I-Sucrose Ingestion ikhupha i-AMPA yokuRhweba ngokuRhweba ngokukhawuleza (2013)

J Neurosci. Umbhalo obhaliweyo ifumaneka kwi-PMC Oct 3, 2013.
Ishicilelwe kwifomu yokugqibela ehleliweyo njenge:
Inguqulelo yokugqibela yomshicileli weli nqaku iyafumaneka simahla kwi J Neurosci
Bona amanye amanqaku ku-PMC Wisdom nqaku epapashwe.

Abstract

Iindlela ezinokuthi ngazo imbuyekezo yendalo ezinje ngeswekile ichaphazele usulelo lwe-synaptic kunye nokuziphatha azivelwanga kwaphela. Apha, siphanda ngokulawula i-nucleus accumbens synapses ngokutya kwe-sucrose. Izifundo zangaphambili zibonise ukuba ukurhweba ngabantu nge-AMPA kuyindlela enkulu yokulawula amandla e-synaptic, kwaye loo nto in vitro, ukurhweba ngabantu ngokufumana i-AMPA receptors equlathe i-GluA1 subunit yenzeka ngamanyathelo amabini abandakanya i-extrasynaptic kunye ne-transaptor receptor yothutho. Sixela ukuba kumgangatho, ukuphinda-phindwa mihla le kwesisombululo se-25% scrose ngokuthe tye okoko kuphakanyiswa okuzenzekelayo kwendawo kunye nokuqokelelwa koqokelelwano olungxamisekileyo olusisiseko ngokufakwa kweCa2+Ii-receptors ezingenakulinganiswa ze-AMPA (ii-CPARs), ezine-GluA1, eziqulethe ii-receptors ze-GluA2. I-Electrophysiological, i-biochemical kunye ne-electron microscopy yophando ityhile ukuba uqeqesho lwe-sucrose (iintsuku ezi-7) lubangele uzinzo (> i-24 hr) ye-GluA1 yabantu, kwaye kwezi gundane i-sucrose stimulus ekhawulezayo (i-5 min) kodwa okwethutyana (<24 hr) iphakanyisiwe I-GluA1 kwiindawo ezingaphandle. Ii-CPAR kunye ne-dopamine D1 receptors zazifuneka in I-vivo yendawo ephakanyisiweyo emva kokufakwa kwe-sucrose. Ngokubalulekileyo, iprotocol yemihla ngemihla ye-7 yokutsalwa kwe-3% isisombululo se-saccharin, i-non-caloric sweetener, ifaka i-synaptic GluA1 ngokufanayo kwi-25% sucrose ingestion. TIziphumo ze-hese zichonga ukurhweba ngabantu nge-GluA1, kuchazwe ngaphambili in vitro, njengesixhobo sommiselo obalaseleyo wokuhanjiswa kwe-synaptic kwi vivo ngomvuzo wendalo. Ukurhweba kukhuselwa yindlela ye-chemosensory engaxhomekekanga kwixabiso le-caloric ye-sucrose.

intshayelelo

Ukusetyenziswa gwenxa kwesiyobisi gwenxa kuyingxaki ebalulekileyo yezempilo yoluntu (Hu kwaye Malik, 2010), kodwa iindlela zendalo, okanye imbuyekezo ye-orosensory enje nge-sucrose elawula ukuhanjiswa kwe-synaptic ukuphembelela isimilo ayaziwa. I-Synaptic plasticity kwi-nucleus accumbens, eyinxalenye ebalulekileyo yokujikeleza komvuzo wobuchopho (I-Sesack kunye neGrace, 2010), negalelo kwiindlela ezininzi zokuziphatha ezixhasayo, kubandakanya nokufunda ngomvuzo (Usuku kunye neCarelli, 2007), iimpendulo zoxinzelelo lwentlalo (I-LaPlant et al., 2010), kunye ne-pathologies yokulutha (ILischer neMalenka, 2011). Ukuvezwa kwe-cocaine okuphindiweyo kubangela ukuba kubekho i-synaptic plasticity kwii-neurons ze-accumbens kunye ne-ventral tegmental area (VTA)UBrebner et al., 2005; Grueter et al., 2010; UMameli et al., 2009; IPascoli et al., 2012; Thomas et al., 2001; Ungless et al., 2001). Emva kokufikelela okwandisiweyo kolawulo lobuqu lwe-cocaine elandelwa kukurhoxiswa kwexesha elide, ii-synapses zinokwenzeka ngokufakwa kwi-Ca2+-Ipermeable, i-GluA2-engekho-uhlobo lwe-AMPA receptors receptors (CPARs), olutyikityo luphawu lokungena ekufuneni i-cocaine (UConrad et al., 2008; McCutcheon et al., 2011a). Ifana necocaine, imbuyekezo ye-orosensory efana ne-sucrose eyinyusa ngokunyuka i-ascobbens dopamine (USmith, 2004), kodwa ukufunyanwa komvuzo we-orosensory (i-orosensory).

