Finanziamenti forniti dal NIMH, NARSAD e la Fondazione Gatsby.

Attrezzature installazione Il primo passo per l&#39;esecuzione e l&#39;esperimento fMRI è la creazione e verifica delle attrezzature. Questi passaggi possono essere eseguiti in qualsiasi ordine. Condizionamento del segnale - Lo scanner si trova in una stanza elettricamente e magneticamente schermati. Tutti i segnali elettrici che vanno dalla sala di controllo alla sala scanner passare attraverso un pannello di filtro per rimuovere tutte le frequenze che potrebbe creare un artefatto nell&#39;immagine MR. Airpuff - Il dispositivo offre un soffio airpuff controllato di pressione fissa e durata. Quando è ordinata a l&#39;occhio, questo è avverso, ma non traumatica. Il regolatore di pressione (WPI pneumatico PicoPump) ad una fonte di aria compressa, che potrebbe essere un serbatoio d&#39;aria, ma usiamo casa aria. La pressione viene regolata a 30 psi. La tempistica è regolato per offrire una boccata 50 msec. L&#39;elettrovalvola è controllato da un segnale del computer. L&#39;uscita del regolatore di pressione viene consegnato al soggetto attraverso un tubo di plastica 1/6th pollice (Tygon). E &#39;importante che il tubo non sia troppo lungo o ci sarà un ritardo tra l&#39;apertura solenoide e il tempo che l&#39;aria compressa arriva alla fine del tubo. Per questa ragione, il regolatore di pressione si trova all&#39;interno della stanza scanner e segnali di controllo sono alimentati tramite un pannello di filtro per garantire che non ci sono artefatti di imaging. Juice - Il distributore di succo di controllare la dimensione e il numero di gocce di succo consegnato alla bocca da un tubo di plastica. L&#39;erogatore succo è costituito da un serbatoio, un computer controllato elettrovalvola, e un lungo tubo che fornisce succo al soggetto. Tutti i componenti elettronici sono fuori dalla stanza dello scanner e quindi presentano senza artefatti nel segnale MR. L&#39;erogatore succo (Strumenti Crist) è sciacquato due volte compilando serbatoio e apertura della valvola a solenoide. Allora è riempito con la bevanda preferita soggetto. Il sistema è testato prima della scansione eseguendo un programma che causa il computer per generare un flusso di impulsi di prova che si traduce in alcune gocce di succo di essere consegnati al soggetto. Il soggetto verifica che il succo è che scorre. Bite bar - Movimento delle testine crea artefatti diversi nell&#39;immagine fMRI. In questo esperimento, i soggetti ricevono airpuffs che può causare movimenti riflessivo, e il succo che richiede la deglutizione. Per ridurre al minimo movimento della testa, un bitebar viene utilizzato per stabilizzare la testa. Il bitebar è collegato alla bobina testa RF. Ogni soggetto ha una custom-dotato boccaglio in materiale termoplastico. Pulsossimetro - FMRI misure di ossigenazione del sangue (BOLD). Ossigenazione del sangue nel cervello può essere influenzata da attività neuronale, ma può anche essere influenzata dal battito cardiaco. Per misurare variazioni dell&#39;ossigenazione legati al ciclo cardiaco, usiamo un pulsossimetro per misurare l&#39;ossigenazione del sangue nel polpastrello utilizzando un sensore a infrarossi. Il pulsossimetro è compatibile MR e il suo segnale di uscita è alimentata tramite un pannello di filtro per la sala di controllo dove è digitalizzato e memorizzato sul computer. Monitoraggio dei gas respiratori - l&#39;ossigenazione del sangue può anche essere influenzato dalla respirazione. Misuriamo la respirazione con un monitor di gas respiratorio che le misure sono scadute livelli di CO2. La RGM è MR compatibile e il suo segnale di uscita è alimentata