JC would like to acknowledge the David Hay Memorial Fund, The University of Melbourne for financial support in writing this manuscript. Funding for this project was provided by an ARC Linkage grant 100200129 (BVB, AJV, CTON).

Etica dichiarazione: Gli esperimenti sugli animali sono stati condotti in conformità con il Codice australiano per la cura e l&#39;uso di animali a scopi scientifici (2013). approvazione etica animale è stato ottenuto dal Comitato Etico degli animali, Università di Melbourne. I materiali sono qui per esperimenti di laboratorio solo, e non destinati ad uso medico o veterinario. 1. Gli elettrodi Preparazione Nota: Un trasmettitore a tre canali è usato per l&#39;impianto chirurgico che consente 2 ERG e la registrazione 1 VEP essere condotta simultaneamente. I tre attivi e non attivi tre elettrodi devono essere pre-modellato in una forma ad anello prima dell&#39;impianto in modo da collegare ad occhio. Ai fini dell&#39;identificazione, il produttore ha racchiuso elettrodi attivi a metà bianco, guaine in plastica colorata mezzo mentre gli elettrodi inattive sono coperti in guaine colorate piene. L&#39;elettrodo di massa (guaina di plastica trasparente) viene lasciato inalterato. Per tutti elettr attivo e inattivocondotta odi i punti 1.1, 1.2, 1.3 e 1.7. <ol><li> Districare il doppio elettrodo di acciaio inossidabile flessibile con due belle pinze a punta. </li><li> Tagliare uno dei filoni in acciaio inox (circa 1 cm dalla punta), lasciando un unico filo più lungo rettilineo rimanente per modellare l&#39;elettrodo ad anello. </li><li> Piegare il singolo filo di acciaio inossidabile di nuovo su se stesso e torsione, formando un anello liscio sulla punta dell&#39;elettrodo. </li><li> Per elettrodi attivi moda ERG questo ciclo ~ 0,2 - 0,5 mm di diametro ruotando la base del ciclo (per gli scopi descritti qui, modellare due elettrodi attivi in ​​questo modo per registrare ERG da entrambi gli occhi), e per la ERG inattivo e elettrodi PEV rendono il diametro anello ~ 0,8 mm di diametro (in questo esempio, fare questo per un elettrodo VEP attiva e tutte tre elettrodi inattive). </li><li> Agganciare circolare VEP elettrodo attivo intorno una vite acciaio inossidabile (diametro 0,7 mm lunghezza 3 mm) per l&#39;elettrodo appoggia contro la testa della vite. </li><li> Hook i 3 elettrodi inattivi (2 ERG, 1 PEV) intorno una seconda vite in acciaio inox (diametro 0,7 mm lunghezza 3 mm). </li><li> Tirare il manicotto di plastica in avanti sopra le estremità taglienti del filo in acciaio inox a due per ridurre l&#39;irritazione. </li><li> Sterilizzare i trasmettitori telemetrici immergendola in 2% glutaraldeide per più di 10 ore a circa 25 ° C. Risciacquare il trasmettitore con soluzione salina sterile per 3 volte. </li></ol> 2. Trasmettitore impianto <ol><li> Preparazione degli animali <ol><li> Disinfettare l&#39;area chirurgica prima sperimentazione pulendo con il 70% di etanolo. Autoclave tutte apparecchi chirurgici prima dell&#39;uso e mantenere in apparecchiature clorexidina quando non in uso durante la chirurgia. Coprire l&#39;animale con un telo chirurgico durante l&#39;intervento chirurgico per mantenere un ambiente sterile. Assicurarsi che tutti gli sperimentatori indossano maschere chirurgiche, guanti e camici sterili. </li><li> Indurre l&#39;anestesia con 1,5 - 2% isoflurano, ad una portata di 3 l / min e manutenzione suned a 1,5 - 2% a 2 L / min durante l&#39;intervento chirurgico. Confermare la sufficiente profondità di anestesia dall&#39;assenza di un riflesso pedale sul pizzicare il muscolo tra le dita. </li><li> Rasatura a 40 mm Area x 30 mm sopra l&#39;addome dall&#39;alto all&#39;inguine allo sterno. </li><li> Rasatura 30 mm Area x 20 mm sopra la fronte, posteriormente agli occhi e anteriore alle orecchie. </li><li> Disinfettare le due aree rasate. Per la zona della fronte disinfettare con 10% povidone-iodio tre volte (evitare l&#39;uso di antisettici a base alcolica per l&#39;area vicino all&#39;occhio, essere coerente con la norma di condotta stabilito dalla Association of tecnologi chirurgici). Sopra l&#39;addome disinfettare con 10% di iodio povidone e il 70% di etanolo. </li><li> Applicare 1 goccia di proximetacaina alla cornea per ulteriori anestesia topica. </li><li> Applicare 1 goccia di sodio carbossimetilcellulosa alla cornea per evitare l&#39;essiccazione degli occhi. </li></ol></li><li> impianto chirurgico <ol><li> Effettuare una incisione 10 millimetrisulla testa lungo la linea mediana verticale tra le orecchie con un bisturi chirurgico. </li><li> Eseguire un&#39;incisione 5 mm addome attraverso lo strato di pelle lungo la linea mediana sotto lo sterno. </li><li> Tunnel a 5 mm di diametro cannula sottocutanea dall&#39;incisione addome per l&#39;incisione testa. </li><li> Far passare i fili degli elettrodi (3 attivi e 3 inattivo) del trasmettitore attraverso la cannula dall&#39;addome alla testa. </li><li> Lasciare l&#39;elettrodo di riferimento con la base del trasmettitore e coprire la punta dell&#39;elettrodo con garza asettica. </li><li> Coprire le punte degli elettrodi (3 attivi e inattivi 3) con garza asettica. </li><li> Fissare la testa del topo ad una piattaforma stereotassico. </li><li> Estendere l&#39;incisione fronte a 30 mm di lunghezza con le forbici chirurgiche. </li><li> Esporre area chirurgica ritraendo la pelle sciolto con 2 punti di sutura (3 - 0) a ~ 3 e 09:00. </li><li> Raschiare il periostio sovrastante il cranio con garza sterile per esporre Bregma, lambda e della linea mediana suture. </li&gt; <li> Praticare due fori attraverso il cranio al VEP attiva (7 millimetri ventrale a Bregma 3 millimetri laterale a linea mediana) e inattivi (5 mm rostrale al Bregma sulla linea mediana) stereotassica coordinate. </li><li> Fissare VEP elettrodi attivi e inattivi con le viti pre-attached in acciaio