Table Of ContentJ.HYM.RES.
2(1), 1993pp.101-105
NewNeurotoxins FromVenomsOfAculeateHymenoptera:
AContributionToTheKnowledgeOfStingingBehaviour
TomPiek
DepartmentofPharmacology,UniversityofAmsterdam.Meibergdreef15.1105AZAmsterdam.TheNetherlands
Abstract-SeveralHymenopteraproduceavenomwhichcontainsaneurotoxiccomponent.Kinins,presentin
venomsofvespid,scoliid,tiphiid,multillidandformicidHymenoptera,irreversiblyblockthetransmissioninthe
insectCNSpresynapticallybydepletion.Poneratoxin,aneurotoxiccompoundisolatedfromantvenom,affectsthe
ioncurrentinvoltage-dependentchannelsinnerveandmusclefibres.Philanthotoxinsblocktheneuromuscular
transmissionandthesynaptictransmissionintheCNSofinsects.Theresultssupporttheideathatentomophagous
AculeatastingtheirpreyintheCNSandexplaintheeffectsofthestingoninsectsandmammals.
INTRODUCTION intothesonofthewasp,"conveysthenotionthat
thepreywasnotkilledbutwasotherwiseincapaci-
WithinthephylumArthropodaseveralgroups tated.
includewellknownproducersofvenoms:thespi- Aprincipalquestionduringtheeighteenthand
ders,thescorpionsandthehymenopteraninsects. nineteenthcenturieswaswhetherpreyofsolitary
Inthefirstseventyyearsofthiscenturyanumberof aculeatewaspswerekilledorparalysed.Advocates
arthropodvenomshasbeendescribed.Amanualof ofthekillingideawereReaumur(1742)andDufour
informationwaspresentedinavolumeonArthro- (1841). Sympathizers with the ideathat prey of
podVenomseditedbySergioBettini(1978).The solitarywaspswerenotkilledbutparalysedwere,
biochemical,pharmacologicalandbehaviouralas- forexample,Bartram(1744, 1749)andAudouin
pectsofvenomsproducedbyHymenopterawere (1839).ItfelltoFabre(1855)tosettletheconflict
collectedtogetherby Piek (1986). Although the ofviewsbyelectricallystimulatingweevils,stung
latterbookprovidesthereaderwithextensivedocu- toacompleteandirreversibleimmobilitybythe
mentationofobservationsandresearchduringthe sphecidwaspCerceristuberculata(Villers)thus
last100yearsonbee.waspandantvenoms,several demonstratingthatthepreywascapableofmove-
ofthemostinterestingwaspandantneurotoxins mentandthereforeindeepparalysis,notdead.
whichhavearelationtothestingingbehaviourof Solitarywaspvenomscauseparalysisbyneuro-
theseinsectswerenotyetchemicallycharacterized toxic action and we now know that neurotoxic
inthefirsthalfofthe 1980's.Itistheaimofthis principles are also found in bee venoms, social
paper to review the several types ofneurotoxic waspvenomsandantvenoms.
effectsofsolitarywaspsandthuscontributetothe
understandingofstingingbehaviour. KININSFROMVESPIDANDSCOLIIDWASP
VENOMS
HYMENOPTERAASNEUROTOXIN
PRODUCERS Kininsareneurotoxiccomponentsofwaspand
antvenoms,causingintheinsectcentralnervous
Theearliestreportofaneurotoxiceffectonan systemapresynapticblockofthecholinergictrans-
insectbyasolitarywaspmaybeintheErh-yayin missionbymeansofanirreversibledepletionof
t'uorDictionaryofOldTerms,ofwhichtheSung transmittersubstance(acetylcholine),probablyby
illustrationsmayoriginatebetween 500and400 meansofanon-competitiveinhibitionofthepre-
BC(Bodenheimer1928).Theidea,that"inseven synaptic choline-uptake (Pieket al. 1987, 1990;
daysthewormstungbythewaspwastransformed Piek 1991).
