فإن الكتاب أود أن أشكر البروفيسور شواب ME ، الدكتور فرويند P. ، الدكتور ويس A. ، الدكتور بشير S. ، الدكتور مير A. ، الدكتور بلوخ J. ، الدكتور برونيه JF ، والدكتور ج. Aebischer ، الدكتور أ. Meszaros الدكتور خامسا Goetschman لمساهمتهم في التجارب السابقة والتحليلات. وشيدت مجموعة التجريبية المنبثقة من قبل أندريه غايار ، وعملات Aebischer برنارد لوران. في منشأة الحيوان ، وضعت القرود تحت إشراف القائمين الحيوانية المهنية : جوزيف Corpataux ، لوران مايار متسلط وجاك شيراك. وأجريت التجارب السلوكية وتحليل البيانات ، وكذلك الأنسجة ، مع مساهمات قيمة للغاية من فنيي المختبرات : فيرونيك موريه (أيضا ويب ماستر) ، Tinguely فرانسواز ، Roulin كريستين ، Bennefeld مونيكا ، مارتي كريستيان وفيشر جورجيت. وأيد هذا العمل من قبل مؤسسة العلوم الوطنية السويسرية ، والمنح لا 31-61857،00 ، 310000-110005 ، 31003A - 132465 (EMR) ، 310030-118357 ، 31003A - 104061 (TW) ، 310030-120411 (ABS) ، PZ00P3_121646 (ES) ، مؤسسة نوفارتيس ، والمركز القومي للبحوث في اختصاص (NCCR) حول &quot;اللدونة العصبية وإصلاح&quot;.

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الإعلان عن أي تضارب في المصالح.