tramite un pannello di filtro per la sala di controllo dove è digitalizzato e memorizzato sul computer. Stimolazione visiva - Un retroproiezione pannello si trova ad una estremità dello scanner foro vicino alla posizione testa soggetto. Soggetti guardare lo schermo di proiezione posteriore attraverso uno specchio montato sulla parte superiore della bobina testa. Le immagini sono proiettate su schermo di proiezione posteriore da un proiettore a cristalli liquidi situati al di fuori della stanza dello scanner. Il fascio di luce del proiettore è diretto all&#39;interno della stanza scanner attraverso un tubo RF schermato. Eye-tracking - E &#39;importante sapere dove il soggetto è alla ricerca per diverse ragioni. Movimenti oculari influenzare la posizione di stimoli visivi sulla retina, che colpisce le risposte visive del cervello. Movimenti oculari può essere utilizzato anche come un mezzo per avere il soggetto indicano una risposta comportamentale. Noi utilizziamo un metodo chiamato &quot;oculography il video a infrarossi&quot; per monitorare gli occhi. In questo metodo, una telecamera a infrarossi è utilizzato per tenere traccia della pupilla. L&#39;emettitore a infrarossi e telecamera sono costruiti in un paio di occhiali appositamente progettato. Questi sono gli occhiali stessi che forniscono la stimolazione visiva. I dati del occhiali vengono elaborati da un computer dedicato che converte le immagini dell&#39;occhio in segnali analogici per la posizione degli occhi orizzontali e verticali. Preparazione soggetto Documentazione e consenso &amp; ndash; Prima della digitalizzazione, ogni soggetto deve compilare moduli diversi in materia di sicurezza e di consenso. Scheda di Piano sperimentale - Il foglio piano specifica il protocollo sperimentale e la sequenze di impulsi e l&#39;ordine in cui vengono eseguiti. Preparazione fisica - Quando il soggetto è pronto per la scansione, una delle prime cose che fanno è mettere in tappi per le orecchie per proteggere l&#39;udito dai rumori ad alto volume e ad alta frequenza scanner. Hanno anche messo in MR-compatibile cuffie sia per comunicare con i soggetti nello scanner e per fornire istruzioni e feedback durante la sessione di scansione. Il soggetto mette in loro portavoce, che include 1 / 16 ° linee pollici per la consegna di succo e di monitoraggio dei gas respiratori. Il soggetto indossa occhiali e regolare la MR compatibile utilizzato per la stimolazione visiva e il monitoraggio dei movimenti degli occhi. Il soggetto si trova sulla superficie dello scanner e la bobina RF è posizionato sopra la loro testa. Il boccaglio (bitebar) è collegato alla bobina RF per impedire il movimento della testa. Il tubo airpuff (Tygon 1 / 6 di pollice) è posizionato vicino a un occhio utilizzando un tubo flessibile (loc-line). L&#39;impulso-ossimetro clip di dito è collegato a un dito e fissato con del nastro adesivo. Procedura di scansione Sequenze di impulsi: Strutturali: Utilizziamo immagini pesate in T1 (immagini strutturale) che danno una chiara definizione della morfologia del cervello dei soggetti &#39;. Il soggetto si trova passivamente e rimane il più fermo possibile durante questa fase, che dura circa 10 minuti. Funzionale: Utilizziamo T2 sequenze EPI (immagini funzionali) per mostrare variazioni dell&#39;ossigenazione del sangue che sono correlati con l&#39;attività neurale. E &#39;durante questa fase che il soggetto è esposto al succo, airpuff e stimoli uditivi. Connettività: Abbiamo usato la diffusione immagini pesate (DWI) per determinare la connettività strutturale tra le regioni del cervello. Il soggetto si trova semplicemente immobile in questa fase.

