Spor eyeblink klassisk betinging (ECC) ble brukt til å vurdere hippocampus-avhengige associative læring i voksen rotter det ble administrert en høy konsentrasjon (11,9% v/v) alkohol under tidlig neonatal hjernens utvikling. ECC prosedyrer er generelt lyden Diagnoseverktøy for å oppdage brain dysfunction over mange psykologiske og biomedisinsk innstillinger.
Neonatal rotter ble gitt en relativt høy konsentrasjon av etylalkohol (11,9% v/v) under postnatal dager 4-9, en tid da fosterets hjernen gjennomgår rask organisasjonsendringer og ligner akselerert hjernen endringer som oppstår i løpet av tredje trimester hos mennesker. Denne modellen av fetal alkohol spektrum lidelser (FASDs) produserer alvorlig hjerneskade, etterligne beløpet og prøve av overstadig-drikking som forekommer i noen alkoholholdige graviditet. Vi beskriver bruk av spor eyeblink klassisk betinging (ECC), en høyere orden variant av associative læring, å vurdere langsiktige hippocampus dysfunksjon som er vanligvis sett i alkohol-eksponerte voksne avkom. På 90 dager gammel, gnagere var kirurgisk forberedt med opptak og stimulerende elektroder, målt electromyographic (EMG) blinker aktivitet fra venstre øyelokket muskelen og levert mild støt bakenfor venstre øye, henholdsvis. Etter en 5 dagers restitusjonsperiode gjennomgikk de 6 økter med spor ECC å bestemme associative læring forskjellene mellom alkohol-eksponert og kontrollere rotter. Spor ECC er en av mange mulige ECC prosedyrer som kan enkelt endres med samme utstyr og programvare, slik at forskjellige nevrale systemer kan vurderes. ECC prosedyrer, kan brukes som Diagnoseverktøy for å oppdage nevrale patologi i forskjellige hjernen systemer og ulike forhold som fornærmelse hjernen.
It is quite hard to imagine that in today's day and age with better health care and access to health services, alcohol abuse remains a major global health concern. Unfortunately, it has been shown that an expectant mother who drinks a high amount of alcohol can have a child with severe brain damage and neurodevelopmental disorders that last a lifetime, as evident in those afflicted with fetal alcohol syndrome (FAS)1,2,3. In women with some confirmed history of maternal alcohol use, the developing fetus is also susceptible to small amounts of alcohol or different patterns of alcohol consumption that produce varying differences in blood alcohol concentrations. In this latter case, while the children may not exhibit the severe morphological or neurobehavioral disruptions as those with FAS, they may still exhibit lifelong cognitive disabilities and emotional disturbances that range from mild to severe3,4. Altogether, FAS and less severe forms of prenatal alcohol-mediated disruptions constitute a collection of fetal alcohol spectrum disorders (FASDs). It is no surprise that FASDs are completely preventable, but astonishingly estimates show that in populations where alcohol abuse is quite common, they remain the primary non-genetic cause of neural and cognitive disability, affecting about 2% to 5% of young US children and those in European countries such as France and Sweden. With respect to the incidence of FAS alone within the US, the prevalence is 2 to 7 per 1,000 live births5, implying that the overall incidence of FASDs to be much higher than that for FAS.
Neuroimaging studies conducted in children with FASDs have shown that they display brain abnormalities, such as a thinner corpus callosum6, smaller anterior cerebellar vermis7, and smaller hippocampus8. These brain abnormalities underlie some of the long-term neurocognitive disruptions observed in children with FASDs. The exact links that tie variations in maternal alcohol-mediated brain changes and variations in the profile (i.e., type, extent) of particular neurocognitive impairments have yet to be clearly determined. But as a starting point, the hippocampus is an excellent candidate for determining its susceptibility to prenatal alcohol effects. Indeed, children with FASDs exhibit deficits in hippocampal-mediated behaviors such as place learning9,10 and delayed object recall11.
