Gostaríamos de agradecer a Joy Bergmann, Attilio Ceretti, Sarah Cohen, Nina Duncan, Aaron Hall, Hanna Lefebo, Madeleine Salvatore por sua ajuda com o comportamento e Dr. Vinay Parikh, Dr. Brittney Yegla e Rob Cole para seus conselhos sobre os parâmetros de SAT. Gostaríamos de agradecer o Dr. Jill McGaughy dicas sobre o processo de formação. Este trabalho foi financiado pelos institutos nacionais de subsídio de saúde 092438 NIMH e National Science Foundation grant 1552416 IOS para D.A.B.

<ol>
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Déficits na atenção sustentada são relatados em neuropsiquiátrica, desenvolvimento neurológico e Neurodegenerativas Transtornos1,2,3, mas os processos que contribuem para o ideal versus atenção interrompida são Understudy. Isto é, em parte, porque o equipamento especializado limita a fácil adaptação de tarefas de atenção de roedores. No entanto, touchscreen câmaras estão abordando esse assunto, fornecendo a flex...

Distúrbios variando de atenção e hiperatividade (TDAH) a esquizofrenia, a doença de Alzheimer, compartilhar deficiências de atenção como um recurso1,2,3. Déficits na atenção sustentada — a capacidade de monitorar continuamente uma situação para eventos intermitentes e imprevisíveis — são particularmente perturbador, porque atenção sustentada é crucial para a atenção selectiva e dividida, assim como outros cognitivo processos de4,5. Mesmo em pessoas saudáveis, dificuldades em sustentar a atenção negativamente afeta a cognição, prejudicando o diário função6. Assim, a compreender a base Neurobiológica da atenção sustentada e como torna-se prejudicado pode levar a intervenções para melhorar a cognição que beneficiaria muitas pessoas.
A fim de delinear circuitos e processos moleculares que contribuem para a adequada contra atenção interrompida, muitos pesquisadores se transformaram em modelos animais não-humanos, onde a manipulação de populações de células específicas e processos moleculares durante a atenção tarefas é possível. O esforço tem levado ao desenvolvimento de uma variedade de tarefas de atenção operante que pode avaliar a capacidade de sustentar a atenção7,8,9,10. Um tal paradigma, desenvolvida pela McGaughy e Sarter (1995), é conhecida como a tarefa de atenção sustentada (SAT). Roedores treinados em SAT devem distinguir julgamentos de sinal, em que uma luz de sinal pisca brevemente, de ensaios não-sinal. Demandas de atenção podem ser aumentadas através da introdução de um distrator visual (piscando houselight)10. Os parâmetros críticos para avaliar a atenção no SAT estão bem documentados e esta tarefa foi validada em ratos machos e fêmeas e camundongos de11,10,12. Uma versão ainda foi adaptada para os seres humanos, destacando o utilitário translacional de SAT13. O uso desta tarefa importante, ajudou a implicar o sistema de corticopetal do prosencéfalo basal na atenção sustentada4. Especificamente, neurônios colinérgicos do núcleo basalis de Meynert (NBM) / substância inominata (SI) região do prosencéfalo basal que projetam para o córtex pré-frontal são críticos para hits, que são respostas precisas sobre sinal ensaios14, 15. Em contraste, os neurônios gabaérgica nesta região são pensados para mediar o desempenho durante a corretas rejeições (CRs), que são respostas precisas em ensaios não-sinal16. Uma vez que o circuito básico para esta tarefa foi criado, fatores que modulam este circuito para prejudicar a atenção, variando de hormônios do estresse a proteínas neurotóxicas foram identificados17,18. Coletivamente, estes estudos destacam-se o utilitário de SAT.
Uma limitação à implementação SAT no laboratório é que o procedimento original requer operante câmaras permanentemente configuradas para a tarefa, com uma luz de painel de centro e duas patilhas extensíveis, um designado para sucessos e outro para CRs10. Em um julgamento de sinal, o luz do painel acende-se brevemente e estendem para indicar uma janela de resposta s 4 alavancas. Durante essa janela, pressionando a alavanca sucesso designada, então, o rato recebe uma recompensa (comida ou água). Prensas de alavanca incorreto em ensaios de sinal, conhecidos como misses, não são recompensadas. Em ensaios não-sinalizado, a luz do painel permanece desligado e, em seguida, alavancas são estendidas. Enquanto um CR é recompensado, prensas de alavanca incorreto em ensaios não-sinalizado, conhecidos como alarmes falsos, não são recompensadas. Uma falha ao fazer qualquer resposta durante a janela de resposta conta como uma omissão. Para expandir a disponibilidade do SAT, modificamos recentemente a tarefa para câmaras operante touchscreen, em que uma parede é um ecrã táctil que pode tanto exibir estímulos visuais e nariz-poke registro respostas19. Câmaras de touchscreen estão se tornando populares por causa de sua versatilidade para executar uma variedade de tarefas (por exemplo, emparelhado associada aprendizagem, aprendizagem de reversão, etc.) com um pedaço de equipamento20. Semelhante a outros procedimentos de touchscreen20,21,22, nossa adaptação do SAT requer o uso de uma capa de touchscreen de plástico opaco, chamado uma máscara, com furos desviá-la. Um buraco central, circular permite a apresentação de um sinal através da apresentação de um estímulo branco na tela por trás disso, e duas áreas de resposta quadrados, um designado para sucessos e outra para o CRs, permitir que o rato fazer toque respostas19. Nas câmaras tradicionais, a janela de resposta de 4-s é normalmente indicada com a apresentação das alavancas, mas um estudo anterior demonstrou que um tom também pode ser usado para sinalizar a janela de resposta23. Em touchscreen SAT, da mesma forma sinalizarmos a janela de resposta com um tom breve. Outros parâmetros de formação (por exemplo, o número de ensaios, intervalo de intertrial, etc.) são mantidos semelhantes entre o tradicional e touchscreen SAT. Anteriormente avaliamos o desempenho entre as versões tradicionais e tela sensível ao toque do SAT e achei que era comparável, sugerindo que touchscreen SAT é uma forma válida para medir a atenção sustentada19. A modificação de touchscreen torna SAT mais versátil porque pode agora ser adaptada para uso difundido nas caixas operante de touchscreen.
O presente protocolo detalha como executar touchscreen SAT no laboratório. Os horários de treinamento foram ligeiramente alterados do nosso relatório anterior da tarefa19 para otimizar o tempo de aquisição em ratos fêmeas, que, ao contrário dos machos, teve dificuldade, avançando através as programações anteriores24. Resultados ilustrando desempenho típico em ratos machos e fêmeas são incluídos, juntamente com dicas para resolução de problemas com a tarefa.

