1) Preparación <ol><li> Para empezar, la puesta en marcha del sistema NeuroStar, tienes que entrar y realizar la prueba de bobina automatizado. </li><li> A continuación, acceder al archivo adecuado del paciente de la lista de disponibles (todos los pacientes tratados con el sistema NeuroStar están conectados al ordenador central) </li><li> Por último, garantizar la configuración de silla no son restrictivas y levantar el brazo de la silla para que sea fácil para el paciente para entrar y sentarse. </li></ol> 2) Asientos y Alineación de la paciente <ol><li> Cuando el paciente llega, le asiento / ella cómodamente en la silla. </li><li> Asegúrese de que tanto el paciente como los tapones de inserción técnico de la oreja. </li><li> Utilizando la cinta adhesiva, fije la correa de posicionamiento de la cabeza justo por encima de las cejas del paciente. Asegúrese de que el punto central se encuentra por encima de la nasion. </li><li> Reclinar el asiento y ajustar el soporte para la cabeza, según sea necesario. </li><li> Ligeramente colocar la correa de la cabeza superior a la de velcro por encima de nasión del paciente. </li><li> Activar el láser y asegurarse de que biseca nasión del paciente. </li><li> Pida al sujeto que mirar a un punto en la pared y ajustar la barra de A / P hasta que esté al nivel de los globos oculares. </li><li> Asegúrese de que el paciente está totalmente centrado y el uso del velcro para asegurar las correas de la cabeza lado de posicionamiento para la silla de tratamiento. </li><li> Ligeramente la almohadilla de participar cabeza de lado para asegurar la estabilidad del cráneo durante la determinación del umbral motor. </li><li> Por último, anote los valores de soporte para la cabeza del sistema utilizando la interfaz de pantalla táctil. </li></ol> 3) Determinar el umbral del motor <ol><li> Para determinar el umbral de motor, mueva la bobina hasta la cabeza de tal manera que la bobina está en el vértice y la ficha Senstar se alinea sobre la parte superior de la oreja del paciente. </li><li> Mueva el A / P barra hacia adelante hasta que se alinea con el lado de la bobina. </li><li> Alinee la línea central de la bobina con la marca de 0 grados en el indicador de ángulo de la bobina y volver la barra de A / P. </li><li> Una vez en contacto y posición se han establecido, pedir a los sujetos a promulgar la posición autoestopista: el brazo doblado a 90 grados en el codo, el puño en forma no estructurada, pulgar extendido hacia el exterior. </li><li> Empezar la estimulación y se centran en un movimiento del pulgar. </li><li> Poco a poco aumentar el nivel de MT 0,68 a 1,10 hasta que el movimiento del pulgar o el dedo se ve concordantes. </li><li> Una vez que el movimiento se ha establecido, comenzar a buscar el punto de acceso del motor. Mover la bobina alrededor de una rejilla como patrón, ajustando el nivel de MT, según sea necesario. Continuar en esta búsqueda hasta que encuentre una ubicación y un ajuste del nivel de energía que provoca exactamente 5 contracciones del pulgar de cada 10 pulsos consecutivos. </li><li> Una vez que el punto caliente del motor se ha determinado, mueva el A / P barra hacia adelante hasta que, una vez más, se alinea con el lado de la bobina. </li><li> Utilizar la interfaz de pantalla táctil para registrar el ángulo A / P posición de la bobina, posición de SOA, y nivel de potencia. </li><li> Por último, presione el botón de MT se encuentra. Ahora estamos listos para recibir tratamiento. </li></ol> 4) El tratamiento del paciente <ol><li> Para empezar, mover la bobina de 5,5 cm anterior a la ubicación de MT. Este será el lugar de tratamiento. </li><li> Asegúrese de que la bobina está en contacto con la cabeza del paciente y pedir al paciente que permanezca quieto durante la duración del tratamiento. </li><li> Los parámetros de tratamiento están precargados en el dispositivo. La estimulación se genera en el umbral de motor 120% con una secuencia de pulsos de 10 Hz a 4 segundos, seguido por un período de silencio 26 segundos. El tratamiento durará un total de 37,5 minutos, este es un total de 3.000 pulsos. </li><li> Pulse el botón Confirmar la secuencia de pulsos a continuación, pulse en el botón Inicio. </li><li> Durante el tratamiento, permanecer en la sala con el paciente y comprobar periódicamente para asegurarse de que la bobina está en contacto con el paciente. </li><li> Después de la secuencia de pulsos se ha completado, mueva la bobina hacia arriba y lejos de la cabeza del paciente y se alejan de la cabeza de lado cojín de apoyo. </li><li> Levante la parte posterior de la silla y levantar el brazo de la silla. </li><li> Ayudar al paciente como él / ella se pone de pie. </li><li> Quitar tapones para los oídos y tocar el botón Cerrar sesión para finalizar la sesión de tratamiento. </li></ol> 5) Los resultados representativos TMS se aplica normalmente al día durante un periodo de 4-6 semanas. Los datos recientes sugieren que algunos pacientes pueden ser propensos a la recaída sintomática después de 4-6 meses. 8 Si esto ocurre, los médicos pueden considerar sesiones de mantenimiento para prolongar efecto. Este tratamiento ha sido eficaz en aproximadamente el 60% de los pacientes tratados. <img src="/files/ftp_upload/2345/2345fig1.jpg" alt="Tabla 5" />

