Os autores gostariam de agradecer ao Prof ME Schwab, Dr. P. Freund, Dr. A. Wyss, Dr. S. Bashir, Dr. A. Mir, Dr. J. Bloch, Dr. JF Brunet, Dr. J. Aebischer, Dr. A. Meszaros, o Dr. V. Goetschman por sua contribuição para experiências anteriores e análises. O experimental set-ups foram construídas por André Gaillard, Aebischer Bernard Monney e Laurent. Nas instalações animais, os macacos foram colocados sob a supervisão de profissionais cuidadores de animais: Josef Corpataux, Laurent Bossy e Jacques Maillard. Os testes comportamentais e análise dos dados, bem como histologia, foram realizadas com as contribuições de grande valor de técnicos de laboratório: Véronique Moret (também web master), Françoise Tinguely, Christine Roulin, Monica Bennefeld, Christiane Marti e Georgette Fischer. Este trabalho foi financiado pela National Science Foundation suíço, não concede 31-61.857,00, 310000-110005, 31003A-132465 (EMR), 310030-118357, 31003A-104061 (TW), 310030-120.411 (ABS), PZ00P3_121646 (ES), Novartis Foundation; O Centro Nacional de Competência em Pesquisa (NCCR) sobre &quot;A plasticidade neural e reparação&quot;.

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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+recovery+of+dexterity+following+unilateral+lesion+of+the+sensorimotor+cortex+in+adult+monkeys.">Mechanisms of recovery of dexterity following unilateral lesion of the sensorimotor cortex in adult monkeys.</a> Exp. Brain. Res. 128, 149-159 (1999).</li><li>Freund, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nogo-A-specific+antibody+treatment+enhances+sprouting+and+functional+recovery+after+cervical+lesion+in+adult+primates.">Nogo-A-specific antibody treatment enhances sprouting and functional recovery after cervical lesion in adult primates.</a> Nature. Med. 12, 790-792 (2006).</li><li>Freund, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anti-Nogo-A+antibody+treatment+promotes+recovery+of+manual+dexterity+after+unilateral+cervical+lesion+in+adult+primates--re-examination+and+extension+of+behavioral+data.">Anti-Nogo-A antibody treatment promotes recovery of manual dexterity after unilateral cervical lesion in adult primates--re-examination and extension of behavioral data.</a> Eur. J. Neurosci. 29, 983-996 (2009).</li><li>Kaeser, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+Unilateral+Motor+Cortex+Lesion+on+Ipsilesional+Hand's+Reach+and+Grasp+Performance+in+Monkeys:+Relationship+With+Recovery+in+the+Contralesional+Hand.">Effects of Unilateral Motor Cortex Lesion on Ipsilesional Hand's Reach and Grasp Performance in Monkeys: Relationship With Recovery in the Contralesional Hand.</a> J. Neurophysiol. 103, 1603-1645 (2010).</li><li>Kaeser, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autologous+adult+cortical+cell+transplantation+enhances+functional+recovery+following+unilateral+lesion+of+motor+cortex+in+primates:+a+pilot+study.">Autologous adult cortical cell transplantation enhances functional recovery following unilateral lesion of motor cortex in primates: a pilot study.</a> Neurosurgery. 68, 1405-1417 (2011).</li><li>Bashir, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short-term+effects+of+unilateral+lesion+of+the+primary+motor+cortex+(M1)+on+Ipsilesional+hand+dexterity+in+adult+macaque+monkeys.">Short-term effects of unilateral lesion of the primary motor cortex (M1) on Ipsilesional hand dexterity in adult macaque monkeys.</a> Brain Structure and Function. , (2011).</li><li>Hamadjida, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+anti-Nogo-A+treatment+on+the+reorganization+of+callosal+connectivity+of+the+premotor+cortical+areas+following+unilateral+lesion+of+primary+motor+cortex+(M1)+in+adult+macaque+monkeys.">Influence of anti-Nogo-A treatment on the reorganization of callosal connectivity of the premotor cortical areas following unilateral lesion of primary motor cortex (M1) in adult macaque monkeys.</a> , (2011).</li><li>Jenny, A. B., Inukai, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+of+motor+organization+of+the+monkey+cervical+spinal+cord.">Principles of motor organization of the monkey cervical spinal cord.</a> J. Neurosci. 3, 567-575 (1983).</li><li>Schmidlin, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progressive+plastic+changes+in+the+hand+representation+of+the+primary+motor+cortex+parallel+incomplete+recovery+from+a+unilateral+section+of+the+corticospinal+tract+at+cervical+level+in+monkeys.">Progressive plastic changes in the hand representation of the primary motor cortex parallel incomplete recovery from a unilateral section of the corticospinal tract at cervical level in monkeys.</a> Brain Research. 1017, 172-183 (2004).</li><li>Schmidlin, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+the+hand+representation+in+the+ipsilateral+primary+motor+cortex+following+unilateral+section+of+the+corticospinal+tract+at+cervical+level+in+monkeys.">Reduction of the hand representation in the ipsilateral primary motor cortex following unilateral section of the corticospinal tract at cervical level in monkeys.</a> BMC Neuroscience. 6, 56-56 (2005).</li><li>Prescott, M. 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Não há conflitos de interesse declarados.

