Name: behavioral assessment of manual dexterity in non-human primates
Uploaded: 2011-11-11T13:00:00
Duration: 16 min
Description: Watch this Scientific Journal Video about Behavioral Assessment of Manual Dexterity in Non-Human Primates at JoVE.com

저자는 교수 ME Schwab, 박사 P. Freund 박사 A. Wyss 박사 S. 바쉬르 박사 A. 미르 박사 J. Bloch 박사 JF Brunet 박사 J.에게 감사를 표합니다 Aebischer 박사 A. Meszaros, 이전 실험과 분석으로 자신의 기여를위한 Dr V. Goetschman. 실험 설정 UPS는 앙드레 가일 라드, 버나드 Aebischer와 로랑 Monney에 의해 건설되었다. 조세프 Corpataux, 로렌 좋아하고과 자크 Maillard : 동물 시설에서 원숭이 전문 동물 관리 인의 감독하에 놓여 있었다. 베로니크 Moret (또한 웹 마스터), 프랑소와 Tinguely, 크리스틴 Roulin, 모니카 Bennefeld, 크리스티안 마티와 조젯 피셔 :뿐만 아니라 조직학으로 데이터의 동작 테스트 및 분석, 실험실 기술자의 매우 귀중한 기여와 함께 실시되었다. 이 작품은 스위스 국립 과학 재단 (National Science Foundation)에 의해 지원되었다, 어떤 31-61857.00는 310000-110005을 부여하지 31003A - 132465 (EMR), 310030-118357, 31003A - 10406​​1 (TW), 310,030-120411 (ABS), PZ00P3_121646 (ES), 노바티스 재단, &quot;신경 소성 및 수리&quot;에 대한 연구에 경쟁력 국립 센터 (NCCR).