<ol>
	<li>Pascual-Leone, A., Davey, M., Wassermann, E. M., Rothwell, J., Puri, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of Transcranial Magnetic Stimulation. , (2002).</li><li>Walsh, V., Pascual-Leone, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Transcranial Magnetic Stimulation: A Neurochronometrics of Mind. , (2005).</li><li>Horvath, J. C., Perez, J., Forrow, L., Fregni, F., Pascual-Leone, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transcranial+Magnetic+Stimulation:+An+Historical+Evaluation+and+Future+Prognosis+of+Therapeutically+Relevant+Ethical+Concerns.">Transcranial Magnetic Stimulation: An Historical Evaluation and Future Prognosis of Therapeutically Relevant Ethical Concerns.</a> , .</li><li>O'Reardon, J. P., Solvason, H. B., Janicak, P. G., Sampson, S., Isenberg, K. E., Nahas, Z., McDonald, W. M., Avery, D., Fitzgerald, P. B., Loo, C., Demitrack, M. A., George, M. S., Sackheim, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+and+safety+of+transcranial+magnetic+stimulation+in+the+acute+treatment+of+major+depression:+a+multisite+randomized+control+trial.">Efficacy and safety of transcranial magnetic stimulation in the acute treatment of major depression: a multisite randomized control trial.</a> Biological Psychiatry. 62, 1208-1216 (2007).</li><li>George, M. S., Lisanby, S. H., Avery, D., McDonald, W. M., Durkalski, V., Pavlicova, M., Anderson, B., Nahas, Z., Bulow, P., Zarkowski, P., Holtzheimer, P. D., Schwartz, T., Sackheim, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Daily+prefrontal+transcranial+magnetic+stimulation+therapy+for+major+depressive+disorder:+a+sham-controlled+randomized+study.">Daily prefrontal transcranial magnetic stimulation therapy for major depressive disorder: a sham-controlled randomized study.</a> Arch Gen Psychiatry. 67, 507-516 (2010).</li><li>Lisanbym, S. H., Husain, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Daily+Left+Prefrontal+Repetitive+Transcranial+Magnetic+Stimulation+(rTMS)+in+the+Acute+Treatment+of+Major+Depression:+Clinical+Predictors+of+Outcome+in+a+Multisite,+Randomized+Controlled+Clinical+Trial.">Daily Left Prefrontal Repetitive Transcranial Magnetic Stimulation (rTMS) in the Acute Treatment of Major Depression: Clinical Predictors of Outcome in a Multisite, Randomized Controlled Clinical Trial.</a> Neuropsychopharmacology. 34, 522-534 (2008).</li><li>Thase, M., Demitrack, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluating+clinical+significance+of+treatment+outcomes+in+studies+of+resistant+major+depression.">Evaluating clinical significance of treatment outcomes in studies of resistant major depression.</a> Biological Psychiatry. 63, 138-138 (2008).</li><li>Demirtas-Tatlidede, A., Mechanic-Hamilton, D., Press, D. Z., Pearlman, C., Stern, W. M., Thall, M., Pascual-Leone, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+open-label,+prospective+study+of+repetitive+transcranial+magnetic+stimulation+(rTMS)+in+the+long-term+treatment+of+refractory+depression:+reproducibility+and+duration+of+the+antidepressant+effect+in+medication-free+patients.">An open-label, prospective study of repetitive transcranial magnetic stimulation (rTMS) in the long-term treatment of refractory depression: reproducibility and duration of the antidepressant effect in medication-free patients.</a> Journal of Clinical Psychiatry. 69, 930-934 (2008).</li></ol>