functional imaging with reinforcement, eyetracking, and physiological monitoring

Noi usiamo imaging funzionale del cervello (fMRI) per studiare i circuiti neurali che sono alla base del processo decisionale. Per comprendere come i risultati influenzano i processi decisionali, semplici operazioni percettive sono combinati con rinforzi appetitiva e avversione. Tuttavia, l&#39;uso di rinforzi, come succhi di frutta e airpuffs possono creare sfide per fMRI. Consegna rinforzo può causare movimenti della testa, che crea artefatti nel segnale fMRI. Rinforzo può anche portare a cambiamenti nella frequenza cardiaca e la respirazione che sono mediati da percorsi autonomi. Cambiamenti nella frequenza cardiaca e la respirazione può influire direttamente sulla fMRI (BOLD) del segnale nel cervello e possono essere confusi con i cambiamenti del segnale che sono causa di attività neurale. In questa presentazione, ci dimostrano i metodi per la gestione dei rinforzi in modo controllato, per stabilizzare la testa, e per la misurazione del polso e respirazione.

Watch this Scientific Journal Video about Functional Imaging with Reinforcement, Eyetracking, and Physiological Monitoring at JoVE.com

Functional Imaging with Reinforcement, Eyetracking, and Physiological Monitoring

Questa presentazione illustra l&#39;utilizzo di fMRI per studiare i circuiti neurali che sono alla base del processo decisionale. Semplici compiti percettivi sono combinati con rinforzi appetitiva e avversione per studiare come i risultati sui processi decisionali.

Imaging funzionale con rinforzo, eyetracking e monitoraggio fisiologico

We use functional brain imaging (fMRI) to study neural circuits that underlie decision-making.  To understand how outcomes affect decision processes, ...

functional-imaging-with-reinforcement-eyetracking-physiological

Research

JoVE Journal

Biology

11.3K Views.  Columbia University. Questa presentazione illustra l'utilizzo di fMRI per studiare i circuiti neurali che sono alla base del processo decisionale. Semplici compiti percettivi sono combinati con rinforzi appetitiva e avversione per studiare come i risultati sui processi decisionali.

Video: Functional Imaging with Reinforcement, Eyetracking, and Physiological Monitoring

Monitoring Actin Disassembly with Time-lapse Microscopy

Monitoraggio Actina Smontaggio con Time-lapse Microscopia

Ricerca

Biologia

Labeling hESCs and hMSCs with Iron Oxide Nanoparticles for Non-Invasive in vivo Tracking with MR Imaging

In recent years, stem cell research has led to a better understanding of developmental biology, various diseases and its potential impact on regenerative medicine. A non-invasive method to monitor the transplanted stem cells repeatedly in vivo would greatly enhance our ability to understand the mechanisms that control stem cell death and identify trophic factors and signaling pathways that improve stem cell engraftment. MR imaging has been proven to be an effective tool for the in vivo depiction of stem cells with near microscopic anatomical resolution. In order to detect stem cells with MR, the cells have to be labeled with cell specific MR contrast agents. For this purpose, iron oxide nanoparticles, such as superparamagnetic iron oxide particles (SPIO), are applied, because of their high sensitivity for cell detection and their excellent biocompatibility. SPIO particles are composed of an iron oxide core and a dextran, carboxydextran or starch coat, and function by creating local field inhomogeneities, that cause a decreased signal on T2-weighted MR images. This presentation will demonstrate techniques for labeling of stem cells with clinically applicable MR contrast agents for subsequent non-invasive in vivo tracking of the labeled cells with MR imaging.

For the evaluation of new stem cell therapies it is important to non-invasively track the injected cells in vivo. This video will show you how to label human mesenchymal and embryonic stem cells with iron oxide based contrast agents in vivo for subsequent MR imaging in vivo.

Etichettatura hESC e hMSCs con nanoparticelle di ossido di ferro per i non-invasiva in vivo inseguimento con la RM

In recent years, stem cell research has led to a better understanding of developmental biology, various diseases and its potential impact on regenerative ...

Labeling Stem Cells with Fluorescent Dyes for non-invasive Detection with Optical Imaging

Optical imaging (OI) is an easy, fast and inexpensive tool for in vivo monitoring of new stem cell based therapies. The technique is based on ex vivo labeling of stem cells with a fluorescent dye, subsequent intravenous injection of the labeled cells and visualization of their accumulation in specific target organs or pathologies. The presented technique demonstrates how we label human mesenchymal stem cells (hMSC) by simple incubation with the lipophilic fluorescent dye DiD (C67H103CIN2O3S) and how we label human embryonic stem cells (hESC) with the FDA approved fluorescent dye Indocyanine Green (ICG). The uptake mechanism is via adherence and diffusion of the lypophilic dye across the phospholipid cell membrane bilayer. The labeling efficiency is usually improved if the cells are incubated with the dye in serum-free media as opposed to incubation in serum-containing media. Furthermore, the addition of the transfection agent Protamine Sulfate significantly improves contrast agent uptake.