<table>
	<tbody>
		<tr>
			<td>Bussey-Saksida Rat Touch Screen Chambers Easy-Install System</td>
			<td>Lafayette Instrument</td>
			<td>80604-20</td>
			<td>Includes: Touch Screen, Chamber base with perforated floors, Large Feeder Reward Area (44.5mm wide x 92.5mm high), Speaker, Houselight, Tone Generator, SAC; Trapezoidal animal working area is 126mm wide at the Feeder, 240mm wide and 332 mm deep at the Screen, and 300 mm in height</td>
		</tr>
		<tr>
			<td>Sound Attenuation Cubicles (SAC)</td>
			<td>Lafayette Instrument</td>
			<td>80604-20</td>
			<td>Attenuation to approximately 35dB; 38kg in weight; External dimensions: 600mm wide x 670mm tall x 352mm deep; Internal dimensions: 540mm wide x 610mm tall x 532mm deep</td>
		</tr>
		<tr>
			<td>Abet II Software for Touch Screens</td>
			<td>Lafayette Instrument</td>
			<td>89505</td>
			<td>Version 2.19</td>
		</tr>
		<tr>
			<td>Whisker Multimedia</td>
			<td>Lafayette Instrument</td>
			<td>80698-1</td>
			<td>Required for Abet II Touch Operation</td>
		</tr>
		<tr>
			<td>Pellet Dispenser</td>
			<td>Lafayette Instrument</td>
			<td>80209-45</td>
			<td>45mg Interchangeable Pellet Size Wheel</td>
		</tr>
		<tr>
			<td>Camera</td>
			<td>Lafayette Instrument</td>
			<td>80600-CAM</td>
			<td>Filtered and focused for IR light</td>
		</tr>
		<tr>
			<td>Microsoft Windows</td>
			<td>Microsoft</td>
			<td></td>
			<td>Windows 7-64 bit recommended</td>
		</tr>
		<tr>
			<td>Controller PC-Touch Screen Chambers</td>
			<td>Lafayette Instrument</td>
			<td>88530</td>
			<td>Dual Core Pentinum Processor; 2.5 GHz or greater; keyboard, mouse, and monitor; Installed PCI card, cable and expansion; Four RS232 ports; Four VGA ports</td>
		</tr>
		<tr>
			<td>Dustless Precision Pellets</td>
			<td>BioServe</td>
			<td>F0165</td>
			<td>45 mg</td>
		</tr>
		<tr>
			<td>Black Acrylic Mask</td>
			<td>Everything Plastic</td>
			<td>Custom Product</td>
			<td>Black polycarbonate with one matte finish side, 3.18 mm thick; Central circular opening, 28.58 mm diameter, 107.95 mm from the bottom, centered between the left and right side; Two square response areas, 28.57 mm x 28.57mm each positioned below and off center, one 85.725 mm from the left and one 85.725 mm from right side, and both 34.925 mm from the bottom.</td>
		</tr>
		<tr>
			<td>Utility Cart</td>
			<td>Fisher Scientific</td>
			<td>11-954-754</td>
			<td>30.75 in. x 18.38 in x 33 in</td>
		</tr>
		<tr>
			<td>Black towel</td>
			<td></td>
			<td></td>
			<td>Large enough to cover cages for animal transport</td>
		</tr>
		<tr>
			<td>Headlights with red lights</td>
			<td>Energizer</td>
			<td>HDL33A2E</td>
			<td>LED white and red light</td>
		</tr>
	</tbody>
</table>

