牲畜生殖内分泌学课件

ClinicalReproductiveEndocrinologyJackyChanClinicalReproductiveEndocrinClinicalReproductiveEndocrinologyReproductivehormonesAssaymethodsPhysiologyofreproductivehormonesinthefemaleClinicalaspectsofreproductiveendocrinologyGeneralcommentsClinicalReproductiveEndocrinReproductivehormonesDefinitionofhormonesChemicalclassesofreproductivehormonesHormonereceptorsInterconversionofsteroidsintargettissuesSynthesisandclearanceofhormonesReproductivehormonesDefinitioDefinitionofhormonessynthesizedandsecretedbyductlessendocrineglandsminutequantitiesdirectlyintothebloodvascularsystemClassicendocrineglands:pituitary,thyroid,parathyroid,adrenal,pancreas,ovary,testis,placenta,pinealglandDefinitionofhormonessynthesDefinitionofhormonesReproductionpituitary,pinealglands,gonads,placentaadrenalandthyroidglandsuterus,hypothalamusDefinitionofhormonesReproducReproductiveendocrineExtrahypothalamiccentresHypothalamiccentresAnteriorpituritaryGnRHOvaryUterusPinealMelatoninEstrogens/progesteroneFSH/LHEstrogens/progesteronePGF2αEstrogensInhibinGonadotropin-releasinghormoneGonadotropichormoneSexsteroidhormonesReproductiveendocrineExtrahyChemicalclassesofreproductivehormonesPeptideandproteinhormonesReleasinghormones,InhibitoryhormonesHypophysealhormonesNeurohypophysealhormonesPlacentalgonadotropinsSubunitsSteroidhormonesProstaglandinsChemicalclassesofreproductiThehormonessecretedfromtheanteriorpituitaryThehormonessecretedfromthePeptideandproteinhormonesReleasinghormones,(GnRH,TRH,CRH,PRH,GH-RH)PeptidehormonesproducedwithinthehypothalamusAdenohypophysisInhibitoryhormonesGrowthhormone(GIH)Prolactin(PIH)portalveinTSH

Insulin,glucagonPeptideandproteinhormonesReReleasinghormonesGonadotropin-releasinghormone(GnRH)pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2(MWofGnRH1,182)regulatethereleaseofFSHandLHReleasinghormonesGonadotropinhypothalamic-pituitary-ovarianaxishypothalamic-pituitary-ovarianLHsendsthesamemessageandresultsinincreasedtestosteroneproductionbythetestisduetoLHeffectontheLeydigcellsofthetestis.

LHandFSHarereleasedintothesystemiccirculation,travelingtothetargetreproductiveorgans(testesandovary)wheresexsteroidhormonesarecontrolled.LHsendsthesamemessageandReleasinghormonesThyrotropin-releasinghormone(TRH)pGlu-His-Pro-NH2(MWofTRH362)regulatethereleaseofTSH,GH,andprolactinPathwaysofthyroidhormonemetabolismReleasinghormonesThyrotropin-ReleasinghormonesCorticotropin-releasinghormone(CRH)CRHisa41-aminoacidpeptideregulatethereleaseofACTHReleasinghormonesCorticotropiPeptideandproteinhormonesHypophysealhormonesGlycoprotein,FSHandLH,prolactinMWofLH30,000(bovine,ovine,procine,equine)MWofFSH32,000(ovine,equine)MWofprolactin23,000(bovine,ovine,procine)FSH,LH,TSH,ACTH–producedinbasophiliccellsProlactin–localizedinacidophiliccellsPeptideandproteinhormonesHyProlactinLuteotrophichormone

(LTH):acorpusluteum-stimulatingeffectinmouse,rat,dogandcat198a.a,MW23,300anda15-minutehalf-lifeinsheepincreasinginlactation(mamotropin)stimulatingthepigeoncropglandtosecret“cropmilk”ProlactinPeptideandproteinhormonesNeurohypophysealhormones

Oxytocin,Vasopressin(synthesizedinhypothalamus)PosteriorpituitaryreleaseAxoplasmicfluidStimulationoftheNcellPeptideandproteinhormonesNePeptideandproteinhormonesPlacentalgonadotropinsPMSG(eCG,MW70,000),aproteinhormoneinserum,producedbyfetaltrophoblasticcellsofpregnantmaresThefunctionofendometrialcupsbeginsonday36until120daysofpregnancy.hCG(MW36,000),aproteinhormone,extractedfromtheurineofpregnantwomenfrom30-40daysofpregnancyglucocortocoid,estrogen,progestin,relaxin…PeptideandproteinhormonesPlEndometrialcupsEndometrialcupsdevelopfromcellsofthechorionicgirdle,circumferentiallyaroundtheconceptusatapointwherethemembranesoftheallantoisandyolksacmeet.

