based therapies for neurodegenerative diseases given their potential to replace affected neurons. However, directing hPSCs into specific neuronal types is complex and requires an accurate protocol that mimics endogenous neuronal development. Here we describe step-by-step a fast feeder-free neuronal differentiation protocol to direct hPSCs to mature forebrain neurons in 37 days in vitro (DIV). The protocol is based upon a combination of specific morphogens, trophic and growth factors, ions, neurotransmitters and extracellular matrix elements. A human-induced PSC line (Ctr-Q33) and a human embryonic stem cell line (GEN-Q18) were used to reinforce the potential of the protocol. Neuronal activity was analysed by single-cell calcium imaging. At 8 DIV, we obtained a homogeneous population of hPSC-derived neuroectodermal progenitors which self-arranged in bi-dimensional neural tube-like structures. At 16 DIV, we generated hPSC-derived neural progenitor cells (NPCs) with mostly a subpallial identity along with a subpopulation of pallial NPCs. Terminal in vitro neuronal differentiation was confirmed by the expression of microtubule associated protein 2b (Map 2b) by almost 100% of hPSC-derived neurons and the expression of specific-striatal neuronal markers including GABA, CTIP2 and DARPP-32. HPSC-derived neurons showed mature and functional phenotypes as they expressed synaptic markers, voltage-gated ion channels and neurotransmitter receptors. Neurons displayed diverse spontaneous activity patterns that were classified into three major groups, namely "high", "intermediate" and "low" firing neurons. Finally, transplantation experiments showed that the NPCs survived and differentiated within mouse striatum for at least 3 months. NPCs integrated host environmental cues and differentiated into striatal medium-sized spiny neurons (MSNs), which successfully integrated into the endogenous circuitry without teratoma formation. Altogether, these findings demonstrate the potential of this robust human neuronal differentiation protocol, which will bring new opportunities for the study of human neurodevelopment and neurodegeneration, and will open new avenues in cell-based therapies, pharmacological studies and alternative in vitro toxicology.
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Técnicas de Cultivo de Célula/métodos , Cuerpo Estriado/cirugía , Neurogénesis/fisiología , Neuronas/trasplante , Células Madre Pluripotentes/citología , Animales , Línea Celular , Cuerpo Estriado/citología , Humanos , RatonesRESUMEN
Human placenta mesenchymal stromal cells were injected to healthy rats either stereotaxically into the striatum or intra-arterially through the internal carotid artery. Some cells injected into the brain migrated along the corpus callosum both medially and laterally or concentrated around small blood vessels. A small fraction of MSC injected intra-arterially adhered to the endothelium and stayed inside blood vessels for up to 48 hours mostly in the basin of the middle cerebral artery. Neither stereotaxic, nor intra-arterial transplantation of mesenchymal stromal cells modulated the proliferation of neural stem cells in the subventricular zone of the brain, but stereotaxic transplantation suppressed activation of their proliferation in response to traumatization with the needle.
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Cuerpo Estriado/citología , Ventrículos Laterales/citología , Trasplante de Células Madre Mesenquimatosas/métodos , Células Madre Mesenquimatosas/citología , Células-Madre Neurales/citología , Placenta/citología , Animales , Arteria Carótida Interna/citología , Movimiento Celular , Proliferación Celular , Cuerpo Estriado/cirugía , Femenino , Humanos , Inyecciones Intraarteriales , Inyecciones Intraventriculares , Ventrículos Laterales/cirugía , Masculino , Células Madre Mesenquimatosas/fisiología , Arteria Cerebral Media/citología , Células-Madre Neurales/fisiología , Placenta/fisiología , Embarazo , Cultivo Primario de Células , Ratas , Ratas Wistar , Técnicas Estereotáxicas , Trasplante HeterólogoRESUMEN
BACKGROUND: Neural stem cells (NSCs) transplantation is considered a promising treatment for Parkinson's disease. But most NSCs are differentiated into glial cells rather than neurons, and only a few of them survive after transplantation due to the inflammatory environment. METHODS: In this study, neural stem cells (NSCs) and microglial cells both forced with the Nurr1 gene were transplanted into the striatum of the rat model of PD. The results were evaluated through reverse transcription polymerase chain reaction (RT-PCR), Western blot, and immunofluorescence analysis. RESULTS: The behavioral abnormalities of PD rats were improved by combined transplantation of NSCs and microglia, both forced with Nurr1. The number of tyrosine hydroxylase+ cells in the striatum of PD rats increased, and the number of Iba1+ cells decreased compared with the other groups. Moreover, the dopamine neurons differentiated from grafted NSCs could still be detected in the striatum of PD rats after 5 months. CONCLUSIONS: The results suggested that transplantation of Nurr1-overexpressing NSCs and microglia could improve the inhospitable host brain environments, which will be a new potential strategy for the cell replacement therapy in PD.
