Esta pesquisa foi suportada por um Young Investigator Award da Aliança Nacional dos EUA para a Pesquisa sobre Esquizofrenia e Depressão e um prêmio CPRF da Fundação de Pesquisa Psiquiátrica Canadense (para FD). ED foi apoiado por uma Wyeth-CIHR Fellowship de Pós-Doutorado. Os autores gostariam de agradecer a Peter Seres de assistência com fMRI coleta de dados e Suen Kristina de assistência com a análise de dados.

1. Recolha e selecção de memórias, Task Imaged e protocolo experimental Cobrança da AMs Emocional <ol><li> Memórias pessoais são extraídas de cada participante durante uma entrevista realizada antes da sessão de fMRI, semelhante ao procedimento empregado para não-emocional AMs (eg, 11, 12). Ao contrário da maioria das técnicas anteriores, a nossa AMQ é especificamente construída para atingir a avaliação de episódios emocionais pessoais e suas propriedades recordatório. Um método comumente usado em pesquisas comportamentais e adaptado para uso em estudos de neuroimagem envolve a Crovitz Schiffman e técnica de 13-15, onde os sujeitos se lembrar específicas, eventos experimentou pessoalmente em resposta às palavras de sinalização. Uma foto de paradigma tem sido usado também para investigar AMs em ambientes mais controlados em estudos fMRI 16, 17. No entanto, estas técnicas não-alvo tipicamente exame da componente emocional de AMs, que são comparados em termos de qualidades recordatório. </li><li> Nosso AMQ é composto por uma lista de 115 pistas verbais para eventos de vida distintos (por exemplo, o nascimento de um membro da família, a morte de um parente), que é uma combinação e extensão das listas utilizadas por outros 18, 19. Para cada sugestão, os participantes são convidados a se lembrar de um episódio singular de sua vida, que ocorreu em um lugar específico e tempo (por exemplo, um exemplo, quando ele / ela tocou em um jogo de basquete específico), em vez de lembrar eventos em geral ou repetida (por exemplo, , jogar basquete colegial). Importante, as memórias devem ser acompanhados pela lembrança de estar pessoalmente envolvido, ao invés de ouvir falar dos outros. </li><li> Após o recolhimento, os participantes são convidados a fornecer uma breve descrição da memória (ver Figura 1), que será então utilizada como uma pista de memória personalizado durante a digitalização fMRI (participantes são ingênuos para o fim específico da entrevista de pré-digitalizar). </li><li> Cada memória também é datado e avaliado em seis escalas Likert 20, 21, para avaliar suas propriedades fenomenológicas (ver Figura 1). Nossas escalas incluídas valência emocional (utilizando uma escala de 7 pontos: -3 = muito negativo, 0 = neutral, 3 = muito positivo), intensidade emocional, significado pessoal, a quantidade de detalhes contextuais, a quantidade de visuo-perceptual detalhes ( ou seja, nitidez), ea freqüência de recuperação (todas do último usando uma escala de 7 pontos: 1 = nunca, 7 = extremamente). </li></ol><img alt="Figura 1" src="/files/ftp_upload/2396/2396fig1.jpg" /> Figura 1. Ilustração da AMQ Administração. Para cada sugestão, lembre-se participantes e descrever brevemente um evento específico, e então data ea taxa-lo em 6 escalas. Seleção de AMs altamente emocional <ol start="5"><li> Em seguida, os 40 memórias mais emocionais (20 positivos e negativos 20) são selecionados para cada participante, com base nas classificações fornecidas no AMQ (ie, potência nominal de 2 ou 3 e -2 ou -3, respectivamente). Os AMs positivas e negativas são então combinados em termos de idade e as propriedades fenomenológicas, para garantir que quaisquer diferenças na atividade cerebral durante o recolhimento não são confundidos por diferenças nestas propriedades básicas. </li><li> Se necessário, as descrições das pistas memória selecionada são ligeiramente adaptado para ser correspondido o mais próximo