Name: assessing spatial memory impairment in a mouse model of traumatic brain injury using a radial water tread maze
Uploaded: 2017-07-17T14:00:00
Description: Watch this Scientific Journal Video about Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze at JoVE.com

Denne forskningen ble støttet av Institute of Translational Health Sciences pilotprosjektet mulighet (UL1TR000423), University of Washington senter for menneskelig utvikling og funksjonshemming, og University of Washington Animal Behavior Core and Brain Imaging Core. Vi vil gjerne anerkjenne Dr. Warren Ladiges for hans rolle i utviklingen og formidlingen av den originale Radial Water Tread labyrintdesign og protokoll presentert her. Vi takker også Toby Cole for hans hjelp med dette prosjektet.

Alle prosedyrer og dyrehåndtering ble utført i samsvar med retningslinjer for dyrepleie utstedt av National Institutes of Health og ved University of Washington Animal Care and Use Committee. 1. Kirurgi <ol><li> Bedøv musen ved 5% isofluran i en induksjonsboks til bevisstløs. Bekreft anestesi ved reduksjon av pustefrekvensen og fravær av tilbaketrekningsrefleks etter tøffel. </li><li> Oppretthold anestesi via nesekegle ved 2-2,5% gjennom kirurgi. Overvåk pustefrekvensen gjennom kiru...

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RWT labyrintprotokollen som presenteres her, skiller mellom CCI-induserte TBI-mus og sham-kontroller, og representerer et gjennomførbart, musekentrisk alternativ til MWM og Barnes sirkelformede labyrint. Mens resultatene som rapporteres her snakker bare om bruken av RWT-labyrinten i en TBI-musemodell, har dette apparatet blitt brukt vellykket i alderen og transgene modeller hvor stress-indusert ikke-compliance som følge av svømmebasert testing gjort ved hjelp av MWM upraktisk 15 , <...

Minnehemming er blant de vanligste symptomene som rapporteres av pasienter som følger traumatisk hjerneskade (TBI) 1 , 2 . Nøyaktig identifisering og vurdering av analoge minneunderskudd i dyremodeller fra TBI er derfor avgjørende for vår forståelse av denne tilstanden og dens ledelse. Her presenterer vi en protokoll for å teste romlig minne i en musemodell av TBI ved hjelp av en Radial Water Tread (RWT) labyrint. Dette apparat ble tidligere vist å vurdere kognitiv svikt i musemodeller for kontrollert kortikale innvirkning (CCI) -indusert TBI 3, og representerer et potensielt alternativ til rotte-validert, bade-baserte tester av kognisjon. Det voksende mangfoldet og tilgjengeligheten av transgene musemodeller har ført til en nylig økning i bruken av mus over rotter i vitenskapelig forskning 4 . Til tross for dette skiftet fortsetter forskerne å stole på atferdsmessig aNd kognitive oppgaver som ble opprinnelig designet og validert for råttabruk. De vanligste testene som for tiden brukes til å vurdere kognisjon hos mus, Morris Water Maze (MWM) og Barnes sirkelformede labyrint, ble spesielt utviklet for å kapitalisere på instinktuelle oppføringer funnet i rotter 5 , 6 . Med tanke på de genetiske, neuroetologiske og kognitive forskjellene som eksisterer mellom disse to artene 4 , er det ikke overraskende at musene konsekvent underperformer disse oppgavene 7 , 8 . Arteravhengige forskjeller i testevnen er spesielt knyttet til svømmebaserte kognitive tester, som for eksempel MWM. Mens både rotter og mus er dyktige svømmere, har forskere identifisert flere musestammer som utfører bemerkelsesverdig dårlig på svømmebaserte kognitive oppgaver 9 , 10 , </Sup&gt; 11 , 12 , 13 . Selv i villdyr-type dyr utgjør rotter generelt mus 7 , 8 . Selv om dette kunne tolkes som en artsspesifikk forskjell i romlig hukommelse, viste analog oppfølgingstesting ved hjelp av en tørrlands labyrint ingen arteravhengige forskjeller i kognitiv ytelse 8 . En rekke faktorer som ikke er relatert til kognisjon, kan utgjøre dette funnet, inkludert arteravhengige forskjeller i enten svømmemuligheter eller søkestrategi. Faktaanalyse av musespesifikke søkestrategier i MWM viser faktisk at ikke-kognitive faktorer (spesielt tigomotaks og passivitet [ dvs. flytende]) kan spille en mer betydelig rolle i MWM-ytelse enn romlig læring 14 . Her demonstrerer vi bruken av en kognitiv test designet for å kapitalisere på innsatsenMuskels stinktuelle oppførsel, og som ikke krever svømming, for å måle romlig minneverdigelse i en musemodell av CCI-indusert TBI. Mens RWT labyrinten ( Figur 1 A-B ) ble oppfattet som en ny hybrid av MWM og Barnes sirkelformede labyrint, ble den spesielt utviklet for å utnytte den thigmotaktiske oppførselen instinktiv til mus 15 , 16 . Apparatet består av et 32-tommers galvanisert stålkar, hvor ni jevnt fordelte utgangshull har blitt kjedet. Hullene er sentrert 2-1 / 4 tommer over gulvet i karet og er dimensjonert for å passe til vanlige 1-1 / 2-tommers ABS DWV SPG x SJ-felleadaptere. Åtte av utgangene er utkapslet fra utsiden og blindet til en dybde på 1 tommer med gummipropper. Den niende er forbundet med en 90 ° akrylonitrilbutadienstyren (ABS) albue til en ugjennomsiktig plastkasse hvorfra musen lett kan fjernes etter testing. I løpet av aKort oppkjøpsperiode, musen er opplært til å bruke de unike visuelle signalene som ligger på labyrinten for å finne denne fluktboksen. Under testingen er labyrinten fylt med en tomme kaldt vann (12-14 ° C), tilstrekkelig aversive nok til å fremme flukt, men ikke dyp nok til at musen er nødvendig for å svømme. RWT labyrinten representerer et billig, lavt vedlikeholdsalternativ til MWM, og har blitt brukt vellykket i alderen og transgene mus 15 , 17 , 18 , 19 og CCI-induserte musemodeller av TBI 3 . Protokollen som er skissert her representerer en enkel og effektiv metode for måling av romlig minneverdighet uten å kreve opplæringstrening, og det kan lett endres slik at det passer til et laboratoriums spesielle behov.