inox (diametro 0,7 mm lunghezza 3 mm) per il cranio con un piccolo cacciavite a ~ 1 mm di profondità nei fori predefiniti. Questo ancora la vite all&#39;osso senza danneggiare il tessuto corticale sottostante. </li><li> Per impiantare gli elettrodi attivi ERG utilizzare un 8-0 sutura per rientrare temporaneamente la palpebra superiore. </li><li> Inserire un 16 a 21 G cannula sottocutanea da dietro l&#39;occhio attraverso la fornice congiuntivale superiore. </li><li> Rimuovere l&#39;ago guida. </li><li> Feed l&#39;elettrodo attivo attraverso il catetere di plastica accorciata dalla fronte verso l&#39;occhio. Quindi rimuovere il catetere di plastica. </li><li> Utilizzare una sutura temporanea (8 - 0.), che viene infilato attraverso il loop elettrodo, per evitare che i electrode dalla scomparsa di nuovo nel tunnel. </li><li> Eseguire un&#39;incisione 0,5 millimetri sulla congiuntiva superiore a ore 12, 1 mm dietro limbus. Utilizzare smussa per esporre la sclera sottostante. </li><li> Impiantare un 8-0 o 9-0 sutura immediatamente dietro il limbus a metà spessore sclerale. </li><li> Rimuovere sutura temporanea da ERG elettrodo attivo. </li><li> Ancorare l&#39;elettrodo attivo ERG alla metà sclerale sutura spessore legando 3 nodi consecutivi che assicurano la punta dell&#39;elettrodo è situato vicino al limbus. </li><li> Chiudere lo sportello congiuntivale con 1 o 2 punti staccati (8 - 0 a 9 - 0). Assicurarsi che la congiuntiva copre completamente l&#39;elettrodo ERG per migliorare il comfort. </li><li> Rimuovere la palpebra scomparsa di sutura. </li><li> Ripetere la procedura per l&#39;occhio controlaterale. </li><li> Applicare il gel cianoacrilato sul cranio per fissare tutte le viti inox e fili degli elettrodi. Assicurarsi che gli elettrodi attivi ERG non sono tirati troppo stretto prima di fissare a eni movimenti degli occhi capaci. </li><li> Chiudere la ferita alla testa con un non-assorbibile 3-0 di sutura. </li><li> Ruotare roditore per esporre zona addominale. Allungare l&#39;incisione cutanea addominale a 40 mm lungo la linea alba con le forbici chirurgiche. </li><li> Effettuare una incisione 35 millimetri attraverso la parete muscolare interna per esporre la cavità addominale interna. </li><li> Utilizzando due punti di sutura (3 - 0) fissare il corpo del trasmettitore a parete addominale interna lato destro dell&#39;animale. Evitare il contatto con il fegato. </li><li> Loop l&#39;elettrodo di massa e sicuro in questa forma con una sutura (3-0). Posizionarlo libero di fluttuare nella cavità addominale. </li><li> Chiudere il peritoneo con una sutura continua (3-0). </li><li> Chiudere l&#39;incisione pelle con punti staccati (3 - 0). </li></ol></li><li> Cure post-operatorie <ol><li> Monitorare l&#39;animale fino a che non ha ripreso conoscenza sufficiente a mantenere decubito sternale. Casa l&#39;animale singolarmente dopo l&#39;intervento. </li><li> somministrare carprofen sottocutanea per l&#39;analgesia (5 mg / kg) una volta al giorno per 4 giorni. </li><li> Aggiungere antibiotici per via orale profilattici (Enrofloxin, 5 mg / kg) per l&#39;acqua potabile per 7 giorni dopo l&#39;intervento chirurgico. </li><li> Applicare una pomata antinfiammatoria per siti di incisione della pelle per ridurre l&#39;irritazione per i primi 7 giorni dopo l&#39;intervento chirurgico. </li></ol></li></ol> 3. Comportamento ERG e PEV Recordings nel ratto conscio <ol><li> Scuro adattarsi animale per 12 ore prima di registrazioni ERG e PEV </li><li> Condurre tutte le manipolazioni sperimentali sotto illuminazione fioca rosso (17,4 cd.m -2, λ max = 600 nm) </li><li> Applicare anestesia topica (0,5% proximetacaina) e dilatando (0,5% tropicamide) scende alla cornea. </li><li> Guida il roditore consapevole in una misura, restrainer chiaro. Nota: La lunghezza di questo tubo di plastica può essere regolata per accogliere ratti di dimensioni diverse con il diametro complessivo fissato a 60 mm. L&#39;estremità anteriore del dispositivo è rastremato per minimizzare mo testavimento e contiene perforazioni per consentire la respirazione normale. Questo fronte affusolata consente l&#39;allineamento e la stabilizzazione della testa e gli occhi del ratto all&#39;apertura della sfera Ganzfeld. Si noti che il roditore è stato acclimatato al dispositivo di immobilizzazione (da 3 a 5 volte) prima di un intervento chirurgico. </li><li> Posizionare il roditore anteriore della ciotola Ganzfeld con gli occhi allineati con l&#39;apertura del contenitore. </li><li> Accendere il trasmettitore a permanenza passando un magnete all&#39;interno di ~ 5 cm del trasmettitore. Verificare che il trasmettitore è acceso controllando la luce LED di stato sulla base del ricevitore. </li><li> Raccogliere segnali su un intervallo di energie luminose (cioè, -5.6 a 1,52 log cd.sm -2) come descritto in precedenza 21. In breve, più segnali medi a livello commutazione luce (~ 80 ripetizioni), inferiore a energie più luminosi luminosi (~ 1 ripetizione). A poco a poco allungare l&#39;intervallo interstimulus da 1 a 180 secondi dal più debole al livello di luce più brillante. </li><li> Per isolare l&#39;asta ERGe le risposte cono utilizzano un paradigma twin-Flash 8. Ad esempio, avviare due flash a 1,52 log cd.sm -2 con un intervallo di 500 msec inter-stimolo in-between. </li><li> Per registrare i segnali VEP, media di 20 ripetizioni alle energie più luminoso luminosi (cioè 1,52 log cd.sm -2, 5 sec intervallo inter-stimolo). </li><li> Per valutare la stabilità dell&#39;impianto, che viene valutato dalla variabilità del segnale nel tempo, condurre ERG e PEV registrazioni 7, 10, 14, 21 e 28 giorni post-intervento chirurgico. </li><li> Dopo periodo sperimentale, eutanasia topi mediante iniezione intracardial di pentobarbiturate (1,5 ml / kg) dopo ketamina: anestesia xilazina (12: 1 mg / kg). </li></ol>