This video shows techniques for labeling of human embryonic stem cells and mesenchymal stem cells with fluorescent dyes. This technique can be used for an in vivo tracking of transplanted stem cells with optical imaging and for histopathological correlations with fluorescence microscopy.

Etichettatura Cellule staminali con le tinture fluorescenti per la rilevazione non invasiva con Optical Imaging

Optical imaging (OI) is an easy, fast and inexpensive tool for in vivo monitoring of new stem cell based therapies. The technique is based on ex vivo ...

Identification of protein complexes with quantitative proteomics in S. cerevisiae

Lipids are the building blocks of cellular membranes that function as barriers and in compartmentalization of cellular processes, and recently, as important intracellular signalling molecules. However, unlike proteins, lipids are small hydrophobic molecules that traffic primarily by poorly described nonvesicular routes, which are hypothesized to occur at membrane contact sites (MCSs). MCSs are regions where the endoplasmic reticulum (ER) makes direct physical contact with a partnering organelle, e.g., plasma membrane (PM). The ER portion of ER-PM MCSs is enriched in lipid-synthesizing enzymes, suggesting that lipid synthesis is directed to these sites and implying that MCSs are important for lipid traffic. Yeast is an ideal model to study ER-PM MCSs because of their abundance, with over 1000 contacts per cell, and their conserved nature in all eukaryotes. Uncovering the proteins that constitute MCSs is critical to understanding how lipids traffic is accomplished in cells, and how they act as signaling molecules. We have found that an ER called Scs2p localize to ER-PM MCSs and is important for their formation. We are focused on uncovering the molecular partners of Scs2p. Identification of protein complexes traditionally relies on first resolving purified protein samples by gel electrophoresis, followed by in-gel digestion of protein bands and analysis of peptides by mass spectrometry. This often limits the study to a small subset of proteins. Also, protein complexes are exposed to denaturing or non-physiological conditions during the procedure. To circumvent these problems, we have implemented a large-scale quantitative proteomics technique to extract unbiased and quantified data. We use stable isotope labeling with amino acids in cell culture (SILAC) to incorporate staple isotope nuclei in proteins in an untagged control strain. Equal volumes of tagged culture and untagged, SILAC-labeled culture are mixed together and lysed by grinding in liquid nitrogen. We then carry out an affinity purification procedure to pull down protein complexes. Finally, we precipitate the protein sample, which is ready for analysis by high-performance liquid chromatography/ tandem mass spectrometry. Most importantly, proteins in the control strain are labeled by the heavy isotope and will produce a mass/ charge shift that can be quantified against the unlabeled proteins in the bait strain. Therefore, contaminants, or unspecific binding can be easily eliminated. By using this approach, we have identified several novel proteins that localize to ER-PM MCSs. Here we present a detailed description of our approach. 

Here we describe a new quantitative proteomics technique for identifying protein complexes in Saccharomyces cerevisiae. In this study, we have used the SILAC method together with affinity purification followed by tandem mass spectrometry to identify with high specificity the binding partners of an ER protein, Scs2p.

L&#39;identificazione di complessi proteici con la proteomica quantitativa in S. cerevisiae

Lipids are the building blocks of cellular membranes that function as barriers and in compartmentalization of cellular processes, and recently, as ...