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현재 행동 작업 (14,15,17 및보고 다양한 트리 트먼트를 테스트하기 위해 자궁 코드의 병변이나 목표로 모터 피질의 병변에 관한 연구의 맥락에서 우리가 연구실에서 지금까지 고려하고 있지만 <a href="http://www.unifr.ch/neuro/rouiller">http:/을 / www.unifr.ch / 신경 / rouiller</a> &gt; 메뉴 표시줄의 메뉴에서 &quot;연구&quot;를 선택한 다음&gt; &quot;모터 시스템&quot;&gt; &quot;장애 후 복구&quot;수동 손재주도 측면이다 등), 그들은 또한, 광범위한 응용 프로그램이있을 수 있습니다 이러한 파킨슨 질환 (MPTP 원숭이) 또는 감각 드 afferentation 영향을 proprioception 및 / 또는 촉감 (특히 시각적인 컨트롤의 부재에서 테스트 2)의 경우에는 다른 pathologies에 고려해야합니다. 수동 손재주에 관여 여기에 제안된 행동 검사는 forelimb의 말초 운동의 모터 제어를 조사하기 위해 적합합니다. 검사의 특이성이 입증됩니다제어 시스템의 관련 구성 요소를 손상하지 않는 병변의 경우에는 적자의 부재 (며칠 제외) : 사실, 병변의 경우에는 손 근육을 제어 motoneurons보다 꼬리 배치, 아무 적자가 없었습니다. 시험 1의 적절가 가능한 세포 요법 치료 17 관계 25 % 기능 회복 강화에 따라 다릅니다 후 병변 회복 곡선, 두 시간 지점에서 촬영한 비디오 시퀀스에 비교하여 감상할 수 있습니다. 처음에 몇 가지 초기, 비교적 짧은 훈련 단계 (일반적으로 2~3개월 지속)에도 불구하고, 여기에 제안된 행동 시험은 복잡한 (예 : 조건부) 작업에 비해 상대적으로 &quot;자연&quot;과 간단되는 대한 원숭이의 훈련 거의 일년 이상 걸릴 수 있습니다. 긍정적 강화는 물 부족보다는보기의 윤리적 지점에 덜 민감 단단한 음식에 따라 달라집니다, 보통보다 COM에서 사용되는플렉스 작업 23. 안정적이고 일관성있는 결과를 얻기 위해 음식에서 원숭이를 뺏으려고 필요가 없습니다. 작업하는 동안받은 산탄은 행동 세션 (;하지만 추가 음식은 앞의 오늘의 오후의 끝날 때까지 주어진 수있는 원숭이가 앞의 야간 먹지 않는 가정)의 날에 음식을 가장 먼저 액세스를 나타냅니다 . 이것은 각 원숭이 행동 하루 중 같은 시간에 세션뿐만 아니라 주택 룸에서 그룹을 형성 다른 원숭이 사이의 동일 순차 순서를 존중을 수행 한다는것 중요합니다. 원숭이가 외부 교란 이벤트 구분으로 행동 작업이 인접 실이나 실험실에서 오는 잠재적인 교란 소음을 마스킹, 배경 음악의 면전에서 실시한다. 이것은 실험 (마지막 매일 실험 세션을 초기 교육, 몇 개월이 아니라면 년간의 기간)의 전체 기간 동안 주어진 원숭이가되는 중요합니다같은 실험자의 감독하에 매일 배치. 수동 손재주 수치 저희 연구실에서 몇 년 이후에 사용되는 현재의 행동 검사, 최근 24-28 문헌에 보고된 설명서 손재주의 다른 시험에 비해 어느 정도하는 것입니다. 현재 보고서의 임시 목표 다른 실험실 (더 나은 비교)에 걸쳐 테스트를 표준화하기위한 중요한 필요하지만있다. 수요에서 설정 UPS의 세부 특성 1-4 그들을 복제하기 위해, 해당 저자에 의해 제공될 수 있습니다 테스트를 보시려면 여기를 그림. 재생 의학 (척수 혹은 대뇌 피질의 병변 회복)의 문제를 넘어, 테스트의 현재 팔레트 조사, 정상이 아닌 인간의 영장류 발달 문제 (손재주가 움직임의 모터 개발의 예 : 시간 ​​코스)에 주소 적합있을 수 있습니다 C로 lateralization 측면 (손 선호 / 지배)와 해독 진화 질문에인간을 포함한 영장류의 과목 종의 모터 능력을 omparing. 그것이 작업의 성능 영향을 줄 수로 apparatuses의 크기는, 영장류 종의 자리 &#39;크기 (두께와 길이)에 따라 적응해야하지만합니다. 현재 연구에서는, 검사는 2.5에서 8 세에 이르기까지와 2.5 사이의 8kg 무게, macaca fascicularis 원숭이에 실시되었다. 32에서 35 mm까지 검지 손가락의 길이 (알약을 조작하기 위해 먼저 사용)은 범위, 검지 손가락의 말초 헛점 (팁)의 둘레는 22, 우리의 연구에 포함되어있는 원숭이에서 25mm 사이 반면. 마찬가지로 이전의 실험에서 테스트 같은 쥐고 시험뿐만 아니라 macaca mulatta에 적합합니다. 모든 실험은 실험실 동물의 관리 및 사용 (1996)에 대한 가이드에 따라 실시와 지역 (스위스) 수의학 당국에 의해 승인되었습니다. 원숭이의 모든 실험 절차초, 동물 시설에서뿐만 아니라 구류 조건, 저희 연구실에서 최근 보고서에 자세히 설명되었습니다 : 참고 문헌에게 12-18를 참조하십시오.

behavioral assessment of manual dexterity in non-human primates

corticospinal (CS) 넓이는 능숙하게 작은 물체를 조작하는 정교한 모터 능력, 주로 영장류 1 특권의 해부 지원입니다. 그 기원 (모터 피질 영역의 병변)에 또는 그 궤도 (자궁 코드 병변) 따라 CS 프로젝션 시스템에 영향을 미치는 장애의 경우, 같은 tetraplegic 또는 hemiplegic 환자에서 본 설명서 손재주의 극적인 손실 (손에 마비가)있다 . 이러한 병변 후에 자연 회복 기능이 있지만, 그것은 성인에서 매우 제한된 남아 있습니다. 다양한 치료 전략은 현재 제안된 아르 (예 : 세포 치료, 억제 axonal 성장 분자의 중화, 성장 요인, 등 응용 프로그램), 주로 설치류에서 개발하는. 그러나, 임상 응용 프로그램을하기 전에, 그것은 종종 비 인간 영장류의 치료 가능성, 효능 및 보안을 테스트하는 것이 좋습니다. 목표가 후에 수동 손재주를 복원하는 경우 이것은 특히 사실입니다중추 신경계의 병변은 설치류의 모터 시스템의 조직과 같은 영장류 1,2와는 다릅니다. 짧은 꼬리 원숭이 원숭이는 치료가 적용되는 모터 피질이나 예를 들면 자궁 코드의 병변으로 인한 적자, 자발적인 기능 회복의 정도를 측정하고를 반영하기 위해, 영장류의 수동 손재주 수치 적합한 행동 모델로 현재 그림 있으며, 평가 얼마나 그것은 기능 회복을 향상시킬 수 있습니다. 수동 손재주의 행동 평가는 성인 짧은 꼬리 원숭이 원숭이의 초기 훈련을 필요로하는 뚜렷한, 보완, 도달 범위 및 이해 수동 작업 (알약을 파악하기 위해 정밀 그립의 사용)에 따라 달라집니다. 동물의 준비는 물론 설정 행동에 대하여 위치로 증명됩니다. 전형적인 원숭이의 성능은 각 작업에 대한 그림입니다. 관련 매개 변수의 수집과 분석은 정확한 손 manipula을 반영기뿐만 아니라 무력의 통제는 설명과 대표적인 결과와 함께 시연하고 있습니다. 이러한 데이터는 행동 데이터가 척수의 병변이나 모터 피질과 어느 정도 치료가 자발적으로 기능 회복을 향상시킬 수 비교에 의해 병변의 영향을 조사하기 위해 악용될 수있는 방법을 보여주는, 더 넓은 맥락에서 다음 배치됩니다 원숭이의 서로 다른 그룹 (예를 들어 치료를 사기 비교 취급). 행동 검사의 장점과 한계가 논의됩니다. 현재의 행동 방식은 신경계 질환 2,3의 맥락에서가 아닌 인간 영장류 모델의 적절을 강조 이전 리포트와 라인에 있습니다.

