Itoh  Naoki

Osada  Makoto

Takahashi G. Keisuke

English

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Osada, Makoto

Takahashi, G. Keisuke

Itoh, Naoki

Tohoku University Repository (TOUR) / 東北大学機関リポジトリ

Recent Research Topics in Reproduction andInnate System of Bivalve Mollusks著者 Osada  Makoto, Takahashi  G, Keisuke, Itoh Naokijournal orpublication titleTohoku journal of agricultural researchvolume 59number 3-4page range 63-66year 2009-03URL http://hdl.handle.net/10097/44366TohokuJournalofAgricukuralResearchVoL59No,3-4,March2009PrinteainJmpαnRecentResearchTopicsinReproductionandInnateImmuneSystemofBivalveMollusksMakotoOsADA,KeisukeG.TAKAHAsHIandNaokiIToHLab・rαto・y(ilAguacvelturαIBi吻y,(hnduateSchO・1・fAg・icztlturαIS・ience,T・1rokuUniver吻(ReGeived,March2,2009)SummaryBivalvespeciesisanimportantfisheryproduetintheindustryofaquacult皿einAsia,Oceania,Europe,andNorthandSouthAmeriGa.Anadvancedtechnol-ogyforar七ificialseed(juvenile)produc七ion,controlofreproduc七ionandgrowもhinahealthアconditionisnecessarytoachieveane伍cientproductioninbivalve閃uacul七ure.Wefocusonl)asignaltransductionofneuroho皿oneandsexhormoneviareceptOrmechanisminrelationtoreproduotionandendoGrinedisrup一もionand2)acellularandhu皿oraldefense皿eGhanisInasaninnatei皿皿u e アsternassociatedwithnon-selfrecognitionandelimina七iontoaimattheabove-meI1七ionedgoal.GonialMitosisandOogenesisThepat七emofproliferationofgonialcellswasdividedintotwophases:phaseI;oogonia、andsperma七〇goniaslowlypreliferate七hroughthegrowingstage:phaseII;oogoniadevelopintooocytesandspermatogoniastarttoprolif-eraterapidlyfromthematurestagethroughthespawningstage.Theneuronsdetectedwi七hanti-mammalianGnRHantibodyweredistributedillthecentralnervoussystem、(CNS)ofbothsexes.TheneuropeptidefractionfromtheCNSandbloodceUofbothsexespro皿otedproliferationofspermatogoniaintheゼnvitrocultureofthe七esticulartissueaswellasmGnRH,whichwasabolishedbythemGnRHantagonistsandan七i-mGnRHan七ibodア,suggestingthatthesper-matogonialproliferationisregulatedbyGnRH-1ikepeptide.Thesefindingssugges七edthatthespermatogonialproliferationatphaseIIin七hescallopwasunder七heneuroendocrinecon七TolbアGnRH.Theaccumulatedvitellinpエ〇七eininthθoocyteisanessential皿aterialforenergyproductionanddevelopmentoflarval七issueuntiltheonsetoffeeding.However,1ittleinformationhasbeenavai}ableconcerningthevitellogenin(Vtg>6364M.Osα δαetα1,synthesisandendocrinecontrolofvitellogenesisinmolluscs.ThesGallopVtgmRNAsignalwasdetectedintheauxiliarycellclosel)rassociaもedwithgrowingoocytes,suggestingthattheVtgsynthesisoeGursthroughheterosyntheticpathwaywithou七mediationthroughthebloodfiowbutoccurs面noveintheovary.Anovelneuropeptide,vitellogenesispromotingfactor(VPF),from七heCNSstronglypremotesVtgproteinsynthesis,whilethetranscriptionofVむg皿RNAispromotedbアestradiol-17β(E、}viaestrogenreceptor(ER).1七wassuggestedthatvitel-logenesisin『bivalvemollusksisunderthecontrolofE2a七thetranscriptionallevelandVPFfromtheCNSatthetranslationalleve1.Vitellogenesiscon七エ lledbythesesignal七ransduetionsissupposedtobein且uencedbyendocrinedisruptingchemicals(EDC).Weconductedananalysisforeffec七〇fEDConthegeneexpressionsofVtgandERasbiom.arkersinbluemusseLNosigni丘eantchangeinもheexpressionofeiもhertheア勾orj望」2genewasrecordedintheE2-exposedmusselinvivo,suggestingthataregula独〕rym Ghanismexists七hatisabletomain七ainGonstantleveloffreeE2byconvertingexcessE、inむoesterifiedproductwhiGh皿a)・havereduceda粗nitアfoエER.