La producción de vídeo de este artículo fue patrocinado por Neuronetics, que produce instrumentos utilizados en estos estudios.

Al considerar la TMS como una potencial intervención terapéutica para la depresión, es importante reconocer que la FDA aprobó el protocolo es muy específico. Para ser concretos, la aprobación de la FDA se limita a 10 Hz por encima del umbral de estimulación aplicada diariamente durante 4-6 semanas utilizando el dispositivo NeuroStar a los pacientes que han fracasado en lograr una mejoría satisfactoria de uno, pero no más de una, las pruebas suficientes medicamentos antidepresivos durante la depresión actual episodio. En este punto, la utilización de cualquier patrón de estimulación alternativa, por supuesto horaria diferente, dispositivo alternativo, o población de pacientes alternativa se considera fuera de la etiqueta. Como resultados se continúan generando en apoyo de la eficacia de la EMT tratamiento 5,6,7, nuevos datos revelan la posibilidad de recaída sintomática ocurre 4-6 meses después de la interrupción del tratamiento. A partir de esta publicación, no existen protocolos de mantenimiento aprobado. Por lo tanto, las cuestiones relativas a un tratamiento adicional y / o mantenimiento efecto deben ser considerados y monitoreados de cerca por los profesionales individuales.

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the neurostar tms device: conducting the fda approved protocol for treatment of depression

El Neuronetics NeuroStar estimulación magnética transcraneal (TMS) es un dispositivo médico de clase II que produce breve duración, campos magnéticos pulsados. Estos campos alternando rápidamente inducir corrientes eléctricas en regiones localizadas, específicas de la corteza que se asocian con diversos cambios fisiológicos y funcionales del cerebro. 1,2,3 En 2007, O&#39;Reardon et al., Utilizando el dispositivo NeuroStar, publicó los resultados de una patrocinados por la industria, multicéntrico, aleatorizado, la estimulación simulada ensayo clínico controlado en el que 301 pacientes con depresión mayor, que previamente no habían logrado responder en por lo menos un tratamiento adecuado el tratamiento con antidepresivos, fueron sometidos a EMT activa o simulada sobre la corteza prefrontal dorsolateral izquierda (córtex prefrontal dorsolateral). Los pacientes, que fueron sin medicación en el momento del estudio, recibieron TMS cinco veces por semana durante 4-6 semanas. 4 Los resultados demostraron que una sub-población de pacientes (aquellos que eran relativamente menos resistentes a la medicación, al no haber no más de dos buenos ensayos farmacológicos) mostraron una mejoría estadísticamente significativa en la Escala de Depresión de Montgomery-Asberg (MADRS), la Valoración de Depresión de Hamilton Scale (HAMD), y varias otras medidas de resultado. En octubre de 2008, con el apoyo de estos y otros resultados similares 5,6,7, Neuronetics obtenido la Administración de Alimentos y Medicamentos de la primera y única (FDA) la aprobación para el tratamiento clínico de una forma específica de medicamentos depresión resistente con un dispositivo de terapia de TMS ( aprobación de la FDA K061053). En este trabajo, vamos a explorar las especificadas por la FDA aprobó el protocolo NeuroStar tratamiento de la depresión (para ser administrado solamente bajo prescripción y por una profesión médica con licencia, ya sea en una o en ambulatorio).