possível de comprimento e complexidade gramatical, é também recomendado para selecionar algumas memórias adicionais para fins de prática. </li></ol> A Task fMRI <ol start="7"><li> A tarefa fMRI é projetado para permitir a comparação da tarefa AM com uma memória semântica (SM) tarefa de controle; os testes AM e SM recuperação têm uma estrutura geral similar (ver Figura 2A). Usamos CIGAL ( <a href="http://www.nitrc.org/projects/cigal/">http://www.nitrc.org/projects/cigal/</a> ) para apresentação do estímulo no scanner MR, mas outros softwares de apresentação de estímulo também pode ser usado. </li><li> A tarefa AM é baseado no pistas personalizadas coletados antes da digitalização. Cada ensaio começa com uma sugestão que desencadeia lembranças AM, que é indicado pelo participante com um aperto de botão. Então, os participantes continuarão lembrando detalhes do evento até cued novamente para a taxa de recolhimento de memória (Figura 2B). </li><li> A tarefa SM envolve geração de exemplares de diferentes categorias semânticas (por exemplo, instrumentos musicais, esportes), que, como recuperação de AM envolve pesquisa em memória e tempo de recuperação prolongado 22 (Figura 2B). Depois de uma categoria semântica é cued, os participantes pressionar um botão assim que eles começam a recordar exemplares dessa categoria e, em seguida, continue lembrando até cued novamente por avaliações de memória. </li></ol><img alt="Figura 2" src="/files/ftp_upload/2396/2396fig2a.jpg" /> <img alt="Figura 2" src="/files/ftp_upload/2396/2396fig2b.jpg" /> Figura 2. Estrutura do Trials fMRI. A. GeralEstrutura dos ensaios. B. estrutura específica dos ensaios AM e SM. <ol start="10"><li> Além da comparação básica de AM e ensaios SM, outras manipulações podem também estar envolvidos. Por exemplo, o foco da recuperação AM pode ser manipulado por dirigir os participantes a prestar atenção aos aspectos emocionais ou não-emocional dos eventos lembrados (Figura 2B). Esta manipulação permite investigação de como foco a recuperação pode influenciar a experiência de AMs e quaisquer alterações associadas em correlatos neurais associados. </li><li> Cada ensaio AM SM ou é seguido por telas de classificação apresentados em ordem contrabalanceada, usando escalas de Likert de 5 pontos (ver Figura 2B). </li><li> O experimento é dividido em roda / blocos de ambos os ensaios para permitir que os participantes tempo para descansar e evitar perda de dados em caso de mau funcionamento do equipamento. Para executar é contrabalançada entre os participantes. Cada execução começa com seis segundos de fixação, para permitir a estabilização do sinal de MR. As condições de AM e SM são apresentados em uma ordem aleatória separadas por um intervalo inter-julgamento de duração variável (5-9 segundos, média = 7seg.). </li></ol> 2. Preparando o assunto para a digitalização Todos os participantes fornecer consentimento informado por escrito antes da execução do protocolo experimental, que é aprovado por um Conselho de Ética. Normalmente, para evitar confunde na lateralização de ativações do cérebro, os participantes analisados ​​são destros. Antes de entrar no quarto de digitalização <ol><li> No dia da varredura, o estado dos participantes atuais afetiva é avaliado 23, para controlar o efeito de humor em lembrança de AMs emocional. Em conjunto com avaliações pós-digitalização, estas avaliações iniciais também pode ser usado para triagem de alterações no humor, como resultado da participação no estudo, e como covariáveis ​​na análise fMRI para investigar ativações cerebrais influenciada por estados atuais. Da mesma forma, a avaliação de traços de personalidade também pode ser feita (por exemplo, neuroticismo), para investigar suas possíveis vieses na tarefa AM 24 e os correlatos neurais associados. </li><li> Antes do exame, os participantes são informados