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assessing spatial memory impairment in a mouse model of traumatic brain injury using a radial water tread maze

Til tross for den nylige økningen i bruk av musemodeller i vitenskapelig forskning, fortsetter forskere å bruke kognitive oppgaver som opprinnelig ble designet og validert for bruk av rotter. Rødtvannstesten (Radial Water Tread) (RWT) av romlig hukommelse (designet spesielt for mus og krever ingen svømming) har tidligere blitt vist for å skille mellom kontrollerte cortical impact-induserte TBI-mus og sham-kontroller. Her presenteres en detaljert protokoll for denne oppgaven. RWT labyrinten kapitaliserer på den naturlige tendensen til mus for å unngå åpne områder for å klemme sidene på et apparat (thigmotaxis). Veggene i labyrinten er foret med ni rømningshull plassert over apparatets gulv, og musene er utdannet til å bruke visuelle signaler for å finne rømningshullet som fører ut av labyrinten. Labyrinten er fylt med en tomme kaldt vann, tilstrekkelig til å motivere flukt, men ikke dyp nok til å kreve at musen svømmer. Oppkjøpsperioden tar bare fire treningDager med en test av minneoppbevaring på dag fem og en langtidsminnetest på dag 12. Resultatene som er rapportert her, tyder på at RWT labyrinten er et mulig alternativ til rotte-validerte, svømmebaserte kognitive tester i vurderingen av romlige Minneunderskudd i musemodeller av TBI.

RWT labyrinten ( Figur 1 ) ble brukt til å undersøke skadeavhengige romlig minneunderskudd hos mus tilfeldig tildelt for å motta enten kontrollert kortikalstimulert indusert TBI eller sham kirurgi. Skaden ble generert ved hjelp av en magnetisk drevet cortical innvirkning med en 3 mm konvekse spiss og følgende skadeparametere: 6 m / s streikhastighet, 1 mm inntrengningsdybde og 200 ms kontakttid. Mus mottok kognitiv testing som startet ved 35 dager etter ...

Watch this Scientific Journal Video about Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze at JoVE.com

Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze

Her presenterer vi en protokoll for en musespesifikk test av kognisjon som ikke krever svømming. Denne testen kan brukes til å klarere å skille kontrollerte kortikale slag-induserte traumatiske hjerneskade-mus fra skamkontroller.

Vurdering av romlig minneverdigelse i en musemodell av traumatisk hjerneskade ved hjelp av en radial vanntrinn labyrint

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and ...