Watch this Scientific Journal Video about The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression at JoVE.com

The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression

En este artículo, analizamos la metodología y las consideraciones relacionadas con el protocolo aprobado por la FDA con el tratamiento de la depresión Neuronetics NeuroStar dispositivo TMS.

El dispositivo de TMS NeuroStar: Realización de la FDA aprobó el Protocolo para el tratamiento de la depresión

The Neuronetics NeuroStar Transcranial Magnetic Stimulation (TMS) System is a class II medical device that produces brief duration, pulsed magnetic ...

the-neurostar-tms-device-conducting-fda-approved-protocol-for

Research

JoVE Journal

Neuroscience

39.4K vistas.  Beth Israel Deaconess Medical Center. En este artículo, analizamos la metodología y las consideraciones relacionadas con el protocolo aprobado por la FDA con el tratamiento de la depresión Neuronetics NeuroStar dispositivo TMS.

Video: The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression

TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS display abnormalities in the expression of FMR1 - a protein required for typical, healthy neural development.3 Recent data has suggested that the loss of this protein can cause the cortex to be hyperexcitable thereby affecting overall patterns of neural plasticity.4,5
In addition, Fragile X shows a strong comorbidity with autism: in fact, 30% of children with FXS are diagnosed with autism, and 2 - 5% of autistic children suffer from FXS.6
Transcranial Magnetic Stimulation (a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability via the application of localized magnetic field pulses 7,8) represents a unique method of exploring plasticity and the manifestations of FXS within affected individuals. More specifically, Theta-Burst Stimulation (TBS), a specific stimulatory protocol shown to modulate cortical plasticity for a duration up to 30 minutes after stimulation cessation in healthy populations, has already proven an efficacious tool in the exploration of abnormal plasticity.9,10 
Recent studies have shown the effects of TBS last considerably longer in individuals on the autistic spectrum - up to 90 minutes.11 This extended effect-duration suggests an underlying abnormality in the brain's natural plasticity state in autistic individuals - similar to the hyperexcitability induced by Fragile X Syndrome.
In this experiment, utilizing single-pulse motor-evoked potentials (MEPs) as our benchmark, we will explore the effects of both intermittent and continuous TBS on cortical plasticity in individuals suffering from FXS and individuals on the Autistic Spectrum.

In this article, we examine the effects of Theta-Burst TMS stimulation on cortical plasticity in individuals suffering from Fragile X syndrome and individuals on the autistic spectrum.

TMS: Uso del protocolo Theta-Burst para explorar Mechasnism de plasticidad en los individuos con el síndrome de X frágil y el autismo

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS ...

Investigación

Neurociencias

Modeling Neural Immune Signaling of Episodic and Chronic Migraine Using Spreading Depression In Vitro

