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 ønsker å takke 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, Dr. V. Goetschman for deres bidrag til tidligere forsøk og analyser. Den eksperimentelle oppsett ble bygget av André Gaillard, Bernard Aebischer og Laurent Monney. I dyret anlegget ble apene plassert under veiledning av profesjonelle dyr vaktmester: Josef Corpataux, Laurent Bossy og Jacques Maillard. Den atferdsmessige tester og analyser av data, samt histologi, ble gjennomført med svært verdifulle bidrag fra laboratorium teknikere: Véronique Moret (også web-master), Françoise Tinguely, Christine Roulin, Monica Bennefeld, Christiane Marti og Georgette Fischer. Dette arbeidet ble støttet av den sveitsiske National Science Foundation, tilskudd No 31 til 61857,00, 310000-110005, 31003A-132465 (EPJ), 310030-118357, 31003A-104061 (TW), 310 030-120 411 (ABS), PZ00P3_121646 (ES), Novartis Foundation; Nasjonalt kompetansesenter i forskning (NCCR) på &quot;Neural plastisitet og vedlikehold&quot;.

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Ingen interessekonflikter erklært.

Selv om den nåværende atferdsmessige oppgavene har vært vurdert så langt i vårt laboratorium i sammenheng med studier knyttet til lesjon i livmorhalsen ledningen eller lesjon av motor cortex med sikte på å teste ulike behandlinger (se 14,15,17 og <a href="http://www.unifr.ch/neuro/rouiller">http:/ / www.unifr.ch / nevro / rouiller</a> &gt; velg &quot;forskning&quot; i den øverste linjen menyen, deretter&gt; &quot;motor system&quot;&gt; &quot;recovery etter lesjon&quot;), kan de også ha et bredere program, som fingerferdighet er også et aspekt å vurdere i annen patologi, som Parkinsons sykdom (MPTP aper) eller manglende sensoriske de-afferentation påvirker propriosepsjon og / eller følelse av touch (spesielt test 2 i fravær av visuell kontroll). Den atferdsmessige testene foreslått her er egnet til å undersøke motorisk kontroll av distale bevegelser av forbena, som er involvert i fingerferdighet. Spesifisiteten av testene er demonstrert avfravær av underskudd (bortsett fra et par dager) i tilfelle av en lesjon som ikke svekker relevante deler av styresystemet: ja, i tilfelle av en lesjon plassert mer caudal enn motoneurons kontrollerende hånd muskler, var det ingen underskudd. Det relevans av test 1 kan bli verdsatt ved å sammenlikne på videosekvenser tatt på to tidspunkter av post-lesjon utvinning kurve, som avviker fra en forbedring av funksjonell bedring på 25%, eventuelt i forhold til en celleterapi behandling 17. På tross av noen innledende, relativt korte trening fase ved begynnelsen (som varer vanligvis 2-3 måneder), de atferdsmessige testene foreslått her er relativt &quot;naturlig&quot; og grei, i forhold til komplekse (f.eks betinget) oppgaver som opplæring av apekatten kan ta nesten et år eller mer. Den positive forsterkning er basert på fast føde, som er mindre følsom på det etiske synspunkt enn vann deprivasjon, vanligvis brukes i mer comPlex oppgaver 23. Det er ikke nødvendig å frata aper fra mat å få stabile og konsistente resultater. Den pellets mottatt under oppgavene representerer det første tilgangen til mat på dagen for de atferdsmessige økten (forutsatt at apen ikke spiser i løpet av den foregående natten, men ekstra mat kan gis fram til slutten av ettermiddagen dagen før) . Det er avgjørende at hver ape utfører de atferdsmessige økten på samme tid av dagen, samt respektere de samme rekkefølge mellom de forskjellige apene danner en gruppe i bolig rommet. Ettersom apene er følsomme for ytre forstyrrende hendelser, bør de atferdsmessige oppgavene utføres i nærvær av bakgrunnsmusikk, maskering potensielle forstyrrende støy som kommer fra nabolandet rom eller laboratorier. Det er avgjørende at under hele varigheten av forsøket (fra første trening fram til den siste daglige eksperimentelle sesjon, en periode på flere måneder om ikke år), er en gitt apekattplassert daglig under tilsyn av den samme eksperimentator. Den nåværende atferdsmessige tester, brukt siden flere år i vårt laboratorium for å kvantifisere fingerferdighet, er til en viss grad sammenlignes med andre tester av fingerferdighet nylig rapportert i litteraturen 24-28. Det er imidlertid et avgjørende behov for å standardisere tester på tvers av ulike laboratorier (for bedre sammenligning), som er et foreløpig mål for denne rapporten. På etterspørsel, illustrerte den detaljerte egenskaper set-ups her for tester 1-4 kan gis av tilsvarende forfatteren, for å gjenskape dem. Utover spørsmålet om regenerativ medisin (recovery fra lesjon i ryggmarg eller cerebral cortex), kan presentere palett av tester være egnet til adresse i normal ikke-menneskelige primater utviklingsspørsmål (f.eks tid løpet av motorisk utvikling av fingernem bevegelser), for å undersøke lateralization aspekter (hånden preferanse / dominans) og å dechiffrere evolusjonære spørsmål ved comparing motoren evner av forskjellige arter av primater, deriblant mennesker. Merk imidlertid at dimensjonene på apparatene bør tilpasses sifrene &quot;størrelse (tykkelse og lengde) av primater, da det kan påvirke oppgaven ytelse. I den foreliggende studien, ble testene gjennomført på Macaca fascicularis aper, som strekker seg 2,5 til 8 år gammel og veier mellom 2,5 og 8 kg. Lengden på pekefingeren (brukes første til å manipulere pellets) varierer 32-35 mm, mens omkretsen av distale falanks (tuppen) på pekefingeren var mellom 22 og 25 mm i aper inkludert i våre studier. Som testet i tidligere forsøk, det samme fatte testene er egnet for Macaca mulatta også. Alle eksperimentene ble utført i samsvar med Guide for Pleie og bruk av forsøksdyr (1996) og godkjent av lokale (sveitsiske) veterinærmyndigheter. Alle eksperimentelle prosedyrer på apekattens, samt fengsling forholdene i dyret anlegget, ble beskrevet i detalj i senere rapporter fra vårt laboratorium: se referanser 12-18.