em detalhes dos procedimentos de verificação, e são dadas instruções específicas para a tarefa comportamental. Os participantes também concluir uma sessão de treinos curtos, para se familiarizarem com a tarefa. </li></ol> Entrar na sala de digitalização <ol start="3"><li> Os participantes são instruídos a mentir em posição supina na mesa de digitalização, e são fornecidos com amortecimento de cabeça adicionais, para assegurar o conforto durante a verificação e para minimizar o movimento. Para minimizar ainda mais o movimento da cabeça, do lado não-adesivo de um comprimento de fita pode ser enrolado em torno de leve na testa dos sujeitos. Assuntos são dadas a proteção de ouvido, bem como fones de isolamento para se comunicar com o pesquisador durante o exame de ressonância magnética. </li><li> Mão direita sujeito é posicionado confortavelmente na caixa de resposta, permitindo assim que a mão esquerda para ser usado para apoiar ou para outras medidas (por exemplo, as respostas de condutância da pele). Um botão de parada de emergência é colocado nas proximidades, para que o assunto pode indicar uma necessidade urgente para parar o scanner. </li><li> Antes de iniciar a coleta de dados, é fundamental para se certificar de que os indivíduos podem ver a tela de projeção claramente para apresentação do estímulo e que os botões de resposta funcionar corretamente. </li></ol> 3. A gravação de dados e processamento Digitalização Parâmetros Foram coletados dados de MRI utilizando um 1.5 Tesla Siemens Sonata scanner para gravações de ressonância magnética. Imagens anatômicas foram série anatômica 3D MPRAGE (tempo de repetição (TR) = 1.600 ms, tempo de eco (TE) = 3,82 ms; número de fatias = 112; tamanho voxel = 1x1x1 mm), e as imagens funcionais foram série de 28 fatias funcional, adquirido axialmente usando uma seqüência echoplanar (TR = 2000 ms; TE = 40 ms; campo de visão FOV = mm 256x256; tamanho voxel 4x4x4 = mm), permitindo assim full-cérebro cobertura. Análise de Dados Usamos Statistical Parametric Mapping (SPM: <a href="http://www.fil.ion.ucl.ac.uk/spm">http://www.fil.ion.ucl.ac.uk/spm</a> ) em combinação com as ferramentas Matlab baseado em casa. Pré-processamento envolvidos passos típicos: garantia de qualidade, alinhamento TR, a correção de movimento, co-registro, normalização e suavização (8 mm 3 Kernel). Individuais e de grupo em nível de análises estatísticas pode envolver comparações de atividade cerebral de acordo com o tipo de memória (AM vs SM), o foco de valência (positivo vs negativo), e recuperação emocional (conteúdo emocional versus não-emocional). 4. Resultados representante <img alt="Figura 3" src="/files/ftp_upload/2396/2396fig3.jpg" /> Figura 3. Correlatos neurais da Recuperação AM. Validando o presente protocolo, a recuperação de AMs rendeu aumento de atividade na recuperação da rede AM 4, 25, incluindo hippoáreas CAMPAL (a), envolvidos na recuperação da memória em geral, o córtex pré-frontal medial (b), associada ao engajamento pessoal, as regiões cuneus / precuneus e parieto-occipitais de junção (c, e, respectivamente), associados ao processamento de representação visuo-espacial , ea junção fronto-temporal (d), envolvidos na recuperação afetiva AM; efeitos semelhantes a este último também foram encontrados na amígdala (não mostrado). Os &quot;mapas de ativação&quot; são sobrepostos a imagens de alta resolução do cérebro exibido em coronal (painel esquerdo) e sagital (painéis de média e direita) pontos de vista; as barras coloridas indicam o gradiente de valores t dos mapas de ativação (p &lt;0,005, 10 voxels contíguos 26), refletindo a atividade do cérebro em tempo bloqueado para o inícios das pistas de memória. Os gráficos ilustram a linha de cursos de tempo do sinal de fMRI (mudança de sinal%), para cada tipo de tentativa e TR (1 TR = 2 segundos). L = Left, R = Right.

