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SheffieldBioSciencePrograms

TheLangendorffHeart

Acomputersimulationoftheeffectsofdrugsontheisolatedperfusedmammalianheartpreparation

By:DavidDewhurst&MalachyDoherty
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Introduction
Theisolatedperfusedheartisausefulpreparationbothforteachingandresearch.Themethod,first
describedbyLangendorffin1895andsubsequentlymodifiedbymanyotherworkersisthemost
widelyusedandallowsanumberofcardiovascularparameterstobemonitoredsimultaneously:

Coronarybloodflow
Cardiacforce
Heartrate
Electrocardiogram

Theprincipleistorapidlyremovetheheartandperfusethecoronaryvesselsbyforcing,ina
retrogradedirection,anappropriateoxygenatedfluidthroughacannulainsertedintotheascending
aorta.Afterpassingthroughthecoronarycirculationtheperfusatedrainsawayandmaybecollected.
Thechambersoftheheartareessentiallyemptythroughouttheexperiment,theheartmuscledoes
verylittleworkandthereisnorealcardiacoutput
Asateachingpreparationinpharmacologyandphysiologyitmaybeusedtoinvestigatetheactions
ofdrugsorionsontheheartandcanalsobeusedtodemonstratetheeffectsofpreload(Starling's
LawoftheHeart).
MenuOptions
INTRODUCTION:abrieftextualintroductiontotheaimsoftheprogram.
METHOD:abrieftextualdescriptionofthepreparationandexperimentalsetupused.
EXPERIMENTS:allowsyoutoinvestigatetheeffectsofarangeofdrugsorionsontheheartThe
effectsofchangingpreload(Starling'sLaw)mayalsobedemonstrated.Thethreesubsections
DRUGS;IONSandSTARLINGSLAWwillbediscussedmorefullyinamoment.
EXPERIMENTS
SelectingEXPERIMENTSleadstoasubmenuwiththreefurtheroptions.Movethehighlightusing
theupanddownarrowkeystotheoptionyouwishtoselectandpress<Enter>toconfirmyour
choice.
DRUGS
Selectingthismenuoptionleadstoafurthersubmenuofferingachoiceofdifferenttypesofdrugs.
Selectingoneoftheseleadstoasecondsubmenugivingachoiceofdrugsofthattypeandselecting
oneoftheseallowsyoutoinvestigatetheeffectsof:

thedrugalone(DrugOnly)
thedruginthepresenceofanotherdrugwhichmayantagoniseitsaction(Antagonist)
thedruginthepresenceofanotherdrugwhichmayenhanceitsaction(Potentiator)

IfyouselectAntagonistorPotentiatoryouwillbegivenafurtherchoiceofeitherpossible
antagonistsorpossiblepotentiators.
Onconfirmingyourchoiceof"experiment"theprogramwillpresentsimulatedresponses,inhigh
resolutiongraphics,appropriatetoyourchoiceofdrugs.Thescreendisplaymimicsachartrecorder
displayandshowscontractileforce,heartrateandcoronarybloodflow.Ahorizontaltimescaleand
appropriatecalibratedverticalscalesareprovidedifyouwishtoquantifydrugeffects,thoughforthe
mostpartqualitativeeffectsaresufficientforteachingpurposes.Thepointofadministrationofthe
drugisindicatedbyaverticaldottedlineandthenameofthedrugordrugsisconfirmedinatext
windowatthebottomofthescreen.
DrugDosesandAdministration
Inthisprogramalldrugsareadministeredbybolusinjectionintoacannulainsertedintothe
ascendingaortaandarethusrapidlydistributedthroughoutthecoronaryvessels.Thedrugdosesare
thosewhichwillproducetheprimarypharmacologicaleffectsofthedrug.Theresponsesshownare,
forthemostpart,derivedfromrealdataobtainedfromavarietyofsources(seebibliography).
Drug+AntagonistorDrug+Potentiator
Theprogramalsoallowsyoutochoosetoinvestigatetheactionofadruginthepresenceofeithera
potentialantagonistorpotentiator.Thescreendisplayisasforthedrugaloneexceptthatthepointof
administrationofeachdrugisseparatelyindicatedbyaverticaldottedline.Theantagonistor
potentiatorisalwaysgivenimmediatelybeforetheagonist.
Itisuseful,wheninvestigatingtheactionofapotentialantagonistorpotentiator,tobeableto
comparetheresultswiththoseofthedrugalone.Ifyouweredoingthisexperimentforrealyou
wouldprobablyhaveatraceofthedrugalonetodirectlycompare.Inthisprogramthisfacilityisalso
availablebypressingtheCOMPAREbuttonwhichwilldisplaythetracefortheagonist(drugonly).
PressingCOMPAREagainreturnsyoutotheoriginaltracei.e.drug+antagonistordrug+
potentiator.Bymovingfromtracetotraceitismucheasiertoassesstheeffectsofantagonistsor
potentiators.
IONS
Theeffectsofanumberofionsonthepreparationmaybeinvestigatedinmuchthesamewayasthe
drugs.Wheretheeffectsofahighconcentrationofanion(higherthanthenormalperfusionfluid)
aretobeinvestigated,theionisadministeredasabolusinjectioninthesamewayasthedrugs.
Wheretheeffectsofalowconcentrationoftheionaretobeinvestigated(lowerthanthenormal
perfusionfluidconcentration)asubstanceisinjectedwhichwilllowertheconcentrationoftheion
e.g.EGTAisgiventolowercalciumionconcentration.
STARLING'SLAW
Selectingthismenuoptionleadstoasubmenuwithtwooptions:IntroductionandExperiment.
Introduction:presentsbrieftextualinformationasbackgroundtotheexperiment.
Experiment:presentsdatatoillustratetheeffectsofincreasingpreloadonthecontractileforce.The
uppertraceshowsleftventricularpressureandthelowertraceindicatestheenddiastolicpressure.