Ii-receptors ze-AMPA (AMPARs) zizinto eziphambili zokulamla zangaphakathi kwinkqubo yokuhambisa inkanuko, kwaye ukurhweba kwabo kunegalelo kwiinkqubo ezahlukeneyo ze-neural, kubandakanya ukufunda kunye nememori (I-Nedelescu et al., 2010; Rumpel et al., 2005; Whitlock et al., 2006). Ii-AMPAR zidityaniswa ziincinci ezine ezahlukeneyo, iGluA1-4. I-GluA2-ene-AMPAR ene-AMPAR yiCa2+-Ingumbono kunye nokugcwala kwetrafikhi ngokwe-synapses, ngelixa ii-GluA2-receptors receptors (CPARs), ezizii-GluA1 homomers, conduct Ca2+ kwaye ubonise ulungiso lwangaphakathi. I-GluA1 ihamba ngokurhweba ngokuthengiswa kwe-synaptic ngokuxhomekeke kumanyathelo amabini apho i-Ser 845 phosphorylation yi-cAMP-exhomekeke kwiprotein kinase (PKA) kunye ne-cGMP-protein protein exhase ye-kinase II (cGKII) ekhuthaza ukuqokelelwa kwe-receptor kwiindawo ezongezelelweyo kwi-plasma membrane (U-Esteban et al., 2003; USerulle et al., 2007; I-al et al., I-2008; I-al et al., I-2005). Emva kofakelo lwasemva kolungelelwaniso, iphosphorylation yeSer 818 yi-PKC izinza ii-AMPARs ngaphakathi kwe-synapse (Boehm et al., 2006), ebambelele kubuninzi beposti ye-postynaptic (Ehlers et al., 2007; Ewe et al., 2006; USerulle et al., 2007). Ca2+/ i-codphorylation yoxhomekeko lweproteinulin (CaMKII) ye-phosphorylation yeSer 567 kunye neSer 831 nayo inegalelo ekufakweni kwe-synaptic nasekujoliseni i-extrasynaptic (Lu et al., 2010; URoche et al., 1996), ngokulandelanayo. Nangona kunjalo, ayaziwa ukuba ngaba kwi vivo Ukudityaniswa kwee-CPAR zisebenzisa ezi ndlela zixeliweyo zikhawulezayo, kunye namanyathelo ngamanqanaba amaninzi in vitro.

Ukuphonononga iindlela apho imbuyekezo ye-orosensory efana ne-sucrose ilawula i-synapses ezivuselelayo, sisebenzise iparadigm yokufakwa ngokufutshane kwe-sucrose kunye neenguqu zokulinganisa kuthumelo lwe-synaptic kwii-neurons ze-ascens. Siphawula ukuba ukuphindaphindwa kwe-sucrose ingestion potentiates kuzisa i-synapses ngokudibanisa ii-CPAR, kwaye ukuba kwisilwanyana esiqeqeshelwe ukuzimela, isikhuthazo esinye se-sucrose sanele ukuhambisa ngokukhawuleza isiyobisi se-GluA1 kwiindawo ezi-extrasynaptic. Kungenxa yokuba i-saccharin, i-non-caloric sweetener, ebambisa ukurhweba ngokuhanahanisa okufanayo ne-sucrose, ukurhweba ngabantu ngokuphendula yindlela ye-orosensory kunokuba i-caloric pathways. Ngaphaya koko, ibhloko ye-CPAR ithintele ukunyuswa okwenziwe ngumsebenzi ozenzekelayo kwi vivo, nokuchonga ngakumbi ii-CPN zee-CPAR njengabalawuli ababalulekileyo beempendulo kwimbuyekezo yendalo.

Impahla nenkqubo

Izifundo kunye neenkqubo zokuhlinzwa

Izifundo yayingamadoda i-Sprague-Dawley iigundane (iTaconic; iimvavanyo zokuziphatha) ezinobunzima be-150-300 gram ekufikeni kunye neesetyhini ze-E18 ezikhulelweyo i-Sprague-Dawley rats (Taconic ;vavanyo lwenkcubeko yeseli). Iigundane zigcinwe i-2 kwikheji nganye yovavanyo lokuziphatha kwindlela ye-12h / 12h umjikelo wokukhanya omnyama (izibane ngaphandle kwe-18: 00) kwaye yayifikelela ekutyeni nasemanzini ad adum ngamaxesha onke. Zonke iinkqubo zokuvavanywa zavunywa yiKomiti yeZonyango kwiZikolo zaseNew York kwiKlinikhi yokuKhathalelwa kweZilwanyana kunye nokuSebenza kwaye zenziwa ngokuhambelana "neMigaqo yoLondolozo lweLwanyana yeLabhoratri" (inombolo ye-NIH yokushicilela i-85-23).

Gxotha uqeqesho kunye nemilinganiselo ye-locomotor

Iigundane zathunyelwa kwigumbi lokuvavanya i-3 iintsuku ezilandelelanayo ze-2 h / usuku kwizindlu zabo zasekhaya. Ngosuku lwesine iibhotile ezinamanzi okanye i-25% sucrose zaziswa ngesiciko se-5 min. Iibhotile zaye zalulwa. Kuzo zonke iimvavanyo, iigundane kwakufuneka ziphuze ubuncinci be-1 g ye-sucrose ngexesha lokungena kwe-5 min ngexesha leentsuku ze-3 zokuqalisa uqeqesho ukuba zibandakanywe kufundo; enyanisweni, zonke iigundane ziye zahlangabezana nale nto ibekiweyo. Emva kokususwa kwebhotile, iigundane zahlala kwigumbi lokuvavanyelwa i-30 min ngaphambi kokuba zibuyele kwindawo yezilwanyana. Ngomhla wokubingelelwa, iigundane zinikezelwa zingafakwanga zi-CO2, yagqitywa yi-guillotine, kwaye iisampulu zezicubu zaqokelelwa kwi-ice. Uvavanyo lwe-locomotor, iigundane zabekwa kwigumbi lokulinganisa le-locomotor (i-Accuscan, Columbus, OH) iyonke ye35-min. Emva kwe-15-min kwigumbi, ibhotile enesithintelo se-bead yaziswa kumphezulu wegumbi kwaye izinza. Ibhotile isuswe ukusuka kwigumbi eliphezulu emva kwe5-min, kwaye iigundane zasala kwigumbi lokufumana i15-min emva kokukhutshwa kwebhotile. Le nkqubo yaphindwa ngokufanayo kwiintsuku ezilandelelanayo ze7. Ukuhamba umgama kwakulinganiswa kusetyenziswa iVeraMax System (iAccuscan, Columbus, OH), eyayibeka iliso kwimisebenzi yezilwanyana ngegridi ye16 × 16 izibane ezikhanyayo ezinqumla kwikheji yezilwanyana (42 × 42 × 30 cm) ngaphambili ukuya ngasemva nangasekhohlo . Ulwazi malunga nemeko yebheram, ephothiweyo kwisilinganiselo samaxesha e-100 ngomzuzwana, igcinwe kwidiski. Umsebenzi ubonakaliswe njengomgama we-ambulatory umda ulinganiselwa kwi-cm ngexesha le-12 ezahlukeneyo i-3-min bins kwiseshoni ye-35 min (umgqomo wokugqibela yayiyi-2-min).