<ol>
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L., Mauromoustakos, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Xylazine-induced+pulmonary+edema+in+rats.">Xylazine-induced pulmonary edema in rats.</a> Toxicol Appl Pharmacol. 108, 417-427 (1991).</li><li>Charng, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinal+electrophysiology+is+a+viable+preclinical+biomarker+for+drug+penetrance+into+the+central+nervous+system.">Retinal electrophysiology is a viable preclinical biomarker for drug penetrance into the central nervous system.</a> J Ophthalmol. , (2016).</li></ol>

RG and RF are employees to the commercial funder of this research (Pfizer Neusentis and Pfizer Global Research). MI was an employee of Pfizer Global Research during this research and is currently an employee of Proteostasis Therapeutics (Cambridge, USA).

A causa della natura mini-invasiva di elettrofisiologia visiva, ERG e PEV registrazioni in pazienti umani sono condotte in condizioni coscienti e richiedono solo l&#39;uso di anestetici topici per il posizionamento degli elettrodi. Al contrario, elettrofisiologia visiva nei modelli animali è convenzionalmente condotto in anestesia generale per consentire il posizionamento degli elettrodi stabile, eliminando gli occhi del corpo e movimenti volontari. Tuttavia, anestetici generali comunemente utilizzati alterano le rispo...