وإن كانت قد نظرت في المهام السلوكية الحالية حتى الان في مختبرنا في سياق الدراسات المتعلقة الآفة من الحبل عنق الرحم أو آفة من القشرة الحركية بهدف اختبار العلاجات المختلفة (انظر 14،15،17 و <a href="http://www.unifr.ch/neuro/rouiller">http:/ / www.unifr.ch / العصبية / rouiller</a> &gt; حدد &quot;البحث&quot; في شريط القائمة العلوي ، ثم&gt; &quot;النظام المحرك&quot;&gt; &quot;الانتعاش بعد الآفة&quot;) ، قد يكون لديهم أيضا تطبيقها على نطاق أوسع ، والبراعة اليدوية أيضا جانبا للنظر في أمراض أخرى ، مثل مرض باركنسون (القرود MPTP) أو في حالة استقبال الحس العميق دي afferentation الحسية المؤثرة و / أو حاسة اللمس (وخاصة اختبار 2 في غياب المراقبة البصرية). الاختبارات السلوكية المقترحة هنا هي مناسبة للتحقيق في التحكم الحركي للحركات البعيدة للforelimb ، كما تشارك في البراعة اليدوية. وتتجلى خصوصية التجارب التي قامت بهاغياب العجز (باستثناء بضعة أيام) في حالة وجود الآفة التي لا تضر العناصر ذات الصلة في نظام التحكم : في الواقع ، في حالة وجود الآفة وضعت أكثر من الذيلية العصبونات الحركية السيطرة على عضلات اليد ، لم يكن هناك عجز. ويمكن تقدير ملاءمة للاختبار 1 بمقارنة على التسلسلات الفيديو التي اتخذت في الوقت اثنين من نقاط منحنى الانتعاش بعد الآفة ، والتي تختلف عن طريق تعزيز الانتعاش وظيفية بنسبة 25 ٪ ، وربما بالنسبة إلى خلية العلاج العلاج 17. على الرغم من بعض الأولية ، مرحلة تدريب قصيرة نسبيا في بداية (دائم عموما 2-3 أشهر) ، والتجارب السلوكية المقترحة هنا هي نسبيا &quot;الطبيعية&quot; ومباشرة ، بالمقارنة مع المهام (مثل المشروطة) المعقدة التي وتدريب القرد قد يستغرق نحو عام أو أكثر. يستند التعزيز الإيجابي على الطعام الصلب ، والتي هي أقل حساسية على جهة النظر الأخلاقية الحرمان من الماء ، وعادة ما تستخدم في أكثر كومالمهام نوع plex 23. ليست هناك حاجة لحرمان القردة من الطعام للحصول على نتائج مستقرة وثابتة. الكريات المهام التي وردت خلال تمثيل وصول أول من الطعام في يوم من الدورة السلوكية (على افتراض أن لا يأكل القرد خلال الليلة السابقة ، ولكن يمكن إعطاء المزيد من المواد الغذائية حتى نهاية بعد ظهر اليوم السابق) . فمن الأهمية بمكان أن ينفذ كل قرد الدورة السلوكية في نفس الوقت من اليوم ، فضلا عن احترام بنفس الترتيب المتسلسل بين القرود مختلفة تشكيل مجموعة في غرفة السكن. كما القرود حساسة لأحداث مزعجة الخارجية ، ينبغي إجراء المهام السلوكية في وجود موسيقى في الخلفية ، إخفاء الضوضاء المزعجة المحتملة القادمة من الغرف المجاورة أو المختبرات. فمن الأهمية بمكان أن كله خلال فترة التجربة (من التدريب الأولي حتى الدورة الأخيرة التجريبية اليومية ، لمدة عدة أشهر إن لم يكن سنوات) ، قرد معطىتوضع اليومية تحت إشراف المجرب نفسه. التجارب الحالية السلوكية ، تستخدم منذ عدة سنوات في مختبرنا لقياس المهارة اليدوية ، وإلى حد ما مماثلة لغيرها من التجارب من البراعة اليدوية التي نشرت مؤخرا في الأدب 24-28. بيد أن هناك حاجة ماسة لتوحيد الاختبارات عبر مختبرات مختلفة (على سبيل المقارنة بشكل أفضل) ، والذي هو هدف مؤقت لهذا التقرير. بناء على الطلب ، ويتضح خصائص مفصلة للمجموعة ، شكا من هنا لاجراء اختبارات ويمكن توفير 1-4 مقدم البلاغ المقابلة ، من أجل تكرارها. أبعد من مسألة الطب التجديدي (الشفاء من الآفة من الحبل الشوكي أو قشرة الدماغ) ، قد لوح الحالي للاختبارات تكون مناسبة لمعالجة قضايا عادية في الرئيسيات غير البشرية التنموية (مثل دوام التطور الحركي للحركات حاذق) ، للتحقيق الجوانب lateralization (تفضيل اليد / الهيمنة) وعلى الأسئلة فك التطورية التي جomparing القدرات الحركية من الأنواع المختلفة من الثدييات ، بما في ذلك البشر. ومع ذلك نلاحظ أنه ينبغي تكييف أبعاد الأجهزة وفقا لحجم الأرقام &quot;(سمك وطول) من أنواع الرئيسيات ، لأنها قد تؤثر على أداء المهمة. في هذه الدراسة ، أجريت التجارب على القردة المكاك fascicularis ، تتراوح بين 2.5 إلى 8 سنوات من العمر ويزن ما بين 2.5 و 8 كيلوغرامات. طول السبابة (استخدم لأول مرة لمعالجة الكريات) يتراوح 32-35 ملم ، في حين أن محيط السلامية القاصية (نصيحة) من السبابة وكان بين 22 و 25 ملم في القرود المدرجة في دراساتنا. كما تم اختبارها في التجارب السابقة ، فإن نفس الاختبارات هي مناسبة لاستيعاب المكاك الخلاسي كذلك. أجريت جميع التجارب وفقا لدليل لرعاية واستخدام حيوانات المختبر (1996) والتي وافقت عليها السلطات (السويسرية) البيطرية المحلية. جميع الإجراءات التجريبية على قردوقد وصفت ثانية ، فضلا عن ظروف الاحتجاز في منشأة الحيوان ، بالتفصيل في التقارير الأخيرة من مختبرنا : انظر مراجع 12-18.