Dissection of Hippocampal Dentate Gyrus from Adult Mouse

The hippocampus is one of the most widely studied areas in the brain because of its important functional role in memory processing and learning, its remarkable neuronal cell plasticity, and its involvement in epilepsy, neurodegenerative diseases, and psychiatric disorders. The hippocampus is composed of distinct regions; the dentate gyrus, which comprises mainly granule neurons, and Ammon's horn, which comprises mainly pyramidal neurons, and the two regions are connected by both anatomic and functional circuits. Many different mRNAs and proteins are selectively expressed in the dentate gyrus, and the dentate gyrus is a site of adult neurogenesis; that is, new neurons are continually generated in the adult dentate gyrus. To investigate mRNA and protein expression specific to the dentate gyrus, laser capture microdissection is often used. This method has some limitations, however, such as the need for special apparatuses and complicated handling procedures. In this video-recorded protocol, we demonstrate a dissection technique for removing the dentate gyrus from adult mouse under a stereomicroscope. Dentate gyrus samples prepared using this technique are suitable for any assay, including transcriptomic, proteomic, and cell biology analyses. We confirmed that the dissected tissue is dentate gyrus by conducting real-time PCR of dentate gyrus-specific genes, tryptophan 2,3-dioxygenase (TDO2) and desmoplakin (Dsp), and Ammon's horn enriched genes, Meis-related gene 1b (Mrg1b) and TYRO3 protein tyrosine kinase 3 (Tyro3). The mRNA expressions of TDO2 and Dsp in the dentate gyrus samples were detected at obviously higher levels, whereas Mrg1b and Tyro3 were lower levels, than those in the Ammon's horn samples. To demonstrate the advantage of this method, we performed DNA microarray analysis using samples of whole hippocampus and dentate gyrus. The mRNA expression of TDO2 and Dsp, which are expressed selectively in the dentate gyrus, in the whole hippocampus of alpha-CaMKII+/- mice, exhibited 0.037 and 0.10-fold changes compared to that of wild-type mice, respectively. In the isolated dentate gyrus, however, these expressions exhibited 0.011 and 0.021-fold changes compared to that of wild-type mice, demonstrating that gene expression changes in dentate gyrus can be detected with greater sensitivity. Taken together, this convenient and accurate dissection technique can be reliably used for studies focused on the dentate gyrus.

A dissection technique for removal of the dentate gyrus from adult mouse under a stereomicroscope was demonstrated in this video-recorded protocol.

Dissezione di giro dentato ippocampale di topo adulto

The hippocampus is one of the most widely studied areas in the brain because of its important functional role in memory processing and learning, its ...

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

We present various methods to record cardiac function in the larval Drosophila. The approaches allow heart rate to be measured in unrestrained and restrained whole larvae. For direct control of the environment around the heart another approach utilizes the dissected larvae and removal of the internal organs in order to bathe the heart in desired compounds. The exposed heart also allows membrane potentials to be monitored which can give insight of the ionic currents generated by the myocytes and for electrical conduction along the heart tube. These approaches have various advantages and disadvantages for future experiments that are discussed. The larval heart preparation provides an additional model besides the Drosophila skeletal NMJ to investigate the role of intracellular calcium regulation on cellular function. Learning more about the underlying ionic currents that shape the action potentials in myocytes in various species, one can hope to get a handle on the known ionic dysfunctions associated to specific genes responsible for various diseases in mammals.

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

We present various ways to monitor heart function in the larva of Drosophila for assessing questions dealing with the function of gap junctions, ion channel mutations, modulation of pacemaker activity and pharmacological studies.

Il monitoraggio delle funzioni cardiaca in larvali Drosophila melanogaster Per gli Studi fisiologici

We present various methods to record cardiac function in the larval Drosophila. The approaches allow heart rate to be measured in unrestrained  and  ...

Long-term Lethal Toxicity Test with the Crustacean Artemia franciscana

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish water and sediment. The choice of biological indicators must be based on reliable scientific knowledge and, possibly, on the availability of standardized procedures. In this article, we present a standardized protocol that used the marine crustacean Artemia to evaluate the toxicity of chemicals and/or of marine environmental matrices. Scientists propose that the brine shrimp (Artemia) is a suitable candidate for the development of a standard bioassay for worldwide utilization. A number of papers have been published on the toxic effects of various chemicals and toxicants on brine shrimp (Artemia). The major advantage of this crustacean for toxicity studies is the overall availability of the dry cysts; these can be immediately used in testing and difficult cultivation is not demanded1,2. Cyst-based toxicity assays are cheap, continuously available, simple and reliable and are thus an important answer to routine needs of toxicity screening, for industrial monitoring requirements or for regulatory purposes3. The proposed method involves the mortality as an endpoint. The numbers of survivors were counted and percentage of deaths were calculated. Larvae were considered dead if they did not exhibit any internal or external movement during several seconds of observation4. This procedure was standardized testing a reference substance (Sodium Dodecyl Sulfate); some results are reported in this work. This article accompanies a video that describes the performance of procedural toxicity testing, showing all the steps related to the protocol.

Long-term Lethal Toxicity Test with the Crustacean Artemia franciscana

This study concerns the development and standardization of a valuable methodological protocol to determine long-term (14 days) lethal toxicity exerted by chemical substances, industrial wastewater or sewage and liquid environmental samples on the saltwater crustacean, Artemia franciscana.