L&#39;ERG e VEP sono minimamente invasiva in strumenti vivo per valutare l&#39;integrità delle vie della retina e visivi, rispettivamente, sia in laboratorio e clinica. Il tutto campo ERG produce una forma d&#39;onda caratteristica che può essere suddiviso in diverse componenti, con ogni elemento che rappresentano diverse classi di cellule del 1,2 percorso della retina. Il classico pieno campo ERG forma d&#39;onda consiste di una pendenza iniziale negativo (a onda), che ha dimostrato di rappresentare fotorecettori attività post esposizione alla luce 2-4. L&#39;a-onda è seguita da una sostanziale forma d&#39;onda positiva (b-wave), che riflette l&#39;attività elettrica di retina centrale, prevalentemente le cellule ON-bipolari 5-7. Inoltre, si può variare l&#39;energia luminosa e inter-stimolo-intervallo per isolare cono dalle risposte asta 8. Il flash VEP rappresenta potenziali elettrici del tronco della corteccia cerebrale e visiva in risposta alla stimolazione della retina luce9,10. Questa forma d&#39;onda può essere suddiviso in componenti precoce e tardiva, con la componente precoce che riflette l&#39;attività dei neuroni della via retino-genicolo-striata 11-13 e alla fine del componente che rappresenta l&#39;elaborazione corticale esibito in varie lamine V1 nei ratti 11,13. Pertanto la misurazione simultanea del ERG e PEV ritorna valutazione globale delle strutture coinvolte nel percorso visivo. Attualmente, al fine di registrare elettrofisiologia negli animali, anestesia viene impiegato per consentire il posizionamento stabile di elettrodi. Ci sono stati tentativi di misurare ERG e PEV nei ratti coscienti 14-16 ma questi studi impiegata una configurazione cablata, che può essere ingombrante e può portare a stress degli animali, limitando il movimento degli animali e il comportamento naturale 17. Con i recenti progressi nella tecnologia wireless, tra cui una maggiore miniaturizzazione e la durata della batteria, è ora possibile implementare un approccio telemetria per ERG und registrazione VEP, diminuendo lo stress associato con le registrazioni cablate e migliorare la redditività a lungo termine. Implantologia stabili completamente interiorizzato di sonde di telemetria hanno dimostrato di essere efficace per il monitoraggio cronico della temperatura, la pressione del sangue 18, l&#39;attività 19 così come elettroencefalografia 20. Tali progressi nella tecnologia sarà anche aiutare con ripetibilità e stabilità delle registrazioni coscienti, aumentando l&#39;utilità della piattaforma per gli studi cronici.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Bioamplifier</td>
			<td>ADInstruments</td>
			<td>ML 135</td>
			<td>Amplifies ERG and VEP signals</td>
		</tr>
		<tr>
			<td>Carboxymethylcellulose sodium 1.0%</td>
			<td>Allergan</td>
			<td>CAS 0009000-11-7</td>
			<td>Maintain corneal hydration during surgery</td>
		</tr>
		<tr>
			<td>Carprofen 0.5%</td>
			<td>Pfizer Animal Health Group</td>
			<td>CAS 53716-49-7</td>
			<td>Post-surgery analgesia, given with injectable saline for fluid replenishment</td>
		</tr>
		<tr>
			<td>Chlorhexadine 0.5%</td>
			<td>Orion Laboratories</td>
			<td>27411, 80085</td>
			<td>Disinfection of surgical instrument</td>
		</tr>
		<tr>
			<td>Cyanoacrylate gel activator</td>
			<td>RS components</td>
			<td>473-439</td>
			<td>Quickly dries cyanoacrylate gel</td>
		</tr>
		<tr>
			<td>Cyanocrylate gel&#160;</td>
			<td>RS components</td>
			<td>473-423</td>
			<td>Fix stainless screws to skull</td>
		</tr>
		<tr>
			<td>Dental burr</td>
			<td>Storz Instruments, Bausch and Lomb</td>
			<td>E0824A</td>
			<td>Miniature drill head of ~0.7mm diameter for making a small hole in the skull over each hemisphere to implant VEP screws</td>
		</tr>
		<tr>
			<td>Drill</td>
			<td>Bosch</td>
			<td>Dremel 300 series</td>
			<td>Automatic drill for trepanning</td>
		</tr>
		<tr>
			<td>Enrofloxin</td>
			<td>Troy Laboratories</td>
			<td></td>
			<td>Prophylactic antibiotic post surgey</td>
		</tr>
		<tr>
			<td>Ganzfeld integrating sphere</td>
			<td>Photometric Solutions International</td>
			<td></td>
			<td>Custom designed light stimulator: 36 mm diameter, 13 cm aperture size</td>
		</tr>
		<tr>
			<td>Gauze swabs</td>
			<td>Multigate Medical Products Pty Ltd</td>
			<td>57-100B</td>
			<td>Dries surgical incision and exposed skull surface during surgery</td>
		</tr>
		<tr>
			<td>Isoflurane 99.9%</td>
			<td>Abbott Australasia Pty Ltd</td>
			<td>CAS 26675-46-7</td>
			<td>Proprietory Name: Isoflo(TM) Inhalation anaaesthetic. Pharmaceutical-grade inhalation anesthetic mixed with oxygen gas for VEP electrode implant surgery</td>
		</tr>
		<tr>
			<td>Kenacomb ointment</td>
			<td>Aspen Pharma Pty Ltd</td>
			<td></td>
			<td>To reduce skin irritation and itching after surgery</td>
		</tr>
		<tr>
			<td>Luxeon LEDs</td>
			<td>Phillips Lighting Co.</td>
			<td></td>
			<td>For light stimulation, twenty 5 watt and one 1 watt LEDs, controlled by Scope software</td>
		</tr>
		<tr>
			<td>Needle (macrosurgery)</td>
			<td>World Precision Instruments</td>
			<td>501959</td>
			<td>for suturing abdominal and head surgery, used with 3-0 suture, eye needle, cutting edge 5/16 circle Size 1, 15mm</td>
		</tr>
		<tr>
			<td>Needle holder (macrosurgery)</td>
			<td>World Precision Instruments</td>
			<td>500224</td>
			<td>To hold needle during abdominal and head surgery</td>
		</tr>
		<tr>
			<td>Needle holder (microsurgery)</td>
			<td>World Precision Instruments</td>
			<td>555419NT</td>
			<td>To hold needle during ocular surgery</td>
		</tr>
		<tr>
			<td>Optiva catheter</td>
			<td>Smiths Medical International LTD</td>
			<td>16 or 21 G</td>
			<td>Guide corneal active electrodes from skull to conjunctiva</td>
		</tr>
		<tr>
			<td>Povidone iodine 10%</td>
			<td>Sanofi-Aventis</td>
			<td>CAS 25655-41-8</td>
			<td>Proprietory name: Betadine, Antiseptic to prepare the shaved skin for surgery 10%, 500 mL</td>
		</tr>
		<tr>
			<td>Powerlab data acquisition system</td>
			<td>ADInstruments</td>
			<td>ML 785</td>
			<td>Acquire signal from telemetry transmitter, paired to telemetry data converter</td>
		</tr>
		<tr>
			<td>Proxymetacaine 0.5%</td>
			<td>Alcon Laboratories&#160;</td>
			<td>CAS 5875-06-9</td>
			<td>Topical ocular analgesia</td>
		</tr>
		<tr>
			<td>Restrainer</td>
			<td>cutom made</td>
			<td></td>
			<td>Front of the restrainer is tapered to minimize head movement, length can be adjusted to accommodate different rat length, overall diameter is 60 mm.&#160;</td>
		</tr>
		<tr>
			<td>Scapel blade</td>
			<td>R.G. Medical Supplies</td>
			<td>SNSM0206</td>
			<td>For surgical incision</td>
		</tr>
		<tr>
			<td>Scissors (macrosurgery)</td>
			<td>World Precision Instruments</td>
			<td>501225</td>
			<td>for cutting tissue on the abodmen and forhead</td>
		</tr>
		<tr>
			<td>Scissors (microsurgery)</td>
			<td>World Precision Instruments</td>
			<td>501232</td>
			<td>To dissect the conjunctiva for electrode attachment</td>
		</tr>
		<tr>
			<td>Scope Software</td>
			<td>ADInstruments</td>
			<td>version 3.7.6</td>
			<td>Simultaneously triggers the stimulus via the ADI Powerlab system and collects data</td>
		</tr>
		<tr>
			<td>Shaver</td>
			<td>Oster</td>
			<td>Golden A5</td>
			<td>Shave fur from surgical areas</td>
		</tr>
		<tr>
			<td>Stainless streel screws&#160;</td>
			<td>MicroFasteners</td>
			<td>L001.003CS304</td>
			<td>0.7 mm shaft diameter, 3 mm in length&#160;</td>
		</tr>
		<tr>
			<td>Stereotaxic frame</td>
			<td>David Kopf</td>
			<td>Model 900</td>
			<td>A small animal stereotaxic instrument for locating the implantation landmarks on the skull</td>
		</tr>
		<tr>
			<td>Surgical drape</td>
			<td>Vital Medical Supplies</td>
			<td>GM29-612EE</td>
			<td>Ensure sterile enviornment during surgery</td>
		</tr>
		<tr>
			<td>Suture (macrosurgery)</td>
			<td>Ninbo medical needles</td>
			<td>3-0</td>
			<td>for suturing abdominal and head surgery, sterile silk braided, 60cm</td>
		</tr>
		<tr>
			<td>Suture needle (microsurgery)</td>
			<td>Ninbo medical needles</td>
			<td>8-0 or 9-0</td>
			<td>for ocular surgery including, suturing electrode to sclera and closing conjunctival wound, nylon suture, 3/8 circle 1&#215;5, 30cm</td>
		</tr>
		<tr>
			<td>Telemetry data converter&#160;</td>
			<td>DataSciences International</td>
			<td>R08</td>
			<td>allows telemetry signal to interface with data collection software</td>
		</tr>
		<tr>
			<td>Telemetry Data Exchange Matrix</td>
			<td>DataSciences International</td>
			<td></td>
			<td>Gathers data from transmitters, pair with receiver</td>
		</tr>
		<tr>
			<td>Telemetry data receiver</td>
			<td>DataSciences International</td>
			<td>RPC-1</td>
			<td>Receives telemetry data from transmitter</td>
		</tr>
		<tr>
			<td>Telemetry transmitter</td>
			<td>DataSciences International</td>
			<td>F50-EEE</td>
			<td>3 channel telemetry transmitter</td>
		</tr>
		<tr>
			<td>Tropicamide 0.5%</td>
			<td>Alcon Laboratories&#160;</td>
			<td></td>
			<td>Iris dilation</td>
		</tr>
		<tr>
			<td>Tweezers (macrosurgery)</td>
			<td>World Precision Instruments</td>
			<td>500092</td>
			<td>Manipulate tissues during abdominal and head surgery</td>
		</tr>
		<tr>
			<td>Tweezers (microsurgery)</td>
			<td>World Precision Instruments</td>
			<td>500342</td>
			<td>Manipulate tissues during ocular surgery</td>
		</tr>
	</tbody>
</table>