Uqeqesho lwe-Saccharin

Ukuthelekisa ukurhweba ngokurhwebesha kwe-GluA1 kunye neziphumo zokungena kwe-saccharin, i-12 yamadoda amadala amadala (i-250 g) igcinwe kwindawo yezilwanyana kumjikelo weyure / umjikelo wexesha elimnyama. Zonke iigundane ziye zahlaliswa kwigumbi lovavanyo ngokuhanjiswa kwigumbi lovavanyo, zashiywa iiyure ze12, zaze zabuyiselwa kwindawo yezilwanyana. Ngomhla omnye we-2th (emva kweentsuku ze-4 zokuhlala), iigundane zanikwa iibhotile zokufikelela ezinamanzi, i-sucrose, okanye saccharin. Iigundane ze-3 zanikwa ukufikelela kwibhotile enamanzi aphumle kumphezulu wekheji ngokuphuma kokungqinga ukuya kwikheji ngesivalo. Ixesha lokungena yayiyimizuzu ye-4, emva koko ibhotile isuswe, kwaye emva kwe-5 min imizuzu eyongezelelweyo iirhasi zahanjiswa zibuyiselwa kwiziko lezilwanyana. Iigundane ze-15 zanikwa ukufikelela kwisisombululo se-4% kwaye i-25 iigundane yanikwa ukufikelela kwisisombululo se-4% ye-saccharin (i-Sweet'n Low). Umthamo wamanzi atshisiweyo wabalwa. Le nkqubo iphindaphindwe ngeentsuku ze-3. Ngomhla we-7th wokusela, kwangoko emva kokukhutshwa kwebhotile, iigundane zenziwa idini kwaye i-ingqokelela ye-savbens yavunwa kwaye amanqanaba e-GluA7 avavanywa yi-blot Western.

I-Electrophysiology

Iigundane zaqeqeshwa ngokuchazwe ngasentla kwizisele ezicacileyo zeplastiki kwaye, emva kokususwa kwebhotile ngemini ye-7, zafakwa i-ketamine (100 mg / kg ip) kunye ne-xylazine (10 mg / kg ip) kwaye yenziwa i-transbankingly eyenziwe nge-saline ebandayo (uvavanyo lwe-MEPSC) okanye ukumiselwa kwangoko (uvavanyo lokulungisa). Iibreyiks zisuswe ngokukhawuleza kwi-fluid ye-cerebrospinal fluid (ACSF) equlathe oku kulandelayo (kwi-mM): kwiimvavanyo ze-MEPSC: NaCl (118), KCl (2.5), CaCl2 (3), MgCl2 (1), NaHCO3 (26), NaH2PO4 (1), D-glucose (10), i-osmolarity ehlengahlengisiweyo kwi-325 mOsm kunye ne-aerated yi95% O2/ 5% CO2 (pH 7.4); ukulungiselela uvavanyo lokulungiswa kwakhona: 75 sucrose, 87 NaCl, 2.5 KCl, 1.25 NaH2PO4, 0.5 CaCl2, 7 MgCl2 6 H2O, 25 NaHCO3, I-10 dextrose, ibubble nge-95% O2 / 5% CO2 (pH 7.4). Iziqwenga zeCoronal (i-300 thickm ubukhulu) eziqulathe i-nukeli eqokelelweyo zanqunyulwa kumkhenkce obandayo we-ACSF kusetyenziswa i-vibrotome (Leica, VT1200S) kwaye igcinwe ingene kwi-ACSF (ACSF, kwi-mM: 124 NaCl, 2.5 KCl, 1.25 NaH2PO4, 2.5 CaCl2, 1.5 MgSO4 I-7H2O, 26 NaHCO3, kunye ne-10 dextrose) ye <30 min; emva koko zigcinwe kwisilayidi sangaphambi kwesifudumezi kubushushu begumbi ubuncinci iyure e-1 ukuvumela ukubuyela kwimeko yesiqhelo. Uvavanyo lwe-MEPSC: isilayidi esinye emva koko satshintshelwa kwigumbi lokurekhoda apho laligcinelwe khona lingene ngenethi yenayiloni kwi-32 ° C ngesifudumezi sesisombululo se-TC324B kunye nomlawuli (iWarner Instruments, CT). Igumbi laqhubeka lisenziwa yi-ACSF kwinqanaba le-2 ml / min rhoqo. I-Medium spiny neurons ezivela kwingingqi ye-nucleus eqokelelene nengingqi echongiweyo yachongwa phantsi kwesikhokelo esibonakalayo kusetyenziswa ukungafani kokuphazamiseka okwahlukileyo kwimicroscopy yevidiyo (Hamamatsu C5405) nge-Olympus BX50WI imicroscope ethe nkqo ifakwe 40x umgama omde wokuntywila emanzini. Iipatch electrode (4-6 MΩ) ezaliswe sisisombululo sepipette se-intracellular esine- (mM): CsCl (145), HEPES (10), EGTA (0.5), kunye neMgATP (5). I-Osmolarity yahlengahlengiswa yaya kwi-290 mOsm nge-sucrose, kwaye i-pH yahlengahlengiswa yaya kwi-7.4 nge-CsOH. Imijikelezo emincinci ye-post-synaptic currents (mEPSCs) yarekhodwa kubukho be-bicuculline (10μM) kunye ne-tetrodotoxin (1μM) isebenzisa i-Axopatch 200B amplifier (Izixhobo zeMolekyuli, i-CA) kunye ne-Digidata 1322A (Izixhobo zeMolekyuli, CA). Uvavanyo lokulungisa: izilayi zigqithiselwe kwigumbi lokurekhoda zaza zathanjiswa (i-2.0-2.5 ml min-1) nge-oksijini ye-ACSF kwi-33-35 ° C equkethe i-50 μm picrotoxin yokuzahlula i-EPSCs. Ushicilelo lweeSomatic zeseli elipheleleyo lwenziwe kwi-msingi yendawo ephakathi ye-spiny neurons kwi-voltage-clamp ene-Multiclamp 700B amplifier (izixhobo zeMolezo) kusetyenziswa ividiyo ye-IR-DIC microscopy. Iipayipi zepatch (4-6 MΩ) zazaliswa sisisombululo se-intracellular (kwi-mM: 125 Cs-gluconate, 2 CsCl, 5 TEA-Cl, 4 Mg-ATP, 0.3 GTP, 10 phosphocreatine, 10 HEPUM -0.5). Idatha yahlanjululwa kwi-3.5 kHz, yadityaniswa kwi-314 kHz, kwaye yahlalutywa ngeClampfit 2 (iiMolekyyitha izixhobo). Isisusa se-extracellular (10-10 ms, 0.01-1 μA, 5 Hz) safakwa kunye neglasi encinci ye-bipolar electrode 150-0.2 mm ukusuka kwi-electrode yokurekhoda. Emva kwe ~ 0.05 imitha yokurekhodwa kwesiseko, isisombululo esine-Naspm (0.5 μM) sagqitywa ukuhlanjwa kwi-10 min. Utshintsho kwi-amplitude ye-EPSC yalinganiswa ngaphambi nangemva kokusetyenziswa kwesiyobisi ekubambeni okunokwenzeka kwe-−200, −10, −70, 50, + 30, + 0 kunye + 20 mV. Isalathiso sokuhlengahlengisa (ir) ibalwe ngokulungisa naluphi na utshintsho olunokubakho ngokuguqukayo kunye nokubalwa kule meko ilandelayo: ir = ((I-70 / 70) / (I+ 40 / 40), phi I-70 kwaye I+ 40 zii-amplope ze-EPSC ezirekhodwe kwi-−70 mV kunye ne-40 mV, ngokulandelelana.