A lungo termine del test di tossicità letale con il crostaceo Artemia franciscana</em

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish ...

Mapping the After-effects of Theta Burst Stimulation on the Human Auditory Cortex with Functional Imaging

Auditory cortex pertains to the processing of sound, which is at the basis of speech or music-related processing1. However, despite considerable recent progress, the functional properties and lateralization of the human auditory cortex are far from being fully understood. Transcranial Magnetic Stimulation (TMS) is a non-invasive technique that can transiently or lastingly modulate cortical excitability via the application of localized magnetic field pulses, and represents a unique method of exploring plasticity and connectivity. It has only recently begun to be applied to understand auditory cortical function 2. 
An important issue in using TMS is that the physiological consequences of the stimulation are difficult to establish. Although many TMS studies make the implicit assumption that the area targeted by the coil is the area affected, this need not be the case, particularly for complex cognitive functions which depend on interactions across many brain regions 3. One solution to this problem is to combine TMS with functional Magnetic resonance imaging (fMRI). The idea here is that fMRI will provide an index of changes in brain activity associated with TMS. Thus, fMRI would give an independent means of assessing which areas are affected by TMS and how they are modulated 4. In addition, fMRI allows the assessment of functional connectivity, which represents a measure of the temporal coupling between distant regions. It can thus be useful not only to measure the net activity modulation induced by TMS in given locations, but also the degree to which the network properties are affected by TMS, via any observed changes in functional connectivity.
Different approaches exist to combine TMS and functional imaging according to the temporal order of the methods. Functional MRI can be applied before, during, after, or both before and after TMS. Recently, some studies interleaved TMS and fMRI in order to provide online mapping of the functional changes induced by TMS 5-7. However, this online combination has many technical problems, including the static artifacts resulting from the presence of the TMS coil in the scanner room, or the effects of TMS pulses on the process of MR image formation. But more importantly, the loud acoustic noise induced by TMS (increased compared with standard use because of the resonance of the scanner bore) and the increased TMS coil vibrations (caused by the strong mechanical forces due to the static magnetic field of the MR scanner) constitute a crucial problem when studying auditory processing. 
This is one reason why fMRI was carried out before and after TMS in the present study. Similar approaches have been used to target the motor cortex 8,9, premotor cortex 10, primary somatosensory cortex 11,12 and language-related areas 13, but so far no combined TMS-fMRI study has investigated the auditory cortex. The purpose of this article is to provide details concerning the protocol and considerations necessary to successfully combine these two neuroscientific tools to investigate auditory processing. 
Previously we showed that repetitive TMS (rTMS) at high and low frequencies (resp. 10 Hz and 1 Hz) applied over the auditory cortex modulated response time (RT) in a melody discrimination task 2. We also showed that RT modulation was correlated with functional connectivity in the auditory network assessed using fMRI: the higher the functional connectivity between left and right auditory cortices during task performance, the higher the facilitatory effect (i.e. decreased RT) observed with rTMS. However those findings were mainly correlational, as fMRI was performed before rTMS. Here, fMRI was carried out before and immediately after TMS to provide direct measures of the functional organization of the auditory cortex, and more specifically of the plastic reorganization of the auditory neural network occurring after the neural intervention provided by TMS. 
Combined fMRI and TMS applied over the auditory cortex should enable a better understanding of brain mechanisms of auditory processing, providing physiological information about functional effects of TMS. This knowledge could be useful for many cognitive neuroscience applications, as well as for optimizing therapeutic applications of TMS, particularly in auditory-related disorders.

Auditory processing is the basis of speech and music-related processing. Transcranial Magnetic Stimulation (TMS) has been used successfully to study cognitive, sensory and motor systems but has rarely been applied to audition. Here we investigated TMS combined with functional Magnetic Resonance Imaging to understand the functional organization of auditory cortex.

Mappatura delle postumi di Theta Burst stimolazione sulla corteccia uditiva umana con imaging funzionale

Auditory cortex pertains to the processing of sound, which is at the basis of speech or music-related processing1. However, despite considerable recent ...