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

수동 손재주는 주로 영장류의 특권으로 행동 작업은 짧은 꼬리 원숭이 원숭이를 개발하고 있습니다. 네 손으로 조작 능력과 힘을 측정 포착 작업을 도달하고 파악, 중추 신경계의 병변 후 기능 회복을 수립하고 치료의 효과를 테스트할 수 있습니다.

비 인간 영장류에서 수동 손재수의 행동 평가

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

JoVE Journal

Neuroscience

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.

쥐에서 자당의 갈망의 보육을 평가하기위한 일반적인 방법

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 ...

연구

신경과학

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.

쥐에 활동 륜 실행에 녹음 및 Circadian 리듬 분석

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.

쥐의 해마 영역의 치아 이랑 (dentate gyrus)에서 수지상 Arborization의 평가

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.

쥐 두피 뉴런의 사상 액틴 (F - 굴지) puncta의 정량

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.

탐지 기술 현장에서 혁신적인 RNA에 의해 설치류 핵 Accumbens에서 도파민 D1-알파 수용 체의 식별

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.

설치류에서 시간적 처리의 평가 대 한 Interstimulus 간격의 힘

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.

성인 소뇌 슬라이스에 장기 우울증 유도의 평가

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.

뇌 슬라이스와 기본 세포 배양에서 피라미드 뉴런의 탄도 라벨링

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.

쥐 뇌 조직을 위한 소수성 조직 청산 방법

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

A Rat Model of EcoHIV Brain Infection

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. Establishment of the EcoHIV infected rat model for studies of drug abuse and neurocognitive disorders, would be beneficial in the study of neuroHIV and HIV-1 associated neurocognitive disorders (HAND). In the present study, we demonstrate the successful creation of a rat model of active HIV infection using chimeric HIV (EcoHIV). First, the lentiviral construct of EcoHIV was packaged in cultured 293 FT cells for 48 hours. Then, the conditional medium was concentrated and titered. Next, we performed bilateral stereotaxic injections of the EcoHIV-EGFP into F344/N rat brain tissue. One week after infection, EGFP fluorescence signals were detected in the infected brain tissue, indicating that EcoHIV successfully induces an active HIV infection in rats. In addition, immunostaining for the microglial cell marker, Iba1, was performed. The results indicated that microglia were the predominant cell type harboring EcoHIV. Furthermore, EcoHIV rats exhibited alterations in temporal processing, a potential underlying neurobehavioral mechanism of HAND as well as synaptic dysfunction eight weeks after infection. Collectively, the present study extends the EcoHIV model of HIV-1 infection to the rat offering a valuable biological system to study HIV-1 viral reservoirs in the brain as well as HAND and associated comorbidities such as drug abuse.

Here, we present a protocol to establish a new rat model of active HIV infection using chimeric HIV (EcoHIV), which is critical for enhancing our understanding of HIV-1 viral reservoirs in the brain and offering a system to study HIV-associated neurocognitive disorders and associated comorbidities (i.e., drug abuse).

에코HIV 뇌 감염의 쥐 모델

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. ...