Ukwahlulahlula-hlula ngokobuncinci kunye nokucinywa kweNtshona

Ii-Accumbens zaqokelelwa emkhenkceni njengoko kuchaziwe apha ngasentla. Xa isiseko kunye neqokobhe zahlulwa ngokwahlukeneyo, ukwahlula kwaqinisekiswa ngokufunwa kwamaqhezu e-synaptosome ye-dopamine β-hydroxylase, i-enzyme efunyenwe kwiziphelo ze-axons ukuya kwigobolondo kodwa hayi eyona nto iphambili (I-Sesack kunye neGrace, 2010). Iseli yonke, i-synaptosome, kunye namaqhezu ePSD alungiswa njengoko kuchaziwe ngaphambili (IJordani et al., 2004). I-Pellets ye-Synaptosomal yaphinda yatshintshwa kwi-200 μl 25 mM Tris ene-1% iTriton X-100, inyikima nge-4 ° C ye-30-min, kunye ne-centrifuged kwi-13,800 × g ye-15-min kwi-microcentrifuge ukuya kwi-pellet PSDs. Ipellet equlathe ii-PSD ezingalunganga yaphinda yavuselelwa kwi-25 mM Tris nge-2% SDS. Amaqhezu ahlalutywa yi-Western blot kwi-SDS-PAGE gels njengoko kuchaziwe ngaphambili (IJordani et al., 2004). Zasetyenziswa ii-antibodies ezilandelayo: dopamine β-hydroxylase (1: 1,000, Abcam), GluA1 (1: 1,000, Millipore), phosphor-Ser 845 GluA1 (1: 1,000, Millipore), GluA2LUM1LLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLXL (1,000: 1, Sigma).

Imicroscopy ye-elektroni

Ngomhla wokuvunwa kwezicubu (ngemini ye-7 yoqeqesho lwe-sucrose), iigundane ezivela kumaqela ovavanyo e-3 (Amanzi, uSurrose / Amanzi, uSunrose; iigundane ze3 / iqela lokuvavanya) zabekwa kumagumbi okulinganisa e-locomotor kwigumbi le15-min; kwi-15-min ibhotile yaziswa ngegumbi eliphezulu. Iigundane kwiqela laManzi zifumene amanzi, iigundane kwiqela le-ucrose lifumene i-25% sucrose, iigundane kwiqela leSucrose / Amanzi, elalichithe i-25% sucrose kwiintsuku ze-6, lafumana amanzi. Iigundane zazifakwe kakhulu kwi-Nembutal (50 mg / kg ip) kwaye zenziwa zaza zafakwa nge-0.1 M phosphate buffer (pH 7.4) equlathe i-4% paraformaldehyde kunye ne-0.1% glutaraldehyde ngomyinge we-50 ml / min, emva koko kwi-3-min, umyinge we-20 ml / imizuzu ye-7-min elandelayo. I-Tissue yayilungiselelwe i-postembed immunogold (i-PEG) kunye nemifanekiso yathathwa njengoko kuchaziwe ngaphambili (I-Nedelescu et al., 2010). Ii-Immunolabels zahlulwa ngokwezikhundla zazo ezinxulumene ne-PSD kwindawo ye-asymmetric synaptic junctions njenge "cleft," "kufutshane ne-PSD" (ngaphakathi kwe-1 PSD ububanzi kwi-PSD), "kwi-PSD," "intraspinous," okanye "ulwandiso lwe-extrasynaptic." isilwanyana ngasinye, ii-synapses ze-93 zaziisampulu ukusuka kumbindi we-accumbens. Isampuli ezingahleliwe yaqinisekiswa ngokuhlalutya zonke ii-synapses zokuqala ze-93 xa zatshintsha ngokulandelelana kwigridi, saza sakhokelela kwinani elifanayo le-synapses kwisilwanyana ngasinye esinye esathi safumana unyango olufana nolwangoku. Iindidi ezimbini zokwenza intsingiselo kwenziwa. Eyokuqala yayikuvavanya inqanaba le-GluR1-immunoreactivity, ngokufaka inani lamasuntsu e-PEG enzeke kwimimandla yokusebenza yomqolo. Olunye ibikuvavanya umyinge weescraps ezibhalwe kwi-PSD ngalo naliphi na inani lamasuntswana e-PEG. Nokuba ii-synapses ezifakwe ilebhile nje ye-1 PEG particle yaqwalaselwa njengebhalwe kuyo, isekwe kumsebenzi wangaphambili obonisa ubunyani benkqubo ye-GluR1-PEG (I-Nedelescu et al., 2010). Iziphumo zonyango kumyinge kunye nenqanaba le-GluR1-immunolabeling zahlaziywa ngendlela ye-ANOVA yendlela ngokuthelekisa okucwangcisiweyo kwe-hoc (Fisher's LSD). Ukuphelisa ucalucalulo, idatha yenziwa yenziwa ngamehlo kathathu: umvavanyi omnye wenza uqeqesho lwe-sucrose kwaye wagcina iirekhodi zezilwanyana kumaqela ovavanyo amathathu, umvavanyi wesibini wenza i-micrographs ze-elektroni kwaye wanika ikhowudi entsha ye-alphanumeric kwi-micrograph nganye kwaye wagcina ikhowudi evaliweyo , kunye nezilingo ezongeziweyo ezithathu zaskena ii-micrographs zaza zamiswa amasuntsu e-PEG. Emva kokugqitywa kokugqitywa kwe-PEG, abavavanyi badibana ukuze babonise ubuchule be-micrograph nganye.