Embora as tarefas presentes comportamentais têm sido considerados até agora em nosso laboratório no contexto de estudos relacionados à lesão da medula cervical ou à lesão do córtex motor, com o objetivo de testar vários tratamentos (ver 14,15,17 e <a href="http://www.unifr.ch/neuro/rouiller">http:/ / www.unifr.ch / neuro / Rouiller</a> &gt; selecione &quot;pesquisa&quot; na barra de menu superior, então&gt; &quot;motor do sistema&quot;&gt; &quot;recuperação após lesão&quot;), eles também podem ter uma aplicação mais ampla, como destreza manual é também um aspecto a considerar em outras patologias, tais como doença de Parkinson (MPTP macacos) ou em caso de propriocepção de afferentation-sensoriais que afetam e / ou o sentido do tato (especialmente o teste 2 em ausência de controle visual). Os testes comportamentais aqui proposto são adequados para investigar o controle motor dos movimentos distal do membro anterior, como envolvidos na destreza manual. A especificidade dos testes é demonstrado pelaa ausência de déficit (exceto por um par de dias) em caso de uma lesão que não prejudique os componentes relevantes do sistema de controle: na verdade, no caso de uma lesão colocou mais caudal do que o motoneurônios controlando músculos da mão, não houve déficit. A pertinência da prova 1 pode ser apreciada através da comparação de seqüências de vídeo feito em dois momentos da curva de recuperação pós-lesão, que diferem por um aumento da recuperação funcional de 25%, possivelmente em relação a uma terapia de tratamento com células-17. Apesar de alguns fase inicial de formação, relativamente curto no início (com duração geralmente 2-3 meses), os testes comportamentais aqui proposta são relativamente &quot;natural&quot; e direto, em comparação com complexos (por exemplo condicional) tarefas para as quais a formação do macaco pode demorar quase um ano ou mais. O reforço positivo é baseado em alimentos sólidos, o que é menos sensível do ponto de vista ético do que a privação de água, usado normalmente em mais comtarefas complexas 23. Não há necessidade de privar os macacos de alimentos para obter resultados estáveis ​​e consistentes. As pelotas recebidas durante as tarefas de representar o primeiro acesso ao alimento no dia da sessão comportamentais (assumindo que o macaco não come durante a noite anterior, no entanto alimento adicional pode ser dada até o final da tarde do dia anterior) . É fundamental que cada macaco realiza a sessão de comportamento, ao mesmo tempo do dia, bem como respeitando a mesma ordem seqüencial entre os macacos diferentes formando um grupo na sala de habitação. Como os macacos são sensíveis a eventos externos perturbadores, as tarefas de comportamento deve ser conduzida na presença de música de fundo, mascarando o ruído perturbador potencial proveniente de salas vizinhas ou laboratórios. É fundamental que durante toda a duração do experimento (de formação inicial até a última sessão diária experimental, um período de vários meses ou mesmo anos), um macaco é dadacolocada diariamente sob a supervisão do pesquisador mesmo. Os testes comportamentais presentes, usado desde há vários anos em nosso laboratório para quantificar a destreza manual, são em certa medida, comparável a outros testes de destreza manual recentemente relatada na literatura 24-28. Há no entanto uma necessidade crucial para padronizar os testes em laboratórios diferentes (para melhor comparação), que é uma meta provisória do presente relatório. Na demanda, as propriedades detalhadas dos set-ups ilustrado aqui para os testes 04/01 pode ser fornecida pelo autor correspondente, a fim de reproduzi-los. Além da questão da medicina regenerativa (recuperação da lesão de medula espinhal ou do córtex cerebral), a paleta atual de testes podem ser adequados para lidar com questões normais em primatas não-humanos do desenvolvimento (por exemplo, tempo de curso do desenvolvimento motor dos movimentos hábeis), para investigar aspectos lateralização (mão de preferência / dominância) e às questões decifrar evolutiva por cOMPARAÇÃO as habilidades motoras de diferentes espécies de primatas, incluindo seres humanos. Note no entanto que as dimensões dos aparelhos devem ser adaptados de acordo com o tamanho dígitos &quot;(espessura e comprimento) das espécies de primatas, como ele pode influenciar o desempenho da tarefa. No presente estudo, os testes foram realizados em macacos Macaca fascicularis, variando de 2,5 a 8 anos de idade e pesando entre 2,5 e 8 kg. O comprimento do dedo indicador (usado pela primeira vez para manipular os pellets) varia de 32 a 35 mm, enquanto que a circunferência da falange distal (ponta) do dedo indicador era entre 22 e 25 mm nos macacos incluídos em nossos estudos. Como testado em experimentos anteriores, os mesmos testes são adequados para agarrar macaca mulatta também. Todos os experimentos foram conduzidos de acordo com o Guia para o Cuidado e Uso de Animais de Laboratório (1996) e aprovado pelo local (suíço) as autoridades veterinárias. Todos os procedimentos experimentais sobre o macacos, bem como as condições de detenção nas instalações animais, foram descritos em detalhes em relatórios recentes de nosso laboratório: ver referências 12-18.

