Alcohol-specific Transcriptional Dynamics Of Memory Reconsolidation And Relapse Part 2

Cawv-nco retrieval ua rau sai tab sis ib ntus upregulation nyob rau hauv lub mRNA qhia ntawm Arc thiab Egr1, tab sis tsis yog ntawm Bdnf, nyob rau hauv lub DH (Fig. 2C–E, I).

Raws li lub sijhawm dhau mus, tib neeg maj mam nkag siab tshiab ntawm kev paub tsis meej ntawm kev tsis nco qab. Nyob rau hauv cov txheej txheem no, ib tug tshiab technology nco retrieval tau maj ua ib tug kev tshawb fawb hotspot.

Yog li puas muaj kev sib raug zoo ntawm cawv nco retrieval thiab nco kev ua tau zoo? Tuaj saib tam sim no! Ua ntej ntawm tag nrho cov, peb yuav tsum tau ua kom meej: Cawv nco retrieval tsis yog ib tug cais technology. Nws yog ib qho cuab yeej uas cuam tshuam rau lub hlwb neurons los ntawm cawv. Cov txheej txheem no tuaj yeem ua rau qee qhov "pw tsaug zog" nco hauv lub hlwb, tso cai rau peb rov ua kom cov kev nco no.

Qhov thib ob, peb yuav tsum paub tias kev txhim kho kev nco tsis tsuas yog tso cai rau peb "nco" ntau yam tab sis yog txheej txheem ntawm kev rov qab thiab rov tsim dua qhov nco hauv ntau txoj kev. Cov txheej txheem no tuaj yeem pab peb sib sau ua ke zoo dua thiab nkag siab txog kev paub peb tau kawm, yog li txhim kho peb lub cim xeeb.

Yog li ntawd, peb tuaj yeem xaus tias muaj kev sib raug zoo ntawm kev haus cawv nco qab thiab kev nco ua haujlwm. Thaum peb siv cov thev naus laus zis no los rov rho tawm cov kev nco yav dhau los tom qab haus cawv, peb tab tom ua cov txheej txheem ntawm kev rov tsim cov kev nco no, yog li txhim kho peb lub cim xeeb rau qee yam.

Tau kawg, peb yuav tsum nco ntsoov tias kev rov qab haus cawv cawv tsis yog ib txoj hauv kev ntev thiab txhim khu kev qha rau kev txhim kho kev nco. Yog tias peb xav kom muaj lub cim xeeb zoo rau lub sijhawm ntev, peb yuav tsum txuas ntxiv sib sau ua ke thiab txhim kho peb lub cim xeeb los ntawm kev kawm txhua hnub thiab lub neej, ua ke nrog lwm yam kev qhia kev nco.

Hauv cov txheej txheem no, peb yuav tsum muaj tus cwj pwm zoo txhua lub sijhawm, txhawm rau txhawb peb txoj kev kawm kev txhawb siab thiab kev nco muaj peev xwm. Yog li ntawd, cia peb ntsib kev cob qhia kev nco, ua peb tus kheej tsis tu ncua, thiab mus tom ntej ua siab loj ntawm txoj kev txhim kho kev nco tsis tu ncua! Nws tuaj yeem pom tau tias peb yuav tsum txhim kho kev nco, thiab Cistanche deserticola tuaj yeem txhim kho kev nco, vim Cistanche deserticola tseem tuaj yeem tswj hwm qhov sib npaug ntawm cov neurotransmitters, xws li nce qib ntawm acetylcholine thiab kev loj hlob. Cov khoom no tseem ceeb heev rau kev nco thiab kev kawm. Tsis tas li ntawd, Nqaij kuj tuaj yeem txhim kho cov ntshav khiav thiab txhawb nqa cov pa oxygen, uas tuaj yeem ua kom lub hlwb tau txais cov as-ham txaus thiab lub zog, yog li txhim kho lub hlwb tseem ceeb thiab kev ua siab ntev.

Tshwj xeeb, Arc thiab Egr1 mRNA qib peaked 30 min tom qab haus cawv nco qab, thiab rov qab mus rau theem pib hauv 60 min tom qab nco tau rov qab, zoo li NoRetrieval pawg.
Nyob rau hauv mPFC, cawv nco retrieval ua rau hloov pauv hloov pauv hauv Arc thiab Egr1 tab sis tsis yog Bdnf mRNAexpression, zoo ib yam li cov qauv qhia pom hauv DH (Fig.2G, H, J).