Migraine and its transformation to chronic migraine are healthcare burdens in need of improved treatment options. We seek to define how neural immune signaling modulates the susceptibility to migraine, modeled in vitro using spreading depression (SD), as a means to develop novel therapeutic targets for episodic and chronic migraine. SD is the likely cause of migraine aura and migraine pain. It is a paroxysmal loss of neuronal function triggered by initially increased neuronal activity, which slowly propagates within susceptible brain regions. Normal brain function is exquisitely sensitive to, and relies on, coincident low-level immune signaling. Thus, neural immune signaling likely affects electrical activity of SD, and therefore migraine. Pain perception studies of SD in whole animals are fraught with difficulties, but whole animals are well suited to examine systems biology aspects of migraine since SD activates trigeminal nociceptive pathways. However, whole animal studies alone cannot be used to decipher the cellular and neural circuit mechanisms of SD. Instead, in vitro preparations where environmental conditions can be controlled are necessary. Here, it is important to recognize limitations of acute slices and distinct advantages of hippocampal slice cultures. Acute brain slices cannot reveal subtle changes in immune signaling since preparing the slices alone triggers: pro-inflammatory changes that last days, epileptiform behavior due to high levels of oxygen tension needed to vitalize the slices, and irreversible cell injury at anoxic slice centers. 
In contrast, we examine immune signaling in mature hippocampal slice cultures since the cultures closely parallel their in vivo counterpart with mature trisynaptic function; show quiescent astrocytes, microglia, and cytokine levels; and SD is easily induced in an unanesthetized preparation. Furthermore, the slices are long-lived and SD can be induced on consecutive days without injury, making this preparation the sole means to-date capable of modeling the neuroimmune consequences of chronic SD, and thus perhaps chronic migraine. We use electrophysiological techniques and non-invasive imaging to measure neuronal cell and circuit functions coincident with SD. Neural immune gene expression variables are measured with qPCR screening, qPCR arrays, and, importantly, use of cDNA preamplification for detection of ultra-low level targets such as interferon-gamma using whole, regional, or specific cell enhanced (via laser dissection microscopy) sampling. Cytokine cascade signaling is further assessed with multiplexed phosphoprotein related targets with gene expression and phosphoprotein changes confirmed via cell-specific immunostaining. Pharmacological and siRNA strategies are used to mimic and modulate SD immune signaling.

Modeling Neural Immune Signaling of Episodic and Chronic Migraine Using Spreading Depression In Vitro

Migraine and its transformation to chronic migraine are immense healthcare burdens in need of improved treatment options. We seek to define how neural immune signaling modulates the susceptibility to migraine, modeled in vitro using spreading depression in hippocampal slice cultures, as a means to develop novel therapeutic targets.

Modelado neuronal de señalización inmune de la migraña episódica y crónica mediante la depresión propagada In Vitro</em

Migraine and its transformation to chronic migraine are healthcare burdens in need of improved treatment options.  We seek to define how neural immune ...

Telomerase Activity in the Various Regions of Mouse Brain: Non-Radioactive Telomerase Repeat Amplification Protocol (TRAP) Assay

Telomerase, a ribonucleoprotein, is responsible for maintaining the telomere length and therefore promoting genomic integrity, proliferation, and lifespan. In addition, telomerase protects the mitochondria from oxidative stress and confers resistance to apoptosis, suggesting its possible importance for the surviving of non-mitotic, highly active cells such as neurons. We previously demonstrated the ability of novel telomerase activators to increase telomerase activity and expression in the various mouse brain regions and to protect motor neurons cells from oxidative stress. These results strengthen the notion that telomerase is involved in the protection of neurons from various lesions. To underline the role of telomerase in the brain, we here compare the activity of telomerase in male and female mouse brain and its dependence on age. TRAP assay is a standard method for detecting telomerase activity in various tissues or cell lines. Here we demonstrate the analysis of telomerase activity in three regions of the mouse brain by non-denaturing protein extraction using CHAPS lysis buffer followed by modification of the standard TRAP assay.
In this 2-step assay, endogenous telomerase elongates a specific telomerase substrate (TS primer) by adding TTAGGG 6 bp repeats (telomerase reaction). The telomerase reaction products are amplified by PCR reaction creating a DNA ladder of 6 bp increments. The analysis of the DNA ladder is made by 4.5% high resolution agarose gel electrophoresis followed by staining with highly sensitive nucleic acid stain.
Compared to the traditional TRAP assay that utilize 32P labeled radioactive dCTP&#39;s for DNA detection and polyacrylamide gel electrophoresis for resolving the DNA ladder, this protocol offers a non-toxic time saving TRAP assay for evaluating telomerase activity in the mouse brain, demonstrating the ability to detect differences in telomerase activity in the various female and male mouse brain region.

Telomerase Activity in the Various Regions of Mouse Brain: Non-Radioactive Telomerase Repeat Amplification Protocol TRAP Assay

Telomerase is expressed in the neonatal brain and also in distinct regions of the adult brain. We present a non-toxic time saving TRAP assay for the analysis of telomerase activity in various regions of the mouse brain and detection of differences in telomerase activity between male and female mouse brains.