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O delineamento experimental introduzida aqui permite investigação dos correlatos neurais de recordar memórias autobiográficas emocional. Este projeto tem o potencial de aumentar o nosso conhecimento de como o cérebro gera preconceitos afetivos (positivo ou negativo) em recordar memórias pessoais, e como essas tendências podem ser moduladas pelo foco de recuperação (nos aspectos emocionais ou não-emocional). Este protocolo tem benefícios adicionais na medida em que também pode ser utilizado com populações c...

brain imaging investigation of the neural correlates of emotional autobiographical recollection

Recordação de memórias autobiográficas emocional (AMs) é importante para o funcionamento cognitivo e afetivo saudável 1 - lembrando AMs positiva está associada com pessoal aumentaram o bem-estar e auto-estima 2, enquanto que lembrar e ruminando AMs negativas pode levar a transtornos afetivos 3. Embora progressos significativos tenham sido feitos na compreensão dos mecanismos cerebrais subjacentes AM recuperação em geral (revista em 4, 5), menos se sabe sobre o efeito da emoção sobre a subjetiva re-experiência de AMs e os correlatos neurais associados. Esta é, em parte devido ao fato de que, ao contrário das investigações do efeito emoção sobre a memória para laboratório baseado em eventos micro (revisto em 6, 7-9), muitas vezes, AM estudos não têm um foco claro sobre os aspectos emocionais da lembrando eventos pessoais (mas veja 10). Aqui, apresentamos um protocolo que permite investigação dos correlatos neurais da relembrando AMs emocionais usando ressonância magnética funcional (fMRI). Pistas para estas memórias são coletados antes da digitalização por meio de um questionário de memória autobiográfica (AMQ), permitindo assim a seleção adequada de AMs emocional baseado em suas propriedades fenomenológica (isto é, intensidade, vivacidade, significado pessoal). Este protocolo pode ser utilizado em populações saudáveis ​​e clínicos semelhantes.

Watch this Scientific Journal Video about Brain Imaging Investigation of the Neural Correlates of Emotional Autobiographical Recollection at JoVE.com

Brain Imaging Investigation of the Neural Correlates of Emotional Autobiographical Recollection

Apresentamos um protocolo que permite a investigação dos correlatos neurais da relembrando memórias autobiográficas emocional, usando ressonância magnética funcional. Este protocolo pode ser usado com participantes saudáveis ​​e clínicos.

Investigação por imagem do cérebro os correlatos neurais da Recordação Autobiographical Emocional

Recollection of emotional autobiographical memories (AMs) is important to healthy cognitive and affective functioning 1 - remembering positive AMs is ...

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9.7K visualizações.  University of Alberta, Edmonton. Apresentamos um protocolo que permite a investigação dos correlatos neurais da relembrando memórias autobiográficas emocional, usando ressonância magnética funcional. Este protocolo pode ser usado com participantes saudáveis ​​e clínicos.

Vídeo: Brain Imaging Investigation of the Neural Correlates of Emotional Autobiographical Recollection

Brain Imaging Investigation of the Neural Correlates of Observing Virtual Social Interactions

The ability to gauge social interactions is crucial in the assessment of others&#x2019; intentions. Factors such as facial expressions and body language affect our decisions in personal and professional life alike 1. These "friend or foe" judgements are often based on first impressions, which in turn may affect our decisions to "approach or avoid". Previous studies investigating the neural correlates of social cognition tended to use static facial stimuli 2. Here, we illustrate an experimental design in which whole-body animated characters were used in conjunction with functional magnetic resonance imaging (fMRI) recordings. Fifteen participants were presented with short movie-clips of guest-host interactions in a business setting, while fMRI data were recorded; at the end of each movie, participants also provided ratings of the host behaviour. This design mimics more closely real-life situations, and hence may contribute to better understanding of the neural mechanisms of social interactions in healthy behaviour, and to gaining insight into possible causes of deficits in social behaviour in such clinical conditions as social anxiety and autism 3.

This article demonstrates an experimental design in which whole-body animated characters are used in conjunction with functional magnetic resonance imaging (fMRI) to investigate the neural correlates of observing virtual social interactions.