EndometrialcupsEndometrialcu270days1.TherelationshipofeCGandCLduringequinepregnancy2.eCGhasbothFSH-likeandLH-likeactivity,itprovidesthestimulusfortheformationofaccessoryCLsandmaintainaplateauofplasmaprogesteronefromabout50-140days.

270days1.TherelationshipofOneStephCGUrinePregnancyTest

hCGisalsousedtotreatwomenwithcertainovariandisorders.ClinicaluseofhCG

OneStephCGUrinePregnancyTAthletesusehCGtoincreasethebodynaturalproductionoftestosteronewhichisoftendepressedbylongtermsteroiduse.HCGtherapyhasbeenfoundtobeveryeffectiveinthepreventionoftesticularatrophyandtousethebodyownbiochemicalstimulatingmechanismstoincreaseplasmatestosteronelevelduringtraining.ClinicaluseofhCG

AthletesusehCGtoincreasetPeptideandproteinhormoneshCGLHhCG-subunitα:96a.asubunitβ:N-terminalspecificforhCGLH-subunitα:96a.asubunitβ:119a.aspecificforLHFSH-subunitα:96a.asubunitβ:specificforFSHTSH-subunitα:89a.ainhumansubunitβ:112a.ainhumanPeptideandproteinhormoneshCPeptideandproteinhormonesSubunits,

,

-subunit,uniqueforeachhormonewithinaspecies,determinesthebiologicalactivity.PeptideandproteinhormonesSuChemicalclassesofreproductivehormonesSteroidhormonesderivedfromtheprecursor,cholesterolMW<500Estrogens(estradiol)Androgens(testosterone)Progesterones(progesterone)ChemicalclassesofreproducticholesterolC27H46OC21H30O2

C19H28O2

C18H24O2

cholesterolC27H46OC21H30O2C1ChemicalclassesofreproductivehormonesProstaglandinsagroupof20-CunsaturatedfattyacidsMW:300-400localhormones,shortbiologicalhalf-lifewithalocalactionfoundinmammaliantissues,uterusandprostataChemicalclassesofreproductiBiosynthesisofPGfromarachidonicacidperoxidaseBiosynthesisofPGfromarachiSynthesisofprostaglandinaffectedbyestrogen

PhospholipidsArachidonicacidProstaglandinH2PGEPGFPGDInactivatedPhospholipaseA2Estrogen↑oxytocinreceptors↑Progesterone↓ProstaglandinH2synthetaseEstrogen↑Progesterone↓:1.proestrus→estrus2.priortoparturitionActivatedPhospholipaseA2SynthesisofprostaglandinaffPGE2stimulatesvasculardilation

bronchiodilation

gastrointestinalanduterinecontraction

inflammatoryresponsemimicshormonesthatactthroughcyclicAMPinhibitsplateletaggregationPGF2

stimulatesvascularconstriction

bronchioconstriction

smoothmusclecontraction,e.g.intestinaltract,uterus

PGI2inhibitsplateletaggregation(increasescAMP)

vascularconstriction

bronchioconstriction

gastrointestinalanduterinecontractionFunctionofprostaglandins

PGE2stimulatesvasculardilatiovascularuteroovarianpathwaysforPGF2αUterinebranchofOVOvarianbranchofOVvascularuteroovarianpathwaysvascularuteroovarianpathwaysforPGF2αSystemicuteroovarianpathway:Localuteroovarianpathway:Combinedsystemicandlocaluteroovarianpathway:horse,rabbitsheep,cattle,(guineapig,hamsters,swinerats)?vascularuteroovarianpathwaysClinicaluseofPGF2

endometritis/pyometraClinicaluseofPGF2endometriClinicaluseofPGF2

inductionofestrusClinicaluseofPGF2inductionOnsetofestrusOvulationprogesteroneCorpusluteum/NormalestruscycleOnsetofestrusOvulationShortenestruscyclewithPGF2