behavioral assessment of manual dexterity in non-human primates

O trato corticoespinhal (CS) é o suporte anatômico da capacidade motora requintado para habilmente manipular pequenos objetos, uma prerrogativa principalmente de primatas 1. Em caso de lesão afetando o sistema de projeção CS na sua origem (lesão de áreas motoras corticais) ou ao longo de sua trajetória (lesão medular cervical), há uma dramática perda de destreza manual (paralisia da mão), como visto em alguns pacientes tetraplégicos ou hemiplégica . Embora haja alguma recuperação funcional após lesão espontânea tal, continua a ser muito limitada no adulto. Várias estratégias terapêuticas são atualmente propostos (por exemplo, terapia celular, neutralização de moléculas inibidoras do crescimento axonal, a aplicação de fatores de crescimento, etc), que são principalmente desenvolvido em roedores. No entanto, antes da aplicação clínica, é frequentemente recomendada para testar a viabilidade, eficácia e segurança do tratamento em primatas não-humanos. Isto é especialmente verdadeiro quando o objetivo é restaurar a destreza manual apósuma lesão do sistema nervoso central, como a organização do sistema motor de roedores é diferente do de primatas 1,2. Macacos são ilustrados aqui como um modelo adequado de comportamento para quantificar a destreza manual em primatas, para refletir os déficits decorrentes da lesão do córtex motor ou da medula cervical, por exemplo, medir o grau de recuperação funcional espontânea e, quando for aplicado um tratamento, avaliar o quanto ele pode melhorar a recuperação funcional. A avaliação comportamental da destreza manual é baseado em quatro distintos, complementares tarefas, alcançar e agarrar manual (uso de aderência precisão de compreender pellets), exigindo uma formação inicial de macacos adultos. A preparação dos animais é demonstrado, bem como o posicionamento com relação ao comportamento set-up. O desempenho de um macaco típico é ilustrado para cada tarefa. A recolha e análise de parâmetros relevantes refletindo manipulação lado precisação, bem como o controle da força, são explicadas e demonstradas com resultados representativos. Estes dados são colocadas então em um contexto mais amplo, mostrando como os dados de comportamento podem ser exploradas para investigar o impacto de uma lesão da medula espinhal ou de uma lesão do córtex motor e em que medida um tratamento pode melhorar a recuperação funcional espontânea, através da comparação diferentes grupos de macacos (tratados contra sham tratado por exemplo). Vantagens e limitações dos testes comportamentais são discutidas. A presente abordagem comportamental está de acordo com relatórios anteriores enfatizando a pertinência do modelo de primatas não-humanos no contexto das doenças do sistema nervoso 2,3.