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Terapia Genética/métodos , Microglía/trasplante , Células-Madre Neurales/trasplante , Miembro 2 del Grupo A de la Subfamilia 4 de Receptores Nucleares/genética , Trastornos Parkinsonianos/terapia , Trasplante de Células Madre/métodos , Anfetamina , Animales , Conducta Animal , Proteínas de Unión al Calcio/genética , Diferenciación Celular , Cuerpo Estriado/cirugía , Neuronas Dopaminérgicas/trasplante , Encefalitis/terapia , Femenino , Hidroxidopaminas , Masculino , Proteínas de Microfilamentos/genética , Miembro 2 del Grupo A de la Subfamilia 4 de Receptores Nucleares/biosíntesis , Trastornos Parkinsonianos/inducido químicamente , Trastornos Parkinsonianos/psicología , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: The purpose of this study was to investigate the role of deep brain stimulation (DBS) of the globus pallidus internus (GPi) in dopamine and dopamine transporter metabolism and to explore the regulatory role of DBS on dopaminergic neurons in Tourette syndrome by constructing an autoimmune model. METHODS: Serum with high concentrations of antinuclear antibodies or phosphate-buffered saline solution was injected into the striatum of rats by a stereotactic technique and micropump. Then, electrodes were planted in the rats' globus pallidus internus. Concentrations of dopamine and dopamine transporter in the striatum were detected by enzyme-linked immunosorbent assay, immunohistochemistry, and Western blot analysis after 7 days of high-frequency stimulation (130 Hz). RESULTS: The tic behavior score of rats in the Tourette syndrome group was higher than that of rats in the control group (P < 0.01). After high-frequency stimulation, the scores of the Tourette syndrome model group and the control group significantly decreased. The concentration of dopamine in the Tourette syndrome model group and the control group also significantly decreased after electric stimulation (P < 0.05). In addition, immunohistochemical analysis and Western blot test results showed that dopamine transporter in the Tourette syndrome model nonstimulation group was lower than in the Tourette syndrome model stimulation group, and that dopamine transporter in the control nonstimulation group was lower than in the control stimulation group (P < 0.05). CONCLUSIONS: The results of this study show that the mechanism of DBS of the GPi in the treatment of Tourette syndrome involved monoamine neurotransmitters, especially the dopamine system, that affected the metabolism and transport of corresponding neurotransmitters, playing an important role in regulating the concentration of synaptic neurotransmitters and changing the biologic activity of basal ganglia nerve circuits.
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Estimulación Encefálica Profunda , Estimulación Eléctrica , Globo Pálido/fisiopatología , Síndrome de Tourette/fisiopatología , Síndrome de Tourette/cirugía , Animales , Cuerpo Estriado/fisiopatología , Cuerpo Estriado/cirugía , Estimulación Encefálica Profunda/métodos , Modelos Animales de Enfermedad , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática/metabolismo , Globo Pálido/cirugía , Masculino , Neostriado/fisiopatología , Neostriado/cirugía , Ratas Sprague-Dawley , Resultado del TratamientoRESUMEN
Monitoring regional tissue oxygenation in animal models and potentially in human subjects can yield insights into the underlying mechanisms of local O2-mediated physiological processes and provide diagnostic and therapeutic guidance for relevant disease states. Existing technologies for tissue oxygenation assessments involve some combination of disadvantages in requirements for physical tethers, anesthetics, and special apparatus, often with confounding effects on the natural behaviors of test subjects. This work introduces an entirely wireless and fully implantable platform incorporating (i) microscale optoelectronics for continuous sensing of local hemoglobin dynamics and (ii) advanced designs in continuous, wireless power delivery and data output for tether-free operation. These features support in vivo, highly localized tissue oximetry at sites of interest, including deep brain regions of mice, on untethered, awake animal models. The results create many opportunities for studying various O2-mediated processes in naturally behaving subjects, with implications in biomedical research and clinical practice.