Investigação por imagem do cérebro os correlatos neurais da Observando Virtual Interações Sociais

The ability to gauge social interactions is crucial in the assessment of others&#x2019; intentions. Factors such as facial expressions and body language ...

Neurociência

Brain Imaging Investigation of the Neural Correlates of Emotion Regulation

The ability to control/regulate emotions is an important coping mechanism in the face of emotionally stressful situations. Although significant progress has been made in understanding conscious/deliberate emotion regulation (ER), less is known about non-conscious/automatic ER and the associated neural correlates. This is in part due to the problems inherent in the unitary concepts of automatic and conscious processing1. Here, we present a protocol that allows investigation of the neural correlates of both deliberate and automatic ER using functional magnetic resonance imaging (fMRI). This protocol allows new avenues of inquiry into various aspects of ER. For instance, the experimental design allows manipulation of the goal to regulate emotion (conscious vs. non-conscious), as well as the intensity of the emotional challenge (high vs. low). Moreover, it allows investigation of both immediate (emotion perception) and long-term effects (emotional memory) of ER strategies on emotion processing. Therefore, this protocol may contribute to better understanding of the neural mechanisms of emotion regulation in healthy behaviour, and to gaining insight into possible causes of deficits in depression and anxiety disorders in which emotion dysregulation is often among the core debilitating features.

We present a protocol that allows investigation of the neural correlates of deliberate and automatic emotion regulation, using functional magnetic resonance imaging. This protocol can be used in healthy participants, both young and older, as well as in clinical patients.

Investigação por imagem do cérebro os correlatos neurais da regulação emocional

The ability to control/regulate emotions is an important coping mechanism in the face of emotionally stressful situations. Although significant progress ...

Brain Imaging Investigation of the Memory-Enhancing Effect of Emotion

Emotional events tend to be better remembered than non-emotional events1,2. One goal of cognitive and affective neuroscientists is 
to understand the neural mechanisms underlying this enhancing effect of emotion on memory. A method that has proven particularly influential in the 
investigation of the memory-enhancing effect of emotion is the so-called subsequent memory paradigm (SMP). This method was originally used to investigate the 
neural correlates of non-emotional memories3, and more recently we and others also applied it successfully to studies of emotional memory (reviewed in4, 5-7). 
Here, we describe a protocol that allows investigation of the neural correlates of the memory-enhancing effect of emotion using the SMP in conjunction with 
event-related functional magnetic resonance imaging (fMRI). An important feature of the SMP is that it allows separation of brain activity specifically 
associated with memory from more general activity associated with perception. Moreover, in the context of investigating the impact of emotional stimuli, 
SMP allows identification of brain regions whose activity is susceptible to emotional modulation of both general/perceptual and memory-specific processing. 
This protocol can be used in healthy subjects8-15, as well as in clinical patients where there are alterations in the neural correlates of emotion perception 
and biases in remembering emotional events, such as those suffering from depression and post-traumatic stress disorder (PTSD)16, 17.

We present a protocol that uses functional magnetic resonance imaging to investigate the neural correlates of the memory-enhancing effect of emotion. This protocol allows identification of brain activity specifically linked to memory-related processing, contrary to more general perceptual processing, and can be used with healthy and clinical populations.

Investigação de imagens do cérebro do Efeito Memória de aumento da Emoção

Emotional events tend to be better remembered than non-emotional events1,2. One goal of cognitive and affective neuroscientists is 
to understand the ...

Brain Imaging Investigation of the Impairing Effect of Emotion on Cognition

Emotions can impact cognition by exerting both enhancing (e.g., better memory for emotional events) and impairing (e.g., increased emotional distractibility) effects (reviewed in 1). Complementing our recent protocol 2 describing a method that allows investigation of the neural correlates of the memory-enhancing effect of emotion (see also 1, 3-5), here we present a protocol that allows investigation of the neural correlates of the detrimental impact of emotion on cognition. The main feature of this method is that it allows identification of reciprocal modulations between activity in a ventral neural system, involved in 'hot' emotion processing (HotEmo system), and a dorsal system, involved in higher-level 'cold' cognitive/executive processing (ColdEx system), which are linked to cognitive performance and to individual variations in behavior (reviewed in 1). Since its initial introduction 6, this design has proven particularly versatile and influential in the elucidation of various aspects concerning the neural correlates of the detrimental impact of emotional distraction on cognition, with a focus on working memory (WM), and of coping with such distraction 7,11, in both healthy 8-11 and clinical participants 12-14.