Watch this Scientific Journal Video about Behavioral Assessment of Manual Dexterity in Non-Human Primates at JoVE.com

Behavioral Assessment of Manual Dexterity in Non-Human Primates

Como destreza manual é uma prerrogativa principalmente de primatas, tarefas comportamentais têm sido desenvolvidos nos macacos. Quatro alcançar e agarrar a preensão tarefas, medir a capacidade de mão de manipulação e força, permitem estabelecer a recuperação funcional após uma lesão do sistema nervoso central e para testar o efeito de um tratamento.

Avaliação comportamental de destreza manual em primatas não-humanos

The corticospinal (CS) tract is the anatomical support of the exquisite motor ability to skillfully manipulate small objects, a prerogative mainly of ...

behavioral-assessment-of-manual-dexterity-in-non-human-primates

Research

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Neuroscience

22.3K visualizações.  University of Fribourg. Como destreza manual é uma prerrogativa principalmente de primatas, tarefas comportamentais têm sido desenvolvidos nos macacos. Quatro alcançar e agarrar a preensão tarefas, medir a capacidade de mão de manipulação e força, permitem estabelecer a recuperação funcional após uma lesão do sistema nervoso central e para testar o efeito de um tratamento.

Vídeo: Behavioral Assessment of Manual Dexterity in Non-Human Primates

Combining Behavioral Endocrinology and Experimental Economics: Testosterone and Social Decision Making

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in rodents have shown a direct link between testosterone and aggressive behavior1 and folk wisdom adapts these findings to humans, suggesting that testosterone induces antisocial, egoistic or even aggressive behavior2. However, many researchers doubt a direct testosterone-aggression link in humans, arguing instead that testosterone is primarily involved in status-related behavior3,4. As a high status can also be achieved by aggressive and antisocial means it can be difficult to distinguish between anti-social and status seeking behavior.
We therefore set up an experimental environment, in which status can only be achieved by prosocial means. In a double-blind and placebo-controlled experiment, we administered a single sublingual dose of 0.5 mg of testosterone (with a hydroxypropyl-&beta;-cyclodextrin carrier) to 121 women and investigated their social interaction behavior in an economic bargaining paradigm. Real monetary incentives are at stake in this paradigm; every player A receives a certain amount of money and has to make an offer to another player B on how to share the money. If B accepts, she gets what was offered and player A keeps the rest. If B refuses the offer, nobody gets anything. A status seeking player A is expected to avoid being rejected by behaving in a prosocial way, i.e. by making higher offers.
The results show that if expectations about the hormone are controlled for, testosterone administration leads to a significant increase in fair bargaining offers compared to placebo. The role of expectations is reflected in the fact that subjects who report that they believe to have received testosterone make lower offers than those who say they believe that they were treated with a placebo. These findings suggest that the experimental economics approach is sensitive for detecting neurobiological effects as subtle as those achieved by administration of hormones. Moreover, the findings point towards the importance of both psychosocial as well as neuroendocrine factors in determining the influence of testosterone on human social behavior.