touchscreen sustained attention task (sat) for rats

Atenção sustentada é a capacidade de monitorar eventos imprevisíveis e intermitentes durante um período prolongado de tempo. Este processo de atenção subserves outros aspectos da cognição e é interrompido em certo neurológico, neuropsiquiátricos e doenças neurodegenerativas. Assim, é clinicamente importante identificar mecanismos que prejudicam e melhorar a atenção sustentada. Tais mecanismos são frequentemente descobertos usando modelos de roedores. Portanto, foram desenvolvidos vários procedimentos de comportamento para os aspectos de atenção sustentada de teste para os roedores. Um, primeiro descrito por McGaughy e Sarter (1995), chamado a tarefa de atenção sustentada (SAT), treina ratos para distinguir entre o sinal (ou seja, breve apresentação de luz) e ensaios não-sinal. Os sinais são curtos e, portanto, requerem atenção cuidadosa para ser percebida. Demandas de atenção podem ser aumentadas ainda mais através da introdução de um distrator (por exemplo, houselight a piscar). Modificamos a esta tarefa para câmaras operante touchscreen, que são configurados com uma tela sensível ao toque em uma parede que pode apresentar estímulos e respostas de registro. Aqui detalhamos nosso protocolo para sab em câmaras de touchscreen. Além disso, apresentamos as medidas padrão de desempenho em ratos Sprague-Dawley machos e fêmeas. Desempenho comparável sobre esta tarefa em ambos os sexos destaca seu uso para estudos de atenção, especialmente porque mais pesquisadores estão incluindo roedores femininas em seu projeto experimental. Além disso, a fácil implementação do SAT para as câmaras cada vez mais popular touchscreen aumenta sua utilidade.

Para a maioria dos estudos de atenção, ratos são treinados para critérios em sab e então manipulações de interromper ou melhorar a atenção são executadas. Dados de aquisição não é normalmente apresentados, mas fazemo-lo aqui para ilustrar preços de aquisição em ratos Sprague-Dawley machos e fêmeas. Para este fim, podemos quantificar o número médio de dias para atingir os critérios em SAT (começando na fase de cutucar nariz). Observe que alguns ratos nunca chegou a cr...

Watch this Scientific Journal Video about Touchscreen Sustained Attention Task SAT for Rats at JoVE.com

Touchscreen Sustained Attention Task SAT for Rats

Touchscreen sustentada atenção tarefa (SAT) para ratos

Atenção sustentada, ou monitorar continuamente situações para eventos intermitentes e imprevisíveis, é um aspecto crítico da cognição. Aqui detalhamos como teste de atenção sustentada em ratos utilizando câmaras operante de touchscreen. Vamos demonstrar o desempenho comparável em ratos machos e fêmeas, tornando esta tarefa útil para estudar a atenção em ambos os sexos.

Sustained attention is the ability to monitor intermittent and unpredictable events over a prolonged period of time. This attentional process subserves ...

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Research

JoVE Journal

Neuroscience

Touchscreen Sustained Attention Task (SAT) for Rats

9.7K visualizações.  Temple University. Atenção sustentada, ou monitorar continuamente situações para eventos intermitentes e imprevisíveis, é um aspecto crítico da cognição. Aqui detalhamos como teste de atenção sustentada em ratos utilizando câmaras operante de touchscreen. Vamos demonstrar o desempenho comparável em ratos machos e fêmeas, tornando esta tarefa útil para estudar a atenção em ambos os sexos.

Vídeo: Touchscreen Sustained Attention Task SAT for Rats

Assessment of Ultrasonic Vocalizations During Drug Self-administration in Rats

Drug self-administration procedures are commonly used to study behavioral and neurochemical changes associated with human drug abuse, addiction and relapse. Various types of behavioral activity are commonly utilized as measures of drug motivation in animals. However, a crucial component of drug abuse relapse in abstinent cocaine users is "drug craving", which is difficult to model in animals, as it often occurs in the absence of overt behaviors. Yet, it is possible that a class of ultrasonic vocalizations (USVs) in rats may be a useful marker for affective responses to drug administration, drug anticipation and even drug craving. Rats vocalize in ultrasonic frequencies that serve as a communicatory function and express subjective emotional states. Several studies have shown that different call frequency ranges are associated with negative and positive emotional states. For instance, high frequency calls ("50-kHz") are associated with positive affect, whereas low frequency calls ("22-kHz") represent a negative emotional state. This article describes a procedure to assess rat USVs associated with daily cocaine self-administration. For this procedure, we utilized standard single-lever operant chambers housed within sound-attenuating boxes for cocaine self-administration sessions and utilized ultrasonic microphones, multi-channel recording hardware and specialized software programs to detect and analyze USVs. USVs measurements reflect emotionality of rats before, during and after drug availability and can be correlated with commonly assessed drug self-administration behavioral data such lever responses, inter-response intervals and locomotor activity. Since USVs can be assessed during intervals prior to drug availability (e.g., anticipatory USVs) and during drug extinction trials, changes in affect associated with drug anticipation and drug abstinence can also be determined. In addition, determining USV changes over the course of short- and long-term drug exposure can provide a more detailed interpretation of drug exposure effects on affective functioning.

Drug self-administration and ultrasonic vocalizations (USV) are used as behavioral assessments in animal research, but rarely in combination. The purpose of this article is to describe the advantages of recording USVs during drug self-administration procedures to assess affective responses to drug experience.

Avaliação de vocalizações ultra-som durante Drogas auto-administração em ratos

Drug self-administration procedures are commonly used to study behavioral and neurochemical changes associated with human drug abuse, addiction and ...

Neurociência

A Procedure for Studying the Footshock-Induced Reinstatement of Cocaine Seeking in Laboratory Rats