Functional Imaging of Brown Fat in Mice with 18F-FDG micro-PET/CT

Brown adipose tissue (BAT) differs from white adipose tissue (WAT) by its discrete location and a brown-red color due to rich vascularization and high density of mitochondria. BAT plays a major role in energy expenditure and non-shivering thermogenesis in newborn mammals as well as the adults 1. BAT-mediated thermogenesis is highly regulated by the sympathetic nervous system, predominantly via &beta; adrenergic receptor 2, 3. Recent studies have shown that BAT activities in human adults are negatively correlated with body mass index (BMI) and other diabetic parameters 4-6. BAT has thus been proposed as a potential target for anti-obesity/anti-diabetes therapy focusing on modulation of energy balance 6-8. While several cold challenge-based positron emission tomography (PET) methods are established for detecting human BAT 9-13, there is essentially no standardized protocol for imaging and quantification of BAT in small animal models such as mice. Here we describe a robust PET/CT imaging method for functional assessment of BAT in mice. Briefly, adult C57BL/6J mice were cold treated under fasting conditions for a duration of 4 hours before they received one dose of 18F-Fluorodeoxyglucose (FDG). The mice were remained in the cold for one additional hour post FDG injection, and then scanned with a small animal-dedicated micro-PET/CT system. The acquired PET images were co-registered with the CT images for anatomical references and analyzed for FDG uptake in the interscapular BAT area to present BAT activity. This standardized cold-treatment and imaging protocol has been validated through testing BAT activities during pharmacological interventions, for example, the suppressed BAT activation by the treatment of &beta;-adrenoceptor antagonist propranolol 14, 15, or the enhanced BAT activation by &beta;3 agonist BRL37344 16. The method described here can be applied to screen for drugs/compounds that modulate BAT activity, or to identify genes/pathways that are involved in BAT development and regulation in various preclinical and basic studies.

Functional Imaging of Brown Fat in Mice with 18F-FDG micro-PET/CT

A method of functional imaging of mouse brown adipose tissue (BAT) is described in which cold-stimulated uptake of 18F-Fluorodeoxyglucose (FDG) in BAT is non-invasively assessed with a standardized micro-PET/CT protocol. This method is robust and sensitive to detect differences in BAT activities in mouse models.

Imaging funzionale del grasso bruno nei topi con FDG micro-PET/CT

Brown adipose tissue (BAT) differs from white adipose  tissue (WAT) by its discrete location and a brown-red color due to rich  vascularization and high ...

Concurrent EEG and Functional MRI Recording and Integration Analysis for Dynamic Cortical Activity Imaging

Electroencephalography (EEG) and functional magnetic resonance imaging (fMRI) are two of the fundamental noninvasive methods for identifying brain activity. Multimodal methods have sought to combine the high temporal resolution of EEG with the spatial precision of fMRI, but the complexity of this approach is currently in need of improvement. The protocol presented here describes the recently developed spatiotemporal fMRI-constrained EEG source imaging method, which seeks to rectify source biases and improve EEG-fMRI source localization through the dynamic recruitment of fMRI sub-regions. The process begins with the collection of multimodal data from concurrent EEG and fMRI scans, the generation of 3D cortical models, and independent EEG and fMRI processing. The processed fMRI activation maps are then split into multiple priors, according to their location and surrounding area. These are taken as priors in a two-level hierarchical Bayesian algorithm for EEG source localization. For each window of interest (defined by the operator), specific segments of the fMRI activation map will be identified as active to optimize a parameter known as model evidence. These will be used as soft constraints on the identified cortical activity, increasing the specificity of the multimodal imaging method by reducing cross-talk and avoiding erroneous activity in other conditionally active fMRI regions. The method generates cortical maps of activity and time-courses, which may be taken as final results, or used as a basis for further analyses (analyses of correlation, causation, etc.) While the method is somewhat limited by its modalities (it will not find EEG-invisible sources), it is broadly compatible with most major processing software, and is suitable for most neuroimaging studies.

An EEG-fMRI multimodal imaging method, known as the spatiotemporal fMRI-constrained EEG source imaging method, is described here. The presented method employs conditionally-active fMRI sub-maps, or priors, to guide EEG source localization in a manner that improves spatial specificity and limits erroneous results.