The procedure described in this protocol shows that testosterone administration and folk beliefs about testosterone may be associated with directly opposed social behaviors.

Combinando Behavioral Endocrinologia e Economia Experimental: Decisão da testosterona e Social Fazendo

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in ...

Neurociência

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 ...

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, including rats, hamsters, and mice, are active during the night and relatively inactive during the day. Many other behavioral and physiological measures also exhibit daily rhythms, but in rodents, running-wheel activity serves as a particularly reliable and convenient measure of the output of the master circadian clock, the suprachiasmatic nucleus (SCN) of the hypothalamus. In general, through a process called entrainment, the daily pattern of running-wheel activity will naturally align with the environmental light-dark cycle (LD cycle; e.g. 12 hr-light:12 hr-dark). However circadian rhythms are endogenously generated patterns in behavior that exhibit a ~24 hr period, and persist in constant darkness. Thus, in the absence of an LD cycle, the recording and analysis of running-wheel activity can be used to determine the subjective time-of-day. Because these rhythms are directed by the circadian clock the subjective time-of-day is referred to as the circadian time (CT). In contrast, when an LD cycle is present, the time-of-day that is determined by the environmental LD cycle is called the zeitgeber time (ZT).
Although circadian rhythms in running-wheel activity are typically linked to the SCN clock6-8, circadian oscillators in many other regions of the brain and body9-14 could also be involved in the regulation of daily activity rhythms. For instance, daily rhythms in food-anticipatory activity do not require the SCN15,16 and instead, are correlated with changes in the activity of extra-SCN oscillators17-20. Thus, running-wheel activity recordings can provide important behavioral information not only about the output of the master SCN clock, but also on the activity of extra-SCN oscillators. Below we describe the equipment and methods used to record, analyze and display circadian locomotor activity rhythms in laboratory rodents.

Circadian rhythms in voluntary wheel-running activity in mammals are tightly coupled to the molecular oscillations of a master clock in the brain. As such, these daily rhythms in behavior can be used to study the influence of genetic, pharmacological, and environmental factors on the functioning of this circadian clock.

Registro e análise dos ritmos circadianos em Running rodas Atividade em Roedores

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

Assessment of Dendritic Arborization in the Dentate Gyrus of the Hippocampal Region in Mice

Dendritic arborization has been shown to be a reliable marker for examination of structural and functional integrity of neurons. Indeed, the complexity and extent of dendritic arborization correlates well with the synaptic plasticity in these cells. A reliable method for assessment of dendritic arborization is needed to characterize the deleterious effects of neurological disorders on these structures and to determine the effects of therapeutic interventions. However, quantification of these structures has proven to be a formidable task given their complex and dynamic nature. Fortunately, sophisticated imaging techniques can be paired with conventional staining methods to assess the state of dendritic arborization, providing a more reliable and expeditious means of assessment. Below is an example of how these imaging techniques were paired with staining methods to characterize the dendritic arborization in wild type mice. These complementary imaging methods can be used to qualitatively and quantitatively assess dendritic arborization that span a rather wide area within the hippocampal region.

We describe two methods for visualization and quantification of dendritic arborization in the hippocampus of mouse models: real-time and extended depth of field imaging. While the former method allows sophisticated topographical tracing and quantification of the extent of branching, the latter allows speedy visualization of the dendritic tree.

Avaliação de dendríticas Arborização no giro dentado do hipocampo em Ratos Região

Dendritic arborization has been shown to be a reliable marker for examination of structural and functional integrity of neurons. Indeed, the complexity ...