behavioral assessment of manual dexterity in non-human primates

Den corticospinal (CS) tarmkanalen er anatomisk støtte fra utsøkte motor evne utfordri manipulere små objekter, et privilegium hovedsakelig av primater 1. Ved lesjon som påvirker CS projiseringssystem på sitt opphav (lesjon i motor kortikale områder) eller langs dens bane (cervikale ledning lesjon), er det et dramatisk tap av fingerferdighet (hånden lammelser), som sett i noen tetraplegic eller hemiplegisk pasienter . Selv om det er noen spontan funksjonell bedring etter en slik skade, er det fortsatt svært begrenset i den voksne. Ulike terapeutiske strategier er i dag foreslått (f.eks celleterapi, nøytralisering av hemmende aksonal vekst molekyler, anvendelse av vekstfaktorer, etc), som er mest utviklet i gnagere. Men før klinisk anvendelse, er det ofte anbefalt å teste gjennomførbarhet, effekt og sikkerhet av behandlingen i ikke-menneskelige primater. Dette er spesielt sant når målet er å gjenopprette fingerferdighet etteren skade i det sentrale nervesystemet, som organiseringen av motoriske system av gnagere er forskjellig fra primater 1,2. Macaque aper er illustrert her som en egnet atferds modell for å kvantifisere fingerferdighet i primater, for å reflektere underskuddene skyldes lesjon av motor cortex eller livmorhalsen snor for eksempel måle omfanget av spontan funksjonelle utvinning og når en behandling er anvendt, evaluere hvor mye den kan forbedre de funksjonelle utvinning. Den atferdsmessige vurderingen av manuell dyktighet er basert på fire forskjellige, komplementære, rekkevidde og gripe manuelle oppgaver (bruk av presisjon grep til å gripe pellets), krever en innledende opplæring av voksne macaque aper. Utarbeidelsen av dyrene er demonstrert, samt posisjonering med hensyn til de atferdsmessige set-up. Ytelsen til en typisk ape er illustrert for hver oppgave. Innsamling og analyse av relevante parametre som reflekterer presis hånd manipulasjonsjon, samt kontroll av makt, er forklart og demonstrert med representative resultater. Disse dataene er plassert så i en bredere sammenheng, viser hvordan de atferdsmessige data kan utnyttes til å undersøke effekten av en ryggmarg lesjon eller en lesjon i den motoriske hjernebarken og i hvilken grad en behandling kan forsterke den spontane funksjonelle utvinning, ved å sammenligne ulike grupper av aper (behandlet versus humbug behandlet for eksempel). Fordeler og begrensninger av atferdsmessige testene er diskutert. Den nåværende atferdsmessige tilnærmingen er i tråd med tidligere rapporter understreke relevans av de ikke-menneskelige primater modell i sammenheng med nervesystemet sykdommer 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

Som fingerferdighet er et privilegium hovedsakelig av primater, har atferdsmessige oppgaver er utviklet i macaque aper. Fire rekkevidde og gripe prehension oppgaver, måling hånd manipulasjon evne og kraft, la etablere funksjonell bedring etter en skade i det sentrale nervesystemet og for å teste effekten av en behandling.

Behavioral Vurdering av Manuelle Behendighet 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 ...

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

Research

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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 generell metode for evaluering Inkubasjon av Sukrose Begjær i 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 ...

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 av Dendritic arborization i Dentate Gyrus av 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.

Kvantifisering av Trådformede Actin (F-aktin) puncta i Rat kortikale nevroner

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.

Identifikasjon av dopamin D1-Alpha reseptor i gnager Nucleus Accumbens av en nyskapende RNA i Situ oppdagelsen teknologien

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 intervall for vurdering av timelige behandling i gnagere

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 av lang tids depresjon induksjon i voksen lillehjernen 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 merking av pyramidale nevroner i hjerneskiver og i primær cellekultur

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 hydrofob vevsryddingsmetode for rottehjernevev

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 rottemodell av EcoHIV hjerneinfeksjon

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