Actividad telomerasa en las diversas regiones del cerebro de ratón: La telomerasa no radiactivo Repetir Protocolo de Amplificación (TRAP) Ensayo

Telomerase, a ribonucleoprotein, is responsible for maintaining the telomere length and therefore promoting genomic integrity, proliferation, and ...

Preparation of Acute Brain Slices Using an Optimized N-Methyl-D-glucamine Protective Recovery Method

This protocol is a practical guide to the N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. Numerous recent studies have validated the utility of this method for enhancing neuronal preservation and overall brain slice viability. The implementation of this technique by early adopters has facilitated detailed investigations into brain function using diverse experimental applications and spanning a wide range of animal ages, brain regions, and cell types. Steps are outlined for carrying out the protective recovery brain slice technique using an optimized NMDG artificial cerebrospinal fluid (aCSF) media formulation and enhanced procedure to reliably obtain healthy brain slices for patch clamp electrophysiology. With this updated approach, a substantial improvement is observed in the speed and reliability of gigaohm seal formation during targeted patch clamp recording experiments while maintaining excellent neuronal preservation, thereby facilitating challenging experimental applications. Representative results are provided from multi-neuron patch clamp recording experiments to assay synaptic connectivity in neocortical brain slices prepared from young adult transgenic mice and mature adult human neurosurgical specimens. Furthermore, the optimized NMDG protective recovery method of brain slicing is compatible with both juvenile and adult animals, thus resolving a limitation of the original methodology. In summary, a single media formulation and brain slicing procedure can be implemented across various species and ages to achieve excellent viability and tissue preservation.

Preparation of Acute Brain Slices Using an Optimized N-Methyl-D-glucamine Protective Recovery Method

This protocol demonstrates the implementation of an optimized N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. A single media formulation is used to reliably obtain healthy brain slices from animals of any age and for diverse experimental applications.

Preparación de rebanadas cerebrales agudas mediante un optimizado N-metil-D-glucamine recuperación protección método

This protocol is a practical guide to the N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. Numerous recent studies have ...

Protocol for Isolating the Mouse Circle of Willis

The cerebral arterial circle (circulus arteriosus cerebri) or circle of Willis (CoW) is a circulatory anastomosis surrounding the optic chiasma and hypothalamus that supplies blood to the brain and surrounding structures. It has been implicated in several cerebrovascular disorders, including cerebral amyloid angiopathy (CAA)-associated vasculopathies, intracranial atherosclerosis and intracranial aneurysms. Studies of the molecular mechanisms underlying these diseases for the identification of novel drug targets for their prevention require animal models. Some of these models may be transgenic, whereas others will involve isolation of the cerebro-vasculature, including the CoW.The method described here is suitable for CoW isolation in any mouse lineage and has considerable potential for screening (expression of genes, protein production, posttranslational protein modifications, secretome analysis, etc.) studies on the large vessels of the mouse cerebro-vasculature. It can also be used for ex vivo studies, by adapting the organ bath system developed for isolated mouse olfactory arteries.

We describe here a reproducible protocol for isolating the mouse circle of Willis.

Protocolo para el aislamiento del Círculo de Willis Ratón

The cerebral arterial circle (circulus arteriosus cerebri) or circle of Willis (CoW) is a circulatory anastomosis surrounding the optic chiasma and ...

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the molecular and cellular basis of higher brain functions. A whole-mount preparation of adult brains with well-preserved morphology is critical for such whole brain-based studies, but can be technically challenging and time-consuming. This protocol describes an easy-to-learn, one-step dissection approach of an adult fly head in less than 10 s, while keeping the intact brain attached to the rest of the body to facilitate subsequent processing steps. The procedure helps remove most of the eye and tracheal tissues normally associated with the brain that can interfere with the later imaging step, and also places less demand on the quality of the dissecting forceps. Additionally, we describe a simple method that allows convenient flipping of the mounted brain samples on a coverslip, which is important for imaging both sides of the brains with similar signal intensity and quality. As an example of the protocol, we present an analysis of dopaminergic (DA) neurons in adult brains of WT (w1118) flies. The high efficacy of the dissection method makes it particularly useful for large-scale adult brain-based studies in Drosophila.