Quantification of Filamentous Actin (F-actin) Puncta in Rat Cortical Neurons

Filamentous actin protein (F-actin) plays a major role in spinogenesis, synaptic plasticity, and synaptic stability. Changes in dendritic F-actin rich structures suggest alterations in synaptic integrity and connectivity. Here we provide a detailed protocol for culturing primary rat cortical neurons, Phalloidin staining for F-actin puncta, and subsequent quantification techniques. First, the frontal cortex of E18 rat embryos are dissociated into low-density cell culture, then the neurons grown in vitro for at least 12-14 days. Following experimental treatment, the cortical neurons are stained with AlexaFluor 488 Phalloidin (to label the dendritic F-actin puncta) and microtubule-associated protein 2 (MAP2; to validate the neuronal cells and dendritic integrity). Finally, specialized software is used to analyze and quantify randomly selected neuronal dendrites. F-actin rich structures are identified on second order dendritic branches (length range 25-75 &#181;m) with continuous MAP2 immunofluorescence. The protocol presented here will be a useful method for investigating changes in dendritic synapse structures subsequent to experimental treatments.

Quantification of Filamentous Actin F-actin Puncta in Rat Cortical Neurons

Filamentous actin (F-actin) plays an important role in spinogenesis, synaptic plasticity, and synaptic stability. Quantification of F-actin puncta is therefore a useful tool to study the integrity of synaptic structures. This protocol describes the procedures of quantifying F-actin puncta labeled with Phalloidin in low-density primary cortical neuronal cultures.

Quantificação de filamentosas actina (F-actina) puncta no Rat neurônios corticais

Filamentous actin protein (F-actin) plays a major role in spinogenesis, synaptic plasticity, and synaptic stability. Changes in dendritic F-actin rich ...

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

In the central nervous system, the D1-alpha subtype receptor (Drd1&#945;) is the most abundant dopamine (DA) receptor, which plays a vital role in regulating neuronal growth and development. However, the mechanisms underlying Drd1&#945; receptor abnormalities mediating behavioral responses and modulating working memory function are still unclear. Using a novel RNA in situ hybridization assay, the current study identified dopamine Drd1&#945; receptor and tyrosine hydroxylase (TH) RNA expression from DA-related circuitry in the nucleus accumbens (NAc) area and substantia nigra region (SNR), respectively. Drd1&#945; expression in the NAc shows a &#34;discrete dot&#34; staining pattern. Clear sex differences in Drd1&#945; expression were observed. In contrast, TH shows a &#34;clustered&#34; staining pattern. Regarding TH expression, female rats displayed a higher signal expression per cell relative to male animals. The methods presented here provide a novel in situ hybridization technique for investigating changes in dopamine system dysfunction during the progression of central nervous system diseases.

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

Identification of dopamine D1-alpha receptor in the nucleus accumbens is critical for clarifying D1 receptor dysfunction during a central nervous system disease. We performed a novel RNA in situ hybridization assay to visualize single RNA molecules in a specific brain area.

Identificação de Receptor de dopamina D1-alfa dentro roedores núcleo Accumbens por uma inovador RNA In Situ tecnologia da deteção

In the central nervous system, the D1-alpha subtype receptor (Drd1&#945;) is the most abundant dopamine (DA) receptor, which plays a vital role in ...

The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in neurocognitive disorders. Despite the popularization of prepulse inhibition (PPI) in recent years, many current protocols promote using a percent of control measure, thereby precluding the assessment of temporal processing. The present study used cross-modal PPI and gap prepulse inhibition (gap-PPI) to demonstrate the benefits of employing a range of interstimulus intervals (ISIs) to delineate effects of sensory modality, psychostimulant exposure, and age. Assessment of sensory modality, psychostimulant exposure, and age reveals the utility of an approach varying the interstimulus interval (ISI) to establish the shape of the ISI function, including increases (sharper curve inflections) or decreases (flattening of the response amplitude curve) in startle amplitude. Additionally, shifts in peak response inhibition, suggestive of a differential sensitivity to the manipulation of ISI, are often revealed. Thus, the systematic manipulation of ISI affords a critical opportunity to evaluate temporal processing, which may reveal the underlying neural mechanisms involved in neurocognitive disorders.