The most insidious aspect of drug addiction is the high propensity for relapse. Animal models of relapse, known as reinstatement procedures, have been used extensively to study the neurobiology and phenomenology of relapse to drug use. Although procedural variations have emerged over the past several decades, the most conventional reinstatement procedures are based on the drug self-administration (SA) model. In this model, an animal is trained to perform an operant response to obtain drug. Subsequently, the behavior is extinguished by withholding response-contingent reinforcement. Reinstatement of drug seeking is then triggered by a discrete event, such as an injection of the training drug, re-exposure to drug-associated cues, or exposure to a stressor 1.
Reinstatement procedures were originally developed to study the ability of acute non-contingent exposure to the training drug to reinstate drug seeking in rats and monkeys 1, 2. Reinstatement procedures have since been modified to study the role of environmental stimuli, including drug-associated cues and exposure to various forms of stress, in relapse to drug seeking 1, 3, 4.
Over the past 15 years, a major focus of the reinstatement literature has been on the role of stress in drug relapse. One of the most commonly used forms of stress for studying this relationship is acute exposures to mild, intermittent, electric footshocks. The ability of footshock stress to induce reinstatement of drug seeking was originally demonstrated by Shaham and colleagues (1995) in rats with a history of intravenous heroin SA5. Subsequently, the effect was generalized to rats with histories of intravenous cocaine, methamphetamine, and nicotine SA, as well as oral ethanol SA 3, 6.
Although footshock-induced reinstatement of drug seeking can be achieved reliably and robustly, it is an effect that tends to be sensitive to certain parametrical variables. These include the arrangement of extinction and reinstatement test sessions, the intensity and duration of footshock stress, and the presence of drug-associated cues during extinction and testing for reinstatement. Here we present a protocol for footshock-induced reinstatement of cocaine seeking that we have used with consistent success to study the relationship between stress and cocaine seeking.

Animal models of relapse, known as reinstatement procedures, have been used extensively to study the role of stress in relapse to drug seeking. Here, we report on a method for inducing the reinstatement of cocaine seeking in laboratory rats via acute exposures to mild, intermittent electric footshock.

Um Procedimento para o Estudo do Reintegração Footshock Induzida de cocaína Buscando em ratos de laboratório

The most insidious aspect of drug addiction is the high propensity for relapse.  Animal models of relapse, known as reinstatement procedures, have been ...

Correlating Behavioral Responses to fMRI Signals from Human Prefrontal Cortex: Examining Cognitive Processes Using Task Analysis

The aim of this methods paper is to describe how to implement a neuroimaging technique to examine complementary brain processes engaged by two similar tasks. Participants' behavior during task performance in an fMRI scanner can then be correlated to the brain activity using the blood-oxygen-level-dependent signal. We measure behavior to be able to sort correct trials, where the subject performed the task correctly and then be able to examine the brain signals related to correct performance. Conversely, if subjects do not perform the task correctly, and these trials are included in the same analysis with the correct trials we would introduce trials that were not only for correct performance. Thus, in many cases these errors can be used themselves to then correlate brain activity to them. We describe two complementary tasks that are used in our lab to examine the brain during suppression of an automatic responses: the stroop1 and anti-saccade tasks. The emotional stroop paradigm instructs participants to either report the superimposed emotional 'word' across the affective faces or the facial 'expressions' of the face stimuli1,2. When the word and the facial expression refer to different emotions, a conflict between what must be said and what is automatically read occurs. The participant has to resolve the conflict between two simultaneously competing processes of word reading and facial expression. Our urge to read out a word leads to strong 'stimulus-response (SR)' associations; hence inhibiting these strong SR's is difficult and participants are prone to making errors. Overcoming this conflict and directing attention away from the face or the word requires the subject to inhibit bottom up processes which typically directs attention to the more salient stimulus. Similarly, in the anti-saccade task3,4,5,6, where an instruction cue is used to direct only attention to a peripheral stimulus location but then the eye movement is made to the mirror opposite position. Yet again we measure behavior by recording the eye movements of participants which allows for the sorting of the behavioral responses into correct and error trials7 which then can be correlated to brain activity. Neuroimaging now allows researchers to measure different behaviors of correct and error trials that are indicative of different cognitive processes and pinpoint the different neural networks involved.

The goal of our research is to correlate behavior to brain activity. Accurate behavioral measures and imaging techniques allow us to elucidate brain-behavior relationships.

Correlacionando as respostas comportamentais aos sinais de fMRI córtex pré-frontal humano: Examinando Processos Cognitivos Usando Análise de Tarefa

The aim of this methods paper is to describe how to implement a neuroimaging technique to examine complementary brain processes engaged by two similar ...