We present a protocol that allows investigation of the neural mechanisms mediating the detrimental impact of emotion on cognition, using functional magnetic resonance imaging. This protocol can be used with both healthy and clinical participants.

Investigação por imagem do cérebro o efeito prejudicial da emoção sobre a Cognição

Emotions can impact cognition by exerting both enhancing (e.g., better memory for emotional events) and impairing (e.g., increased emotional ...

Detection of Microregional Hypoxia in Mouse Cerebral Cortex by Two-photon Imaging of Endogenous NADH Fluorescence

The brain's ability to function at high levels of metabolic demand depends on continuous oxygen supply through blood flow and tissue oxygen diffusion. Here we present a visualized experimental and methodological protocol to directly visualize microregional tissue hypoxia and to infer perivascular oxygen gradients in the mouse cortex. It is based on the non-linear relationship between nicotinamide adenine dinucleotide (NADH) endogenous fluorescence intensity and oxygen partial pressure in the tissue, where observed tissue NADH fluorescence abruptly increases at tissue oxygen levels below 10 mmHg1. We use two-photon excitation at 740 nm which allows for concurrent excitation of intrinsic NADH tissue fluorescence and blood plasma contrasted with Texas-Red dextran. The advantages of this method over existing approaches include the following: it takes advantage of an intrinsic tissue signal and can be performed using standard two-photon in vivo imaging equipment; it permits continuous monitoring in the whole field of view with a depth resolution of ~50 &mu;m. We demonstrate that brain tissue areas furthest from cerebral blood vessels correspond to vulnerable watershed areas which are the first to become functionally hypoxic following a decline in vascular oxygen supply. This method allows one to image microregional cortical oxygenation and is therefore useful for examining the role of inadequate or restricted tissue oxygen supply in neurovascular diseases and stroke.

Here we describe a method to directly visualize microregional tissue hypoxia in the mouse cortex in vivo. It is based on concurrent two-photon imaging of nicotinamide adenine dinucleotide (NADH) and the cortical microcirculation. This method is useful for high resolution analysis of tissue oxygen supply.

Detecção de hipóxia microrregional no Cortex Cerebral mouse por imagem de dois fótons de fluorescência endógena NADH

The brain's ability to function at high levels of metabolic demand depends on continuous oxygen supply through blood flow and tissue oxygen diffusion. ...

Extinction Training During the Reconsolidation Window Prevents Recovery of Fear

Fear is maladaptive when it persists long after circumstances have become safe. It is therefore crucial to develop an approach that persistently prevents the return of fear. Pavlovian fear-conditioning paradigms are commonly employed to create a controlled, novel fear association in the laboratory. After pairing an innocuous stimulus (conditioned stimulus, CS) with an aversive outcome (unconditioned stimulus, US) we can elicit a fear response (conditioned response, or CR) by presenting just the stimulus alone1,2 . Once fear is acquired, it can be diminished using extinction training, whereby the conditioned stimulus is repeatedly presented without the aversive outcome until fear is no longer expressed3. This inhibitory learning creates a new, safe representation for the CS, which competes for expression with the original fear memory4. Although extinction is effective at inhibiting fear, it is not permanent. Fear can spontaneously recover with the passage of time. Exposure to stress or returning to the context of initial learning can also cause fear to resurface3,4.
Our protocol addresses the transient nature of extinction by targeting the reconsolidation window to modify emotional memory in a more permanent manner. Ample evidence suggests that reactivating a consolidated memory returns it to a labile state, during which the memory is again susceptible to interference5-9. This window of opportunity appears to open shortly after reactivation and close approximately 6hrs later5,11,16, although this may vary depending on the strength and age of the memory15. By allowing new information to incorporate into the original memory trace, this memory may be updated as it reconsolidates10,11. Studies involving non-human animals have successfully blocked the expression of fear memory by introducing pharmacological manipulations within the reconsolidation window, however, most agents used are either toxic to humans or show equivocal effects when used in human studies12-14. Our protocol addresses these challenges by offering an effective, yet non-invasive, behavioral manipulation that is safe for humans.
By prompting fear memory retrieval prior to extinction, we essentially trigger the reconsolidation process, allowing new safety information (i.e., extinction) to be incorporated while the fear memory is still susceptible to interference. A recent study employing this behavioral manipulation in rats has successfully blocked fear memory using these temporal parameters11. Additional studies in humans have demonstrated that introducing new information after the retrieval of previously consolidated motor16, episodic17, or declarative18 memories leads to interference with the original memory trace14. We outline below a novel protocol used to block fear recovery in humans.