PGF2

injectionforinductionofestrusOnsetofestrusOvulationOnsetofestrusprogesteroneCorClinicaluseofPGF2

inductionofabortion/parturitionClinicaluseofPGF2inductionClinicaluseofPGF2

inductionofabortion/parturitionClinicaluseofPGF2inductionClinicaluseofPGF2

inductionofabortion/parturitionClinicaluseofPGF2inductionClinicaluseofPGF2

misallianceinductionofabortionendometritis/pyometraonlyinthecaseofopenpyometra!!ClinicaluseofPGF2misallianHormonereceptorsSteroidhormonesfatsoluble,abletoenterallcellslowconcentration(estrogen:10-150pmol/Linnon-pregnantfemale)Receptorsenablethetargetcells

-toconcentratethehormonewithinthecell-toelicitparticularcellularresponses-tocreateabiologicalresponseonspecificbindingHormonereceptorsSteroidhormoThesteroidhormonereceptorismadeupofthreedomains:amodulatingN-terminaldomain(transcription-activationdomain),aDNA-bindingdomainaC-terminalsteroid-bindingdomain.

ThesteroidhormonereceptoriHormonereceptorspassivediffusionTargetcellsreceptorsnon-protein-boundhormoneprotein-boundsteroidhormonesteroid-receptorcomplexRNAsynthesis↑proteinsynthesis↑TranslocationofcomplexintonucleusE2:mammaryglandP4:endometriumT:sertolicellHormonereceptorspassivediffuHormonereceptorsTargetcellsreceptorsactionsofFSHandLHingonadsProteinhormoneprotein-receptorcomplexcyclicAMPproteinkinaseactionsofcholesterolGproteinHormonereceptorsreceptorsacti牲畜生殖内分泌学课件TissuesresponsivetoandrogensTissuesresponsivetoandrogenInterconversionofsteroidsintargettissuestestosterone5α-reductase5α-dihydrotesterone(5α-DHT)TargetcellsInterconversionofsteroidsinProgesteroneconversionintheplacentaCholesterolPregnenoloneProgesterone17α-HydroxyprogesteroneAndrostenedioneEstrone17α-HydroxylaseCortisolTestosteroneEstradiol17βPGF2α↑parturitionProgesteroneconversionintheSynthesisandclearanceofhormonesSynthesisandclearancearemainthefactors,whichinfluencethehormoneconcentrations.Theconcentrationofahormonereflectsitsrateofsynthesisorsecretion.metabolizedbyliverdetectionofthebodyfluids(plasma,urine,saliva,feces…)reproductivecondition,exwildanimalsSynthesisandclearanceofhorSynthesisandclearanceofhormonesunderthecontrolofgonadotropinsinpulsatilemode♂Thesecretionoftestosterone

changesinpulsatilerate.varyingfrom3.5–20nmol/Lwithinhours♀ThepulserateofE2andP4

remainsstable.SynthesisandclearanceofhorSynthesisandclearanceofhormonesPlacentagonadotropins(hCG,eCG)producedinhighconcentrationlongerhalf-lifethanthepituitarygonadotropinsFSH,LH≒10-30minuteshCG≒1.5dayseCG≒6daysequineLH>otherspeciesSynthesisandclearanceofhorSynthesisandclearanceofhormonesPGF2α15-keto-13,14-dihydro-PGF2αHalflifeprimaryPGF2α<20seconds15-keto-13,14-dihydro-PGF2α≒8minutes25mgPGF2α,IMincows>90%excretedintheurineandfeces(2:1)over48hoursSynthesisandclearanceofhorAssayMethodsQuantificationofspecifichormonesisnecessaryfortheendocrinologicalevents.Weight-gainchangechemicaldeterminationsforsteroidhormonesurineanalysisAssayMethodsQuantificationofImmunoassaytechniqueAgAbImmunoassaytechniqueAgAbAssayMethodsImmunoassayRadio-immunoassay(Bersonetal.,1959),Sen:10-9Enzymemultipledimmunoassaytechnique,Sen:10-6Fluoescineimmunoassay,Sen:

10-9EIA(Engvalletal.,1970s),Sen:10-12Chemiluminescenceimmunoassay,Sen10-15AssayMethodsImmunoassayRadioimmunoassayBecauseoftheeasewithwhichiodineatomscanbeintroducedintotyrosineresiduesinaprotein,theradioactiveisotopes125Ior131Iareoftenusedastheradioactiveantigen.RadioimmunoassaySandwichEIASandwichEIAcompetitiveEIAcompetitiveEIAsemiquantitativeELISAtestsemiquantitativeELISAtestThechemicalluminolisoxidisedbyH2O2,toproduceamoleculeinanexcitedstate.Whentheexcitedelectronsinthemoleculesfallbacktotheirgroundstates,lightisgivenout.LuminolExcitedState

GroundStateThechemicalluminolisoxidisAssayMethodsImmunoassayRadio-immunoassay(Bersonetal.,1959),Sen:10-9CompetitiveproteinbindingEIA(Engvalletal.,1970s),Sen:10-12Chemiluminescenceimmunoassay,Sen10-15DiagnosticaidsTherapy-monitoringAssayMethodsImmunoassayNorthernsemi-sphereSubtropicalTaiwanMonitoringtheseasonalestrusactivityofmareNorthernsemi-sphereSubtropicaPhysiologyofreproductivehormonesinthefemaleEstrouscycleControlofthecorpusluteumEarlypregnancyPregnancyandparturitionPhysiologyofreproductivehorEndocrinecontrolofcyclicalreproductiveactivityEndocrinecontrolofcyclicalThedurationofestrus,timeofovulationanddurationofCLfunctionindomesticanimalsThedurationofestrus,timeoInfluenceofthelengthofdaylightontheovaryfunctioninmaresPhotoreceptorsintheeyesPinealHypothalamusAnteriorpituitaryOvariesincreasingdaylightdecreasingdaylightneurotransmissionportalsystembloodcirculationMelatoninMelatoninGnRHGnRHGonadotropineGonadotropineInfluenceofthelengthofdayestrousbehaviorofmaresestrousbehaviorofmaresLargeamountsofequineLHarereleasedduringan8-to9-dayperiodwithovulationoccurringonthe3rdto5thday.

Controlofthecorpusluteum

8-9daysLargeamountsofequineLHare牲畜生殖内分泌学课件thecainternahasLHreceptorandproducestestosterone.granulosacellhasFSHreceptorwhichcausescelltotaketestosteronefromthecaandconvertittoestradiol.dominantfolliclehasLHreceptorsongranulosawhichcanrespondtoLHtodriveprogesteroneproduction.

Eachgranulosacellhasapproximately1500FSHreceptors

P4+feedbackLH

thecainternahasLHreceptorOvariansteroidogenesisFSHcausesincreasedaromataseactivity,whichconvertstestosteronetoestradiolProductionRateofSexSteroidsinWomenatDifferentStagesoftheMenstrualCycle

DAILYPRODUCTIONRATESEXSTEROIDS*Early

Follicular

Preovulatory

MidlutealProgesterone(mg)142517Hydroxyprogesterone(mg)0.5

4

4

Dehydroepiandrosterone(mg)77

7

Androstenedione(mg)2.64.73.4Testosterone(mg)144

171

126

Estrone(mg)50

350

250

Estradiol(mg)36380250OvariansteroidogenesisFSHcaucumuluscellsproducehyaluronicacidandproteinstocausecumulusexpansionincreasedbloodflowtofollicle(vascularpermeability↑)ovulationcumuluscellsproducehyaluron牲畜生殖内分泌学课件Triggersincreaseproteinandsteroidsynthesis,andcellulardifferentiationwithinfolliclePlasminogenisconvertedtoplasminbyplasminogenactivator(activecollagenase↑).Prostaglandinstimulatessmoothmuscletocontractandforcetherupturedfollicletoempty.