Qhov nce hauv Arc thiab Egr1 mRNA kev qhia hauv DHand mPFC tau ua ntej los ntawm kev nce phosphorylation ntawm qhov kev hloov pauv ntawm cAMP cov lus teb cov ntsiab lus-binding protein (CREB) (Daim duab S2), yav dhau los qhia los tswj cov kev qhia ntawm cov noob no [53].

Ua ke, cov txiaj ntsig tau pom tias qhov kev rov qab los ntawm kev nco txog cawv ua rau muaj kev hloov pauv raws sijhawm nyob rau hauv kev qhia ntawm Arc thiab Egr1 tab sis tsis yog ntawm Bdnf hauv DH thiab mPFC, tsa qhov muaj peev xwm hloov pauv ntawm cov noob no tuaj yeem koom nrog hauv kev sib koom ua ke ntawm cawv. nco.

Kev rov qab los ntawm kev nco txog cawv ua rau nce qib ARCprotein hauv DH

ARC muaj lub luag haujlwm tsim tau zoo hauv synaptic plasticity [54] andneuronal kev sib txuas lus [55], thiab nws tau pom yav dhau los ua lub luag haujlwm hauv kev sib koom ua ke ntawm ntau yam kev nco [5, 22, 46, 47].

Qhov tseem ceeb tshaj plaws, peb yav dhau los tau pom tias kev tshawb fawb txog cawv tau nce qib ARC protein ntau hauv amygdalaand mPFC [5]. Tam sim no peb nug seb ARC cov protein ntau puas tau nce ntxiv hauv DH, muab kev txhawb nqa ntawm Arc mRNAinduced los ntawm kev haus cawv nco qab (Daim duab 2C).

Yog li, cov nas tau raug cob qhia los qhia txog cawv-CPP, raws li tau piav qhia saum toj no (FigureS3A, B). Ib hnub tom qab qhov kev xeem CPP, cov dej cawv nco tau rov qab, thiab cov ntaub so ntswg hauv hlwb tau sau 60, 120, lossis 360 feeb tom qab.

Pom tau tias ARC cov protein ntau hauv DH nce 60 feeb tom qab haus cawv nco qab, rov qab mus rau theem pib hauv ib teev tom ntej (Daim duab S3). Ua ke, cov txiaj ntsig no qhia tias kev rov qab haus cawv nce ntxiv rau Arc mRNA thiab ARC proteinexpression hauv DH.

Kev txo qis ntawm ARC kev qhia hauv DH cuam tshuam txog kev haus cawv rov ua dua tshiab

Yog tias qhov nce ntawm ARC kev qhia hauv DH tom qab kev rov qab haus cawv yog qhov tseem ceeb rau kev haus cawv nco rov qab, tom qab ntawd nws qhov kev txwv tsis pub rov qab haus cawv yuav tsum cuam tshuam xws li kev nco, ua rau kev tshem tawm ntawm cawv-CPP qhia.

Txhawm rau txo qis ARC qib hauv DH thaum lub sijhawm haus cawv nco rov qab, peb siv cov tshuaj antisenseoligodeoxynucleotides (AS-ODN) qhia tawm tsam Arc mRNA [56].Knockdown ntawm ARC hauv lub hlwb thaj tsam cuam tshuam nrog kev nco txog kev sib koom ua ke thiab kev sib koom ua ke siv Arc AS-ODN yav dhau los qhia tias cuam tshuam cov kev sib koom ua ke. ntawm aversive andappetitive memories [46, 47, 56, 57] thiab reconsolidation offear memories [46, 47, 57], as well as impair morphine related memory reconsolidation [22].

Txhawm rau kuaj seb ARC downregulation puas cuam tshuam txog cawv nco rov qab thiab tshem tawm kev nrhiav cawv, peb tau cob qhia cov nas kom pom cawv-CPP (Daim duab 3B, saib daim duab S4 rau cov tib neeg cov ntaub ntawv).