implantation and recording of wireless electroretinogram and visual evoked potential in conscious rats

L&#39;elettroretinogramma pieno campo (ERG) e potenziali evocati visivi (PEV) sono strumenti utili per valutare retina e l&#39;integrità via visiva sia in ambito clinico e di laboratorio. Attualmente, le misure ERG e PEV preclinici sono eseguiti con l&#39;anestesia per garantire posizionamento degli elettrodi stabili. Tuttavia, la presenza di anestesia ha dimostrato di contaminare le normali risposte fisiologiche. Per superare questi confonde anestesia, sviluppiamo una nuova piattaforma per saggiare ERG e PEV nei ratti coscienti. Gli elettrodi sono impiantati chirurgicamente sotto-conjunctivally sull&#39;occhio per saggiare la ERG e epidurale sopra la corteccia visiva per misurare il VEP. Una gamma di ampiezza e di sensibilità / parametri di temporizzazione sono analizzati sia per la ERG e PEV ad aumentare le energie luminose. I segnali ERG e PEV sono dimostrato di essere stabile e ripetibile per almeno 4 settimane impiantazione post-chirurgico. Questa capacità di registrare i segnali ERG e PEV senza anestesia confonde nelle s precliniciEtting dovrebbe fornire la traduzione superiore ai dati clinici.

La risposta fotorecettore viene analizzato montando una gaussiana ritardata al bordo anteriore del tratto discendente iniziale della risposta ERG gli ultimi 2 energie luminose (1,20, 1,52 log CSM -2) per ciascun animale, in base al modello di agnello e Pugh 22, formulato da Hood e Birch 23. Questa formula restituisce una ampiezza ed un parametro di sensibilità, (Figura 1C e 1D, rispettivamente). Una funzione iperbolica f...

