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DearEditor,
Wehavestudiedthevaluablecommentsfromyou,theassistanteditorandreviewerscarefully,andtriedourbesttorevisethemanuscript.Thepointtopointrespondstothereviewer’scommentsarelistedasfollowing:
Respondstothereviewer’scomments:
Reviewer1
Comment1:inpage3,line40,wefedrats..."changedtoratswerefedwith...
Response:Accordingtothereviewer’scomment,wehavecorrectedthesentence.Furthermore,wehavehadthemanuscriptpolishedwithaprofessionalassistanceinwriting.
Comment2:page25.Thestyleofreference40isnotright(usinginitialsforthefirstnames).Sincethispaperhasbeenpublished,thevolumeandpageNosshouldbeprovided.
Response:Thankyouforyourcarefulwork.Wehaveaddedthevolumeandpagenumbersforreference40.
Reviewer2
Comment:Iwouldliketothanktheauthorsfortheireffortsinaddressingthecriticismswithadditionalexperiments.Theonecriticismthattheydidnotaddresswasrelatingtoenergyexpenditureasthereasonthattheanimalsonthelowcalciumdietgainedmoreweight.WhileIunderstandthatperformingthisexperimentwillnotaffecttheconclusionofthismanuscript,IdobelievethatthispointcouldbediscussedintheDiscussionsection.
Response:Thankyouforyourvaluableadvice.Basedonthepreviousrevision,wefurtheraddresstherelationshipbetweenlowcalciumdietandenergyexpenditureinthesectionofdiscussionaccordingtoyourthoughtfulcomments.
Reviewer3
Comment1:Inthetextyouoftenwrite:“Aspreviouslydescribed”.Unlessthatpaperisfromyourlaboroneofthemethodpaperco-authorsisonthepresentMSthisisnotquitepropersincethestatementinfersmethoddevelopmentfromyourlab.Therearenumerousinstanceslikethatinthemethodssection;theseshouldallbechanged“accordingtothosedescribedby…..”
Response:Wearesorryforthislanguagemistake.Wehavecarefullycorrectedthisphrasethroughoutthemanuscriptaccordingtoyourcomment.
Comment2:Therearestillsomewording,sentencestructureandgrammaticalissueseveninthisbasicallywellputtogetherMS.Forexample,whileauthorsmayhavebeenexcitedaboutthedatayoucannotstartasentencewith“Excitedly”inline418or“Whatever”inline395.
Response:Thankyouverymuchtopointoutthesentencestructureandgrammaticalissuesinourmanuscript.Accordingtothecommentsfromyouandtheeditors,wepolishedthemanuscriptwithaprofessionalassistanceinwriting,conscientiously.
Comment3:Inmyviewabigomissioninthisworkisignoringtheanabolicsideoflipidmetabolismaswellasthermogenesisissues.ForexampleallanimalsconsumedthesameamountoffeedbutwehadextrafatstorageinthelowCadietgroups.Sowheredidtheextraenergygo?Zemeletal(citation34)insimilarworkindicatethatincreasedthermogenesisonthehighCadietexplainsthedissipationofdietaryenergy.FurthereventhoughZemeletal(#34)indicatedlipogenesiswasenhancedinthelowCadietsthatwasin2000andyoushouldhavemonitoredexpressionofFASandUCPeitherasmRNAabundanceoractualFAS/UCPchangesviaproteomicsorblottingtechniques.InanycasethesecontrolsaremissinghereandnotemphasizedintheMS.CasualreadingofthispaperwouldleadtotheconclusionthatthedietaryCaeffectonfatdepositionisstrictlyafunctionofincreasedordecreasedlipolysis.Whilelipolysisappearstobeamajorplayer,lipogenesisandthermogenesiscannotbeignoredforcompleteness.InFig8youalsoshowadeclineincAMPforthelowCadiet.WellbetaagonistsorcAMPenhancersregulatetranscriptionofadiposeandliverFAS(inrats(JBiolChem271:2307,1996)andrecentlywithlargeanimalmodels(HausmanetalJAnimalScience87:1218,2009andHalseyetalJAnimalScience89:1011,2011).Inaddition
Comment7:Line150-153.TheqRT-PCRmethodologyisnotatallunderstandableasyouciteaTexasA&Mpublishedpaper.ThisiscompletelyinsufficientwiththenewlyestablishedstandardsongeneexpressionviaqRT-PCR.Thereisnomentionofefficienciesofamplificationsinthesedatanorhowtheuseofthereferencegenewasestablishedetc.IthinkPfafflandBustinhaverecentlywrittenanarticleonthis;pleasetotallyrevise150-153inlinewithwhatyoudidandapplyingthenewstandards.
Response:Thankyouverymuch.BecausetheJNrestrictsthenumberoftotalwordsofmanuscript,wecitedtheTexasA&Mpublishedpaper.Inthenewlysubmittedmanuscript,wedescribethedetailedprotocolsinourlab.
Comment8:Line179onNotclearasinsentencestalkaboutdifferentATcellsourcesetc..revise.
Response:Wearesorryfornotaddressingtheadiposetissuecellsourcesclearly.Wehaverewrittenthemethods.
Comment9:AnypreviousdocumentableworkwithsiRNA?
Response:Yes,wehavedocumentableworkwithsiRNAinourresearchteam.TheresultswerepublishedinthejournalofBiochemBiop
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