102 JournalofHymenopteraResearch
Waspkininsarepolypeptidesof9-18amino- etal.1987).ItwasshownthatThr6BKirreversibly
acidresiduescontainingabradykinin-likesequence blockssynaptictransmissionfromthecerealnerve
as partofthe molecule. The bradykinin-like se- XIofthecockroachtoagiantinterneuroninthe
quenceiseitheridenticaltothevertebratebradyki- sixth abdominal ganglion (Hue and Piek 1988.
nin, ordiffers in a single OH-group (prolin be- 1989).
comeshydroxyprolin,Hyp'-bradykinin)ordiffers Thepresenceofkininsasproducersofneuro-
inasingleCH3-group(Thrh-bradykinin).Abrady- toxicinsecticidaleffectsinthevenomsofVespidae,
kinin-like substance wasdiscoveredas avenom Scoliidae, Tiphiidae, Mutillidae and Formicidae
constituentofthewaspParavespulavulgaris(L.) mightberesolvedasatoxinologicalargumentfor
(JacquesandSchachter,1954)andwascalledwasp thephylogeneticrelationshipofthesegroups(Piek
kinin (SchachterandTain 1954). Although the 1991, 1992). However, part of this view is in
chemicalcharacterizationofthiswaspkininisstill conflictwithotherevidence(Brothers 1975).
unknown,othersocialwaspkininshavebeenchar-
acterized chemically (for reviews see Nakajima PONERATOXIN,ANEUROTOXICANTVENOM
1986andPiek1991). COMPONENT
Despite the fact that wasp kinins have been
knownformore than 35 years, theirneurotoxic Comparedtotheinformationavailableforbee
actionshadnotbeendiscoveredbeforethedemon- andwaspvenoms,knowledgeaboutneurotoxicant
strationofthepresenceofkininsinthevenomofthe venomsislimited.Nevertheless,antvenomshave
solitarywaspMegascoliaflavifrons(F.)(Pieketal. providednewtoolsforthestudyofneurophysiol-
1983, 1984b, Yasuhara et al. 1987). A point of ogy,suchaspiperidinealkaloidsinfireantvenoms
interestisthatthiswaspstingslarvaeoftheEuro- (review:Schmidt1986)andponeratoxin(Pieketal.
peanrhinocerosbeetleOryctesnasicornisL.pro- 1991a,b).
ducing an irreversible paralysis, by successively Followingenvenomationofmanbytheponerine
penetratingtheventralsideofallsegments,which antParaponeraclavata(F.),Schmidtetal.(1984)
containnerveganglia(Pieketal.1983).Thisscoliid reporteduncontrollabletremblingwhichwasnot
wasp probably injects its venom into the nerve causedbypainalone.Thevenomcontainsatleast
ganglia,aphenomenonwhichhascommonlybeen two fractions which block specifically neuronal
observedformanysolitaryaculeatewasps(seealso signalsintheinsectcentralnervoussystem(CNS)
section on philanthotoxins). The idea that these (Pieketal.1991a).Oneofthefractionsofthecrude
waspsstingintothenervegangliapromptedstudy venomwaspharmacologicallycharacterizedasa
oftheeffectofthevenomanditsfractions,andof kinin.Althoughthechemicalstructureofthisant
purified or synthesized toxins, by means of kininisstillunknown,itspharmacologicalactionis
microperfusionofaninsectganglion.Thisbecame comparable to the wasp kinins described in the
thestartofaseriesofexperimentsontheneurotoxic precedingsection.
aaccttiivoenokifntihnrienontihneev-e6-nbormadoyfkMi.nfilna(vTihfirJo,BnsK()Y,tahseumhoasrta mosTthpeotseenctonbdloncekuerrootfoxCiNcSfrfaucntcitoniopnrso.vIetdcotnotbaeintehde
etal.1987)andofColpointerrupta(F.)(Pieketal. averypotentneurotoxicpeptideof25aminoacid
1990). residues, called poneratoxin (Pieketal. 1991a).