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Suministros de Energía Eléctrica , Oximetría/instrumentación , Prótesis e Implantes , Tecnología Inalámbrica/instrumentación , Animales , Sustitutos Sanguíneos/análisis , Cuerpo Estriado/metabolismo , Cuerpo Estriado/cirugía , Hipoxia/metabolismo , Ratones , Ratones Endogámicos C57BL , Modelos Animales , Oxígeno/análisis , Ratas , Ratas Sprague-Dawley , Materiales InteligentesAsunto(s)
Cuerpo Estriado/patología , Dopamina/metabolismo , Gastrectomía , Enfermedad de Parkinson/patología , Edad de Inicio , Anciano de 80 o más Años , Cuerpo Estriado/cirugía , Femenino , Gastrectomía/métodos , Humanos , Masculino , Enfermedad de Parkinson/metabolismo , Enfermedad de Parkinson/cirugíaRESUMEN
BACKGROUND: Parkinson's disease (PD) is the second most common neurodegenerative disease, presenting with midbrain dopaminergic neurons degeneration. A number of studies suggest that microglial activation may have a role in PD. It has emerged that inflammation-derived oxidative stress and cytokine-dependent toxicity may contribute to nigrostriatal pathway degeneration and exacerbate the progression of the disease in patients with idiopathic PD. Cell therapies have long been considered a feasible regenerative approach to compensate for the loss of specific cell populations such as the one that occurs in PD. We recently demonstrated that erythropoietin-releasing neural precursors cells (Er-NPCs) administered to MPTP-intoxicated animals survive after transplantation in the recipient's damaged brain, differentiate, and rescue degenerating striatal dopaminergic neurons. Here, we aimed to investigate the potential anti-inflammatory actions of Er-NPCs infused in an MPTP experimental model of PD. METHODS: The degeneration of dopaminergic neurons was caused by MPTP administration in C57BL/6 male mice. 2.5 × 105 GFP-labeled Er-NPCs were administered by stereotaxic injection unilaterally in the left striatum. Functional recovery was assessed by two independent behavioral tests. Neuroinflammation was investigated measuring the mRNAs levels of pro-inflammatory and anti-inflammatory cytokines, and immunohistochemistry studies were performed to evaluate markers of inflammation and the potential rescue of tyrosine hydroxylase (TH) projections in the striatum of recipient mice. RESULTS: Er-NPC administration promoted a rapid anti-inflammatory effect that was already evident 24 h after transplant with a decrease of pro-inflammatory and increase of anti-inflammatory cytokines mRNA expression levels. This effect was maintained until the end of the observational period, 2 weeks post-transplant. Here, we show that Er-NPCs transplant reduces macrophage infiltration, directly counteracting the M1-like pro-inflammatory response of murine-activated microglia, which corresponds to the decrease of CD68 and CD86 markers, and induces M2-like pro-regeneration traits, as indicated by the increase of CD206 and IL-10 expression. Moreover, we also show that this activity is mediated by Er-NPCs-derived erythropoietin (EPO) since the co-injection of cells with anti-EPO antibodies neutralizes the anti-inflammatory effect of the Er-NPCs treatment. CONCLUSION: This study shows the anti-inflammatory actions exerted by Er-NPCs, and we suggest that these cells may represent good candidates for cellular therapy to counteract neuroinflammation in neurodegenerative disorders.
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Encefalitis/etiología , Encefalitis/cirugía , Eritropoyetina/uso terapéutico , Células-Madre Neurales/trasplante , Trastornos Parkinsonianos/complicaciones , Recuperación de la Función/fisiología , 1-Metil-4-fenil-1,2,3,6-Tetrahidropiridina/farmacología , Animales , Técnicas de Cocultivo , Cuerpo Estriado/metabolismo , Cuerpo Estriado/cirugía , Citocinas/metabolismo , Modelos Animales de Enfermedad , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática/metabolismo , Eritropoyetina/genética , Eritropoyetina/metabolismo , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Fuerza Muscular/efectos de los fármacos , Fuerza Muscular/fisiología , Células-Madre Neurales/fisiología , Trastornos Parkinsonianos/etiología , Trastornos Parkinsonianos/patología , Recuperación de la Función/efectos de los fármacos , Recuperación de la Función/genética , Olfato/efectos de los fármacos , Olfato/fisiología , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
BACKGROUND: Parkinson's disease (PD) is caused by the gradual loss of dopamine-producing cells in the brain. This study evaluated the potential neuroprotective role of puerarin (PR) on dopamine (DA)-producing cells in vitro and in vivo. METHOD: In vitro, the effects of PR on proliferation and differentiation and DA releases of mesenchymal stem cells (MSCs) were assayed by CCK-8, flow cytometry, real-time PCR and ELISA respectively. Then the differentiated cells were labeled with enhanced green fluorescent protein (EGFP) and administrated into PD animal models induced by 6-OHDA. The proliferation and differentiation of labeled cells were identified by fluorescence microscopy and immunostaining. RESULTS: In vitro, after being treated with different concentrations of PR for 1 week, the TUJ1, TH and DAT protein and mRNA expression and DA releases increased significantly. In vivo, after transplantation of PR-treated DA-producing cells, the symptoms of PD improved significantly from the second week after transplantation; more transplanted cells survived and migrated to wider region along injection line; more transplanted cells proliferated and differentiated into TH+ cells; more DA was detected in the striatum during 6 weeks' observation. CONCLUSION: The results suggest that PR promote DA neuron survival, proliferation and differentiation.