ovulationTriggersincreaseproteinand牲畜生殖内分泌学课件ControlofthecorpusluteumCLisresponsiblefortheestrouscyclicity.UteruscontrolsthelifespanoftheCLinthecow,ewe,sowandmare.EstrogensinitiateuterustosynthesizesPGF2α.RegressionofCLisaccomplishedwithin48hrs.ControlofthecorpusluteumCLControlofthecorpusluteumEstrogensendometrialoxytocinreceptorsinitiatethereleaseofPGF2α,whichcausesthereactionoflutealregression.Inruminants,releasingofPGF2αisinducedbyoxytocinsecretedbytheCL.Indogandcat,PGF2α

CL?ControlofthecorpusluteumEsControlofthecorpusluteumLHstimulatesthesecretionofprogesteronefromsmalllutealcellswhilePGF2αinhibitssecretionofprogesteronefromlargelutealcellsandcausesdegenerationofthesecells.ControlofthecorpusluteumLHEarlypregnancyMaternalrecognitionmodificationofPGF2α

bytheluteotropicproductsfromtheconceptusanduterusTrophoblastproteins(cow,ewe,doe)Estrogens(sow,mare)intrauterinemobilityoftheembryo(mare)?(bitch,queen)antiluteolyticactionorluteotrophiceffecttoextendthelutealactivityEarlypregnancyMaternalrecognEarlypregnancyTrophoblastproteins(cow,ewe,doe)

oTP-1secretedbetween10and21-24daysbTp-1secretedbetween16-19dayscTP-1secretedonDay17tobinduterineoxytocinreceptorstoblocktheuterinePGF2αsecretionEarlypregnancyTrophoblastproTrophoblastproteinsOvinetrophoblastprotein(oTP-1,IFN-τ)MW18,000,producedonDay10inhibitingtheuterineoxytocinreceptorsinhibitingthesynthesisofPGF2αPGF2↓,PGE2↑TrophoblastproteinsOvinetropTrophoblastproteinoxytocinreceptorTrophoblastproteinoxytocinreEarlypregnancyConceptus-derivedestrogens(sow,mare)estroneandestradiol17-β

convertedfromprogesteroneonDay11,Day14-18rapidenlongationoftheblastocystreleasingofcalcium,specificproteinsalteringanendocrineofPGtoanexocrinePGmetabolizedtoinactivePGFMPGF2PGE2EarlypregnancyConceptus-derivThesecretionofestrogensbytheblastocyststimulatesthereleaseofcalcium,specificpolypeptidesandproteins.alteringthePGF2fromtheendocrinetoanexocrineintrauterinePGF2metabolisedtoinactivePGFMbyfetalmembranesThesecretionofestrogensbyThereisextensivemobilityoftheequineconceptuswithinthehornsanduterinebodybeforefixationoccursbetweendays16and18.??EndometrialProstaglandinSynthesisInhibitor,(EPSI)fromendometriumThereisextensivemobilityofTimeofmaternalrecognitionofpregnancy

DayofDayofdefiniterecognitionattachmentMare14-1636-38Cow16-1718-22Sow1218Ewe12-1316Goat17TimeofmaternalrecognitionoCorpusluteum&PregnancyP4fromCL:cow,sow,goat,dogandcatP4fromCL+fetoplacentalunit:mare,sheep

Corpusluteum&PregnancyP4frPregnancy1.Thesheepplacentaproducesenoughprogesteronebyday60-70.2.Lutealprogesteroneisrequiredthroughoutgestationincattleandgoatsbecausetheirplacentasecretemuchsmallerquantitiesofprogesterone.Pregnancy1.Thesheepplacenta牲畜生殖内分泌学课件Fetal-PlacentalUnitDihydroepiandrosteroneFetal-PlacentalUnitDihydroepiParturitionwithin12hoursParturitionProgesterone

Parturitionwithin12hoursParParturitionParturitionFetalhypothalamusFetalpituitaryACTHFetaladrenalAdrenalcorticosteroidsFeto-placentalestrogensCotyledons/myometriumReleaseofPGF2