Ib hnub tom qab CPP xeem 1, cov nas tau txais ib qho intra-hippocampalinfusion ntawm Arc AS-ODN los yog tswj scrambled (SCR)-ODN. Txij li thaum lub AS-ODN txo qis ARC cov protein ntau 5 h tom qab infusion [57] (Daim duab S5, AS-ODN validation), cawv nco tau rov qab 4 hafter infusion, tso cai rau qhov downregulation tshwm sim ib teev tom qab retrieval, nyob ib ncig ntawm lub ncov ntawm nce nyob rau hauv ARC. protein ntau ntau los ntawm kev haus cawv nco qab (Daim duab S3C).

Thaum qhov chaw nyiam raug sim rau hnub tom qab, peb pom tias cawv-CPP raug tshem tawm hauv cov nas uas tau txais Arc AS-ODN, qhov SCRODN-kho nas tseem qhia CPP (Fig. 3B).

Peb qhov kev tshawb pom no pom zoo tias intra-hippocampal infusion ntawm Arc AS-ODN cuam tshuam qhov kev rov ua dua ntawm cawv nco los ntawm kev tiv thaiv qhov nce ntawm ARC protein ntau ntau los ntawm kev nco qab.

Txhawm rau kuaj ntxiv seb puas muaj kev cuam tshuam lub cim xeeb no yog vim muaj kev cuam tshuam ntawm kev xa rov qab ARC induction tshwj xeeb, peb xaiv lub sijhawm thib ob hauv qhov theoretical 5–6h [9, 10] "reconsolidation window" rau Arc AS-ODN infusion.

Muab hais tias qhov kev nthuav qhia ntawm ARC protein nce siab tshaj 1 teev tom qab kev nco qab thiab rov qab mus rau hauv paus 2 h tom qab kev nco rov qab (raws li tau teev tseg hauv daim duab S3C), peb tsom rau ARC proteinexpression 3 h tom qab kev nco qab.

Yog li, peb infused Arc ASODN los yog tswj SCR-ODN rau hauv DH 2 h ua ntej nco retrieval, uas yuav tsum downregulate ARC qib 5 h tom qab, piv txwv li, 3 hafter nco retrieval (Daim duab S5B).

Nyob rau hauv ib qho chaw nyiam kev sim ua ib hnub tom qab, peb pom tias ob pawg neeg pheej tsis pom cawv-CPP (Fig. 3C).

Cov kev tshawb pom no qhia tau hais tias kev txo qis ntawm ARC qib dhau los ntawm nws qhov kev rov qab-dependentinduction tsis cuam tshuam nrog kev txuas ntxiv ntawm kev haus cawv tsis tu ncua. Ntxiv mus, ntawm no peb qhia tau hais tias ARCdependent "reconsolidation window" kav tsis ntev tshaj 3 teev tom qab kev nco qab.

"reconsolidation window", piv txwv li, ntau tshaj 5-6 teev tom qab nco rov qab [9, 10], yuav tsis cuam tshuam rau kev nco tom ntej.

Txhawm rau kuaj qhov kev xav no, peb infused Arc AS-ODN lossis tswj SCRODN rau hauv DH 4 h tom qab kev nco rov qab (Daim duab 3D), uas xav tias yuav cuam tshuam rau ARC protein qhia 9 h tom qab nco rov qab (ie, 5 h tom qab; Daim duab S5).

Kev ntsuam xyuas ib hnub tom qab tau qhia tias ob pawg neeg tau pom tias muaj kev nyiam nyiam rau cov khoom siv cawv. Cov kev tshawb pom no qhia tau hais tias kev txo qis ntawm ARC protein qhia ntau teev tom qab kev nco qab rov qab (piv txwv li, sab nraum ARC-raws li kev sib koom ua ke qhov rais) tsis cuam tshuam rau qhov nco qab qhov qhia txog cawv-CPP.

Ua ke, cov kev tshawb pom no qhia tias hippocampalupregulation ntawm ARC protein qhia tau pom sai sai tom qab haus cawv nco qab yog qhov yuav tsum tau ua kom rov qab haus dej haus cawv, vim tias inhibition ntawm cov retrieval-induced nce qib ntawm ARCprotein ua rau poob ntawm cawv-nrhiav, yuav cuam tshuam cov txheej txheem nco rov qab.