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Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Cuerpo Estriado/cirugía , Dopamina/metabolismo , Neuronas Dopaminérgicas/efectos de los fármacos , Neuronas Dopaminérgicas/trasplante , Isoflavonas/farmacología , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/efectos de los fármacos , Fármacos Neuroprotectores/farmacología , Trastornos Parkinsonianos/cirugía , Animales , Células Cultivadas , Cuerpo Estriado/metabolismo , Cuerpo Estriado/patología , Modelos Animales de Enfermedad , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática/genética , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática/metabolismo , Neuronas Dopaminérgicas/metabolismo , Neuronas Dopaminérgicas/patología , Relación Dosis-Respuesta a Droga , Masculino , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/patología , Oxidopamina , Trastornos Parkinsonianos/inducido químicamente , Trastornos Parkinsonianos/metabolismo , Trastornos Parkinsonianos/patología , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Factores de Tiempo , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismo , Tirosina 3-Monooxigenasa/genética , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
Many genes are expressed in embryonic brains, and some of them are continuously expressed in the brain after birth. For such persistently expressed genes, they may function to regulate the developmental process and/or physiological function in neonatal brains. To investigate neurobiological functions of specific genes in the brain, it is essential to inactivate genes in the brain. Here, we describe a simple stereotaxic method to inactivate gene expression in the striatum of transgenic mice at neonatal time windows. AAV-eGFP-Cre viruses were microinjected into the striatum of Ai14 reporter gene mice at postnatal day (P) 2 by stereotaxic brain surgery. The tdTomato reporter gene expression was detected in P14 striatum, suggesting a successful Cre-loxP mediated DNA recombination in AAV-transduced striatal cells. We further validated this technique by microinjecting AAV-eGFP-Cre viruses into P2Foxp2fl/fl mice. Double labeling of GFP and Foxp2 showed that GFP-positive cells lacked Foxp2 immunoreactivity in P9 striatum, suggesting the loss of Foxp2 protein in AAV-eGFP-Cre transduced striatal cells. Taken together, these results demonstrate an effective genetic deletion by stereotaxically microinjected AAV-eGFP-Cre viruses in specific neuronal populations in the neonatal brains of floxed transgenic mice. In conclusion, our stereotaxic technique provides an easy and simple platform for genetic manipulation in neonatal mouse brains. The technique can not only be used to delete genes in specific regions of neonatal brains, but it also can be used to inject pharmacological drugs, neuronal tracers, genetically modified optogenetics and chemogenetics proteins, neuronal activity indicators and other reagents into the striatum of neonatal mouse brains.
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Encéfalo/cirugía , Cuerpo Estriado/cirugía , Animales , Cuerpo Estriado/metabolismo , Expresión Génica , Ratones , Ratones TransgénicosRESUMEN
Parkinson's disease is characterized by the progressive loss of dopaminergic neurons from the substantia nigra, a process that leads to a dopamine deficiency in the striatum. This deficiency is responsible for the development of motor symptoms, including resting tremor, bradykinesia, rigidity and postural instability. Based on the observation of substantial neuronal death, alternatives to Parkinson's disease treatment have been studied, including cell-based therapies. The present study aimed to assess the therapeutic potential of intravenous and intrastriatal transplant of bone marrow mononuclear cells in a mouse model of Parkinson's disease. Animals underwent stereotaxic surgery and received an injection of 6-hydroxydopamine into their medial forebrain bundle. Three weeks later, mice were injected with bone marrow mononuclear cells or saline through the caudal vein or directly into their right striatum. Motor function was assessed using the rotarod and apomorphine-induced rotation tests. Our results showed that intrastriatal bone marrow mononuclear cells, but not intravenous, have a short-term therapeutic effect on dopaminergic response in this mice model of parkinsonism assessed by the apomorphine-induced rotation test. This phenomenon was not identified on the rotarod test, showing dissociation between dopaminergic response and motor behavior. Further experiments are needed to elucidate the precise mechanisms involved in these effects.
