1) Preparazione <ol><li> Per iniziare, avviare il sistema NeuroStar, accedere ed eseguire il test automatico della bobina. </li><li> Quindi, accedere al file appropriato paziente dall&#39;elenco disponibile (tutti i pazienti trattati con il sistema NeuroStar sono collegati al computer centrale) </li><li> Infine, garantire le impostazioni sedia non sono restrittive e sollevare il braccio di sedia per rendere più facile per il paziente di entrare e sedersi. </li></ol> 2) posti a sedere e Allineamento del paziente <ol><li> Quando il paziente arriva, lui sedile / lei comodamente in poltrona. </li><li> Assicurarsi che sia il paziente che le spine inserire nell&#39;orecchio tecnico. </li><li> Utilizzando la striscia adesiva, collegare la cinghia posizionamento della testa appena sopra le sopracciglia del paziente. Assicurarsi che il punto centrale è situato sopra la nasion. </li><li> Reclinare la sedia e regolare il poggiatesta, se necessario. </li><li> Leggermente attaccare la cinghia superiore testa al velcro sopra nasion del paziente. </li><li> Attivare il laser e assicurarsi che taglia in due nasion del paziente. </li><li> Chiedere al soggetto di fissare un punto sul muro e regolare la barra A / P, fino a quando non è a livello dei bulbi oculari. </li><li> Assicurarsi che il paziente è completamente centrata e utilizzare il velcro per fissare le cinghie testa posizionamento lato alla sedia trattamento. </li><li> Leggermente coinvolgere il pad testa laterale per garantire la stabilità del cranio durante la determinazione della soglia del motore. </li><li> Infine, registrare le impostazioni di sistema di supporto testa utilizzando l&#39;interfaccia touch screen. </li></ol> 3) Determinazione della soglia del motore <ol><li> Per determinare la soglia di motore, spostare la bobina fino alla testa in modo che la bobina è al vertice e la scheda SenStar è allineata sopra la parte superiore dell&#39;orecchio del paziente. </li><li> Spostare la A / P barra in avanti finché non si allinea con il lato della bobina. </li><li> Allineare la linea centrale sulla bobina con il segno 0 gradi l&#39;indicatore angolo di bobina e tornare al bar A / P. </li><li> Una volta che il contatto e la posizione sono stati stabiliti, chiedere al soggetto di mettere in atto la posizione autostoppista: braccio piegato a 90 gradi al gomito, pugno tiene in modo impreciso, pollice esteso verso l&#39;esterno. </li><li> Iniziare la stimolazione e concentrarsi su ogni movimento del pollice. </li><li> Aumentare gradualmente il livello di MT 0,68-1,10 fino a un dito pollice o concordanti movimento è visto. </li><li> Una volta che il movimento è stabilito, avviare la ricerca del punto di attivazione del motore. Spostare la bobina in giro in una griglia come modello, la regolazione del livello MT, se necessario. Continuare questa ricerca fino a trovare una posizione e un livello di regolazione della potenza che suscita esattamente 5 contrazioni pollice su 10 impulsi consecutivi. </li><li> Una volta che il motore hotspot è stata determinata, spostare l&#39;A / P barra in avanti finché non, ancora una volta, si allinea con il lato della bobina. </li><li> Utilizzare l&#39;interfaccia touch screen per registrare l&#39;angolo di bobina A / P posizione, posizione SOA, e livello di potenza. </li><li> Infine, premere il pulsante MT Trovato. Siamo ora pronti per il trattamento. </li></ol> 4) Il trattamento del paziente <ol><li> Per iniziare, spostare il 5,5 centimetri bobina anteriore alla posizione MT. Questo sarà il luogo di trattamento. </li><li> Assicurarsi che la bobina è in contatto con la testa del paziente e chiedere al paziente di rimanere immobile per tutta la durata del trattamento. </li><li> I parametri di trattamento sono precaricati nel dispositivo. Stimolazione saranno generati a motore soglia del 120%, con una sequenza di impulsi di 10 Hz per 4 secondi, seguito da un periodo di 26 secondi tranquillo. Il trattamento durerà per un totale di 37,5 minuti questo è un totale di 3.000 impulsi. </li><li> Premere il pulsante Conferma sequenza di impulsi, quindi premere il pulsante Start. </li><li> Durante il trattamento, rimane nella stanza con il paziente e controllare periodicamente per assicurare la bobina è in contatto con il paziente. </li><li> Dopo la sequenza di impulsi è stata completata, spostare la bobina in alto e lontano dalla testa del paziente e allontanare il capo pad lato supporto. </li><li> Sollevare la parte posteriore della sedia e sollevare il braccio della sedia. </li><li> Assistere il paziente come lui / lei si alza. </li><li> Togliere tappi per le orecchie e toccare il pulsante Logout per terminare la sessione di trattamento. </li></ol> 5) Rappresentante Risultati TMS è tipicamente applicata ogni giorno per un periodo di 4-6 settimane. I dati recenti suggeriscono alcuni pazienti possono essere soggette a recidiva sintomatica dopo 4-6 mesi. 8 Se ciò si verifica, gli operatori possono prendere in considerazione sedute di mantenimento per prolungare effetto. Questo trattamento è stato efficace in circa il 60% dei pazienti trattati. <img src="/files/ftp_upload/2345/2345fig1.jpg" alt="Tabella 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 produzione video di questo articolo è stato sponsorizzato da Neuronetics, che produce strumenti utilizzati in questi studi.