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

Um método geral para a avaliação de Incubação de Craving Sacarose em Ratos

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

Mapping Cortical Dynamics Using Simultaneous MEG/EEG and Anatomically-constrained Minimum-norm Estimates: an Auditory Attention Example

Magneto- and electroencephalography (MEG/EEG) are neuroimaging techniques that provide a high temporal resolution particularly suitable to investigate the cortical networks involved in dynamical perceptual and cognitive tasks, such as attending to different sounds in a cocktail party. Many past studies have employed data recorded at the sensor level only, i.e., the magnetic fields or the electric potentials recorded outside and on the scalp, and have usually focused on activity that is time-locked to the stimulus presentation. This type of event-related field / potential analysis is particularly useful when there are only a small number of distinct dipolar patterns that can be isolated and identified in space and time. Alternatively, by utilizing anatomical information, these distinct field patterns can be localized as current sources on the cortex. However, for a more sustained response that may not be time-locked to a specific stimulus (e.g., in preparation for listening to one of the two simultaneously presented spoken digits based on the cued auditory feature) or may be distributed across multiple spatial locations unknown a priori, the recruitment of a distributed cortical network may not be adequately captured by using a limited number of focal sources.
Here, we describe a procedure that employs individual anatomical MRI data to establish a relationship between the sensor information and the dipole activation on the cortex through the use of minimum-norm estimates (MNE). This inverse imaging approach provides us a tool for distributed source analysis. For illustrative purposes, we will describe all procedures using FreeSurfer and MNE software, both freely available. We will summarize the MRI sequences and analysis steps required to produce a forward model that enables us to relate the expected field pattern caused by the dipoles distributed on the cortex onto the M/EEG sensors. Next, we will step through the necessary processes that facilitate us in denoising the sensor data from environmental and physiological contaminants. We will then outline the procedure for combining and mapping MEG/EEG sensor data onto the cortical space, thereby producing a family of time-series of cortical dipole activation on the brain surface (or "brain movies") related to each experimental condition. Finally, we will highlight a few statistical techniques that enable us to make scientific inference across a subject population (i.e., perform group-level analysis) based on a common cortical coordinate space.

We use magneto- and electroencephalography (MEG/EEG), combined with anatomical information captured by magnetic resonance imaging (MRI), to map the dynamics of the cortical network associated with auditory attention.

Mapeamento Dynamics cortical MEG simultâneas / EEG e anatomicamente constrangido mínima norma-Estimativas: um exemplo Atenção Auditiva

Magneto- and electroencephalography (MEG/EEG) are neuroimaging techniques that provide a high temporal resolution particularly suitable to investigate the ...

Generation of Topically Transgenic Rats by In utero Electroporation and In vivo Bioluminescence Screening

In utero electroporation (IUE) is a technique which allows genetic modification of cells in the brain for investigating neuronal development. So far, the use of IUE for investigating behavior or neuropathology in the adult brain has been limited by insufficient methods for monitoring of IUE transfection success by non-invasive techniques in postnatal animals. 
 For the present study, E16 rats were used for IUE. After intraventricular injection of the nucleic acids into the embryos, positioning of the tweezer electrodes was critical for targeting either the developing cortex or the hippocampus. 
 Ventricular co-injection and electroporation of a luciferase gene allowed monitoring of the transfected cells postnatally after intraperitoneal luciferin injection in the anesthetized live P7 pup by in vivo bioluminescence, using an IVIS Spectrum device with 3D quantification software. 
 Area definition by bioluminescence could clearly differentiate between cortical and hippocampal electroporations and detect a signal longitudinally over time up to 5 weeks after birth. This imaging technique allowed us to select pups with a sufficient number of transfected cells assumed necessary for triggering biological effects and, subsequently, to perform behavioral investigations at 3 month of age. As an example, this study demonstrates that IUE with the human full length DISC1 gene into the rat cortex led to amphetamine hypersensitivity. Co-transfected GFP could be detected in neurons by post mortem fluorescence microscopy in cryosections indicating gene expression present at &ge;6 months after birth.
 We conclude that postnatal bioluminescence imaging allows evaluating the success of transient transfections with IUE in rats. Investigations on the influence of topical gene manipulations during neurodevelopment on the adult brain and its connectivity are greatly facilitated. For many scientific questions, this technique can supplement or even replace the use of transgenic rats and provide a novel technology for behavioral neuroscience.

