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

Forfatterne vil gerne takke professor 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, Dr. V. Goetschman for deres bidrag til tidligere forsøg og analyser. Den eksperimentelle set-ups blev konstrueret af André Gaillard, Bernard Aebischer og Laurent Monney. I Dyrefacilitet var aber under tilsyn af professionelle dyre viceværter: Josef Corpataux, Laurent Bossy og Jacques Maillard. De adfærdsmæssige test og analyse af data, samt histologi, blev udført med de meget værdifulde bidrag af laboranter: Véronique Moret (også web-master), Françoise Tinguely, Christine Roulin, Monica Bennefeld, Christiane Marti og Georgette Fischer. Dette arbejde blev støttet af den schweiziske National Science Foundation, tilskud Nej 31-61857,00, 310000-110005, 31003A-132465 (EMR), 310030-118357, 31003A-104061 (TW), 310030-120.411 (ABS), PZ00P3_121646 (ES), Novartis Foundation, Det Nationale Center for Kompetence i forskning (NCCR) om &quot;Neural plasticitet og reparation&quot;.

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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. 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Ingen interessekonflikter erklæret.

Selv om den nuværende adfærdsmæssige opgaver har været betragtet som så vidt i vores laboratorium i forbindelse med undersøgelser vedrørende læsion af de cervikale ledningen eller læsion af den motoriske hjernebark med det formål at afprøve forskellige behandlinger (se 14,15,17 og <a href="http://www.unifr.ch/neuro/rouiller">http:/ / www.unifr.ch / neuro / rouiller</a> &gt; vælg &quot;forskning&quot; i øverste bjælke menuen, og derefter&gt; &quot;motor system&quot;&gt; &quot;helbredelse efter læsion&quot;), kan de også have en bredere anvendelse, da håndelag er også et aspekt at overveje i andre patologier, såsom Parkinsons sygdom (MPTP aber) eller i tilfælde af sensorisk de-afferentation påvirker proprioception og / eller følesansen (især testen 2 i fravær af visuel kontrol). De adfærdsmæssige undersøgelser foreslås her, er egnede til at undersøge motorisk kontrol af distal bevægelser af forelimb, som er involveret i håndelag. Specificiteten af ​​testene fremgår afmangel af underskuddet (bortset fra et par dage) i tilfælde af en læsion, som ikke forringer relevante dele af kontrolsystemet: ja, i tilfælde af en læsion placeret mere caudale end motoneurons kontrollerende hånd musklerne, var der ingen underskud. Relevansen af test 1 kan blive værdsat ved at sammenligne på video sekvenser taget på to tidspunkter i den post-læsion opsving kurve, som adskiller sig ved en forbedring af funktionel genopretning af 25%, eventuelt i forhold til en celle terapi behandling 17. På trods af nogle indledende, relativt korte uddannelse fase primo (varighed normalt 2-3 måneder), er de adfærdsmæssige test her foreslåede relativt &quot;naturlige&quot; og ligetil, i forhold til komplekse (fx betinget) opgaver, som uddannelse af aben kan tage næsten et år eller mere. Den positive forstærkning er baseret på fast føde, der er mindre følsomme på etisk synspunkt end vand afsavn, der normalt bruges i mere comkomplekse opgaver 23. Der er ingen grund til at fratage aber fra mad til at opnå stabile og ensartede resultater. De træpiller modtaget i løbet af opgaver repræsenterer den første adgang til foder på dagen for den adfærdsmæssige session (forudsat, at aben ikke spiser i løbet af den foregående nat, men yderligere mad kan gives indtil udgangen af ​​eftermiddagen den foregående dag) . Det er afgørende, at begge aben udfører de adfærdsmæssige møde på samme tidspunkt af dagen, samt respektere den samme rækkefølge mellem de forskellige aber at danne en gruppe i huset rummet. Da aber er følsomme over for eksterne foruroligende begivenheder, bør de adfærdsmæssige opgaver skal udføres i overværelse af baggrundsmusik, maskering potentielle forstyrrende støj fra tilstødende rum eller laboratorier. Det er afgørende, at der under hele varigheden af ​​eksperimentet (fra grunduddannelse op til den sidste daglige eksperimentelle session, en periode på flere måneder, hvis ikke år), en given aben erplaceret dagligt under tilsyn af den samme eksperimentator. Den nuværende adfærdsmæssige test, der anvendes i flere år inden vores laboratorium for at kvantificere fingerfærdighed, er til en vis grad kan sammenlignes med andre tests af håndelag for nylig rapporteret i litteraturen 24-28. Der er dog et afgørende behov for at standardisere tests på tværs af forskellige laboratorier (for bedre sammenligning), hvilket er et foreløbigt mål for denne rapport. On demand, illustrerede den detaljerede egenskaber af set-ups her for test 1-4 kan leveres af den tilsvarende forfatter, for at kopiere dem. Ud over spørgsmålet om regenerativ medicin (recovery fra læsion af rygmarven og hjernebarken), kan denne palet af test være egnet til at løse i almindelige ikke-menneskelige primater udviklingsmæssige problemer (f.eks Tidsforløbet for motoriske udvikling af behændige bevægelser), at undersøge lateralization aspekter (hånd præference / dominans) og til at afkode evolutionære spørgsmål af comparing den motoriske evner af forskellige arter af primater, herunder menneskelige forsøgspersoner. Bemærk dog, at dimensioner af apparater bør tilpasses i overensstemmelse med cifrene &#39;størrelse (tykkelse og længde) af primat arter, da det kan påvirke opgaveløsningen. I nærværende undersøgelse, blev forsøgene udført på Macaca fascicularis aber, fra 2,5 til 8 år gammel og vejer mellem 2,5 og 8 kg. Længden af ​​pegefingeren (bruges først til at manipulere pellets) varierer fra 32 til 35 mm, mens omkredsen af ​​den distale phalanx (spids) af pegefingeren var mellem 22 og 25 mm i aber indgår i vores undersøgelser. Som afprøvet i tidligere forsøg, den samme grådige tests er velegnede til Macaca mulatta så godt. Alle forsøgene blev udført i overensstemmelse med vejledningen for pleje og brugen af forsøgsdyr (1996) og godkendt af de lokale (schweiziske) veterinære myndigheder. Alle eksperimentelle procedurer på abens, samt fængselsforholdene i dyret anlægget, blev beskrevet i detaljer i de seneste rapporter fra vores laboratorium: se referencer 12-18.