Quando si considera TMS come intervento potenziale terapeutico per la depressione, è importante riconoscere la FDA ha approvato il protocollo è altamente specifica. Per essere concreto, l&#39;approvazione della FDA è limitato a 10 Hz stimolazione suprathreshold applicata ogni giorno per 4-6 settimane di utilizzare il dispositivo NeuroStar su pazienti che non sono riusciti a raggiungere un miglioramento soddisfacente da uno, ma non più di uno, adeguati studi farmaci antidepressivi durante la depressione in corso episodio. A questo punto, l&#39;utilizzo di qualsiasi modello di stimolazione alternativa, naturalmente orario alternativo, dispositivo alternativo, o popolazione di pazienti alternativo è considerato off-label. I risultati continuano ad essere generati a sostegno dell&#39;efficacia del trattamento TMS 5,6,7, nuovi dati rivela la possibilità di recidiva sintomatica si verifica 4-6 mesi dopo la cessazione del trattamento. A partire da questa pubblicazione, non ci sono protocolli di manutenzione approvata. Pertanto, le questioni concernenti il ​​trattamento aggiuntivo e / o manutenzione effetto deve essere considerato e controllato da vicino da singoli professionisti.

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

Il Neuronetics NeuroStar Stimolazione Magnetica Transcranica (TMS) System è un dispositivo medico di classe II che produce breve durata, campi magnetici pulsati. Questi campi alternarsi rapidamente indurre correnti elettriche all&#39;interno localizzata, regioni oggetto della corteccia che sono associati a diversi cambiamenti fisiologici e funzionali del cervello. 1,2,3 Nel 2007, O&#39;Reardon et al., Utilizzando il dispositivo NeuroStar, ha pubblicato i risultati di un settore sponsorizzato, multisito, randomizzato, sham-stimolazione studio clinico controllato in cui 301 pazienti con depressione maggiore, che in precedenza aveva fallito a rispondere ad almeno uno studio adeguato trattamento antidepressivo, sono stati sottoposti sia TMS attiva o falsa sulla sinistra della corteccia prefrontale dorsolaterale (DLPFC). I pazienti, che erano senza farmaci al momento dello studio, ha ricevuto TMS cinque volte alla settimana per 4-6 settimane 4. I risultati hanno dimostrato che una sottopopolazione di pazienti (coloro che erano relativamente meno resistenti ai farmaci, non essendo riuscito non più di due buone prove farmacologiche) hanno mostrato un miglioramento statisticamente significativo di Montgomery-Asberg Depression Scale (MADRS), l&#39;Hamilton Depression Rating Scale (HAMD), e varie altre misure di esito. Nell&#39;ottobre del 2008, sostenuta da questi e altri risultati simili 5,6,7, Neuronetics ottenuto l&#39;Amministrazione prima e unica Food and Drug Administration (FDA) l&#39;approvazione per il trattamento clinico di una specifica forma di depressione refrattaria ai farmaci utilizzando un dispositivo TMS Terapia ( Approvazione FDA K061053). In questo articolo, esploreremo specificato FDA ha approvato NeuroStar protocollo di trattamento della depressione (per essere somministrato solo sotto prescrizione medica e da una professione medica in licenza sia un in-o ambulatoriale).

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

In questo articolo prendiamo in esame la metodologia e le considerazioni relative al protocollo di trattamento della depressione FDA ha approvato con il Neuronetics NeuroStar dispositivo TMS.

Il dispositivo TMS NeuroStar: Condurre la FDA ha approvato protocollo per il trattamento della depressione

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

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Research

JoVE Journal

Neuroscience

39.4K Views.  Beth Israel Deaconess Medical Center. In questo articolo prendiamo in esame la metodologia e le considerazioni relative al protocollo di trattamento della depressione FDA ha approvato con il 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: Utilizzo del Theta Burst protocollo Esplora Mechasnism di plasticità in individui con Sindrome X Fragile e 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 ...

Ricerca

Neuroscienze

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.

Modellistica neurale segnalazione immunitario di emicrania episodica e cronica utilizzo Depressione Diffusione 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.

La telomerasi attività nelle varie regioni del cervello di topo: telomerasi non radioattivo Repeat Amplification Protocol (TRAP) Assay

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.

Preparazione di fettine di cervello acuto utilizzando un' ottimizzata N-metilico-D-metodo di ripristino protettivo glucamine

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.

Protocollo per isolare il circolo di Willis mouse

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 semplice passo-dissezione Protocollo per tutto il montaggio Preparazione per adulti<em&gt; Drosophila</em&gt; Brains

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 protocollo per l'amministrazione di 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 ...

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.

Utilizzando un dispositivo di microfluidica per stimolazione meccanica e alta risoluzione Imaging di 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 protocollo per la terapia di fotobiomodulazione Transcranial in topi

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.

Valutazione dell'induzione della depressione a lungo termine nelle fette di cerebellar adulto

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