Generation of Topically Transgenic Rats by In utero Electroporation and In vivo Bioluminescence Screening

Genes can be manipulated during development of the cortex or the hippocampus of the rat via in utero electroporation (IUE) at E16, to enable fast and targeted modifications in neuronal connectivity for later studies of behavior or neuropathology in adult animals. Postnatal in vivo imaging for control of IUE success is performed by bioluminescence of activating co-transfected luciferase.

Geração de topicamente transgênicas Rats por<em&gt; In utero</em&gt; Eletroporação e<em&gt; In vivo</em&gt; Triagem Bioluminescência

 In utero electroporation (IUE) is a technique which allows genetic modification of cells in the brain for investigating neuronal development. So far, the ...

Unilateral Pyramidotomy of the Corticospinal Tract in Rats for Assessment of Neuroplasticity-inducing Therapies

The corticospinal tract (CST) can be completely severed unilaterally in the medullary pyramids of the rodent brainstem. The CST is a motor tract that has great importance for distal muscle control in humans and, to a lesser extent, in rodents. A unilateral cut of one pyramid results in loss of CST innervation of the spinal cord mainly on the contralateral side of the spinal cord leading to transient motor disability in the forelimbs and sustained loss of dexterity. Ipsilateral projections of the corticospinal tract are minor. We have refined our surgical method to increase the chances of lesion completeness. We describe postsurgical care. Deficits on the Montoya staircase pellet reaching test and the horizontal ladder test shown here are detected up to 8 weeks postinjury. Deficits on the cylinder rearing test are only detected transiently. Therefore, the cylinder test may only be suitable for detection of short term recovery. We show how, electrophysiologically and anatomically, one may assess lesions and plastic changes. We also describe how to analyse fibers from the uninjured CST sprouting across the midline into the deprived areas. It is challenging to obtain &#62;90% complete lesions consistently due to the proximity to the basilar artery in the medulla oblongata and survival rates can be low. Alternative surgical approaches and behavioural testing are described in this protocol. The pyramidotomy model is a good tool for assessing neuroplasticity-inducing treatments, which increase sprouting of intact fibers after injury.

The corticospinal tract, one of the major sensorimotor tracts, can be lesioned unilaterally in the rodent brainstem in order to test neuroplasticity-inducing therapies for the central nervous system. This surgical procedure (&#8220;pyramidotomy&#8221;) and postoperative assessments are described in this protocol.

Unilateral Pyramidotomy do Corticospinal Tract em Ratos de Avaliação de Terapias de indução de neuroplasticidade

The corticospinal tract (CST) can be completely severed unilaterally in the medullary pyramids of the rodent brainstem. The CST is a motor tract that has ...

Conducting Concurrent Electroencephalography and Functional Near-Infrared Spectroscopy Recordings with a Flanker Task

Concurrent EEG and fNIRS recordings offer an excellent opportunity to gain a full understanding of the neural mechanism of cognitive processing by inspecting the relationship between the neural and hemodynamic signals. EEG is an electrophysiological technology that can measure the rapid neuronal activity of the cortex, whereas fNIRS relies on the hemodynamic responses to infer brain activation. The combination of EEG and fNIRS neuroimaging techniques can identify more features and reveal more information associated with the functioning of the brain. In this protocol, fused EEG-fNIRS measurements were performed for concurrent recordings of evoked-electrical potentials and hemodynamic responses during a Flanker task. In addition, the critical steps for setting up the hardware and software system as well as the procedures for data acquisition and analysis were provided and discussed in detail. It is expected that the present protocol can pave a new avenue for improving the understanding of the neural mechanisms underlying various cognitive processes by using the EEG and fNIRS signals.

The present protocol describes how to perform concurrent EEG and fNIRS recordings and how to inspect the relationship between the EEG and fNIRS data.&#160;

Realização de Eletroencefalografia simultânea e gravações de espectroscopia quase infravermelha funcional com uma tarefa flanker

Concurrent EEG and fNIRS recordings offer an excellent opportunity to gain a full understanding of the neural mechanism of cognitive processing by ...