Conditioned fear can be diminished through an inhibitory process called extinction, but can resurface under conditions such as the passage of time or exposure to stress. Our protocol presents a novel way of preventing fear recovery by introducing extinction during the reconsolidation window (the re-storage phase of a reactivated memory).

Formação extinção durante a janela de reconsolidação impede a recuperação do Medo

Fear is maladaptive when it persists long after circumstances have become safe. It is therefore crucial to develop an approach that persistently prevents ...

Co-analysis of Brain Structure and Function using fMRI and Diffusion-weighted Imaging

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when studying the brain, as the functional role that a brain area fulfills may be largely defined by its connections to other brain areas. In this report, we describe a novel, non-invasive approach for relating brain structure and function using magnetic resonance imaging (MRI). This approach, a combination of structural imaging of long-range fiber connections and functional imaging data, is illustrated in two distinct cognitive domains, visual attention and face perception. Structural imaging is performed with diffusion-weighted imaging (DWI) and fiber tractography, which track the diffusion of water molecules along white-matter fiber tracts in the brain (Figure 1). By visualizing these fiber tracts, we are able to investigate the long-range connective architecture of the brain. The results compare favorably with one of the most widely-used techniques in DWI, diffusion tensor imaging (DTI). DTI is unable to resolve complex configurations of fiber tracts, limiting its utility for constructing detailed, anatomically-informed models of brain function. In contrast, our analyses reproduce known neuroanatomy with precision and accuracy. This advantage is partly due to data acquisition procedures: while many DTI protocols measure diffusion in a small number of directions (e.g., 6 or 12), we employ a diffusion spectrum imaging (DSI)1, 2 protocol which assesses diffusion in 257 directions and at a range of magnetic gradient strengths. Moreover, DSI data allow us to use more sophisticated methods for reconstructing acquired data. In two experiments (visual attention and face perception), tractography reveals that co-active areas of the human brain are anatomically connected, supporting extant hypotheses that they form functional networks. DWI allows us to create a "circuit diagram" and reproduce it on an individual-subject basis, for the purpose of monitoring task-relevant brain activity in networks of interest.

We describe a novel approach for simultaneous analysis of brain function and structure using magnetic resonance imaging (MRI). We assess brain structure with high-resolution diffusion-weighted imaging and white-matter fiber tractography. Unlike standard structural MRI, these techniques allow us to directly relate anatomical connectivity to functional properties of brain networks.

Co-análise de estrutura e função cerebral usando ressonância magnética funcional e imagem ponderada em difusão

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when ...

A Computer-assisted Multi-electrode Patch-clamp System

The patch-clamp technique is today the most well-established method for recording electrical activity from individual neurons or their subcellular compartments. Nevertheless, achieving stable recordings, even from individual cells, remains a time-consuming procedure of considerable complexity. Automation of many steps in conjunction with efficient information display can greatly assist experimentalists in performing a larger number of recordings with greater reliability and in less time. In order to achieve large-scale recordings we concluded the most efficient approach is not to fully automatize the process but to simplify the experimental steps and reduce the chances of human error while efficiently incorporating the experimenter's experience and visual feedback. With these goals in mind we developed a computer-assisted system which centralizes all the controls necessary for a multi-electrode patch-clamp experiment in a single interface, a commercially available wireless gamepad, while displaying experiment related information and guidance cues on the computer screen. Here we describe the different components of the system which allowed us to reduce the time required for achieving the recording configuration and substantially increase the chances of successfully recording large numbers of neurons simultaneously.