Ukufakwa kweCannula kunye nenaliti ye-intracranial

Inaliti ye-Intracranial yaqeshwa ukuhambisa i-Naspm kunye ne-APV kwisiseko se-accumbens. Ukufakwa kwe-cannula, njengoko kuchaziwe ngaphambili (UCarr et al., 2010), iigundwane zazingafakwanga kangako ngetepamine (100 mg / kg ip) kunye ne-xylazine (10 mg / kg ip) kwaye yajoveka emva kokungasebenzi kakuhle nge-analgesic benamine (1 mg / kg subcutaneous). Iigundane zazifakwe stereotaxically zafakwa ezimbini ze-26-geji yesikhokelo se-cannulae (PlasticsOne, Roanoke, VA) ngokulandelelana kumanqanaba aqokeleleneyo kunye nezihlanganisi: i-1.6 mm ingaphandle kwe-bregma; I-2.9 mm kamva kwi-sagittal suture, iingcebiso zifakwe kwi-8 ° ukuya kumbindi, i-5.6 mm ventral ukuya kumphezulu wecala. I-Cannulae yayibanjelwe endaweni yi-acrylic zamazinyo kwaye i-patency yagcinwa ngesitayile sokuhambisa. Kwisitofu se-intracranial, i-Naspm kunye nezisombululo ze-APV zalayishwa kwi-30 cm ubude be-PE-50 tubing eqhotyoshelwe kwelinye icala ukuya kwi-25-μl Hamilton syringes ezaliswe ngamanzi afakiweyo kwaye kwelinye icala kwi-31-gauge injin cannulae, eyandisa i-2.0 mm ngaphaya kwezikhokelo ezifakiweyo. Iirinji zibekwe kumawele abephethe i-harvard 2272 microliter syringe pump ehambise umthamo we-0.5 μl inaliti ngaphezulu kwexesha le-100 sec. Umzuzu omnye emva kokugqitywa kwenaliti, i-injnulae ye-injnulae yasuswa kwizikhokelo, izitayile zatshintshwa, kunye nezilwanyana zabekwa kumagumbi okuvavanya afunwayo. Ukulandela ukubingelela kwezilwanyana, amacandelo obuchopho be-cryogenic ahlalutya ukuphuculwa kwendawo ye-cannula; Izilwanyana ze-2 ngaphandle kwe-15 izilwanyana azifakwanga kufundo ngenxa yokubekwa okungafanelekanga kwe-cannula.

Uhlalutyo lwesatisatisti

I-ANOVA yendlela enye elandelwa ngovavanyo lwe-hoc post ye-hoc yayisetyenziselwa i-elektroni microscopy, immunocytochemistry kunye novavanyo lwe-biotinylation. Iimvavanyo ezimbini zovavanyo lwe-t zabafundi zasetyenziselwa i-electrophysiology. Uvavanyo lwe-sucrose tester, iindlela ezimbini ze-ANOVA zasetyenziswa, zilandelwa luvavanyo lwe-hoc post lwe-hoc.