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Cuerpo Estriado/cirugía , Neuronas Dopaminérgicas/efectos de los fármacos , Trasplante de Células Madre Mesenquimatosas , Actividad Motora , Enfermedad de Parkinson/prevención & control , Administración Intravenosa , Animales , Apomorfina/administración & dosificación , Modelos Animales de Enfermedad , Agonistas de Dopamina/administración & dosificación , Neuronas Dopaminérgicas/patología , Proteínas Fluorescentes Verdes/metabolismo , Masculino , Haz Prosencefálico Medial/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Ratones Endogámicos C57BL , Oxidopamina/toxicidad , Enfermedad de Parkinson/fisiopatología , Prueba de Desempeño de Rotación con Aceleración Constante , Sustancia Negra/efectos de los fármacos , Sustancia Negra/patologíaRESUMEN
Following the failure of a Phase II clinical study evaluating human retinal pigment epithelial (hRPE) cell implants as a potential treatment option for Parkinson's disease, speculation has centered on implant function and survival as possible contributors to the therapeutic outcomes. We recently reported that neonatal hRPE cells, similar to hRPE cells used in the Phase II clinical study, produced short-lived in vitro and limited in vivo trophic factors, which supports that assumption. We hypothesize that the switch from fetal to neonatal hRPE cells, between the Phase I and the Phase II clinical trial may be partly responsible for the later negative outcomes. To investigate this hypothesis, we used two neonatal hRPE cell lots, prepared in a similar manner to neonatal hRPE cells used in the Phase II clinical study, and compared them to previously evaluated fetal hRPE cells for behavioral changes following unilateral striatal implantation in 6-hydroxydopamine-lesioned rats. The results showed that only fetal, not neonatal, hRPE cell implants, were able to improve behavioral outcomes following striatal implantation in the lesioned rats. These data suggest that fetal hRPE cells may be preferential to neonatal hRPE cells in restoring behavioral deficits.
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Trasplante de Células , Trastornos Parkinsonianos/cirugía , Epitelio Pigmentado de la Retina/citología , Epitelio Pigmentado de la Retina/embriología , Anfetamina/farmacología , Animales , Supervivencia Celular , Senescencia Celular , Estimulantes del Sistema Nervioso Central/farmacología , Cuerpo Estriado/cirugía , Células Epiteliales/trasplante , Femenino , Humanos , Recién Nacido , Masculino , Actividad Motora/efectos de los fármacos , Actividad Motora/fisiología , Oxidopamina , Trastornos Parkinsonianos/fisiopatología , Distribución Aleatoria , Ratas Sprague-Dawley , Epitelio Pigmentado de la Retina/crecimiento & desarrollo , Caminata/fisiologíaRESUMEN
The choroid plexus (CP) has been explored as a cellular therapeutic due to its broad-ranging secretome and demonstrated longevity in a variety of encapsulation modalities. While the CP organ is normally involved in disease repair processes in the brain, the range of indications that could potentially be ameliorated with exogenous CP therapy is widespread, including diseases of the central nervous system, hearing loss, chronic wounds, and others. The CP can be isolated from animal sources and digested into a highly purified epithelial culture that can withstand encapsulation and transplantation. Its epithelium can adapt to different microenvironments, and depending on culture conditions, can be manipulated into various three-dimensional configurations with distinct gene expression profiles. The cocktail of proteins secreted by the CP can be harvested in culture, and purified forms of these extracts have been evaluated in topical applications to treat poorly healing wounds. When encapsulated, the epithelial clusters can be maintained for extended durations in vitro with minimal impact on potency. A treatment for Parkinson's disease utilizing encapsulated porcine CP has been developed and is currently being evaluated in a Phase I clinical trial. The current chapter serves to summarize recent experience with CP factor delivery, and provides a description of the relevant materials and methods employed in these studies.