behavioral assessment of manual dexterity in non-human primates

Den corticospinal (CS) tarmkanalen er den anatomiske støtte fra den udsøgte motorens evne til at dygtigt manipulere små objekter, et prærogativ hovedsageligt af primater 1. I tilfælde af læsion påvirker CS projektion system på dens oprindelse (læsion af motor kortikale områder) eller langs sin bane (livmoderhalskræft ledning læsion), er der et dramatisk tab af håndelag (hånd lammelse), som ses i nogle tetraplegiker eller hemiplegisk patienter . Selvom der er en vis spontan funktionel genopretning efter sådanne forandringer, er det stadig meget begrænset i den voksne. Forskellige terapeutiske strategier er i øjeblikket foreslået (fx celleterapi, neutralisering af hæmmende axonal vækst molekyler, anvendelse af vækstfaktorer, osv.), som er mest udviklet sig i gnavere. Men før klinisk anvendelse, er det ofte anbefales at afprøve, effekt og sikkerhed af behandlingen hos ikke-menneskelige primater. Dette gælder især, når målet er at genoprette håndelag efteren læsion af det centrale nervesystem, som tilrettelæggelsen af det motoriske system af gnavere er forskellig fra den, primater 1,2. Makak-aber er illustreret her som en passende adfærdsmæssige model til at kvantificere fingerfærdighed med primater, at afspejle de underskud som følge af læsion af den motoriske hjernebark eller livmoderhals ledning for eksempel måle omfanget af spontane funktionel genopretning, og når en behandling er anvendt, evaluere hvor meget det kan forbedre den funktionelle opsving. De adfærdsmæssige vurdering af håndelag er baseret på fire forskellige, komplementære, nå og gribe manuelle opgaver (brug af præcision greb til at forstå pellets), hvilket kræver en grunduddannelse af voksne makak aber. Forberedelsen af ​​dyrene er påvist, samt positionering i forhold til adfærdsmæssige set-up. Udførelsen af ​​en typisk abe er illustreret for hver opgave. Indsamling og analyse af relevante parametre, der afspejler præcis hånd manipulationtion, samt kontrol af magt, er forklaret og demonstreret med repræsentative resultater. Disse data placeres derefter i en bredere sammenhæng, der viser, hvordan de adfærdsmæssige data kan udnyttes til at undersøge virkningen af ​​en rygmarven læsion eller en læsion af den motoriske hjernebark og i hvilket omfang en behandling kan forstærke den spontane funktionelle opsving, ved at sammenligne forskellige grupper af aber (behandlet versus simuleret behandlet for eksempel). Fordele og begrænsninger af de adfærdsmæssige test diskuteres. Den nuværende adfærdsmæssige tilgang er i overensstemmelse med tidligere rapporter, der understreger relevansen af ikke-menneskelige primat model i forbindelse med sygdomme i nervesystemet 2,3.

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Behavioral Assessment of Manual Dexterity in Non-Human Primates

Som håndelag er et prærogativ hovedsageligt af primater, har adfærdsmæssige opgaver er udviklet i makak aber. Fire nå og gribe prehension opgaver, måle hånd manipulation evne og kraft, gør det muligt at oprette funktionelle bedring efter en læsion af det centrale nervesystem og for at afprøve effekten af ​​en behandling.

Adfærdsmæssige Vurdering af håndelag i ikke-menneskelige primater

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

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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.

En generel metode til evaluering Inkubation af Saccharose Savner hos rotter

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.

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.

Vurdering af dendritiske arborization i gyrus dentatus i hippocampus-regionen i mus

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.

Kvantificering af Filamentøs actin (F-actin) puncta i Rat kortikale neuroner

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.

Identifikation af dopamin D1-Alpha Receptor inden for gnavere kernen Accumbens af en innovativ RNA In Situ opdagelse teknologi

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.

Kraften i Interstimulus Interval for vurdering af tidsmæssige forarbejdes i Rodents

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.

Vurdering af langvarig depression induktion i voksne cerebellar skiver

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.

Ballistisk mærkning af pyramideformede neuroner i hjerneskiver og i primærcellekultur

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.

En hydrofobisk vævsclearingmetode til rottehjernevæv

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).

En rottemodel af EcoHIV-hjerneinfektion

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