IINKCUKACHA

Ukuchazwa kwparadigm yokuntywila

Sisebenzise i-sucrose ingestion paradigm ukuze siphande ngeziphumo zendalo, umvuzo we-orosensory kuthumelo lwe-synaptic (Umzobo 1A). Iigundane ezingabantu abadala zahanjiswa kwigumbi lovavanyo ngeentsuku ezintathu ezilandelelanayo. Ngosuku lwesine (usuku lokuqala loqeqesho), iigundane zabekwa kwigumbi lokulinganisa lendawo. Emva kwemizuzu ye-15 yokulinganisa umsebenzi we-locomotor kwigumbi, iibhotile ezinamanzi (elezilwanyana zaManzi) okanye isisombululo se-25% sucrose (kwizilwanyana zeSurrose) zaziswa kwigumbi lokulinganisa ngemingxunya ekwi-insimbi yegumbi. Iibhotile zisuswe emva kwemizuzu ye-5 kunye nomsebenzi we-locomotor ulinganiswe ngemizuzu eyongezelelweyo ye-15 ngaphambi kokuba izilwanyana zibuyele ezindlini. Siphindaphinde le nkqubo ngeentsuku ze-7 zilandelelana. Kolunye uvavanyo, uqeqesho lwe-sucrose lwandiswa kwi-8th usuku. Ukufikelela okufutshane, ngaphandle kokuphikiswa kwesisombululo esinokubakho kakhulu kusivumele ukuba siphande zombini kunye neziphumo ezibandayo zokungena kwe-sucrose, njengoko izilwanyana zithembeke ngokufanelekileyo ukuzinyanzela ngamandla ngexesha leefestile zokungena ngaphakathi kweentsuku ezintathu zoqeqesho (Umzobo 1B). Ezi meko zovavanyo zenza ukuba ukuthelekiswa kwamaqela ovavanyo kwangoko emva kokuphelisa ukungena. Inkqubo yethu yokubandakanyeka kolu phononongo yayikukuba iigundwane ziqala ukusebenzisa ubuncinci igram ye-sucrose ngexesha lokufikelela ngaphakathi kwiintsuku ezintathu zokuqalisa koqeqesho; Akukho zilwanyana zakhutshelwa ngaphandle kwesi sifundo ngokubhekisele kule meko.

Umzobo 1  

Ukufakwa kwakhona kwe-sucrose okuphindaphindiweyo kubangela ukuphakanyiswa kwexeshana kwendawo eyenzekelayo.

Siphawule ukuba kwisithuba seentsuku ezintathu zoqeqesho, izilwanyana zeSukrose zitya kakhulu isisombululo sokuzimela kunokuba izilwanyana zamanzi zichithe amanzi (Umzobo 1B). Ukongeza, ngelixa kungekho mahluko ubalulekileyo kwindawo ezenzekelayo eyayiqwalaselweyo ngeentsuku zoqeqesho 1-6 (idatha ayiboniswanga), siqwalasele ukuphakama okuphezulu komgama ohanjiwe kwizilwanyana zeSucrose xa kuthelekiswa nezilwanyana zamanzi kwimizuzu emithathu emva kokukhutshwa kwebhotile ngemini ye-7 (Umzobo 1D), kwaye lo mahluko wawukhona nakwi-8 ngosuku (Umzobo 1E). Akukho mahluko kumgama ewonke ohanjiweyo wabonwa phakathi kwamanzi kunye nezilwanyana zeSuprose kwimizuzu emithathu yangaphambi kokungeniswa kwebhotile ngalo naluphi na usuku lweentsuku zovavanyo (Umzobo 1C), ukucebisa indawo ephakanyisiweyo yayiyimpendulo ebanzi yokufakwa kwe-sucrose ngokuthe ngqo kumgangatho oqeqeshwe nge-sucrose, endaweni yempendulo enemeko kwigumbi locomotor. Ukugcina oku, kungenzeka ukuba kukho unxibelelwano oluqinisekileyo phakathi kwenani le-sucrose echithiweyo kunye nomgama ewonke ohanjiweyo (Umzobo 1F). Kwakungekho mahluko kwizisindo zezilwanyana phakathi kwamaqela eSuprose kunye naManzi ngaphambi okanye emva kweentsuku ze-7 zoqeqesho (idatha ayiboniswanga).

Ukufakwa ekutyeni kufaka ukufakwa kweCPAR

Uqeqesho lokuphuma kwangaphambi kokukhokelela ekunyukeni okwexeshana kwesahlo ngosuku lokugqibela loqeqesho. Ukufumanisa ukuba ngaba esi siphumo sokungena kwe-sucrose sihamba kunye notshintsho lwe-electrophysiological kwii-nucleus accumbens, ummandla olawula indlela yokuziphatha, salungiselela i-nucleus accumbens slices kwangoko emva kokususwa kwebhotile ngemini ye-7 kwaye yarekhodwa kwi-neurons ye-accumbens core (Umzobo 2A). Owona ndoqo uphambili uphendulisiwe kwiimpendulo zendlela yokufumana isivumo (I-Sesack kunye neGrace, 2010). Sifumanise ukuba zombini i-amplitude kunye nobuninzi be-spontaneous miniaturevustive currents (ii-MEPSCs) zazinkulu kakhulu kwindawo encinci ye-Sucrose yezilwanyana xa kuthelekiswa nezilwanyana zamanzi (Umzobo 2B). Oku kubonise ukuba ukusetyenziswa okuphindaphindiweyo kwe-sucrose kunokulawula ngokuqinisekileyo ukuhanjiswa kwe-synaptic kwimbonakalo ye-nucleus accumbens. Ukufumanisa ukuba ukubandakanywa kwe-CPAR kudlale indima yokudlala emva kokuphuma, sazimisela ukulungiswa kweendawo ezinomtsalane ngokulinganisa ii-EPSCs kwizixhobo ezahlukileyo zolusu (Umzobo 2C, 2D kunye 2E). I-CPARs zilungisa ngaphakathi izinto ezinokubakho ngenxa ye-endo native polyamine blockade. Siphawule ukuphinda senziwe urekhodo olusuka kwi-neurons yezilwanyana zeSucrose, njengoko kuboniswe kukungangqinelani kubudlelwane be-I / V, ngokuthelekiswa nezilwanyana zamanzi (Umzobo 2E), ukongeza ukonyuka okukhulu kwesalathiso sokulungiswa (Umzobo 2F).

Umzobo 2  

Ii-synapses eziphambili ze-Accumbens zinokwenzeka emva kokungena okuphindaphindiweyo kwe-sucrose.