behavioral assessment of manual dexterity in non-human primates

و(CS) هو السبيل القشري التشريحية للدعم القدرة الحركية الرائعة للتلاعب بمهارة الأشياء الصغيرة ، وهي أساسا من اختصاص الرئيسات 1. في حالة الآفة التي تؤثر على نظام الإسقاط CS من مصدره (آفة المناطق القشرية الحركية) أو على طول مساره (عنق الرحم آفة النخاع) ، وهناك خسارة كبيرة من البراعة اليدوية (شلل اليد) ، كما رأينا في بعض المرضى tetraplegic أو مفلوج . على الرغم من أن هناك بعض التعافي العفوية وظيفية بعد هذه الآفة ، فإنها لا تزال محدودة جدا في البالغين. ويقترح استراتيجيات علاجية مختلفة في الوقت الحاضر (مثل العلاج بالخلايا ، تحييد الجزيئات المثبطة محواري النمو ، وتطبيق عوامل النمو ، الخ) ، والتي يتم تطويرها في الغالب في القوارض. ومع ذلك ، قبل التطبيق السريري ، يوصى غالبا ما لاختبار جدوى وفعالية وأمان العلاج في الرئيسيات غير البشرية. هذا صحيح خصوصا عندما يكون الهدف هو استعادة البراعة اليدوية بعدآفة النظام العصبي المركزي ، وتنظيم نظام المحرك من القوارض وهو يختلف عن ذلك من قرود 1،2. قرود المكاك وتتضح هنا نموذجا سلوكية مناسبة لقياس المهارة اليدوية في المقدمات ، لتعكس العجز الناجم عن الآفة من القشرة الحركية أو الحبل عنق الرحم على سبيل المثال ، وقياس مدى الانتعاش وظيفية عفوية ، وعندما يتم تطبيق العلاج وتقييم إلى أي مدى يمكن أن تعزز الانتعاش وظيفية. ويستند التقييم السلوكي للالبراعة اليدوية على أربعة ، مكملة متميزة وفهم المهام الوصول ، والخط (استخدام قبضة الدقة في فهم الكريات) ، مما يتطلب التدريب الأولي للقرود المكاك الكبار. وأظهرت استعدادا للحيوانات ، وكذلك تحديد المواقع فيما يتعلق السلوكية انشاء. ويتضح من أداء قرد نموذجية لكل مهمة. جمع وتحليل المعايير ذات الصلة التي تعكس دقة manipula اليدوأوضح نشوئها ، فضلا عن السيطرة على القوة ، وأظهرت النتائج مع الممثل. توضع هذه البيانات بعد ذلك في سياق أوسع ، والتي تبين كيف يمكن استغلال البيانات السلوكية للتحقيق في تأثير آفة في الحبل الشوكي أو آفة من القشرة الحركية ، وإلى أي مدى يمكن علاج تعزيز التعافي العفوية الفنية ، التي تقارن مجموعات مختلفة من القرود (يعامل معاملة مقابل صورية على سبيل المثال). وتناقش مزايا وقيود من الاختبارات السلوكية. النهج الحالي السلوكية يتماشى مع التقارير السابقة التأكيد على ملاءمة نموذج الرئيسيات غير البشرية في سياق أمراض الجهاز العصبي 2،3.

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Behavioral Assessment of Manual Dexterity in Non-Human Primates

والبراعة اليدوية هي أساسا من اختصاص الرئيسيات ، وضعت في المهام السلوكية قردة المكاك. ويصل أربعة فهم المهام الإمساك ، وقياس قدرة جهة التلاعب والقوة ، والسماح بإنشاء الانتعاش وظيفية بعد آفة النظام العصبي المركزي ولاختبار أثر العلاج.

السلوكية تقييم البراعة اليدوية في الثدييات العليا غير البشرية

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

22.2K Views.  University of Fribourg. والبراعة اليدوية هي أساسا من اختصاص الرئيسيات ، وضعت في المهام السلوكية قردة المكاك. ويصل أربعة فهم المهام الإمساك ، وقياس قدرة جهة التلاعب والقوة ، والسماح بإنشاء الانتعاش وظيفية بعد آفة النظام العصبي المركزي ولاختبار أثر العلاج.

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

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.

طريقة عامة لتقييم الحضانة لالتوق السكروز في الجرذان

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.

تسجيل وتحليل إيقاعات الساعة البيولوجية في تشغيل عجلة النشاط في القوارض

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.

تقييم شجيري تشجر في التلفيف المسنن من منطقة الحصين في الفئران

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.

الكمي لشعيري أكتين (F-الأكتين) نقاط وفي الجرذ العصبونات القشرية

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.

تحديد مستقبلات الدوبامين D1-ألفا داخل Accumbens نواة القوارض بالحمض النووي الريبي مبتكرة في الموقع تكنولوجيا الكشف عن

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.

قوة إينتيرستيمولوس الفاصل الزمني للتقييم لتجهيز الزمانية في القوارض

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.

تقييم الحث علي الاكتئاب علي المدى الطويل في شرائح البالغين المخيخار

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.

وضع العلامات البالستية للخلايا العصبية الهرمية في شرائح الدماغ وفي ثقافة الخلايا الأولية

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.

طريقة إزالة الأنسجة الكارهة للماء لأنسجة دماغ الفئران

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).

نموذج الفئران من عدوى الدماغ EcoHIV

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