Multi-electrode patch-clamp recordings constitute a complex task. Here we show how, by automating of many of the experimental steps, it is possible to accelerate the process leading to qualitative improvement in performance and number of recordings.

A Multi-eletrodo Sistema patch-clamp assistida por computador

The patch-clamp technique is today the most  well-established method for recording electrical activity from individual  neurons or their subcellular ...

Analysis of Brain Mitochondria Using Serial Block-Face Scanning Electron Microscopy

Human brain is a high energy consuming organ that mainly relies on glucose as a fuel source. Glucose is catabolized by brain mitochondria via glycolysis, tri-carboxylic acid (TCA) cycle and oxidative phosphorylation (OXPHOS) pathways to produce cellular energy in the form of adenosine triphosphate (ATP). Impairment of mitochondrial ATP production causes mitochondrial disorders, which present clinically with prominent neurological and myopathic symptoms. Mitochondrial defects are also present in neurodevelopmental disorders (e.g. autism spectrum disorder) and neurodegenerative disorders (e.g. amyotrophic lateral sclerosis, Alzheimer&#39;s and Parkinson&#39;s diseases). Thus, there is an increased interest in the field for performing 3D analysis of mitochondrial morphology, structure and distribution under both healthy and disease states. The brain mitochondrial morphology is extremely diverse, with some mitochondria especially those in the synaptic region being in the range of &#60;200 nm diameter, which is below the resolution limit of traditional light microscopy. Expressing a mitochondrially-targeted green fluorescent protein (GFP) in the brain significantly enhances the organellar detection by confocal microscopy. However, it does not overcome the constraints on the sensitivity of detection of relatively small sized mitochondria without oversaturating the images of large sized mitochondria. While serial transmission electron microscopy has been successfully used to characterize mitochondria at the neuronal synapse, this technique is extremely time-consuming especially when comparing multiple samples. The serial block-face scanning electron microscopy (SBFSEM) technique involves an automated process of sectioning, imaging blocks of tissue and data acquisition. Here, we provide a protocol to perform SBFSEM of a defined region from rodent brain to rapidly reconstruct and visualize mitochondrial morphology. This technique could also be used to provide accurate information on mitochondrial number, volume, size and distribution in a defined brain region. Since the obtained image resolution is high (typically under 10 nm) any gross mitochondrial morphological defects may also be detected.

Mitochondrial visualization and analysis from mammalian brain tissue is a challenging task. Here, we describe how three dimensional (3D) reconstruction analysis from the serial block-face scanning electron microscopy (SBFSEM) can be used to gain insights on the morphological and volumetric analysis of this critical energy generating organelle.

Análise do Cérebro mitocôndrias Usando Serial Block-cara Scanning Electron Microscopy

Human brain is a high energy consuming organ that mainly relies on glucose as a fuel source. Glucose is catabolized by brain mitochondria via glycolysis, ...

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in adult mammalian animals remains elusive. In this study, we describe an experimental procedure for measuring calcium dynamics through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy. This method enables recordings and quantifications of neural activity in bilateral mouse brain regions one at a time in the same experiment for a prolonged period in vivo. Key aspects of this method, which can be completed within an hour, include minimally invasive surgery procedures for creating dual optical windows, and the use of 2P imaging. Although we only demonstrate the technique in the S1 area, the method can be applied to other regions of the living brain facilitating the elucidation of structural and functional complexities of brain neural networks.

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

We describe an experimental procedure for measuring neuronal activity through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy in vivo.

Imagem de atividade Neural no córtex somatossensorial primário usando Thy1-GCaMP6s de ratos transgênicos

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in ...