Watch this Scientific Journal Video about Implantation and Recording of Wireless Electroretinogram and Visual Evoked Potential in Conscious Rats at JoVE.com

Implantation and Recording of Wireless Electroretinogram and Visual Evoked Potential in Conscious Rats

Mostriamo le procedure di impianto e di registrazione chirurgiche per misurare i segnali elettrofisiologici visivi dall&#39;occhio (elettroretinogramma) e il cervello (visivo potenziale evocato) nei ratti coscienti, che è più simile alla condizione umana in cui le registrazioni sono effettuate senza anestesia confonde.

L&#39;impianto e la registrazione di Wireless Elettroretinogramma e potenziale visivo evocato ratto conscio

The full-field electroretinogram (ERG) and visual evoked potential (VEP) are useful tools to assess retinal and visual pathway integrity in both ...

implantation-recording-wireless-electroretinogram-visual-evoked

Research

JoVE Journal

Behavior

8.6K Views.  University of Melbourne.  Mostriamo le procedure di impianto e di registrazione chirurgiche per misurare i segnali elettrofisiologici visivi dall'occhio (elettroretinogramma) e il cervello (visivo potenziale evocato) nei ratti coscienti, che è più simile alla condizione umana in cui le registrazioni sono effettuate senza anestesia confonde. 

Video: Implantation and Recording of Wireless Electroretinogram and Visual Evoked Potential in Conscious Rats

The Successive Alleys Test of Anxiety in Mice and Rats

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones. Handley, Mithani and File et al. performed the first studies on the plus-maze design we use today, and in 1987 Lister published a design for use with mice. 
Time spent on, and entries into, the open arms are an index of anxiety; the lower these indices, the more anxious the mouse is. Alternatively, a mouse that spends most of its time in the closed arms is classed as anxious. 
One of the problems of the plus-maze is that, while time spent on, and entries into, the open arms is a fairly unambiguous measure of anxiety, time in the central area is more difficult to interpret, although time spent here has been classified as &#x201C;decision making&#x201D;. In many tests central area time is a considerable part of the total test time.
Shepherd et al. produced an ingenious design to eliminate the central area, which they called the &#x201C;zero maze&#x201D;. However, although used by several groups, it has never been as widely adopted as the plus-maze. 
In the present article I describe a modification of the plus-maze design that not only eliminates the central area but also incorporates elements from other anxiety tests, such as the light-dark box and emergence tests. It is a linear series of four alleys, each having increasing anxiogenic properties. It has given similar results to the plus-maze in general. Although it may not be more sensitive than the plus-maze (more data is needed before a firm conclusion can be reached on this point), it provides a useful confirmation of plus-maze results which would be useful when, for example, only a single example of a mutant mouse was available, as, for example, in ENU-based mutagenesis programs.

The plus-maze measures anxiety-like behaviour in rodents. There are two opposite closed and two opposite open arms; anxious rodents avoid the open arms. The central area is neither completely open nor closed, so time spent here is ambiguous and difficult to interpret. Here a modification of the plus-maze protocol eliminating this area is described.

Le successive Vicoli test di ansia nei topi e nei ratti

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones.  ...

Ricerca

Comportamento

Automated Visual Cognitive Tasks for Recording Neural Activity Using a Floor Projection Maze

Neuropsychological tasks used in primates to investigate mechanisms of learning and memory are typically visually guided cognitive tasks. We have developed visual cognitive tasks for rats using the Floor Projection Maze1,2&#160;that are optimized for visual abilities of rats permitting stronger comparisons of experimental findings with other species.
In order to investigate neural correlates of learning and memory, we have integrated electrophysiological recordings into fully automated cognitive tasks on the Floor Projection Maze1,2. Behavioral software interfaced with an animal tracking system allows monitoring of the animal's behavior with precise control of image presentation and reward contingencies for better trained animals. Integration with an in vivo electrophysiological recording system enables examination of behavioral correlates of neural activity at selected epochs of a given cognitive task.
We describe protocols for a model system that combines automated visual presentation of information to rodents and intracranial reward with electrophysiological approaches. Our model system offers a sophisticated set of tools as a framework for other cognitive tasks to better isolate and identify specific mechanisms contributing to particular cognitive processes.

We describe protocols for training rats for chronic electrophysiological recordings in fully automated cognitive tasks on a Floor Projection Maze.

Automated compiti cognitivi visivi per la registrazione Neural all&#39;attività utilizzando una proiezione Piano Maze

Neuropsychological tasks used in primates to investigate mechanisms of learning and memory are typically visually guided cognitive tasks. We have ...

Methods to Explore the Influence of Top-down Visual Processes on Motor Behavior

Kinesthetic awareness is important to successfully navigate the environment. When we interact with our daily surroundings, some aspects of movement are deliberately planned, while others spontaneously occur below conscious awareness. The deliberate component of this dichotomy has been studied extensively in several contexts, while the spontaneous component remains largely under-explored. Moreover, how perceptual processes modulate these movement classes is still unclear. In particular, a currently debated issue is whether the visuomotor system is governed by the spatial percept produced by a visual illusion or whether it is not affected by the illusion and is governed instead by the veridical percept. Bistable percepts such as 3D depth inversion illusions (DIIs) provide an excellent context to study such interactions and balance, particularly when used in combination with reach-to-grasp movements. In this study, a methodology is developed that uses a DII to clarify the role of top-down processes on motor action, particularly exploring how reaches toward a target on a DII are affected in both deliberate and spontaneous movement domains.