Ukuqinisekisa ukuphakama kwamanqanaba e-CPAR ngenye indlela, sarekhoda i-accumbens ii-neurons eziphambili emva kokufakwa kwisibambisi esithile se-CPAR, i-1-Naphthylacetyl spermine (Naspm) kwindawo yokuhlambela. Sifumene ukuba i-Naspm iyanciphise kakhulu i-EPSC amplitude ekurekhodeni kwii-neurons ezivela kwi-Sucrose kodwa hayi izilwanyana zamanzi (Amanani 3A-C). Ukongeza, emva konyango lwe-Naspm, ubudlelwane be-I / V kwii-neurons ezivela kwizilwanyana zeSucrose baba ngumtya, babonisa ukunqunyulwa kwee-CPARs kwi-synapses yesilwanyana ye-Sucrose, ngelixa kungekho mpembelelo iphambili kubudlelwane be-I / V yabonwa emva konyango lwe-Naspm kwii-neurons ezivela kwizilwanyana zamanzi (Imifanekiso 3D). Ezi ziphumo zibonisa ukuba ukuphindaphindeka kokufakwa kwe-sucrose kukhuthaza ukufakwa kwee-CPAR kwii-synapses zesiseko se-accumbens.

Umzobo 3  

Ukufakwa kwe-inhibion ​​ye-Sucrose ukubanjwa kwe-Ca2 + -permeable AMPA receptors.

Ukucutha ukuntywila kufaka i-GluA1 ukurhweba

Ii-CPAR zii-receptors ze-AMPA ezingekhoyo kwi-GluA2 AMPA receptor subunit. Ke, ukudityaniswa kwe-synaptic kwee-CPARs kuhlala kubandakanya umsebenzi oxhomekekileyo wokurhweba ngokurhweba kwi-GluA1 subunit (U-et al., 2009; UIsake et al., 2007; U-Liu kunye noZukin, 2007; Ukutyala et al., 2006). Ukuqinisekisa ukufakwa kweCPAR ye-synaptic emva koqeqesho lwe-sucrose, saphanda ukuba ingaba intsebenzo ye-sucrose intenion yonyusa i-synaptic expression yeGluA1. Iigundane zanikwa ukufikelela kwi-sucrose njengoko kuchaziwe apha ngasentla ngeentsuku ze-7 zilandelelana. Ngeentsuku i-1, i-3, i-5, kunye ne-7, sahlula yonke iseli, i-synaptosome kunye ne-postynaptic density (PSD) iziqwenga ezivela kwimimandla emithathu yobuchopho: i-accumbens core (imbindi), iqokobhe lokuqokelela (iqokobhe) kunye ne-somatosensory cortex (cortex). Sihlalutye yonke i-cell kunye ne-PSD ngamaqhezu yi-Western blot ebonisa i-GluA1 kunye neGluA2.

Asifumananga zinguqu kwi-GluA1 okanye iGluA2 kuwo onke amaqhezu eseli amaqhekeza eentsuku zokugcina kwiintsuku zovavanyo ezivavanyiweyo, ephakamisa ukusetyenziswa okuphindaphindiweyo kwe-sucrose akulawuli amanqanaba apheleleyo ezi proteni (Amanani 4A-C). Kwiqokelela lamaqonga ePSD, nangona kunjalo, iGluA1 inyuke kakhulu ngemini ye-7 kumbindi kodwa hayi kwibholam ngelixa i-GluA2 ingatshintshanga kakhulu kwelinye iqhezu (Amanani 4D-4F nedatha ayiboniswanga). Asiboni sonyuso lubalulekileyo kwi-GluA1 kwii-ingqokelela zeeprayimari ze-PSD kwiintsuku zovavanyo zangaphambili (Amanani 4D-F) kunye neGluA1 okanye iGluA2 ayitshintshanga kumaqhezu ePSD e-cortex nangaluphi na usuku lweentsuku zovavanyo (idatha ayiboniswanga). Ukunyusa i-GluA1, ngakumbi ehambelana neGluA2, kwii -PSD eziphambili emva kokungena okuphindaphindiweyo kwe-sucrose kukugcina ukulungiswa okwandileyo okubonwe kwi-accumbens core neurons, njengoko kuchaziwe ngasentla.

Umzobo 4  

Ukuxinana kwe-postsynaptic GluA1, kodwa ingeyiyo i-GluA2, inyuka kwi-nucleus accumbens core emva kokungena kwe-sucrose.

Umsebenzi oxhomekeke ekuthengiseni iGluA1 ubonakalisile ukuba negalelo leplastiki ye-synaptic in vitro kwaye kwakhona kwi vivo (Lu kunye noRoche, 2011). Indlela ekhawulezayo, kunye neninzi yamanyathelo okurhweba ngeGluA1 ibonakalisiwe in vitro (USerulle et al., 2007; I-al et al., I-2008; I-al et al., I-2005). Ukuza kuthi ga ngoku, igalelo lale nkqubo yamanyathelo ngamanqanaba amaninzi ekuqokeleleni i-synaptic ye-GluA1 kwi vivo khange ihlolwe. Ukufumanisa ukuba uqeqesho lwe-sucrose lubangela ukurhweba nge-GluA1 ngokupheleleyo ngumatshini we-multistep, safaka indawo ekuthiwa yi-GluA1 kwindawo ye-nucleus ingqokelela i-synapses ye-sucrose- kunye nezilwanyana eziqeqeshiweyo ngamanzi yi-microscopy yobuninzi. I-Acumbens basic tishu yavunwa ngomhla wesixhenxe woqeqesho lwe-sucrose ukusuka kumaqela ovavanyo e-3 kwiigundane. Ezi yayiziigundane: I-1) isela amanzi ngeentsuku ze-7 (Amanzi), i-2) etshisa i-sucrose yeentsuku ze-7 (iSurrose), kunye ne-3) itya i-sucrose yeentsuku ze-6 kunye namanzi ngosuku lwe-7 (i-Sucrose / Amanzi). Iigundane kwenziwa idini kwi-7th Usuku, imizuzu ye-5 emva kokusetyenziswa kwe-sucrose okanye emanzini. Ke ngoko, uthelekiso lwamaqela amabini kuvavanyo, uSuprose / Amanzi kunye nezilwanyana zeSukrose, omnye komnye nakwiZilwanyana zaManzi zityhila amaxesha oguquko lweposynicic ebangelwa kukusetyenziswa kwe-sucrose kwiigundane eziqeqeshelwe ukuzimela. Silinganise i-postembed immunogold (PEG) -labeled GluA1 kwi-5 amacandelo ahlukeneyo e-postynaptic: dendritic spine cytosol (intraspinous), i-extrasynaptic plasma membrane (membrane), PSD, kufutshane ne-PSD, kunye ne-syaptic ezintathu kunye neqela le-PSD '((Umzobo 5A). Ukuphelisa ukuvavanywa kokukhetha, uvavanyo lwee-elektroni zombane zeempawu zenziwa zabonwa ngamehlo kathathu.