Starling'sLaw(Heterometricautoregulation)isbestdemonstratedbyplottingcontractileforce
(systolicpressurediastolicpressure)againstdiastolicpressure.Thegraphoftheexperimental
resultsmayalsobedisplayed.
TUTORS'NOTES
ThePreparation
Langendorffpreparationsusingtheheartofrat,guineapig,rabbitandcathavebeendescribed
frequentlythoughtherabbitorguineapigpreparationsseemtobethemostwidelyusedinteaching.
Theanimalispretreatedwiththeanticoagulantheparin,anaesthetised,andatrachealcannula
insertedforventilation.Thethoraxisopened,thepericardiumremovedandtheascendingaortafreed
fromsurroundingconnectivetissue.Acannulaisinsertedintotheaortaandcoronaryperfusion
initiated.Theinferiorvenacavaisclampedandthepulmonaryarteryincised.Retrogradeperfusion
closestheaorticvalveandtheperfusatepassesintothecoronaryvesselsandoutviathecoronary
sinusandinferiorvenacava.
Finallytheheart,whichisnowattachedtothecannula,isremovedfromtheanimalandtransferred
totheconstantpressureperfusionapparatus.Itshouldthenbeleftfor1530minutestoadjusttothe
invitroconditions.Severalperfusionfluidshavebeenusede.g.KrebsHenseleit,TyrodeandLocke
salinesolutions,and
McEwensRingersolution.Thefluidsareoxygenated,filtered,equilibratedtopH7.4andmaintained
at37C.Thismethodgreatlyreducesthepossibilityofmyocardialdamageduetoischaemiaandis
preferabletoremovaloftheheartfromafreshlykilledanimal.
Thefollowingcardiovascularparametersaremeasured:
ContractileForce
Either:insertasmallhookorattachaspringloadedcliptotheventricleandconnecttoabalanced
isotoniclever(tensiontransducer).
or:insertasmall(latex)balloon,fixedtoacathetertipandfilledwithwater,intotheleftventricle
viaoneofthepulmonaryveins,theleftatriumandmitralvalve.Attachtheendofthecathetertoa
pressuretransducer.Optimiseventricularpreloadbyexpandingtheballoon.
HeartRate
Instantaneousheartrateismeasuredusingaratemeterortachographtriggeredfromthetension
transducer.
CoronaryBloodFlow(CBF)
Iftheheartisperfusedatconstantpressure(normally5060mmHginguineapig)themeanvolume
ofperfusionfluiddrainingfromtheheartisameasureofthecoronarybloodflow.Thismaybe
collectedperunittime,orthenumberofdropsofperfusatefallingofftheheartmaybecountedusing
anelectricaldropcounter.Inguineapigsthenormalcoronarybloodflowisapproximatelyl0
ml/min;inrabbitsapproximately20ml/min.Ifavasoconstrictordrugisaddedthencoronaryvessels
willconstrictandbloodflowwillbereduced;likewiseacoronaryvasodilatorwillincreaseCBF.
Itshouldberememberedthatbloodflowthroughcoronaryvesselsisnormallyphasicbeinghighest
duringdiastoleandmuchreducedduringsystole.Drugswhichaffecttheforceofcontractionand/or
theheartratewillthusalsoaffectCBFeventhoughtheymayhavenodirecteffectoncoronary
vessels.Inadditionmyogenicautoregulationofthecoronaryvascularsystemwilloccurinresponse
toeitheranincreaseoradecreaseinCBF.

Bibliography
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PhysiologyandPharmacology,BiologicalMeasurementTechniquesV.BiomesstechnikVerlag
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Bowman,W.C.&Rand,M.J.(1980)TextbookofPharmacology(2ndedition)BlackwellScientific
Publications.
Burn,J.H.&Walker,J.M.(1954)Anticholinesterasesintheheartlungpreparation.J.Physiol.
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Burn,J.H.,Gunning,AJ.&Walker,J.M.(1957)Effectsofnoradrenalineandadrenalineontheatrial
rhythmintheheartlungpreparation.J.Physiol.137,141153.
Flack,M.&Hill,L.(1919)Atextbookofphysiology.Arnold
Harper,B.&Hughes,I.E.(1977)Acomparisoninrabbitisolatedheartsofthedysrrhythmogenic
potentialofamitriptyline,maprotilineandmianserininrelationtotheirabilitytoblocknoradrenaline
uptake.Br.J.Pharmac.59,651660.
Katz,LN.,etal(1938)Effectsofvariousdrugsonthecoronarycirculationofthedenervatedisolated
heartofthedogandcat.Pharmacol.Rev.399415.
Kitchen,I.(1984)textbookofinvitropracticalpharmacology.BlackwellScientificPress.
McDowall,R.J.S.(1946)Thestimulatingactionofacetylcholineontheheart.J.Physiol.104,392
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Acknowledgements
WewouldliketoacknowledgethefinancialsupportofTheLordDowdingFundandtheadviceand
contributionsmadebyDrG.G.S.CollinsandDrI.E.Hughestowardsthesuccessfulcompletionof
thispackage.
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