ρOocyteMaturationandSpawningSero七〇nin(5-hydroxytriptamine;5-HT)isamajorneurotransmitterthattriggersoocアtematurationandsequentialspawninginbivalve皿ollusks.How-ever,ineffec七ualityand/ori皿dividualvariationefexogenous5-HT-inducedspaw-ning伽 麗 ㊨o,leadingtounstablear七ificialinduGtion、ofspawningandseedpエodu.etion,whichisthebottleneGkofaquacultureindustr)rinbivalves,havebeenreported.Anexistenceofinhibitortoregulate5一且Tfunctionwasexpec七ed.Recentlアwedemonstratedanewpa七hwayfromthecentralnervoussyste皿(CNS)t()エegulateoocytematuration.Wefoundanovelneuroproteinwithamolecularmassof60kDa,namedoocytematurationarrestingfactor(OMAF),whichinhibitSareinitia七ionofoocアteIneiosisinducedbア5-HT.TheOMAF,whichisauniversalsubstanceforbivalvespecies,istransportedfromtheCNStotheovarアthroughbloodflowandprohibit5-HT-inducedoocytemaもuration.duetothein士erferenceofextracellularCa2+in且uxintooocytes,eventuallyresultingintheinhibitionofspawning.Ontheoもherhand,itseemsthatPGF2、inhibits5-HT-inducedtransportofmatureeggs七hroughthegonoduc七,whichisresultedfromanin、hibi七ionofthecilioexcitatoryac七ivityof5-HTintheciliatedepitheriumofthegonoduct.Weconcludedthaむoocytematurationandsubsequenteggreleasetriggeエedby5-HTaresuppエessivelγcontroUedb)・OMAFandPGF2,respectively.Repuo・o〔加o認o?zαn(llnnαteSgystem(ゾBivαlve65GriticalComponentsinHumoralDefenseMechanisms:PRPsandLysozymesInordertocombatinfectionbアbacteria,fungiandparasi七es,bivalvemollusesrelyoninnateimmunitycomprisedofnon-speGificphagocytosisbyhemocytesandhumoraldefensefaGtors.Foraprincipleofhumoraldefensemechanisms,tアpicalmicエobialpathogen-associatedmolecularpatters(PAMPs)areエecognizedanddistinguishedbymultiplepatternrecognitionproteins(PRPs),suchaslectinsandpeptidoglycanrecognitionproteins(PGRPs),andsubsequentlyvariousimmunereactionsareinduGed,Carbohydratespecificbindingproteins,lecもins,areknowntoreGognizenon-selfmoleculesandaGtasopsoninforphagocytosis,andthusconsideredasanim、porbantPRPs.Sofar,identificationofC一もアpelectins,galectinsaswellaslectincDNAsequencesan(i七he廿tissueexp1essionanalysishavebeenconductedinvariousspeciesofbivalvemollusks.0皿previousworksdemonstrated七ha七MσL-4,anovelisofor皿lectinpuri且edfro皿theplasmaoftheManilaclaln,Rudit〔4)espUhilipupuinαoiem,significant1アfa、cilit{Ltes巧b幅ocellsphagocアtosedbyc1amhemocytes,inadditiontobacteriostaticabili七y.Wealsodiscoveredpres-enceofmultiplePGRPsinthePaeificoyst}er,arαssostreαgigαs,thus,meGhanismstorecognizePAMPsapPeartobecruciallyimportantforbivalvemollusks.Lアsoz)・mes(EC3.2ユユ7>,cleavingtheglアcosidicbondofbacterialcellwallsinagreatvarietyoforganisms,areconsideredasoneof七hLerepresenthu皿oraldefensefactorsinbivalvemollusGs.Ourrecentmolecular-basedworksdemon-stratedthatthePacificoアsterpossessesthree七ypesoflysoz)τ]皿es,CGL-1,CGL-2andCGL-3,andeachofthemh{msdifferentbiochemicalpropertiesanddifferenttissueexpression.CGL-1,identifiedfrom七hehemecytes,isexpressedinmultipletissues,especiallymantle,hemocアtes,anddigestivegland,andrecom-binantCGL-1showedgreatactivityinrelativelアhighsa}tconcentra七ionsandpll.Basedonthesecharacteエistics,itisconsidered七hatCGL-1isbe七tersuitedtofunctioninthemantletissuesandthehemolymph,wherepHandsaltconcentra一七ionsareusuaUアhigh,forself-defense,althoughitsexpressioninthedigestivediverticulamayindicateanadditionalfunction,digestion.Anotherlysozyme,CGI」-2,isexpressedinthedigestivθcellsofthedigestivetubule,anditsisoelectricpointislowerもhantheotherlysozymes.Moreover,thenumberofarginineresiduesandproteasecut七ingsitesinCGL-2arefewer,sugges七ingthatCGL-2hasanincreasedresistance七〇proteolyticdigestiveenzymes,andfunctionfordiges-tion.Theもhirdlysozyme,CGL-3ismainlアsynthesizedinthemantlebutlessinthediges七ivediverticula,and七heCGL-・3seems七〇besuitedinhigherpHcondi一もion.ThesepropertiessuggestthatむhepエedictedfunctionofCGI、-3isself-defenseratherthandigestiveenzyme.66.Z14.Osαdαetα1,ReferencesItoh,N.,andTakahashi,K,(},Distribu七ionofmul七iplepep七idoglycanrθcogni-tionproteinsinthetissuesofPaci且coアster,Crαssostreαgi9αs,Oomp.Biochem.PhysinL,150B:409-417,2008.Itoh,N.,andTakahashi,K.G.GDNAGIoningandinsituhybridiza七ionofanovellysozym・ein七hePacificoアster,σrassostreagigas・σomp・Bぢoo瘤賜.Physiol.,148B:160-166,2eO7.Ma七su皿oto,T.,Nakamura,A.M.,andTakahashi,K,G,Clo且ingofcDNAsandh)Tbridizationanalysisof1)rsozymesfrom七wooys七erspecies,Oo'assos-treαgigαsandOstreαedulis,Oomp.BtOchem,Physiol,,145B=325-330,2006.Nakamura,S.,Osada,M.,andKilima,A.hvolvemenもofGnRHneuronin七hespermatogonialproliferationofbhesGallop,Pα`伽(}pectenyessoensis,!レfo乙、Ue写}rod.1)enリ.,74,108-115,2007.Osada,M.,Nakamura,S.,andKiji皿a,A.QuantitativeanalysisofthepatternofgonialproliferatienduringsexualmaturationinむheJapanesescallopPαtinopecten軍essoens哲s.Pish,Se盛,73,1318-1324,2007.Osada,M.,Tawara)rama,H.,andMori,KEstrogensアn七hesisinrelationtOgonadaldevelopmen七〇fJapanesescallop,.Pαtin()pectenyessoensis:GonadalprG丘leandimmunoloGalizationofP450aromataseandestrogen.Oompa.BfOchem.PhysbOl.PartB,139,123-128,2004.Osada,M,,Harata,M.,Kishida,M.,andKijima,A.Molecu工arcloningandexpressionanalアsisofvitellogenininsca・llop,Pαtinof)eot8nyessoensis(Bivalvia,Mollusca).Mol.Reproa.Dev.67,273-281,2004.Osada,M.,Takamura,T.,Satr〕,H。,andMori,K.Vitellogeninsynthesisintheovaryofscallop,Pα拓9z(rpectengessoensis:Controlb)Testradiol-17βandGen七ralnervoussアste皿.」,Exp.Zool.,299A,172-179,2003.Puinean,M,,Labadie,P.,Hil1,E.M.,Osada,M.,Kishi{ia,M.,Nakao,R,Novillo,A.,Callard,1.P.,andRobchell,J.]M.Laboratorアexposureto17β一esbradiolfailstoinduGevitellogeninandestrogenreceptorgeneexpressionin七hemarineinver七ebrate.Mシ拡伽se房ulis.・4guat.Tox-icol.,79,376-383,2006.Tanabe,T.,Osada,M.,K)Tozuka,K,Inaba,K,andKijima,A.Anoveloocytema加ra七ionarrestingfactorin七heGentralnervoussアstemofscallopsinhibitSserotonin-inducedooGアtema加rationandspawningofbivalvemollusks.aen.σompU.Encl・・翻oい47,352-361,2GO6・Takahashi,K.G.,Kuroda,T.,andMuroga,KPurifica七ionandantibacterialeharaGterizationofanovelisofoエmoftheManilacla皿lectin(MCL-4)fromtheplasmaoftheM乱nilaclam,Rteaitapesphilapptnαeum.σo?η多.BtOchem.」PhysCk)1.,150B,45-52,2008.Ya皿aura,K,,Takahashi,K.G.,andSuzuki,T.Idenbificationandtissueexpres-sionanal)TsisofC-typelectinandgalectininthePa(}ificoys七er,σゲαssos加θαgigαs.aomp.Bioc]tem.PhysCk)よ,149B,168-175,2008.

Recent Research Topics in Reproduction and Innate System of Bivalve Mollusks

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Recent Research Topics in Reproduction and Innate System of Bivalve Mollusks

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