It is unclear how top-down signals from the ventral visual stream affect movement. We developed a paradigm to test motor behavior towards a target on a 3D depth inversion illusion. Significant differences are reported in both deliberate, goal-directed movements and automatic actions under illusory and veridical viewing conditions.

Metodi per Esplora l&#39;influenza di Top-down processi visivi sul comportamento motorio

Kinesthetic awareness is important to successfully navigate the environment. When we interact with our daily surroundings, some aspects of movement are ...

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

The Hole Consiglio Modified - Behavior, Cognition e l&#39;interazione sociale in topi e ratti di misura

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

Conscious and Non-conscious Representations of Emotional Faces in Asperger's Syndrome

Several neuroimaging studies have suggested that the low spatial frequency content in an emotional face mainly activates the amygdala, pulvinar, and superior colliculus especially with fearful faces1-3. These regions constitute the limbic structure in non-conscious perception of emotions and modulate cortical activity either directly or indirectly2. In contrast, the conscious representation of emotions is more pronounced in the anterior cingulate, prefrontal cortex, and somatosensory cortex for directing voluntary attention to details in faces3,4. Asperger&#39;s syndrome (AS)5,6 represents an atypical mental disturbance that affects sensory, affective and communicative abilities, without interfering with normal linguistic skills and intellectual ability. Several studies have found that functional deficits in the neural circuitry important for facial emotion recognition can partly explain social communication failure in patients with AS7-9. In order to clarify the interplay between conscious and non-conscious representations of emotional faces in AS, an EEG experimental protocol is designed with two tasks involving emotionality evaluation of either photograph or line-drawing faces. A pilot study is introduced for selecting face stimuli that minimize the differences in reaction times and scores assigned to facial emotions between the pretested patients with AS and IQ/gender-matched healthy controls. Information from the pretested patients was used to develop the scoring system used for the emotionality evaluation. Research into facial emotions and visual stimuli with different spatial frequency contents has reached discrepant findings depending on the demographic characteristics of participants and task demands2. The experimental protocol is intended to clarify deficits in patients with AS in processing emotional faces when compared with healthy controls by controlling for factors unrelated to recognition of facial emotions, such as task difficulty, IQ and gender.

An EEG experimental protocol is designed to clarify the interplay between conscious and non-conscious representations of emotional faces in patients with Asperger&#39;s syndrome. The technique suggests that patients with Asperger&#39;s syndrome have deficits in non-conscious representation of emotional faces, but have comparable performance in conscious representation with healthy controls.

Rappresentazioni coscienti e non coscienti della Facce emotivi nella sindrome di Asperger

Several neuroimaging studies have suggested that the low spatial frequency content in an emotional face mainly activates the amygdala, pulvinar, and ...

Manipulation of Epileptiform Electrocorticograms (ECoGs) and Sleep in Rats and Mice by Acupuncture

Ancient Chinese literature has documented that acupuncture possesses efficient therapeutic effects on epilepsy and insomnia. There is, however, little research to reveal the possible mechanisms behind these effects. To investigate the effect of acupuncture on epilepsy and sleep, several issues need to be addressed. The first is to identify the acupoints, which correspond between humans, rats, and mice. Furthermore, the depth of insertion of the acupuncture needle, the degree of needle twist in manual needle acupuncture, and the stimulation parameters for electroacupuncture (EA) need to be determined. To evaluate the effects of acupuncture on epilepsy and sleep, a feasible model of epilepsy in rodents is required. We administer pilocarpine into the left central nucleus of the amygdala (CeA) to simulate focal temporal lobe epilepsy (TLE) in rats. Intraperitoneal (IP) injection of pilocarpine induces generalized epilepsy and status epilepticus (SE) in rats. Five IP injections of pentylenetetrazol (PTZ) with a one-day interval between each injection successfully induces spontaneous generalized epilepsy in mice. Recordings of electrocorticograms (ECoGs), electromyograms (EMGs), brain temperature, and locomotor activity are used for sleep analysis in rats, while ECoGs, EMGs, and locomotor activity are employed for sleep analysis in mice. ECoG electrodes are implanted into the frontal, parietal, and contralateral occipital cortices, and a thermistor is implanted above the cerebral cortex by stereotactic surgery. EMG electrodes are implanted into the neck muscles, and an infrared detector determines locomotor activity. The criteria for categorizing vigilance stages, including wakefulness, rapid eye movement (REM) sleep, and non-REM (NREM) sleep are based on information from ECoGs, EMGs, brain temperature, and locomotor activity. Detailed classification criteria are stated in the text.

Manipulation of Epileptiform Electrocorticograms ECoGs and Sleep in Rats and Mice by Acupuncture

This paper demonstrates the performance of acupuncture, epilepsy models, and the analysis of sleep in rodents. The acupuncture procedure and the identification of acupoints are described. Pilocarpine or pentylenetetrazol (PTZ) is used to induce epilepsy. Electrocorticogram (ECoG), electromyogram (EMG), brain temperature, and locomotor activity recordings are employed for sleep analysis.

Manipolazione di epilettiforme Electrocorticograms (ECoGs) e sonno in ratti e topi con l&#39;agopuntura

Ancient Chinese literature has documented that acupuncture possesses efficient therapeutic effects on epilepsy and insomnia. There is, however, little ...