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Células Inmovilizadas/citología , Células Inmovilizadas/trasplante , Plexo Coroideo/citología , Células Epiteliales/citología , Células Epiteliales/trasplante , Alginatos/química , Animales , Cápsulas/química , Técnicas de Cultivo de Célula/métodos , Separación Celular/métodos , Tratamiento Basado en Trasplante de Células y Tejidos , Células Cultivadas , Células Inmovilizadas/metabolismo , Plexo Coroideo/metabolismo , Cuerpo Estriado/citología , Cuerpo Estriado/cirugía , Medios de Cultivo Condicionados/metabolismo , Células Epiteliales/metabolismo , Ácido Glucurónico/química , Ácidos Hexurónicos/química , Humanos , Neuronas/citología , Neuronas/metabolismo , Enfermedad de Parkinson/terapia , Ratas , PorcinosRESUMEN
Erythropoietin-releasing neural precursor cells (Er-NPCs) are a subclass of subventricular zone-derived neural progenitors, capable of surviving for 6 hr after death of donor. They present higher neural differentiation. Here, Er-NPCs were studied in animal model of Parkinson's disease. Dopaminergic degeneration was caused by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine intraperitoneal administration in C57BL/6 mice. The loss of function was evaluated by specific behavioral tests. Er-NPCs (2.5 × 105) expressing the green fluorescent protein were administered by stereotaxic injection unilaterally in the left striatum. At the end of observational research period (2 weeks), most of the transplanted Er-NPCs were located in the striatum, while several had migrated ventrally and caudally from the injection site, up to ipsilateral and contralateral substantia nigra. Most of transplanted cells had differentiated into dopaminergic, cholinergic, or GABAergic neurons. Er-NPCs administration also promoted a rapid functional improvement that was already evident at the third day after cells administration. This was accompanied by enhanced survival of nigral neurons. These effects were likely promoted by Er-NPCs-released erythropoietin (EPO), since the injection of Er-NPCs in association with anti-EPO or anti-EPOR antibodies had completely neutralized the recovery of function. In addition, intrastriatal administration of recombinant EPO mimics the effects of Er-NPCs. We suggest that Er-NPCs, and cells with similar properties, may represent good candidates for cellular therapy in neurodegenerative disorders of this kind.
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Tratamiento Basado en Trasplante de Células y Tejidos/métodos , Cuerpo Estriado/cirugía , Eritropoyetina/metabolismo , Intoxicación por MPTP/terapia , Células-Madre Neurales/trasplante , Recuperación de la Función/fisiología , Animales , Antígenos/metabolismo , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/metabolismo , Colina O-Acetiltransferasa/metabolismo , Modelos Animales de Enfermedad , Eritropoyetina/genética , Intoxicación por MPTP/metabolismo , Intoxicación por MPTP/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Trastornos del Movimiento/etiología , Trastornos del Movimiento/terapia , Fuerza Muscular/fisiología , Proteínas del Tejido Nervioso/metabolismo , Células-Madre Neurales/metabolismo , Proteoglicanos/metabolismo , Tirosina 3-Monooxigenasa/genética , Tirosina 3-Monooxigenasa/metabolismo , Ácido gamma-Aminobutírico/metabolismoRESUMEN
BACKGROUND AND OBJECTIVE: Pediatric basal ganglia germ cell tumors (GCTs) represent a rare subset of tumors about which little is known. We aimed to summarize the clinical features and radiological findings of this special subgroup of GCTs. METHODS: From January 2010 to January 2015, 12 pediatric patients with basal ganglia GCTs were treated in our hospital. The clinical features, radiologic findings, diagnosis, treatment, and outcome of these patients were analyzed retrospectively. Our institutional diagnostic principle and treatment strategy of this disease were discussed. RESULTS: GCTs accounted for 25.5% of all the pediatric basal ganglia tumors treated in our hospital. There were 9 male and 3 female patients with a mean age of 11.5 ± 2.1 years. The most common symptom was progressive hemiparesis (n = 9, 75%). The radiologic findings showed that the lesions predominately located in caput of caudate nucleus (n = 9, 75.0%), followed by lenticular nucleus (n = 3, 25.0%). Hemiatrophy was commonly observed (n = 8, 66.7%). Eight patients were diagnosed as having germinomas, and 4 patients as having nongerminomatous germ cell tumors. During the follow-up period, preoperative neurologic dysfunctions improved in 7 patients and remained stable in 3. Two patients developed new onset of neurologic dysfunction after the treatment. Two patients suffered from tumor recurrence. CONCLUSIONS: GCTs are not as rare as considered in pediatric basal ganglia tumors. They bear some distinctive clinical and radiologic features, which can help with the accurate diagnosis and successful management of such tumors.