The overall goal of this Radial Water Tread Maze protocol is to test spatial memory in a mouse model of traumatic brain injury using a mouse centric cognitive task that does not require swimming. This method can be especially useful for testing cognition in mouse models, where motor deficits and or a lack of motivation makes swimming based cognitive testing protocols such as the Morris Water Maze impractical. Though this method could provide insight into group differences and cognition in a controlled cortical impact model of traumatic brain injury in mice, it can also be useful for measuring cognition in aged, and transgenic mouse models that are known to perform poorly in swimming based tests.On all testing days, perform the following preparatory steps. First, allow the mice to acclimate to the test room, while in their home cages for at least 30 minutes. The round maze features visual cues along the inner walls.The movable escape box positioned at one of nine different exit holes. While the mice acclimate, sanitize the exits and the box using a 70%ethanol spray. Next, fill the maze with roughly one inch of cold water.Keeping the water temperature consistent between trials is very important. So measure accurately, and adjust as needed. Now, position the escape box at one of the nine exit holes.Using a heating source to keep the box warm, ideally using a heating pad. Also, seal the box from light. The escape box should always be dark, warm and relatively odorless.The light source is also likely to be a cue so do not change its position between trials. Remove the test mouse from its cage gently by the tail, and place it in the center of the apparatus. Then immediately start timing how long it takes for the mouse to find the exit.If a mouse climbs into a terminating hole, and does not spontaneously reenter the maze after 20 seconds, do not stop the trial. Instead, guide the mouse back into the center of the maze by hand, and let the mouse complete the trial. If after three minutes, the mouse hasn't exited the maze, score the trial as a failure, and record the time as three minutes.Then by hand, carefully guide the animal along the straightest path to the exit. Once in the escape box, allow the mouse to remain there for one minute. Then transfer it back to its home cage while the maze gets reset.Between trials, thoroughly sanitize the escape box and exits with a 70%ethanol spray to prevent the mouse from using old factory cues to locate the correct escape route. This can be done pretty quickly. Now, repeat the trial on the same mouse twice more for a total of three consecutive trials.Every testing day consists of three trials per mouse. No more, and no less. In addition to sanitizing, drain the maze and refresh the water between testing different mice.Be sure to keep the water temperature between 12 and 14 degrees Celsius. Use containers of ice or colder hot water to adjust the tap water temperature into this range. Between testing days, it is important to make certain that the visual cues in the maze, and in the space surrounding the maze remain consistent.A complete testing regime includes five consecutive testing days all run in the same manner. The fifth day is considered the measure for memory retention. Provided no tests are performed between day six and 11, a test on day 12 can be used to measure long term memory.The radial water maze was used to investigate injury dependence spatial memory deficits in mice receiving either controlled cortical impact induced traumatic brain injury or a sham injury. The TBI surgery is described in detail in the text protocol and in a JoVE article by Romin, Gao and Chen from 2014. Briefly put, the injury was generated using a solenoid driven cortical impact with a three millimeter convex tip.The shams were operated on but impacted by the device. 35 days post surgery, the described testing regime was performed. All the mice passed the criteria test for a desire to escape the maze.So all were included in the analysis. The results show a clear difference between the TBI mice and sham controls. Escape latency was significantly longer for injured mice on the first day of training, and on both of the two memory test days.Thus, the described TBI affected the animal's capacity to learn. After watching this video, you should have a good understanding of how to properly perform cognition testing using a mouse specific test of spatial cognition. Once mastered, this technique can be done in less than 15 minutes per animal if it is performed properly.While attempting this procedure, it is important to remember that the positioning of visual cues and the water temperature must remain consistent across all trials.

Research

JoVE Journal

Behavior

Shallow Water (Paddling) Variants of Water Maze Tests in Mice

Shallow Water Paddling Variants of Water Maze Tests in Mice

Grunt vann (Padling) Varianter av Water Maze Tester i Mus

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to ...

Use of an Eight-arm Radial Water Maze to Assess Working and Reference Memory Following Neonatal Brain Injury

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food ...

The Double-H Maze: A Robust Behavioral Test for Learning and Memory in Rodents

Double-H Maze: En Robust Behavioral Test for læring og hukommelse i Gnagere

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their ...

Morris Water Maze Test: Optimization for Mouse Strain and Testing Environment

Morris Water Maze Test: Optimalisering for Mouse Sil og testing miljø

 The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, ...

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury

Oppdage atferdsmessige underskudd i rotter etter traumatisk hjerneskade

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, ...

Stress-Enhanced Fear Learning, a Robust Rodent Model of Post-Traumatic Stress Disorder

Stress-forbedret frykt læring, en Robust gnager modell av posttraumatisk stresslidelse

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders ...

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget

Romanen objekt gjenkjenning og objektet plassering opptreden i mus på et budsjett

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various ...

Analysis of Learning and Memory Ability in an Alzheimer's Disease Mouse Model using the Morris Water Maze

Analyse av læring og hukommelse evne i en Alzheimers sykdom Mouse modell ved hjelp av Morris Water Maze

A Morris water maze (MWM) experiment forces experimental animals to swim and learn to find a platform hidden in the water. It is widely used in scientific ...

Inducing Post-Traumatic Epilepsy in a Mouse Model of Repetitive Diffuse Traumatic Brain Injury

Indusere posttraumatisk epilepsi i en musemodell av repeterende diffus traumatisk hjerneskade

Traumatic brain injury (TBI) is a leading cause of acquired epilepsy. TBI can result in a focal or diffuse brain injury. Focal injury is a result of ...

Utilizing a Reconfigurable Maze System to Enhance the Reproducibility of Spatial Navigation Tests in Rodents

Bruk av et rekonfigurerbart labyrintsystem for å forbedre reproduserbarheten av romlige navigasjonstester hos gnagere

Several maze shapes are used to test spatial navigation performance and behavioral phenotypes. Traditionally, each experiment requires a unique maze ...