The impact of exposure to serum lipids during in vitro culture on the transcriptome of bovine

Theriogenology81(2014)712–722

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Theriogenology

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Theimpactofexposuretoserumlipidsduringinvitrocultureonthetranscriptomeofbovineblastocysts

GaelCagnone,Marc-AndréSirard*

CentredeRechercheenBiologiedelaReproduction,DépartementdesSciencesAnimales,InstitutdesNutraceutiquesetdesAlimentsFonctionnels,UniversitéLaval,QuébecCity,Qc,Canada

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Articlehistory:

Received23June2013

Receivedinrevisedform27October2013Accepted11December2013Keywords:Fattyacid

MitochondrionEnergy

Largeoffspringsyndrome

AssistedreproductivetechnologyIn ammation

abstract

Invitroculturehasadetrimentalimpactonearlyembryonicdevelopment,andserumadditiontoIVCisrecognizedtocompromiseblastocystquality.Particularly,serumfattyacidsaffectembryoniclipidcompositionandreducecryopreservationsurvival.Tounder-standthemolecularpathwaysofserum-inducedembryonicstress,thisstudyexaminedtheearlydevelopmentofbovineembryosproducedindifferentprotein-orlipid-supplementedculturemedia:BSAalone(control),BSAþserumlipidfraction(SELF),delipidatedserumandtotalserum.Theseprotein–lipidtreatmentswereappliedfromtheeightto16cellstagestotheblastocyststage.Asplanned,SELFtreatmentincreasedthefattyacidconcentrationinthemediumcomparedwithcontrolmediumbutdidnotinduceembryotoxicity.However,microarraycomparisonbetweenblastocystsculturedinBSAwithoutorwithSELFrevealeddifferentialtranscriptomicpro leassociatedwithceramide-inducedoxidativestressandin ammation.Moreover,theSELFtreatmenthadasigni cantimpactongenesinvolvedincholesterolmetabolism(LDLR,HMGCS1),withthepotentialupstreamcontrolofthetran-scriptionfactorsSREBPandPPARA,twomajorregulatorsofcholesterolmetabolism.Inaddition,theexpressionofpluripotence-relatedgenes(APEX,CLDN6)wasdownregulatedinblastocystssubjectedtoeitherSELFortotalserum.Takentogether,theseresultsillustratehowtheearlyembryonictranscriptomerespondstoincreasedlipidexposurethroughanin ammatoryandmetabolicsignature.

Ó2014ElsevierInc.Allrightsreserved.

1.Introduction

Asuccessfulpregnancyrequiresanembryoofhighdevelopmentalcompetence.However,assistedreproduc-tivetechnologies,especiallyprolongedculture,mayhavenegativeeffectsonembryoquality.Forexample,IVCcon-ditionshavebeenshowntoimpactblastocystdevelopmentandalsofetal,placental,andpostnatalgrowth[1–3],asdemonstratedbythehighincidenceofthelargeoffspringsyndromeincattle[4].Comparedwiththenaturalcondi-tionsintheoviduct,thesuboptimalIVCenvironmentgeneratesstressandcompromisestheviabilityoftheearly

*Correspondingauthor.Tel.:þ14186567359;fax:þ14186563423.E-mailaddress:Marc-Andre.Sirard@fsaa.ulaval.ca(M.-A.

Sirard).0093-691X/$–seefrontmatterÓ2014ElsevierInc.Allrightsreserved.doc.xuehai.net

embryo.AlthoughnumerouschangeshavebeenmadetoIVCprotocolstodecreaseembryonicstress,culturecondi-tionsarestillsuboptimalforproperembryodevelopment.Therefore,embryonicstressduringIVCneedstobechar-acterizedatthemolecularleveltooptimizeIVCconditionstoproduceblastocystsofhigherquality[5].

WiththedevelopmentofmicroarraytechnologiesandtheabilitytouseminuteamountsofmRNA,transcriptomicanalysishasemergedasarelevanttooltostudytheem-bryonicresponsetocultureconditions[6].Aftertheearlycleavagestages,transcriptionoftheembryonicgenomeisactivatedandthischaracterizesthematernaltoembryonictransition(8–16cellstagesinbovine)[7].Embryonicgenomeactivationiscrucialtosupportsubsequentdevel-opment[8]andseveralanalysesofglobalRNAcontentrevealedsigni cantmodi cationsingeneexpressioninIVC

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