When avenom solution, preparedofisolated This has been synthesized by ProfessorTerumi
tvheenhoamermeosleryvmopirhsooffaMn.fOlrayvcitfersonnsaswiacsorinnijsecltaervdai,notro N1a0k8atjoim1a0-(6TMokytoh)e.sAytntchoentciecntproanteiornastovxairnyi(ngPofTrXo)m
whenafemaleM.flavifronswasforcedtostingthe blockssynaptictransmissionintheinsectCNSina
larva,noparalysisoccured(Pieketal.1983).How- concentration-dependent way and it depolarizes
ever,thevenompreparationwasveryactive,asa giantinterneurons.Atcomparableconcentrations
blockerofsynaptictransmission,wheninjected(by PoTXaffectstheelectricalactivityofisolatedcock-
microperfusion)intothesixthabdominalganglion roach axons, as well as ofisolated frog and rat
ofthecockroach,PeriplanetaamericanaL.(Piek skeletal muscle fibres. The explanation ofthese
Volume2,Number1,1993 103
actionsisthefindingthat PoTXprolongsaction PTX) and -philanthotoxin (PTX-4.3.3) respec-
potentialsandthusinducesslowautomaticactivity. tively (Karst et al. 1990, 1991, Karst and Piek,
Thisisaconsequenceofaslowsodiumion-current 1991).Thesephilanthotoxinsarepolyamine-like
activationatunusualnegativevaluesofpotential structuresthatblockboththeinsectneuromuscular
anddue to slow deactivation (Pieket al. 1991b transmissionandthenicotinicsynaptictransmis-
Duvaletal. 1991,1992).Thisexplainstheinsecti- sionintheinsectcentralnervoussystem(Hueand
cidalactionofastingoftheantintoaninsectprey, Piek 1989,Karstetal. 1990.1991,KarstandPiek
andalsothefibrillationofskeletalmusclesofman 1991).
(Schmidtetal.1980)andinsects(Pieketal.1991b).
Asavenomreservoirofasingleantspecimen STINGINGBEHAVIOUR
{P. clavata)maycontainabout 1 ugponeratoxin
(Piek et al. 1991a), injection ofits content may InhisSouvenirsEntomologiquesFabre(1879-
result,forsmallinsects,inafinalconcentrationof 1910)presentedtheviewthatsolitarywaspssting
rbeestewreveoinr1con5taenndt1inj4Mec.teEdveinntoattehnethCoNfStheimvmeendoim- athceoinrcveincttriamtseidnCtNheSC(NbSup.reHsetiodbbseeertlveesd,pwreeeyvihlasvainndg
aFtoerlyvearntdebirrarteevserasibboluytb3l0ocksstinngesuroofnalPafruanpcotinoenrsa. sthoamteprbeeyetwliethlaarvmaoer)etodibfefussteuCngNoSncweeraensdtoubnsgemrovreed
cSlcahvmaitdatpeetralk.g19c8a0n).bIeftlheitsharlesu(lLtsD5i(n):a3b3oustt3in0gsu/gkgo;f twhhainchonsctei.ngAcmriocknegtshtihsreeextaimmpelse,sonwceereinSepahcehxofsptph.e
poneratoxin per kg, the final concentration of three main nerve ganglia, and Ammophila spp.
poneratoxin will be inthe orderof 10'8T0~7M, a whichstingcaterpillarsineverysegmentcontain-
levelwhichisprobablylethal.Theuncontrollable ingamajornerveganglion.
tremblingandunbearablepaincausedbyastingby Basedonobservationsovermorethantwenty
P.clavataisexplainedbythehyperactivityofthe years,Steiner(1986)testedinsphecidwaspssix
nervousnetworkcausedbyprolongationofaction differenthypothesesconcerningstingnumber,lo-
potentials. cationofsting,stingdirectionandothercharacter-
It is concluded that poneratoxin alone could istics.Conceivably,thenumber,locationanddirec-
explainthereversibleneurotoxicityofthevenom tionofsuccessivestingscouldhavebeenaffected
ofP.clavata,butthekininpresentinthevenom, by (1) soft spots in the integument ofthe prey
maycontributetotheinsecticidalactionconsider- (Ferton'ssoftspotshypothesis;Steiner1986),(2)
ably by blocking irreversibly the synaptic neu- bodysegmentation,(3)legbases,(4)completeset
rotransmission. ofgangliaor(5)gangliainvolvedin locomotion
anddefence.Thenullhypothesiswouldberandom
PARALYSISOFPREYBYPHILANTHOTOXINS stinging.Steiner(1986)concludedthatalthougha
widespectrumofstingingmethodscouldnoteasily
Thetwofirstexamplesofneurotoxins(kinins be encapsulated in a single simple formula, the
and poneratoxin) are polypeptides produced by locomotor ganglia hypothesis ofstinging is the
vespoidHymenoptera.Ourknowledgeofvenoms best-fitting one fora numberofaculeate wasps
ofthosewaspswhichbelongtoaquitedifferent which prey on large or powerful insects. These
group,theApoideaspheciformes(orsphecidwasps) wasps give at least one different sting for each
isrelativelypoor.Thevenomanditsconstituent clearlyseparatenervecentreinvolvedinlocomo-
neurotoxins has been well described for only a tion,attack,defenceorresistanceoftheprey.