Thaum muab rov qab, appetitive alcohol- thiab non-alcohol-associatedmemories qhia zoo xws li Arc thiab Egr1 transcriptional dynamics

Peb qhov kev tshawb pom cuam tshuam txog Arc thiab Egr1 kev qhia nyob rau hauv kev sib koom ua ke ntawm cawv nco yog nyob rau hauv txoj kab nrog cov kev tshawb fawb yav dhau los uas qhia tias cov IEGs cuam tshuam rau hauv kev sib koom ua ke ntawm ntau hom kev nco [19, 22, 46, 47, 57].

Peb xav tias qhov kev hloov pauv thiab kev txhais lus ntawm IEGs no tsis yog kev haus dej cawv tshwj xeeb, thiab tuaj yeem ua ib feem ntawm cov txheej txheem hauv paus rau kev ua cov kev nco rov qab, suav nrog kev nco qab [16]. Txhawm rau tshawb nrhiav qhov muaj peev xwm ntxiv no, wetested seb qhov kev rov qab los ntawm cov dej cawv tsis muaj cawv, sucrose-koom nrog kev nco los ntawm CPP raws tu qauv zoo sib xws yuav hloov Arc thiab / lossis Egr1mRNA qhia, raws li nws tau ua nrog kev nco txog cawv.

Rau qhov no, peb thawj zaug tau cob qhia cov nas hauv cov txheej txheem sucrose-CPP zoo ib yam li cov txheej txheem cawv-CPP siv saum toj no, ua ke ib qho chaw ntawm CPP apparatus nrog yeem noj sucrose pellets (Fig. 4A, Kev sim tsim).

Tom qab plaub qhov kev sib koom ua ke, cov nas pom tau tias muaj kev nyiam nyiam rau sucrose-sociated compartment (Fig.4B). Tom qab ntawd, lub cim xeeb suav nrog sucrose tau muab rov qab los ntawm kev nthuav tawm mus rau sucrose-paired compartment. Cov ntaub so ntswg ntawm lub hlwb los ntawm Kev Tshawb Fawb thiab Tsis Muaj Kev Tswj Xyuas Cov Pab Pawg tau sau 10, 30, lossis 60 min tom qab nco tau rov qab. Arc thiab Egr1 mRNA theem hauv DH thiab mPFC tau raug soj ntsuam.

Peb pom tias sucrose nco retrieval ua rau sai thiab hloov pauv hloov pauv ntawm Arc thiab Egr1 mRNA qhia hauv DH thiab mPFC (Daim duab 4C–F). Raws li tau piav qhia los ntawm cov kab dashed hauv Fig.

4C-F, cov qauv ntawm mRNA qhia upregulation nyob rau hauv ob lub hlwb cheeb tsam, induced los ntawm retrieval ntawm sucrose nco, zoo li lub upregulation ntawm cov noob tau soj ntsuam raws li dej cawv nco retrieval.

Cov kev tshawb pom no yog li qhia tias kev hloov pauv sai thiab hloov pauv hauv Arc thiab Egr1 mRNA kev qhia hauv DH thiab mPFC cuam tshuam nrog kev rov qab los ntawm ob qho tib si cawv thiab sucrose-nco nco.

Cov cwj pwm ntawm cov cawv tshwj xeeb transcriptional dynamics rau nco retrieval: ib tug transcriptomic tsom xam

Muab peb qhov kev tshawb pom tias Arc thiab Egr1 cov ntaub ntawv hloov pauv tau hloov pauv thaum khaws cov cawv cawv- thiab tsis muaj cawv nrog kev nco, tom ntej no peb nrhiav kev txheeb xyuas cov ntawv sau npe tshwj xeeb rau kev nco txog cawv los ntawm kev ua RNA-seqanalysis ntawm DH thiab mPFC (Fig. 5A, Exper. tsim).

Txog rau qhov no, tom qab kev cob qhia cawv-CPP (Fig. 5B), cov kev nco txog cawv tau raug muab rov qab (nrog pab pawg tswj tsis tau rov qab). TheDH thiab mPFC tau sau 30 feeb tom qab thiab ua tiav rau RNAseq tsom xam (Fig. 5C, D).

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