Rodent models of FASDs have proven to be invaluable in elucidating the mechanisms leading to neurocognitive disruptions seen in children with FASDs. A well-established binge-exposure model that we have adopted involves delivering alcohol to rats during postnatal days 4-912,13, a period when the brain undergoes rapid synapse and dendritic contact formation, comparable to human fetal week 24 and extending into the 3rd trimester14,15,16,17. This particular model induces significant loss of hippocampal neurons18,19 and neurons in many other brain regions such as the cerebellum12,13,14,15,16,17,18,19,20,21,22,23, accompanied by severe impairments in cognitive functions spanning different domains21,24,25. Cognitive disruption from early alcohol exposure in rats may be assessed in different ways, particularly with eyeblink classical conditioning (ECC). ECC is a paradigm that has been utilized for more than a century to scientifically investigate the fundamental basis of learning26,27 and as such, provides a useful method to better understanding the adverse neurocognitive consequences resulting from fetal alcohol exposure. It is a very flexible paradigm that allows investigators to use a variety of different ECC procedures, any of which can be examined across many mammalian species ascending the phylogenetic scale (from mice to humans) and over different courses of brain development28,29,30,31. Furthermore, the fundamental neural circuits that mediate associative learning in this paradigm are supported by experimental and neuropsychological reports in these same species26,32,33,34,35,36,37.
One form of ECC, trace ECC is demonstrated in this paper (Figure 1). To provide context, it is compared against the more traditional form – delay ECC. The ECC paradigm was modeled after classical conditioning using dogs, first carried out by the Nobel-Prize winning physiologist, Ivan Pavlov. Pavlov discovered that certain stimuli such as tones do not naturally elicit salivation, but when it precedes and overlaps with the delivery of food, the salivary response can be strengthened from repeated presentations of the two, provided that this tone-food contingency is maintained. This is an example of delay ECC, with the notion that associative strength is mediated by immediate temporal contiguity between the two stimuli, thus making learning conditions optimal for an animal. He also tested other variations of the tone-food contingency, such as turning the tone off and leaving a "trace" period before delivering the food. When these two stimuli were discontiguous enough, it became much harder for the dogs to emit salivation responses prior to the delivery of the food. The discontiguity between the tone being turned off and the delivery of the food is thus an example of trace ECC. As rodents do not naturally salivate to the presence of food, more species-relevant stimuli such as mild shock are used instead; they also do not naturally emit defensive eyeblink responses to tones. With this backdrop, rodent ECC procedures involve presenting a tone at a given decibel level and pairing it in some fashion with mild shock to either the eyelid muscle (orbicularis oculi) or the temporalis muscle to elicit an eyeblink response. The tone is considered a conditioned stimulus (CS) while the shock is considered an unconditioned stimulus (US). In delay ECC, the CS is presented first; this stimulus remains on for a given duration. Afterwards, the US is delivered. These two stimuli overlap for a given duration, and then both terminate simultaneously; the resultant eyeblink response emitted due to the US is considered an unconditioned response (UR). In this procedure, rodents learn to emit eyeblink responses sometime after the CS is presented, but just before the US, in order to anticipate this aversive stimulus. The learned eyeblink response is referred to as a conditioned response (CR). For trace ECC, the CS and US are separated by a period of time that is void of stimuli known as a trace interval; they do not overlap in time as in delay ECC. During this interval, the animal is tasked to resolve the associational requirements between stimuli. Similar to delay ECC, learning occurs when the animal consistently emits a blink response after the CS turns off, but immediately before delivery of the US. Over some amount of acquisition training (CS paired with US), learning curves (i.e., based on different CR measurements) develop. Lesion and neuroimaging studies show that successful learning in delay ECC is dependent on having intact cerebellar-brain stem neuro-circuitry38,39,40, whereas trace ECC is a higher-order procedure that requires additional neural engagement from the hippocampus41,42,43,44 and other cortical structures45,46. Because of the timing-related requirements needed in order to acquire trace CRs successfully, this task is also more difficult to learn (even for normal subjects).
Figure 1: Trace eyeblink classical conditioning. An actual waveform is shown that is representative of an adult rat in the unintubated-control (UC) group. The tone CS (85 dB, 2.8 kHz) is first presented for 380 ms. A trace interval of 500 ms ensues, where no stimuli are present. Afterwards a shock US (1.6 mA) is delivered for 100 ms. Successful learning in this task occurs when the frequency (%) or amplitude (in volts) of eyeblinks during the conditioned response (CR) time window (Total CR period) increases over many sessions of training. In particular, rodents with an intact hippocampus will usually emit more well-timed CRs (Adaptive CRs) just prior to the onset of the shock US (within a 200-ms window). Startle responses (SRs) during the first 80 ms after tone CS onset and unconditioned responses (URs) are also measured. Non-associative SRs are typically low or nonexistent in well-trained rodents, while URs are expected to be high in frequency and amplitude. This task requires that the rodent learn to bridge the association between the CS offset and US onset (during the trace interval), therefore making it inherently more difficult to acquire compared to delay ECC. Please click here to view a larger version of this figure.