single species, i.e. that of the sphecid wasp Inasinglecasethelocalizationinacrosssection
Philanthustriangulum{¥.).Thevenomcontainsat ofthemesothoraxofMitscadomesticaL.ofradio-
leasttwodifferenttoxinsthatareantagonistsforthe actively labelled venom of the sphecid wasp
glutamatergic neuromusculartransmission in in- Mellinusan'ensisL.hasbeendemonstrated(Piek
sects(PiekandSpanjer1986).Thetwotoxinsare 1978,seealsoSteiner 1986).Theextensivedocu-
chemicallycharacterizedas B-philanthotoxin(-B- mentationonstingingbehaviourinrelationtothe
104 JournalofHymenopteraResearch
effectsofstingingonprey(Steiner1986,Piekand LITERATURECITED
Spanjer1986)suggestsageneralrule:entomopha-
gousAculeatastingintotheCNSoftheprey.The Audouin, V. 1839. Deuxieme lettre pour servir de
blockingactionintheinsectCNSofthevenomsof materiaux a 1'histoire des insectes contenant des
PhiIanthustriangulum(HueandPiek1989)andof observationssurles moersdes Odyneres.A/»irt/e.v
MpoergtSasesvScetorelaiilneaurf'nlpsauvcbiolfnircsolnhusesdi(oPpnii.leoktesttuald.ie1s9i8n7)ouarlsloabsourpa-- BartdnTerersasatmnsS,scaimJc.eatnid1co7een4ss4o.NofaftActunhlreaeyaRlcloiceynsoaulP2n:eStn1os0coi4ifl-evts1ay1on3iom.afe.LocPnuhrdiioloonuss4o3pw:ha3is6cp3as-l
toriesindicatethatthevenomsofsphecidwasps 365.
ontehuerromtuhasncuPl.artrtiraanngsumliusmsidoind.nWoteafmfaecyttshpeeciunlsaetcet Batrfarmo,mJ.Pe1n7s4i9l.vaAniad.esPchriipltoisoonphoifcathleTgrreaantsablcatcikonwsasopf
thattheabilityof/3,triangulumtoparalyseitsprey theRoyalSocietyofLondon46:278-279.
(workersofthehoneybee)byaperipheraleffecton Bettini,S. 1978.ArthropodVenoms.SpringerVerlag.
musclecontractionhasbeendevelopedbecauseof Bodenheimer,F.S.1928.MaterialenzurGeschichteder
bWthrheoeundg,ahntgfteoorrgoeeutxshaemdrpelfweei,ntshiavteefnebmheaohlnaeevyiPbo.euertrwoiofarnktgehuerlsu,pmretyhi.es BrotaSEhccneuitrleosenm.acotDele.oBJguH.liyl1eme9eth7nii5nos.p5LPti0hen:yrn4lea8o,.3g-eV6Ton4hly8.e.aI.nUdnIIi.cvlWear.sssiiJtfuiynckao,tfiBoKenraolnfisnta.hse
waspissometimeskilledbyoneofthelastbeesto Dufour,L. 1841.Observationssurlesmetamorphoses
beattacked.Therefore,itmaybesaferforthewasp duCercerisbupresticidaetsurl'industrieet1"instinct
togivearandomsting,whichquicklybutrevers- entomologiquedecetHymenoptere. Annatesdes
iblyincapacitatesthebee.Subsequentlyastingis SciencesNaturelles2:353-370.