MyometrialcontractionsPosteriorpituitaryOxytocinFetalmaturationDecreaseinplacentalP4LuteolysisoftheCLofpregnancyDecreaseinserumP4IncreasedmyometrialexcitabilityAdrenalineRelaxationandsofteningofcervixpelvicligaments,perineumRelaxin?ExpulsionoffetusStimulatesvaginaandcervixAbdominalstrainingCortisol

ParturitionFetalhypothalamusFetalpituitParturitionProstaglandinsaresolubleinfatandwater,ProstaglandinspassfromcelltocellActions:smoothmusclecontractionluteolysissofteningofcervicalcollagendevelopinggapjunctionsmovementofCa2+betweenthemyofibrilsPGF2α,PGE↑,PGI2↓ParturitionProstaglandinsareSmoothmusclecellsSmoothmusclecellsmolecules(<1000daltons)thatcanpassincludecalciumions,cAMP,ATP,glutathione,andlargemoleculeslikeaminoacidsandsugarsetc.

molecules(<1000daltons)thaSmoothmusclecontractionMuscleprotein:myosinandactinCalmodulinactivatesMLCkinaseCalcium-CalmodulinMLCkinaseinactivemyosinlightchaincontractionsactomyosincomplex

(M*-ATP)+A<---->(M*-ADP-A)+Pi(M*-ADP-A)<----->(M-A)+ADP

SmoothmusclecontractionMusclSmoothmusclecontraction1.Calciumionscomefromoutsideofthecell.2.Calciumionsbindtocalmodulin-MLCK

complexonmyosin.3.TheenzymecomplexbreaksupATPintoADPandtransfersthePidirectlytomyosin.4.ThisPitransferactivatesmyosin.5.Myosinformscross-bridgeswithactin.6.Whencalciumispumpedoutofthecell,thePigetsremovedfrommyosinbyphosphatase.7.Themyosinbecomesinactive,andthemusclerelaxes.Smoothmusclecontraction1.CThetopviewshowsarelaxedsmoothmusclecell.

Notethefocaldensitiesandthenetworkofactinandmyosinfilaments.

Whencontracted,thefilamentsslidetogetherandpullthecelltoamoreroundedappearance.SmoothmusclecontractionThetopviewshowsarelaxedsCalmodulinisoneoftheprimaryreceptorsofintracellularcalcium.4Ca2+bindtocalmodulin.Ateachbindingsite,Cainteractswithoxygenatoms,mainlyglutamateandaspartaseside-chaincarboxylgroup.Calmodulinmediatesprocessessuchasinflammation,metabolism,apoptosis,musclecontraction,nervegrowth,immuneresponse….CalmodulinisoneoftheprimaParturitionActionsofestrogenscontractileproteinsynthesis↑receptorsofoxytocinandPGF2α

↑calmodulinsynthesis↑MLCkinaseactivity↑gapjunctions↑ParturitionActionsofestrogenParturitionActionsofprogesteronegapjunctions↓receptorsofoxytocinandPGF2α↓synthesisofoxytocinandPGF2α↓calmodulinbinding↑ParturitionActionsofprogesteParturitionActionsofoxytocindependentonprogesterone↓,estrogens↑

stimulationofreceptorsintheanteriorvaginaand

cervixoxytocinreleaseincreasingPGreleaseincreasingCa2+releaseParturitionActionsofoxytocinClinicalaspectsofreproductiveendocrinologySteroidhormoneconcentrationsarelowerinanimalsthaninhuman.(estrogens<1/10)Progesteroneanalysisgivethemostusefulinformationonthereproductivestatus.Blood,milk,urine,salivaandfecesaresourcesforendocrineinformation.ClinicalaspectsofreproductiCow:P4forearlydiagnosisofpregnancyNegativeforecast(98%)isbetterthanpositiveforecast(67-90%).P4forpostpartumovarianactivityEstronesulfatefordiagnosisofpregnancyfromDay120,96%accuracyrateBovinepregnancyspecificproteinBforearlydiagnosisofpregnancyonDay30ClinicalaspectsofreproductiveendocrinologyCow:ClinicalaspectsofreprodP4forpostpartumovarianactivityorearlydiagnosisofpregnancySamplingtimeP4forpostpartumovarianactiFollicularcystOvariancystP4forpostpartumovarianactivityFollicularcystOvariancystP4P4forpostpartumovarianactiv