Here we demonstrate the adverse functional consequences of neonatal alcohol exposure that is delivered in a binge-like manner, as assessed by a trace ECC procedure that delivers an 85 dB tone CS (2.8 kHz) which remains on for 380 ms, followed by a 1.6 mA shock US which remains on for 100 ms, and these stimuli are separated by a trace period of 500 ms. We have reported on the utility of this behavioral assay in previous studies examining choline intervention and iron supplementation in mitigating the effects of neonatal alcohol exposure18,47. Indeed, trace ECC can be used as a diagnostic tool to assess neonatal alcohol-induced hippocampal pathology. The advantage it has over delay ECC is that it is more sensitive to detecting disturbances in hippocampal function, which is compromised in humans with FASDs.
Demonstration of ECC extends far outside the fetal alcohol field. Many variants of ECC (e.g., delay, trace, compound, reversal) can be used to elucidate ontogenetic differences in learning across development, the neurobiological basis of associative learning in normal mammals, as well as the vulnerabilities of different brain systems to many challenges, including (but not limited to) teratogens, environmental toxins, traumatic brain injury, neurodegenerative diseases, and psychiatric conditions.
Neonatal rotte unger mottatt etylalkohol postnatal dagene 4-9 utstilt spor eyeblink condition impairments i voksen alder. Disse resultatene støtter ideen om at alkohol er en teratogen med varig skadevirkninger hippocampus funksjon. Samlet, betinget svarer i spor prosedyren var lavere for rotter utsatt for alkohol sammenlignet med rotter i begge kontroll grupper. Associative læring nedskrivningen i alkohol-eksponerte rotter var ikke påvirket av motiverende eller motor forskjeller (dvs., ingen forskjeller i blinker til sjokk U.S. intensitet).
Mens spor ECC er et nyttig diagnostisk verktøy for Klargjørende utfordring-indusert hippocampus neuropathology, må resultatene fra denne metoden plasseres i riktig sammenheng. Først involvert de prosessuelle nøkkelelementene i denne demonstrasjonen målrettet levering av alkohol under et kjent vindu på å utvikle hjernen, fabrikasjon av elektroden maskinvare som tillater registrering av electromyographic aktivitet og leverer støt, kirurgisk implantasjon av nevnte maskinvare og påfølgende dyreforsøk bruker en læring paradigme som vurderer en kognitiv funksjon av interesse. På hvert trinn i prosessen, må være forsiktig å ikke forårsaker unødvendig/utilsiktet skade gnager fagene og å overvåke deres helse tegn regelmessig. Atferdsmessige resultatene gir “vinduet” erkjennelse, en psykologisk konstruere som er bare nøyaktig beskrevet når deres helse er ikke svekket av eksperimentelle feil omfatter alkohol dosering, maskinvare eller kirurgisk implantasjon. Dermed skal hvert fremgangsmåter for element i forskningsprosessen gjennomføres på en forsvarlig måte for å sikre at resultatene fra ECC kan ekstrapolert funn hos mennesker. Dernest ECC paradigmet gir innsikt om natur associative lære, men hensyn må tas ikke å forlenge funn ved hjelp av denne tilnærmingen og bredt tilskrive dem til andre kognitive domener – som arbeidshukommelsen, kort/lang sikt minner og bevissthet – med mindre en har innlemmet noen fasett av disse domenene i en ECC studie av eksperimentell design. For eksempel denne demonstrasjonen undersøkt oppkjøpet fasen av spor ECC læring, men undersøke ikke minne bevaring i rotter når de gjennomført opplæring. Minne er dermed en uavhengig psykologisk prosess som bør vurderes i tillegg til læring. Ved design, kan en innlemme et minne bevaring intervall for å vurdere enten kortsiktige eller langsiktige minne evne. For det tredje, anerkjennelse at det er parallell minne systemer54 som kan arbeide samtidig med motiverende, eksperimentelle og hormonelle faktorer som atferd, er avgjørende for forståelse som associativity (under ECC), men en av mange prosesser som avslører hva er “god” eller “dårlig” om læring. Til slutt, spor ECC er ikke rent hippocampus-avhengige aktiviteten som andre områder av hjernen mekle noen del av CR. Dermed må en forståelse av samspillet mellom