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer scale. However, current methods mostly rely on tethered cable recordings or only use unidirectional systems, allowing either recording or stimulation of neural activity, but not at the same time or same target. Here, a new wireless, bidirectional device for simultaneous multichannel recording and stimulation of neural activity in freely behaving rats is described. The system operates through a single portable head stage that both transmits recorded activity and can be targeted in real-time for brain stimulation using a telemetry-based multichannel software. The head stage is equipped with a preamplifier and a rechargeable battery, allowing stable long-term recordings or stimulation for up to 1 h. Importantly, the head stage is compact, weighs 12 g (including battery) and thus has minimal impact on the animal&#180;s behavioral repertoire, making the method applicable to a broad set of behavioral tasks. Moreover, the method has the major advantage that the effect of brain stimulation on neural activity and behavior can be measured simultaneously, providing a tool to assess the causal relationships between specific brain activation patterns and behavior. This feature makes the method particularly valuable for the field of deep brain stimulation, allowing precise assessment, monitoring, and adjustment of stimulation parameters during long-term behavioral experiments. The applicability of the system has been validated using the inferior colliculus as a model structure.

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

A wireless, bidirectional system for multi-channel neural recordings and stimulation in freely behaving rats is introduced. The system is light and compact, thus having minimal impact on the animal&#180;s behavioral repertoire. Moreover, this bidirectional system provides a sophisticated tool to assess causal relationships between brain activation patterns and behavior.

Una connessione Wireless, interfaccia bidirezionale per registrazione In Vivo e la stimolazione dell'attività neurale a comportarsi liberamente ratti

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer ...

Using Electroencephalography Measurements and High-quality Video Recording for Analyzing Visual Perception of Media Content

This article explores a method to detect differences in visual perception in humans. The method used is based on the psychological (or "cognitive") function of eyeblinks. Participants' eyeblinks are detected and acquired while watching videos specifically created for the investigation. The detection and acquisition of eyeblinks are carried out with the help of a 20-channel electroencephalographic (EEG) wireless device. The international 10-20 system for electrode placement is followed. A high-definition (HD) video camera is used to record participants' facial expressions, for contrast purposes. Instead of using pre-existing media content, purpose-made video content has been created following specific criteria of interest for this investigation, with stimuli enabling researchers to manage the precise parameters of interest. Otherwise, results could be contaminated with uncontrolled variables. The synchronization of the presentation of video stimuli with EEG recordings needs to be done in milliseconds. Analysis of collected data is performed with robust software for working with big matrices. Statistically significant differences in eyeblink rate related to media professionalization and editing style are found with the reported experimental procedures.

We present detection, acquisition, and analysis of eyeblink rates while watching media content.

Utilizzando misure di elettroencefalografia e registrazione Video di alta qualità per l'analisi di percezione visiva del contenuto multimediale

This article explores a method to detect differences in visual perception in humans. The method used is based on the psychological (or "cognitive") ...

Eye Tracking During Visually Situated Language Comprehension: Flexibility and Limitations in Uncovering Visual Context Effects

The present work is a description and an assessment of a methodology designed to quantify different aspects of the interaction between language processing and the perception of the visual world. The recording of eye-gaze patterns has provided good evidence for the contribution of both the visual context and linguistic/world knowledge to language comprehension. Initial research assessed object-context effects to test&#160;theories of modularity in language processing. In the introduction, we describe how subsequent investigations have taken the role of the wider visual context in language processing as a research topic in its own right, asking questions such as how our visual perception of events and of speakers contributes to&#160;comprehension informed by&#160;comprehenders&#39; experience. Among the examined aspects of the visual context are actions, events, a speaker&#39;s gaze, and emotional facial expressions, as well as spatial object configurations. Following an overview of the eye-tracking method and its different applications, we list the key steps of the methodology in the protocol, illustrating how to successfully use it to study visually-situated language comprehension. A final section presents three sets of representative results and illustrates the benefits and limitations of eye tracking for investigating the interplay between the perception of the visual world and language comprehension.

The present article reviews an eye-tracking methodology for studies on language comprehension. To obtain reliable data, key steps of the protocol must be followed. Among these are the correct set-up of the eye tracker (e.g., ensuring good quality of the eye and head images) and accurate calibration.

Eye Tracking durante la comprensione del linguaggio visivo situato: Flessibilità e limitazioni a scoprire gli effetti di contesto visivo

The present work is a description and an assessment of a methodology designed to quantify different aspects of the interaction between language processing ...

Visual Classical Conditioning in Wood Ants

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, studies implementing classical conditioning paradigms with harnessed insects have been important for investigating the exact cues that individuals learn and the neural mechanisms underlying learning and memory formation. Here we present a protocol for evoking visual associative learning in wood ants through classical conditioning. In this paradigm, ants are harnessed and presented with a visual cue (a blue cardboard), the conditional stimulus (CS), paired with an appetitive sugar reward, the unconditional stimulus (US). Ants perform a Maxilla-Labium Extension Reflex (MaLER), the unconditional response (UR), which can be used as a readout for learning. Training consists of 10 trials, separated by a 5-minute intertrial interval (ITI). Ants are also tested for memory retention 10 minutes or 1 hour after training. This protocol has the potential to allow researchers to analyze, in a precise and controlled manner, the details of visual memory formation and the neural basis of learning and memory formation in wood ants.

We present a protocol for the classical conditioning of harnessed ants that permits researchers to study visual learning and memory formation at a level of analysis not possible with freely moving individuals.

Visual condizionamento classico in legna formiche

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, ...