givenintothethoracicganglioncomplexandthe Duval,A..C.O.Malecot,M.PelhateandT.Piek.1991
process is often completed by a sting into the Poneratoxinconvertsfastvoltage-dependentsodium
suboesophagealganglion.Thiscompletestinging channelsintoslowoperatingonesatnegativepoten-
seqAuednicfeferreesnutl,tsbuitntaolaocnegr-ttaeirnmdpeagrraeleysciso.mparable DuvatPiloa,nlse.Ar.aP,teoCs.xtiiOcn.i,dMaeanSleceicweotnto,cxeiMn3.2f:Pr3eol7mh4a-at3ne7a5an.ntdvTe.noPime,k.re1v9e9a2l.s
cstoimnpgriengssbaehFa.vainodurLihraissnbiegernadFe.sAcrsitbiendgfinortAhempthuolreaxx NanaiCnhtaerncnoenlvserinsifornogbestkewleeetnaltmwuosgcalteinfigbrmeos.dePsflioifgtehres
ofthecockroachtransientlyparalysesthevictim. Archiv420:239-247.
During that short-lived immobility ofthe cock- Fabre,J.H.1855.ObservationssurlesmoersdeCerceris
roach, the wasp stings carefully into the etsurlacausedelalongconservationdescoleopteres
suboesophagealganglion.Onlythislatter dont ils approvisionment leurs larves. Amies de
stingresultsinadelayedandirreversiblechangeof ScienciasNaturaes4:129-150.
behaviourofthecockroach(Steiner, 1986,Pieket Fabre, J. H. 1879-1910. Souvenirs Entomologiques.
taalot.y,bpIeti1c9sma8uli4pg,pvhoet1r9nt8boe9emd).bccyloemasprtousdtiiheattsiootnfhevoefenPxo.pmltasrniaoatfniogotunhleuormfbhetaehs-e HuegDpI.rneeeBplG.sta.sigadrGSnea.edsvreLoTiu,.ennsPtPsai8,yre3niek2asd.p:..t2,1i79Nc-8e3t8u3r.r.aoEntfsofmxeic'st8ss8i.oonfInkintientrihnneasitanisnoendcatrleCClNaotSne,-d
huntingorwasp-huntingsphecidwasps.Itmaybe Jaques.R. andM. Schachter. 1954.Thepresenceof
achallengetostudentsofthebiologyofthesolitary histamine,5-hydroxytryptamineandapotentslow-
waspstocollectargumentsinfavouroragainstthe contractingsubstanceinwaspvenom.BritishJour-
abovementionedview. nalofPharmacology99:53-57.
Karst,H.andT.Piek.1991.Structure-activityrelation-
shipofphilanthotoxins-II.Effectoftheglutamatergic
gatedionchannelsofthelocust-musclefibremem-
brane. ComparativeBiochemistryandPhysiology
98C:479-489.
Volume2,Number1,1993 105
Karst,H..R.H.Fokkens,N.deHaan,G.Heuver.B.Hue. Piek,T.,B.Hue.A.Lind,P.Mantel.J.VanMarieandJ.
C.Kruk,N.M.M.Nibbering,T.Piek,W.Spanjer,Y. H.Visser.1989.ThevenomofAmpulexcompressa
C. Tong and W. van der Vliet. 1990. Beta- -effectsonbehaviourandsynaptictransmissionof
philanthotoxin,anovelglutamatergicantagonistfor cockroaches.ComparativeBiochemistryandPhysi-
insectneuromusculartransmission.ComparativeBio- ology92C:175-183.
chemistryandPhysiology97C:317-327. Piek,T.,B.Hue,P.Mantel,T.Nakajima,M.Pelhateand
Karst, H.,T. Piek,J. vanMarie,A. LindandJ. van T. Yasuhara. 1990. Threonine6-bradykinin in the
Weeren-Kramer. 1991. Structure-activityrelation- venomofthewaspColpainterrupta(F.)presynapti-
shipofphilanthotoxins-I.Pre-andpostsynapticinhi- callyblocksnicotinicsynaptictransmissioninthe
bition of the locust neuromuscular transmission. insectCNS.ComparativeBiochemistryandPhysiol-
ComparativeBiochemistryandPhysiology98C:471- ogy96C:157-162.