forskjellige nevrale kretser og/eller typen stimulans parametere som benyttes i en studie, tas i betraktning når implikasjoner basert på diskret resultater. Lillehjernen, for eksempel bidrar også til spor ECC, der det påvirker topografiske kjennetegner CR og CR timing, spesielt når ISI er kortvarig. Spor ECC påvirkes ikke hos mennesker med lillehjernen skade som er testet med en lang spor intervall (1000 ms), men påvirkes i de som mottar en kortere spor intervall (400 ms)34. Bilaterale lesjoner av dorsal mediale prefrontal cortex (mPFC) som mål fremre cingulate og mediale agranular regionene i mus, hindre videre oppkjøp av spor CRs55, mens ødeleggelse av den caudal mPFC i kaniner produserer lignende resultater46. Disse funnene også markere betydningen av å vurdere arten forskjeller i prefrontal bidrag til lillehjernen-hjerne stamme drevet associative læring, som spor ECC. Mens neonatal alkohol eksponering under PD 4-9 negativt påvirket oppkjøpet av 500-ms spor CRs for voksen rotter i denne studien og andre47,56, er dette ikke samme sak for neonatal alkohol-eksponerte rotter ha en 300-ms spor intervall, selv når utfordret på en relativt høy dose av alkohol (5 g/kg)57, antyder at spor svekkelse i alkohol-eksponerte rotter er avhengig av varigheten av spor intervallet.
I denne studien var hippocampus understreket som blir avgjørende for formidling spor ECC, og når utfordret av neonatal alkohol eksponering, viser neural-relaterte skader som gjenspeiles av nedsatt i oppkjøpet av spor CRs. Det må imidlertid advarte at lillehjernen-hjerne stilk krets, spesielt interpositus kjernen, er avgjørende for mange fasetter av ECC, inkludert kjøp, uttrykk og topografiske kjennetegn for CR, avhengig av ECC aktiviteten inkludert spor ECC36,40,55,58,59. Faktisk samhandler denne nevrale kretsen med hippocampus for kjøring uttrykk for CRs under høyere orden former for ECC, for eksempel spor ECC60. Om alkohol eksponering under tidlig hjernens utvikling spesielt påvirker hippocampus funksjon i spor ECC er ikke helt klart. Mange forskjellige hjernen regioner er utsatt for tidlig alkohol fornærmelse, inkludert mPFC, lillehjernen og hippocampus18,19,23,47,61,62, og det er svært sannsynlig at alkohol forstyrrer funksjon av disse strukturene i varierende grad og varierende, men funksjonelt viktige forskjeller over mange ECC prosedyrer. På tross av fallgruver vedrørende tolkningen av resultater fra spor ECC studier, har vellykkede oppkjøpet av spor CRs vist minst stole på en intakt hippocampus, som støttes av dyr lesjon studier42,44,63,64,65. Denne fremgangsmåten er dermed en svært verdifull tilnærming for å vise koblingene mellom utviklingsmessige alkohol eksponering å spore betinget svarer fordi nerve kretssystem underliggende det, er mye bedre forstått enn andre hippocampus-avhengige aktiviteter, for eksempel sted læring i Morris vann labyrinten, romanen objekt gjenkjenning og kontekstuelle og frykt condition.
ECC som atferdsdata til “analysen” erkjennelse, har omfattende anvendelse innen utviklingsmessige neuroteratology. Faktisk støtter nylig resultatene fra vår lab ideen om at utvikling hippocampus er svært følsom for alkohol-effekter, som kan begrenses ved forskjellige intervensjonsradiologi strategier18,47. Den store fordelen her er at en bedre forståelse av alkohol-indusert spor ECC læring underskudd, de kan være intelligent andre problemer i hippocampus-baserte funksjoner utenfor associative læring – særlig de kjent for å være formidlet av den samme hippocampus neurocircuitry.
Anvendelse av spor ECC og andre variantene (f.eks, forsinkelse, tilbakeføring, diskriminering, sammensatte) å belyse den nevrobiologiske mekanismer og nevrale systemer involvert i associative læring, kan utvides utover området fetal alkohol forskning. For eksempel har dette paradigmet fått mye oppmerksomhet i menneskelig tilfeller og dyr modeller av psykiatriske tilstander som schizofreni66,67, nevrodegenerative sykdommer som Alzheimers sykdom68,69, og narkotika misbruk70,71,72. Dens fordeler som forskning å vurdere nevrokognitive funksjon og dysfunksjon er dermed tydelig over mange psykologiske og biomedisinsk disipliner, inkludert nevrovitenskap.
