Trace Blink klassisk betingning (ECC) användes för att bedöma Hippocampus-beroende associativ inlärning hos vuxna råttor som administreras en hög koncentration (11,9% v/v) av alkohol under tidig neonatal hjärnans utveckling. ECC förfaranden är i allmänhet ljud diagnostiska verktyg för att upptäcka dysfunktion i hjärnan över många psykologiska och biomedicinska inställningar.
Neonatala råttor administrerades en relativt hög koncentration av etylalkohol (11,9% v/v) under postnatal dagar 4-9, en tid då fostrets hjärnan genomgår snabb organisationsförändring och liknar accelererade förändringar i hjärnan som uppstår under den tredje trimestern hos människor. Denna modell av fetala alkoholeffekter spektrum (FASDs) producerar allvarlig hjärnskada, härma det beloppet och mönster av berusningsdrickande som uppstår i vissa gravida alkoholhaltiga mödrar. Vi beskriver användningen av trace Blink klassisk betingning (ECC), en högre ordningens variant av associativ inlärning, att bedöma långsiktiga Hippocampus dysfunktion som vanligtvis ses i alkohol-exponerade vuxna avkommor. Vid 90 dagars ålder, gnagare var kirurgiskt beredda med inspelning och stimulerande elektroder, som mätte elektromyografisk (EMG) blinkar aktivitet från den vänstra ögonlock muskeln och levererade mild chock posteriort vänster öga, respektive. Efter en 5 dagars återhämtningsperiod genomgick de 6 sessioner av trace ECC att bestämma associativ inlärning skillnader mellan alkohol-exponerade och styra råttor. Trace ECC är ett av många möjliga ECC förfaranden som kan enkelt ändras med samma utrustning och programvara, så att olika neurala system kan bedömas. ECC förfaranden i allmänhet kan användas som diagnosverktyg för att upptäcka neurala patologi i olika hjärnsystem och olika förhållanden som förolämpar hjärnan.
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 råttungar som fått etylalkohol postnatal dagarna 4-9 uppvisade spår Blink luftkonditionering nedskrivningar i vuxen ålder. Dessa fynd stöder tanken att alkohol är en teratogen med bestående negativa effekter på Hippocampus funktion. Övergripande, konditionerat svara i förfarandet för trace var lägre för honråttor som exponerats för alkohol jämfört med råttor i båda kontrollgrupper. De associativa lärande nedskrivningar i alkohol-exponerade råttor inte påverkades av motiverande eller motoriska skillnader (dvs., inga skillnader i blinkar till chock U.S. intensitet).
Trace ECC är ett användbart diagnostiskt verktyg för belysa utmaningen-inducerad Hippocampus neuropatologi, måste resultaten från denna metod placeras i rätt sammanhang. Först, de processuella nyckelelement i denna demonstration skedde riktade leverans av alkohol under en känd sårbarhet för hjärnans utveckling, tillverkning av elektroden hårdvara som tillåter inspelning av elektromyografisk aktivitet och levererar chock, kirurgisk implantation av ovannämnda maskinvara och efterföljande djurförsök med ett lärande paradigm som bedömer kognitiv funktion av intresse. Vid varje steg i processen, måste vara försiktig att inte orsaka onödiga/oavsiktlig skada gnagare frågor och att regelbundet kontrollera sin hälsa tecken. Deras beteende resultat ger fönstret’ ‘ att kognition, en psykologisk konstruktion som är bara exakt beskrivs när deras hälsa inte äventyras av experimentella fel omfattar alkohol dosering, hårdvara defekter eller kirurgisk implantation. Således måste varje procedurmässiga element i forskningsprocessen genomföras på ett sätt för att säkerställa att resultaten från ECC kan extrapoleras till fynd hos människa. För det andra, ECC paradigm ger insikt om arten av associativ inlärning, men försiktighet måste iakttas inte att förlänga resultaten med hjälp av denna metod och i stort sett tillskriver dem andra kognitiva domäner – såsom arbetsminne, kort/lång sikt minnen och medvetande – om inte en har införlivat vissa aspekt av dessa domäner inom en ECC studie av experimentell design. Exempelvis denna demonstration undersökte fasen förvärv av trace ECC lärande, men prövade inte minne lagring i råttor efter att de avslutat utbildningen. Minnet är således en oberoende psykologiska process som bör utvärderas förutom lärande. Av design, kan en införliva ett minne retention intervall för att bedöma antingen kortvarigt eller långvarigt minne förmåga. För det tredje är erkännande att det finns parallella minne system54 som kan arbeta samtidigt tillsammans med motiverande, upplevelsebaserat och hormonella faktorer som bidrar till beteende, viktigt för förståelse som associativitet (under ECC) är men en av många processer som avslöjar vad som är ”bra” eller ”dålig” om lärande. Slutligen, trace ECC är inte en rent Hippocampus-beroende uppgift, som andra regioner i hjärnan kan medla