477. Piek,T.. A. Duval. B. Hue, H. Karst, B. Lapied, P.
Nakajima,T. 1986. Pharmacologicalbiochemistryof Mantel,T.Nakajima,M.PelhateandJ.O.Schmidt.
vespid venoms. In: T. Piek, ed.. Venoms ofHy- Poneratoxin,anovel peptideneurotoxinfromthe
menoptera,pp.309-327. AcademicPress,London. venomoftheant,Paraponeraclavata.Comparative
Piek,T.1978.Wespegifalsnatuurlijkinsecticide.Natuur BiochemistryandPhysiology99C:487-495.
enTechniek46:l5S-l73. Reaumur(Ferchauld,R.A.).1742.Memoirespourservir
Piek.T. 1986.VenomsofHymenoptera:Biochemical, aI'histoiredesinsectes,T.VI.ImprimerieRoyale,
Pharmacological andBehaviouralAspects. Aca- Paris.
demicPress,London. Schachter,M. andE. M.Thain. 1954.Chemical and
Piek,T. 1991. Neurotoxickininsfromwaspandant pharmacologicalpropertiesofthepotentslowcon-
venoms.Toxicon29:139-149. tractingsubstance(kinin)inwaspvenom.British
Piek,T.1992.Atoxinologicalargumentinfavourofthe JournalofPharmacology9:353-359.
closerelationshipofVespidae.Scoliidae.Tiphiidae, Schmidt, J. O. 1986. Chemistry, pharmacology and
MutillidaeandFormicidaetHymenoptera).Proceed- chemicalecologyofantvenoms.In:T.Piek,ed.,
ingsofExperimentalandAppliedEntomology3:99- VenomsoftheHymenoptera,pp.425-508.Academic
104. Press,London.
Piek,T.andW.Spanjer.1986.Chemistryandpharma- Schmidt,J.O.,M.S. BlumandW.L.Overal. 1984.
cologyofsolitarywaspvenoms.In:T.Piek,ed.. Hemolyticactivitiesofstinginginsectvenoms.Ar-
Venoms ofHymenoptera, pp. 161-307. Acadmic chivesforInsectBiochemistryandPhysiology1:155-
Press,London. 160.
Piek,T.,A.Buitenhuis,R.T.Simonthomas,J.G.R. Steiner,A.1986.Stingingbehaviourofsolitarywasps.
UfkesandP.Mantel.1983.Smoothmusclecontract- In:T.Piek.ed.,VenomsofHymenoptera,pp.63-160.
ingcompoundsinthevenomofMegascoliaflavifrons AcademicPress,London.
(Hym.-Scoliidae) with notes on the stinging Yasuhara,T.,P.Mantel,T.NakajimaandT.Piek.1987.
behaviour.ComparativeBiochemistryandPhysiol- Twokininsisolatedfromanextractofthevenom
ogy75C:145-152. reservoirsofthesolitarywaspMegascoliaflavifrons.
Piek,T„J. H. VisserandR. L. Veenendaal. 1984a. Toxicon25:527-535.
Changeinbehaviourofthecockroach.Periplaneta
americana,afterbeingstungbythewaspAmpulex
compressa.EntomologiaExperimentalisetApplicata
35:192-203.
Piek,T.,P.MantelandC.J.W. vanGinkel. 1984b.
Megascoliakinin,abradykinin-likecompoundinthe
venomofMegascoliaflavifrons(Fab.)(Hymenoptera:
Scolidae).ComparativeBiochemistryandPhysiol-
ogy78C473-474.
Piek,T.,B.Hue.L.Mony.T.Nakajima,M.Pelhateand
T.Yasuhara.1987.Blockofsynaptictransmissionin
insect CNS by toxins from venom of the wasp
Megascoliaflavifrons(Fab.).ComparativeBiochem-
istryandPhysiology87C287-295.