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

The adult Drosophila brain is a valuable system for studying neuronal circuitry, higher brain functions, and complex disorders. An efficient method to dissect whole brain tissue from the small fly head will facilitate brain-based studies. Here we describe a simple, one-step dissection protocol of adult brains with well-preserved morphology.

Un simple de un solo paso para la disección Protocolo Todo el montaje Preparación de adulto<em&gt; Drosophila</emLos cerebros&gt;

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the ...

A Protocol for the Administration of Real-Time fMRI Neurofeedback Training

Neurologic disorders are characterized by abnormal cellular-, molecular-, and circuit-level functions in the brain. New methods to induce and control neuroplastic processes and correct abnormal function, or even shift functions from damaged tissue to physiologically healthy brain regions, hold the potential to dramatically improve overall health. Of the current neuroplastic interventions in development, neurofeedback training (NFT) from functional Magnetic Resonance Imaging (fMRI) has the advantages of being completely non-invasive, non-pharmacologic, and spatially localized to target brain regions, as well as having no known side effects. Furthermore, NFT techniques, initially developed using fMRI, can often be translated to exercises that can be performed outside of the scanner without the aid of medical professionals or sophisticated medical equipment. In fMRI NFT, the fMRI signal is measured from specific regions of the brain, processed, and presented to the participant in real-time. Through training, self-directed mental processing techniques, that regulate this signal and its underlying neurophysiologic correlates, are developed. FMRI NFT has been used to train volitional control over a wide range of brain regions with implications for several different cognitive, behavioral, and motor systems. Additionally, fMRI NFT has shown promise in a broad range of applications such as the treatment of neurologic disorders and the augmentation of baseline human performance. In this article, we present an fMRI NFT protocol developed at our institution for modulation of both healthy and abnormal brain function, as well as examples of using the method to target both cognitive and auditory regions of the brain.

The ability to induce and/or control neural plasticity may be critical in future treatments for neurologic disorders and the recovery from brain injury. In this paper, we present a protocol on the use of neurofeedback training with functional magnetic resonance imaging to modulate human brain function.

Un protocolo para la administración de Real-Time fMRI entrenamiento de Neurofeedback

Neurologic disorders are characterized by abnormal cellular-, molecular-, and circuit-level functions in the brain. New methods to induce and control ...

Using a Microfluidics Device for Mechanical Stimulation and High Resolution Imaging of C. elegans

One central goal of mechanobiology is to understand the reciprocal effect of mechanical stress on proteins and cells. Despite its importance, the influence of mechanical stress on cellular function is still poorly understood. In part, this knowledge gap exists because few tools enable simultaneous deformation of tissue and cells, imaging of cellular activity in live animals, and efficient restriction of motility in otherwise highly mobile model organisms, such as the nematode Caenorhabditis elegans. The small size of C. elegans makes them an excellent match to microfluidics-based research devices, and solutions for immobilization have been presented using microfluidic devices. Although these devices allow for high-resolution imaging, the animal is fully encased in polydimethylsiloxane (PDMS) and glass, limiting physical access for delivery of mechanical force or electrophysiological recordings. Recently, we created a device that integrates pneumatic actuators with a trapping design that is compatible with high-resolution fluorescence microscopy. The actuation channel is separated from the worm-trapping channel by a thin PDMS diaphragm. This diaphragm is deflected into the side of a worm by applying pressure from an external source. The device can target individual mechanosensitive neurons. The activation of these neurons is imaged at high-resolution with genetically-encoded calcium indicators. This article presents the general method using C. elegans strains expressing calcium-sensitive activity indicator (GCaMP6s) in their touch receptor neurons (TRNs). The method, however, is not limited to TRNs nor to calcium sensors as a probe, but can be expanded to other mechanically-sensitive cells or sensors.