Event Related Potentials (ERPs) and other EEG Based Methods for Extracting Biomarkers of Brain Dysfunction: Examples from Pediatric Attention Deficit/Hyperactivity Disorder (ADHD)

Neuropsychiatric diagnoses like ADHD are based on subjective methods like interviews, rating scales and observations. There is a need for more brain-based supplements. Stimulant medication is the most common treatment for ADHD. Clinically useful predictors of response have so far not been reported. The aim of this paper is to describe the EEG based methods we apply to extract potential biomarkers for brain dysfunction. Examples relate to biomarkers for pediatric ADHD, and prediction of medication response. The main emphasis is on Event Related Potentials (ERPs).
A nineteen channel EEG is recorded during a 3 min eyes-opened task, a 3 min eyes-closed task, and a 20 min cued visual GO/NOGO task (VCPT). ERPs are recorded during this task. The goal of the ERP protocol is to extract biomarkers of assumed brain dysfunctions that significantly differentiate between a patient group and healthy controls. The protocol includes recording during standard conditions and artifact correction. ERP waves can be used or transformed into latent components. The components of the patient group are compared with controls, empathizing components that, when compared, show relatively high effect sizes. Sub-groups of the patients are selected on the basis of the cluster analysis in the space of the components. Treatment procedure (such as medication, tDCS or neurofeedback protocol) can be applied and the changes in components related to treatment in the subgroups are observed, forming the basis for clinical recommendations.
The methods described were applied in a study of 87 pediatric ADHD patients. The index of medication response discriminated significantly between responders and non-responders with a large, and clinically meaningful effect size (d = 1.84). In an ongoing study comparing ADHD children with matched controls, several variables discriminate significantly between patients and controls. The global index will exceed d = .8. The EEG based methods described here could be clinically meaningful.

Event Related Potentials ERPs and other EEG Based Methods for Extracting Biomarkers of Brain Dysfunction: Examples from Pediatric Attention Deficit/Hyperactivity Disorder ADHD

EEG-methods are applied for extracting biomarkers of brain dysfunctions. The focus is on multi-channel event-related potentials (ERPs) recorded in a cued GO/NOGO task. Non-brain artifacts are corrected and ERPs are compared with the normative data. Examples relate to biomarkers for ADHD diagnosis and prediction of medication response.

Potenciais Relacionados a Eventos (ERPs) e outros métodos baseados em EEG para extrair biomarcadores de disfunção cerebral: Exemplos do Transtorno de Déficit de Atenção Pediátrica/Hiperatividade (TDAH)

Neuropsychiatric diagnoses like ADHD are based on subjective methods like interviews, rating scales and observations. There is a need for more brain-based ...

Preparation of Peripheral Nerve Stimulation Electrodes for Chronic Implantation in Rats

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. Several recent studies have demonstrated the effectiveness of chronic VNS in enhancing central nervous system plasticity to improve motor rehabilitation, extinction learning, and sensory discrimination. Construction of chronically implantable devices for use in such studies is challenging due to rats&#8217; small size, and typical protocols require extensive training of personnel and time-consuming microfabrication methods. Alternatively, commercially available implantable cuff electrodes can be purchased at a significantly higher cost. In this protocol, we present a simple, low-cost method for construction of small, chronically implantable peripheral nerve cuff electrodes for use in rats. We validate the short and long-term reliability of our cuff electrodes by demonstrating that VNS in ketamine/xylazine anesthetized rats produces decreases in breathing rate consistent with activation of the Hering-Breuer reflex, both at the time of implantation and up to 10 weeks after device implantation. We further demonstrate the suitability of the cuff electrodes for use in chronic stimulation studies by pairing VNS with skilled lever press performance to induce motor cortical map plasticity.

Existing approaches for constructing chronically implantable peripheral nerve cuff electrodes for use in small rodents often require specialized equipment and/or highly trained personnel. In this protocol we demonstrate a simple, low-cost approach for fabricating chronically implantable cuff electrodes, and demonstrate their effectiveness for vagus nerve stimulation (VNS) in rats.

Preparação de Eletrodos de Estimulação Nervosa Periférica para Implantação Crônica em Ratos

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. ...