The authors have nothing to disclose.
Dette arbeidet ble støttet av et stipend til TDT fra den alkohol drikke Medical Research Foundation (ABMRF).
Neonatal Alcohol Exposure | |||
190 Proof Ethyl Alcohol (USP) | Pharmco-AAPER | 225-36000 [ECU Medical Storeroom] | Can be substituted; should be USP; avoid using 200 proof ethyl alcohol |
Container/Basket for Pups | Any | ||
Corn Oil | Any | Food grade | |
Heated Water Therapy Pump w/ Pads | Gaymar | TP-500 | To keep pups warm; can be substituted |
Hypodermic Needles 22G x 1 in, Sterile | Any | ||
Hypodermic Needles 30G x 1/2 in, Sterile | Any | ||
Isopropyl Alcohol 70% | EMD Millipore | PX1840-4 [Fisher Scientific] | Can be substituted; reagent grade www.fishersci.com |
Long-Evans Rats (Female and Male Breeders) | Charles River Laboratories | N/A [ECU Dept. of Comparative Medicine] | Age and weight need to be specified; pricing varies by these factors www.criver.com |
Micro Dissecting Scissors, 3.5 in, 23 mm Blades | Biomedical Research Instruments | 11-2200 | For cutting PE tubing brisurgical.com |
Polyethylene 10 Tubing (0.011 in. I.D.; 0.024 in. O.D.) | BD Diagnostic Systems | 22-204008 [Fisher Scientific] | Can be substituted www.fishersci.com |
Polyethylene 50 Tubing (0.023 in. I.D.; 0.038 in. O.D.) | BD Diagnostic Systems | 22270835 [Fisher Scientific] | Can be substituted www.fishersci.com |
Regulated water heater or baby milk bottle warmer | Any | Optional; helps with warming up cold milk solutions | |
Tuberculin Syringes, Sterile, 1.0 ml | Any | ||
Tuberculin Syringes, Sterile, 10 ml | Any | Can be used to draw out ethyl alcohol or use appropriate size micropipet | |
Weigh Scale | Any | Should have good resolution (in gram units) | |
Name | Company | Catalog Number | Comments |
EMG Headstage Fabrication and Bipolar Electrode Modification | |||
Bipolar Electrode, 2 Channel SS Twisted | Plastics One, Inc. | MS303/2-B/SPC ELECT SS 2C TW .008" | Must specify custom length of 20 mm below pedestal www.plastics1.com |
Centi-Loc Strip Socket Insulator (aka, Micro Strip) | ITT Cannon / ITT Interconnect Solutions | CTA4-IS-60* or CTA4-1S-60* | *Depends on vendor; see www.onlinecomponents.com or www.avnetexpress.avnet.com |
Dental Pliers, Serrated | CMF Medicon | 390.20.05 | Can be substituted; use to crimp wires to male contact pins www.medicon.de |
Micro Dissecting Scissors, 3.5 in, 23 mm Blades | Biomedical Research Instruments | 11-2200 | Only use to cut 3T wires; cutting 10T wires will damage the blade – use the blade of the wire stripper instead brisurgical.com |
PTFE-Coated Stainless Steel Wire, 10T (Bare Diameter .010 in) | Sigmund Cohn-Medwire | 316SS10T | www.sigmundcohn.com |
PTFE-Coated Stainless Steel Wire, 3T (Bare Diameter 0.003 in) | Sigmund Cohn-Medwire | 316SS3T | www.sigmundcohn.com |
Razor Blade | Any | To strip 1 mm from prongs of bipolar electrode | |
Relia-Tac Socket Contact Pin, Male | Cooper Interconnect | 220-P02-100 | See Allied Electronics Cat # 70144761 www.alliedelec.com |
Tweezers, High Precision, Serrated, 4 3/4 in | Electron Microscopy Sciences | 78314-00D | To grasp 10T wire firmly while stripping PTFE with smooth tweezers www.emsdiasum.com |
Tweezers, High Precision, Smooth, 4 3/4 in | Electron Microscopy Sciences | 78313-00B | www.emsdiasum.com |
Tweezers, Ultra Fine Tips, 4 3/4 in | Electron Microscopy Sciences | 78510-0 | To strip 1 mm of PTFE from one end of 3T wire; grasp shielded portion with smooth tweezers www.emsdiasum.com |
Wire Stripper, 16-26 AWG | Any | Use the blade end to cut micro strips | |
Name | Company | Catalog Number | Comments |