Temporal processing, a preattentive process, may underlie deficits in higher-level cognitive processes, including attention, commonly observed in neurocognitive disorders. Using prepulse inhibition as an exemplar paradigm, we present a protocol for manipulating interstimulus interval (ISI) to establish the shape of the ISI function to provide an assessment of temporal processing.

O poder do intervalo de Interstimulus para a avaliação do processamento Temporal em roedores

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in ...

Assessment of Long-term Depression Induction in Adult Cerebellar Slices

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

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

Avaliação da indução de depressão em longo prazo em fatias Cerebelares adultas

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

Ballistic Labeling of Pyramidal Neurons in Brain Slices and in Primary Cell Culture

It has been reported that the size and shape of dendritic spines is related to their structural plasticity. To identify the morphological structure of pyramidal neurons and dendritic spines, a ballistic labeling technique can be utilized. In the present protocol, pyramidal neurons are labeled with DilC18(3) dye and analyzed using neuronal reconstruction software to assess neuronal morphology and dendritic spines. To investigate neuronal structure, dendritic branching analysis and Sholl analysis are performed, allowing researchers to draw inferences about dendritic branching complexity and neuronal arbor complexity, respectively. The evaluation of dendritic spines is conducted using an automatic assisted classification algorithm integral to the reconstruction software, which classifies spines into four categories (i.e., thin, mushroom, stubby, filopodia). Furthermore, an additional three parameters (i.e., length, head diameter, and volume) are also chosen to assess alterations in dendritic spine morphology. To validate the potential of wide application of the ballistic labeling technique, pyramidal neurons from in vitro cell culture were successfully labeled. Overall, the ballistic labeling method is unique and useful for visualizing neurons in different brain regions in rats, which in combination with sophisticated reconstruction software, allows researchers to elucidate the possible mechanisms underlying neurocognitive dysfunction.

We present a protocol to label and analyze pyramidal neurons, which is critical for evaluating potential morphological alterations in neurons and dendritic spines that may underlie neurochemical and behavioral abnormalities.

Rotulagem Balística de Neurônios Piramidais em Fatias Cerebrais e na Cultura Celular Primária

It has been reported that the size and shape of dendritic spines is related to their structural plasticity. To identify the morphological structure of ...

A Hydrophobic Tissue Clearing Method for Rat Brain Tissue

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered tissue samples. Traditional immunolabeling procedures require cutting the sample into thin sections, which restricts the ability to label and examine intact structures. However, if brain tissue can remain intact during processing, structures and circuits can remain intact for the analysis. Previously established clearing methods take significant time to completely clear the tissue, and the harsh chemicals can often damage sensitive antibodies. The iDISCO method quickly and completely clears tissue, is compatible with many antibodies, and requires no special lab equipment. This technique was initially validated for the use in mice tissue, but the current protocol adapts this method to image hemispheres of control and transgenic rat brains. In addition to this, the present protocol also makes several adjustments to preexisting protocol to provide clearer images with less background staining. Antibodies for Iba-1 and tyrosine hydroxylase were validated in the HIV-1 transgenic rat and in F344/N control rats using the present hydrophobic tissue clearing method. The brain is an interwoven network, where structures work together more often than separately of one another. Analyzing the brain as a whole system as opposed to a combination of individual pieces is the greatest benefit of this whole brain clearing method.

Here we present a hydrophobic tissue clearing method that allows for the viewing of target molecules as part of intact brain structures. This technique has now been validated for F344/N control and HIV-1 transgenic rats of both sexes.

Um método de limpeza de tecido hidrofóbico para tecido cerebral de rato

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered ...