Trace eyeblink klassisk konditionering (ECC) blev brugt til at vurdere hippocampus-afhængige associative læring i voksne rotter, der blev indgivet en høj koncentration (11,9% v/v) af alkohol i den tidlige neonatal hjerne udvikling. I almindelighed, er ECC procedurer lyd diagnostiske redskaber til påvisning af dysfunktion i hjernen på tværs af mange psykologiske og biomedicinsk indstillinger.
Neonatal rotter blev administreret en forholdsvis høj koncentration af ethanol (11,9% v/v) under postnatal dage 4-9, når de fostrets hjerne gennemgår hurtig organisatorisk forandring og ligner accelereret hjernen ændringer, der opstår under tredje trimester hos mennesker. Denne model af føtale alkohol spektrum forstyrrelser (FASDs) producerer svær hjerneskade, efterligne beløb og mønster af binge-drinking der opstår i nogle alkoholholdige gravide. Vi beskriver brugen af trace eyeblink klassisk konditionering (ECC), en højere orden variant af associative læring, at vurdere langsigtede hippocampus dysfunktion, der typisk ses i alkohol-udsatte voksne afkom. På 90 dage i alder, gnavere var kirurgisk rede med optagelse og stimulerende elektroder, som målte Elektromyografi (EMG) blinker aktivitet fra venstre øjenlåg muskel og leveret mild chok posteriort for venstre øje, hhv. Efter en 5 dages tilbagebetalingsperioden gennemgik de 6 sessioner af trace ECC at bestemme associative læring forskelle mellem alkohol-eksponerede og kontrol rotter. Trace ECC er en af mange mulige ECC procedurer, der kan nemt ændres ved hjælp af det samme udstyr og software, således at forskellige neurale systemer kan vurderes. ECC procedurer i almindelighed, kan bruges som diagnostiske redskaber til påvisning af neurale patologi i forskellige hjernen systemer og forskellige betingelser, der fornærmer 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, der modtog ethylalkohol under postnatal dage 4-9 udstillet trace eyeblink condition funktionshæmninger i voksenalderen. Disse resultater støtter tanken om, at Alkohol er en teratogent med varige skadelige virkninger på Hippocampus funktion. Samlede, konditioneret reagerer i proceduren spor var lavere for rotter udsat for alkohol i forhold til rotter i begge kontrolgrupper. Associative læring svækkelser i alkohol-eksponerede rotter blev ikke påvirket af motiverende eller motor forskelle (dvs., ingen forskelle i blinker til chok U.S. intensitet).
Mens spor ECC er et nyttigt diagnostisk redskab for informative udfordring-induceret hippocampus neuropatologiske, skal resultaterne fra denne metode placeres i rette sammenhæng. Først, de centrale proceduremæssige elementer i denne demonstration involveret målrettet levering af alkohol under en kendt vindue af sårbarhed over for udviklingslandene hjernen, fabrikation af elektrode hardware, der giver mulighed for optagelse af Elektromyografi aktivitet og leverer chok, kirurgisk implantation af førnævnte hardware og efterfølgende dyreforsøg ved hjælp af en læring paradigme, der vurderer en kognitiv funktion af interesse. På hvert trin i processen, skal pleje tages ikke medføre unødvendige/utilsigtede skader for de gnavere emner og til at overvåge deres sundhed tegn regelmæssigt. Deres adfærdsmæssige resultater give kognition “vinduet”, en psykologisk konstruktion, der er kun præcist beskrevet Hvornår deres helbred ikke bringes i fare af eksperimentelle fejl omfatter alkohol dosering, hardware fejl eller kirurgisk implantation. Således, hvert proceduremæssige element i forskningsprocessen skal gennemføres på en forsvarlig måde for at sikre, at resultaterne fra ECC kan ekstrapoleres til resultater i mennesker. For det andet, ECC paradigme giver indsigt om arten af associative læring, men skal sørges for ikke at udvide resultaterne ved hjælp af denne fremgangsmåde og bredt tilskrive dem til andre kognitive domæner – som arbejdshukommelsen, kort/lang sigt erindringer og bevidsthed – medmindre man har indarbejdet nogle facet af disse domæner i en ECC undersøgelse af eksperimentelle design. For eksempel, denne demonstration undersøgt erhvervelse fase af trace ECC læring, men undersøgte ikke hukommelse opbevaring i rotter, når de afsluttet uddannelsen. Hukommelse er således en selvstændig psykologisk proces, der bør evalueres ud over læring. Ved design, kan man optage en hukommelse opbevaring interval for at vurdere enten kortsigtet eller langsigtet hukommelse evne. For det tredje er anerkendelse, der er parallelle memory systemer54 , kan arbejde samtidig med motiverende, erfaringsbaseret og hormonelle faktorer, som bidrager til adfærd, afgørende for forståelse for, at Associativitet (under ECC) er, men en af mange processer, der afslører, hvad der er “god” eller “dårlig” om læring. Endelig er trace ECC ikke et rent hippocampus-afhængige opgave, som andre områder af hjernen kan mægle nogle komponent af CR. Dermed, forståelse af samspillet mellem forskellige neurale kredsløb og/eller type af stimulus parametre, der er udnyttet i en undersøgelse, skal tages i betragtning, når de foretager konsekvenser baseret på diskrete resultater. Lillehjernen, eksempelvis bidrager også til trace ECC, hvor det påvirker de topografiske karakteristika af CR og CR timing, især når ISI er korte varighed. Trace ECC påvirkes ikke i mennesker med cerebellare skader, der er testet med en lang spore interval (1.000 ms), men påvirkes i dem, der modtager en kortere trace interval (400 ms)34. Derudover forhindre bilaterale læsioner af den dorsale mediale præfrontale cortex (mPFC), der er målrettet de forreste cingulate og mediale agranular regioner i mus, erhvervelse af trace CRs55, mens ødelæggelse af den caudale mPFC i kaniner producerer lignende resultater46. Disse resultater også fremhæve betydningen af overvejer arter forskelle i præfrontal bidrag til cerebellare-hjernen stammer drevet associative læring, såsom trace ECC. Mens udsættelse for neonatal alkohol under PD 4-9 negativt påvirket erhvervelse af 500-ms spor CRs for voksne rotter i denne undersøgelse og andre47,56, er dette ikke den samme sag for neonatal alkohol-eksponerede rotter, der oplever en 300-ms spor interval, selv når udfordret på en relativt høj dosis af alkohol (5 g/kg)57, tyder på, at spore værdiforringelse i alkohol-eksponerede rotter er afhængig af varigheden af trace interval.
I denne undersøgelse, blev hippocampus fremhævet som er livsvigtige for mægle trace ECC, og når udfordret af neonatale alkohol eksponering, udstiller neurale-relaterede skader, som reflekteres af svækkelser i erhvervelse af trace CRs. Det skal dog advaret om, at lillehjernen-hjernen stilk kredsløb, især interpositus kernen, er afgørende for mange facetter af ECC, herunder erhvervelse, udtryk og topografiske egenskaber af CR, afhængigt af typen ECC opgave herunder trace ECC36,40,55,58,59. Faktisk interagerer denne neurale kredsløb med hippocampus for kørsel udtryk for CRs under højere-ordens former af ECC, såsom trace ECC60. Om udsættelse for alkohol i den tidlige hjerne udvikling specielt påvirker hippocampus funktion i trace ECC er ikke helt klart. Mange forskellige hjerneregioner er sårbare over for tidlig alkohol fornærmelse, herunder mPFC, cerebellum og hippocampus18,19,23,47,61,62, og det er meget sandsynligt, at Alkohol forstyrrer driften af disse strukturer i varierende grad og varierende, men funktionelt vigtige forskelle på tværs af mange ECC procedurer. På trods af faldgruberne vedrørende fortolkning af resultaterne fra spor ECC undersøgelser, har vellykket erhvervelse af trace CRs vist sig at mindst stole på en intakt hippocampus, som understøttes af animalske læsion undersøgelser42,44,63,64,65. Denne procedure er således fortsat en meget værdifuld tilgang for at vise sammenhængen mellem udsættelse for udviklingsmæssige alkohol at spore aircondition reagerer fordi den neurale kredsløb underliggende, er meget bedre forstået end andre hippocampus-afhængige opgaver, såsom læring i Morris vand-labyrint, Roman objekt anerkendelse, og kontekstuelle og spore frygter conditioning.
ECC som adfærdsmæssige metode til “analyse” kognition, har udbredt anvendelse inden for udviklingsmæssige neuroteratology. Ja, de seneste resultater fra vores lab støtter tanken om at udvikle hippocampus er meget følsomme over for alkohol effekter, som kan mindskes ved forskellige interventionel strategier18,47. Den vigtigste fordel her er, at med en bedre forståelse af alkohol-induceret trace ECC læring underskud, de kan være intelligent af andre problemer i hippocampus-baserede funktioner uden for associative læring – især dem, der vides at være medieret af den samme hippocampus neurocircuitry.
Anvendelse af trace ECC og dens andre varianter (fx, forsinkelse, tilbageførsel, diskrimination, sammensatte) at belyse de neurobiologiske mekanismer og neurale systemer involveret i associative læring, kan forlænges ud over føtale alkohol forskningsfelt. For eksempel, har dette paradigme fået megen opmærksomhed i menneskelige tilfælde og dyremodeller for psykiatriske lidelser som skizofreni66,67, neurodegenerative sygdomme såsom Alzheimers sygdom68,69, og narkotika misbrug70,71,,72. Dens fordele som en Forskningsmetode til at vurdere neurokognitive funktion og dysfunktion er således tydeligt på tværs af mange psykologiske og biomedicinsk discipliner, herunder neurovidenskab.
The authors have nothing to disclose.
Dette arbejde blev støttet af en bevilling til TDT fra den alkohol drikkevarer medicinske 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 |