Eyelid Surgery | |||
Surgical Instruments (High Quality Stainless Steel) | |||
2 x Dressing Forceps, 4 in Serrated | Biomedical Research Instruments | 30-1205 | Can be substituted; extra forceps for grasping electrodes/screws outside of surgery tray brisurgical.com |
Dressing Forceps, 3 in Serrated | Biomedical Research Instruments | 30-1200 | Can be substituted brisurgical.com |
Instrument Tray | Biomedical Research Instruments | 24-1355 | Can be substituted brisurgical.com |
Knife Handle No. 3, 5 in | Biomedical Research Instruments | 26-1000 | Can be substituted brisurgical.com |
Micro Dissecting Forceps, 3.5 in, Fine Points | Biomedical Research Instruments | 10-1630 | Can be substituted brisurgical.com |
Micro Dissecting Forceps, 3.5 in, Smooth Platform (0.3 x 5 mm) | Biomedical Research Instruments | 10-1720 | brisurgical.com |
Micro Dissecting Scissors, 3.5 in, Extremely Delicate, 15 mm Blades | Biomedical Research Instruments | 11-2000 | Can be substituted brisurgical.com |
Plain Splinter Forceps, 3.5 in | Biomedical Research Instruments | 30-1600 | Can be substituted brisurgical.com |
#10 Stainless Steel Surgical Blade for #3 Handle, Sterile | Any | Can be substituted | |
0-80 x 0.125 in Stainless Steel Screws | Plastics One, Inc. | 0-80 x 0.125 | Can be substituted www.plastics1.com |
Alcohol Prep Pads, Sterile | Fisher Scientific | 22-363-750 [Fisher Scientific | Can be substituted www.fishersci.com |
Betadine Povidone-Iodine | Purdue Frederick Co. | 6761815101 [Fisher Scientific] | Can be substituted www.fishersci.com |
Betadine Povidone-Iodine Prep Pads | Moore Medical | 19-898-946 [Fisher Scientific] | Can be substituted www.fishersci.com |
Cotton-Tipped Swabs, Autoclavable | Any | Typically 7.6 cm or 15.2 cm length | |
Drill Bit for Pin Vise, #55 (0.052 in) | Any | Metal should resist rusting and corrosion | |
Gauze Pads, 2 in x 2 in | Fisher Scientific | 22-362-178 [Fisher Scientific] | Can be substituted www.fishersci.com |
General Purpose Latex/Nitrile/Vinyl Gloves | Any | ||
Glass Bead Sterilizer | Any | Sterilize instruments between surgeries | |
Heated Water Therapy Pump w/ Pads x 2 | Gaymar | TP-500 | Can be substituted; separate pumps are recommended – 1 for surgery, 1 for recovery |
Hypodermic Needles 26G x 3/8 in, Sterile | Any | ||
Isoflurane | Vedco | NDC 50989-150-12 | Manfacturer can be substituted; veterinary approval may be required |
Isoflurane Vaporizer System, Tabletop, Non-Rebreathing | Parkland Scientific | V3000PK | Can be substituted www.parklandscientific.com |
Jewelers Screwdriver w/ 1.8-2 mm Blade | Any | Metal should resist rusting and corrosion | |
Ortho-Jet BCA Package (Dental Cement) | Lang Dental | B1334 | Contains powder (1 lb) and liquid www.langdental.com |
Oxygen Tank with Pressure Regulator, Large | Local supplier | ||
Porcelain Crucible, High-Form, Glazed, 10 ml | CoorsTek, Inc. | 07-965C [Fisher Scientific] | Can be substituted with Fisher FB-965-I Wide-Form Crucible www.fishersci.com |
Puralube Veterinary Ophthalmic Ointment, Sterile | Henry Schein Company | NC0144682 [Fisher Scientific] | Can be substituted www.fishersci.com |
Quatricide PV-15 | Pharmacal | PV-15 | Antimicrobial disinfectant; can be substituted www.pharmacal.com |
Rat Gas Anesthesia Masks for Stereotaxic Surgery | Stoelting Company | 51610 | www.stoeltingco.com |
Rat Stereotaxic Apparatus w/ Ear Bars (45 Degree) | Any | 45 degree bars are recommended to prevent damaging eardrums | |
Roboz Surgical Instrument Milk | Roboz Surgical | NC9358575 [Fisher Scientific] | Can be substituted; for lubricating instruments during autoclaving www.fishersci.com |