Using a Microfluidics Device for Mechanical Stimulation and High Resolution Imaging of C. elegans

New tools for mechanobiology research are needed to understand how mechanical stress activates biochemical pathways and elicits biological responses. Here, we showcase a new method for selective mechanical stimulation of immobilized animals with a microfluidic trap allowing high-resolution imaging of cellular responses.

Usando un dispositivo de microfluidos para la estimulación mecánica y alta resolución de imagen de C. elegans

One central goal of mechanobiology is to understand the reciprocal effect of mechanical stress on proteins and cells. Despite its importance, the ...

A Protocol for Transcranial Photobiomodulation Therapy in Mice

Transcranial photobiomodulation is a potential innovative noninvasive therapeutic approach for improving brain bioenergetics, brain function in a wide range of neurological and psychiatric disorders, and memory enhancement in age-related cognitive decline and neurodegenerative diseases. We describe a laboratory protocol for transcranial photobiomodulation therapy (PBMT) in mice. Aged BALB/c mice (18 months old) are treated with a 660 nm laser transcranially, once daily for 2 weeks. Laser transmittance data shows that approximately 1% of the incident red light on the scalp reaches a 1 mm depth from the cortical surface, penetrating the dorsal hippocampus. Treatment outcomes are assessed by two methods: a Barnes maze test, which is a hippocampus-dependent spatial learning and memory task evaluation, and measuring hippocampal ATP levels, which is used as a bioenergetics index. The results from the Barnes task show an enhancement of the spatial memory in laser-treated aged mice when compared with age-matched controls. Biochemical analysis after laser treatment indicates increased hippocampal ATP levels. We postulate that the enhancement of memory performance is potentially due to an improvement in hippocampal energy metabolism induced by the red laser treatment. The observations in mice could be extended to other animal models since this protocol could potentially be adapted to other species frequently used in translational neuroscience, such as rabbit, cat, dog, or monkey. Transcranial photobiomodulation is a safe and cost-effective modality which may be a promising therapeutic approach in age-related cognitive impairment.

Photobiomodulation therapy is an innovative noninvasive modality for the treatment of a wide range of neurological and psychiatric disorders and can also improve healthy brain function. This protocol includes a step-by-step guide to performing brain photobiomodulation in mice by transcranial light delivery, which can be adapted for use in other laboratory rodents.

Un protocolo de Transcranial Photobiomodulation terapia en ratones

Transcranial photobiomodulation is a potential innovative noninvasive therapeutic approach for improving brain bioenergetics, brain function in a wide ...

Assessment of Long-term Depression Induction in Adult Cerebellar Slices

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from parallel fibers (PF) to Purkinje cells (PC) is considered the basis for motor learning, and deficiencies of both LTD and motor learning are observed in various gene-manipulated animals. Common motor learning sets, such as adaptation of the optokinetic reflex (OKR), the vestibular-ocular reflex (VOR), and rotarod test were used for evaluation of motor learning ability. However, results obtained from the GluA2-carboxy terminus modified knock-in mice demonstrated normal adaptation of the VOR and the OKR, despite lacking PF-LTD. In that report, induction of LTD was only attempted using one type of stimulation protocol at room temperature. Thus, conditions to induce cerebellar LTD were explored in the same knock-in mutants using various protocols at near physiological temperature. Finally, we found stimulation protocols, by which LTD could be induced in these gene-manipulated mice. In this study, a set of protocols are proposed to evaluate LTD-induction, which will more accurately allow examination of the causal relationship between LTD and motor learning. In conclusion, experimental conditions are crucial when evaluating LTD in gene-manipulated mice.

In some gene-manipulated animals, using a single protocol may fail to induce LTD in cerebellar Purkinje cells, and there may be a discrepancy between LTD and motor learning. Multiple protocols are necessary to assess LTD-induction in gene-manipulated animals. Standard protocols are shown.

Evaluación de la inducción a largo plazo de la depresión en rodajas de cerebelosores adultos

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from ...