vissa komponent av CR. Således, en förståelse för samspelet mellan olika neurala kretsar eller typ av stimulans parametrar som används i en studie, måste beaktas när du gör konsekvenser utifrån diskret resultat. Lillhjärnan, exempelvis bidrar också till att spåra ECC, där det påverkar de topografiska egenskaperna av CR och CR timing, särskilt när ISI är kort varaktighet. Trace ECC påverkas inte i människor med cerebellär skada som testas med långa spår intervall (1000 ms), men påverkas i dem som får ett kortare spår intervall (400 ms)34. Dessutom, bilaterala lesioner av dorsala mediala prefrontala cortex (mPFC) som är inriktade på de främre cingulum och mediala agranular regionerna i möss, förhindra förvärv av trace CRs55, medan förstörelsen av den kaudala mPFC i kaniner ger liknande resultat46. Dessa fynd också belysa vikten av arter skillnader i prefrontala bidrag till lillhjärnan-brain stem driven associativ inlärning, såsom trace ECC. Medan neonatal alkohol exponering under PD 4-9 påverkas negativt förvärv av 500-ms spår CRs för vuxen råtta i denna studie och andra47,56, är detta inte samma ärende för neonatala alkohol-exponerade råttor som upplever en 300-ms spår intervall, även när utmanas på en relativt hög dos av alkohol (5 g/kg)57, tyder på att spåra nedskrivning i alkohol-exponerade råttor är anhörigen på varaktighet av trace-intervallet.
I denna studie betonades hippocampus som är mycket viktigt för att förmedla trace ECC, och när utmanas av neonatal alkohol exponering, uppvisar neurala-relaterade skador som reflekteras av nedskrivningar i förvärvet av trace CRs. Det måste dock varnade att lillhjärnan-hjärnan stjälken kretsar, särskilt interpositus kärnan, är avgörande för många aspekter av ECC, inklusive förvärv, uttryck och topografiska dragen av CR, beroende på vilken typ av ECC uppgift inklusive trace ECC36,40,55,58,59. Ja, denna neural krets interagerar med hippocampus för körning uttrycket av CRs vid högre ordningens former av ECC, såsom trace ECC60. Om exponering för alkohol under tidig hjärnans utveckling påverkar särskilt Hippocampus funktion i spårningen ECC är inte helt klart. Många olika hjärnregioner är utsatta för tidig alkohol förolämpning, inklusive mPFC, lillhjärnan och hippocampus18,19,23,47,61,62, och det är mycket troligt att alkohol stör funktionen hos dessa strukturer i varierande grad och varierande, men funktionellt viktiga skillnader över många ECC förfaranden. Trots fallgroparna när det gäller tolkningen av resultat från trace ECC studier, har framgångsrika förvärv av trace CRs visat att åtminstone förlita sig på en intakt hippocampus, som stöds av animaliskt lesion studier42,44,63,64,65. Detta förfarande således återstår ett värdefullt tillvägagångssätt för att påvisa sambanden mellan utvecklingsmässiga alkohol exponering att spåra luftkonditionerade svara eftersom den neurala kretsar underliggande är mycket bättre förstås än andra Hippocampus-beroende uppgifter, till exempel plats lärande i Morris vatten labyrinten, romanen objekt erkännande, och kontextuella och spåra fruktar luftkonditionering.
ECC som beteendevetenskaplig metod till ”assay” kognition, har utbredd tillämpning inom utvecklande neuroteratology. Ja, senaste rön från vårt labb stöd för uppfattningen att utveckla hippocampus är mycket känslig för alkohol effekter, som kan minskas genom olika interventionella strategier18,47. Den viktigaste fördelen här är att med en bättre förståelse av alkohol-inducerad trace ECC lärande underskott, de kan vara förutsägande av andra problem i hippocampus-baserade funktioner utanför associativ inlärning – särskilt de kända för att vara medierade av den samma Hippocampus neurocircuitry.
Tillämpningen av trace ECC och dess andra varianter (t.ex., dröjsmål, återföring, diskriminering, sammansatta) att klarlägga de neurobiologiska mekanismer och neurala system inblandade i associativ inlärning, kan utvidgas utöver fetala alkohol forskningsområdet. Till exempel har detta paradigm fått mycket uppmärksamhet i människans fall och djurmodeller av psykiatriska tillstånd såsom schizofreni66,67, neurodegenerativa sjukdomar som Alzheimers sjukdom68,69, och narkotika missbruk70,71,72. Dess fördelar som en forskningsmetod att bedöma neurokognitiv funktion och dysfunktion är således uppenbart över många psykologiska och biomedicinska discipliner, inklusive neurovetenskap.
The authors have nothing to disclose.
Detta arbete stöds av ett bidrag till TDT från den alkohol dryck 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 |
|
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> |
|
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 |
|
Webcams x 4, Surveillance Software | Any | ||
PC Computer Running MS Windows OS | Any |