Rodent Hair Trimmer | Any | ||
Sodium Chloride | Fisher Scientific | S641-500 [Fisher Scientific] | To make 0.9% saline; reagent grade; USP www.fishersci.com |
Stainless Steel Microspatula (Blade: 0.75 L x 0.18 in. W) | Fisher Scientific | 21-401-15 [Fisher Scientific] | Can be substituted www.fishersci.com |
Starrett Pin Vise, 0.000 in – 0.055 in | Any | Nickel-plated or equivalent recommended to resist rusting and corrosion | |
Sterile Surgical Gloves | Any | ||
Sterilization Wraps, 20 in x 20 in, Autoclavable | Propper Manufacturing | 11-890-8C [Fisher Scientific] | Useful for wrapping autoclavable supplies and on sterile field during surgery www.fishersci.com |
Surgical Drape, Sterile/Autoclavable | Any | May need to cut to size for rats | |
Surgical Gown* | Any | *If required by IACUC | |
Surgical Mask | Any | ||
Tuberculin Syringes, Sterile, 1.0 ml | Any | ||
Weigh Scale | Any | Should have good resolution (in gram units) | |
Name | Company | Catalog Number | Comments |
Eyeblink System and Components (assuming 4-rodent system) | |||
5 Channel Commutator x 4 | Plastics One, Inc. | SL2 + 3C | www.plastics1.com |
Bipolar Electrode Cable, Dual 305 x 4 | Plastics One, Inc. | 305-305 80CM TT2 (C) | Provides plug end to bipolar electrode on rat and to commutator; must be modified www.plastics1.com |
Cable, 5 Channel, Shielded, 26 AWG x 4 | Any | To fabricate commutator cable; this must be made from scratch | |
Chamber for Operant Test Box (Inside: 24 H x 23 W x 14 D in) x 4 | Med-Associates | Can be substituted; inner dimensions should fit operant test box comfortably, with room for acoustical foam; fit with fan – 55-60 dB www.med-associates.com |
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Eyeblink System and Software | JSA Designs | N/A | Proprietary and customized for research lab |
Heat Shrink Tubing (3/16 in, 1/4 in, 3/8 in, 1/2 in Diameters) | Any | To protect modified commutator cable soldered ends and splices | |
Melamine Triple Peak Acoustical Foam w/Black Hypalon (24 x 48 in) | McMaster-Carr | 9162T5 | Can be substituted; cut to fit 4 housing chambers www.mcmaster.com |
Operant Test Box (Exterior 12.5 L x 10 W x 13.5 in H), Complete x 4 | Med-Associates | ENV-007 Custom Package | With stainless steel grid floor and custom top (3 in hole in center for commutator cable) www.med-associates.com |
Oscilloscope (Optional) | Any | Recommended minimum specs: 200 MHz analog bandwidth, 1 GS/s real-time sampling, 4 channels; see www.picotech.com /td> |
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Piezo Tweeters (Speakers) x 4 (7 x 3 in) | MCM Electronics | 53-805 | Must match frequency range specifications for eyeblink system (2500 Hz – 25 KHz) www.mcmelectronics.com |
Soldering Station, Solder, Flux, Tinner | Any | For soldering 26 AWG cables to female sockets (that fit male relia-tac contact pins) and bipolar plugs | |
Stimulus Isolators x 4 | WPI International | A365 | These units run on 16-9V alkaline batteries; a suitable rechargeable version (A365R) is available www.wpiinc.com |
Tripolar Electrode Cable for SL3C Commutator x 4 | Plastics One, Inc. | 335-335 80cm TT3 C | Provides plug end to EMG headstage on rat and to commutator; must be modified www.plastics1.com |
USB LED Lights x 4 | Any | USB-based lights do not cause electrical "noise" with the EMG signals from the rats www.plastics1.com |
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Webcams x 4, Surveillance Software | Any | ||
PC Computer Running MS Windows OS | Any |