Umzobo 5  

I-microncopy ye-elektroni ityhila ungeniso lokurhweba ngamanyathelo amaninzi e-GluA1 ngokutsalwa kwe-sucrose.

Zombini ii-Sucrose kunye ne-Sucrose / izilwanyana zamanzi ezibonisiweyo ziphakanyiswe kakhulu kwi-intraspinous GluA1 ezihambelana nezilwanyana zamanzi (Umzobo. 5B kunye 5C). Oku kuphakamisa ukuba ukusetyenziswa kwe-sucrose engapheliyo kunyusa ichibi elingena ngaphakathi kwe-GluA1-equlathe ii-receptors ze-AMPA ezikufuphi neziza ezi-synaptic, ii-receptors ezinokuthi zifumaneke ngokurhweba ngokuthengisa, kwaye, okubaluleke kakhulu, ukuba eli chibi le-intracellular linokuqhubeka kwiiyure ze-24 emva kokusetyenziswa kokugqibela kwe-sucrose . Siphinde sajonga umbuzo obalulekileyo wokuba ingaba ukunyusa okunyanzelekileyo kwe-sucrose kunokubangela ukurhweba nge-GluA1 ngokukhawuleza. Saqaphela ukuba i-extrasynaptic plasma membrane GluA1 yonyuswe kakhulu kwizilwanyana zeSucrose xa kuthelekiswa nezilwanyana zeSucrose / zamanzi nezamanzi (Amanani 5B kunye 5D). Olu luvo lubonisa ukuba umvuzo wendalo, orosensory obonelelwa sisusus esisodwa unokukhawuleza (<5 min) kodwa okwethutyana (ixesha lokubola <24 h) kuphakamisa inani labemi be-extrasynaptic ye-GluA1 equlathe ii-receptors ze-AMPA, esenza ichibi lelebile apho abafumana khona inokugcwala kwi-synapse.

Ngokuphawulekayo, in vitro Izifundo ziphakamise ukuba ukudibanisa i-synaptic kwee-AMPA receptors zenzeka ngamanyathelo e-2. Kwindawo yokuqala, i-glutamate- okanye i-dopamine-exhomekeke kwi-phosphorylation ye-dopamine iphakamisa amanqanaba ee-receptors kwiindawo ezongezelelweyo kwi-plasma membrane (U-Esteban et al., 2003; USerulle et al., 2007; I-al et al., I-2008; I-al et al., I-2005), ngelixa yesibini, i-phosphorylation ye-Ser 818 ikhuthaza ukufakwa kwe-synaptic (Boehm et al., 2006). Ukuthelekiswa kwethu ngemicroscopy yezilwanyana zeSucrose kunye ne-Sucrose / Amanzi kunye nezilwanyana zaManzi kubonisa ukuba inyathelo lokuqala lokurhweba ngeGluA1 in vitro (IMakino neMalinow, 2009), ukurhweba ngokurhweba ngokukhawuleza kwi-membrane ye-extrasynaptic, nako kuyenzeka kwi vivo ulungiselelo lomvuzo we-orosensory.

Ukugcina kunye ne-electrophysiology kunye neziphumo ze-biochemical ezichazwe apha ngasentla, i-PeG EM ibonise ukuba ukuthathwa kwe-sucrose nako kwafaka inyathelo lesibini kwi-GluA1 yokurhweba nge-GluA1 i-receptor yokungena kwi-syrude ukususela kwinqanaba le-GluA1-immunoreactivity kwi-PSD yayinkulu kakhulu kwi-Sucrose xa kuthelekiswa neempuku zamanzi, kwaye kwabakho indlela eya kunyuko lweGluA1 eSucrose / Amanzi ngokuthelekiswa neegundane zamanzi (Amanani 5B kunye 5E). Ukunyuka kwezilwanyana ezi-Sucrose / Amanzi kuyahambelana nokuba nokudityaniswa okukhawulezileyo kwe-GluA1 egqiba nge-synaptic half-life of ~ 24 hr ,, okanye ngokufakwa ngokukhawuleza kwe-GluA1 kunye nokufakwa endaweni ye-synaptic GluA1 / 2 ngaphezulu kwexesha elifanayo. Ipesenti yezivumelwano ezibonisa i-GluA1 kwi-PSD nayo ibinkulu kakhulu kwiirats zeScrrose xa kuthelekiswa namagundane Amanzi (Umzobo 5F), Ukucebisa ukuba i-GluA1 ithengiselwe kwi-synapses ebikade ingenayo iGluA1. Oku kuphakamisa ukuba ukonyuka kokuphakama kwe-mEPSC okujongwa kwiziphumo zeempuku ze-Sucrose ukusuka kunyuko lwe-synaptic GluA1, kunye nokonyuka kokuphindaphindeka kwe-mEPSC kungakhokelela ekufunyanweni kwe-GluA1 ukuya kwii-synapses zangaphambili ezazithule, nangona ukukhutshwa kwe-glutamate kungenakugxothwa. Silinganise nenani leesynapses kwiqela ngalinye lokuvavanya ukujonga ukuba ngaba i-sucrose ingestion incased synaptogenesis; kwakungekho mahluko phakathi kwamaqela amathathu ovavanyo (idatha ayiboniswanga). Sigqiba kwelokuba ukuphinda kungene ngaphakathi kwe-sucrose kuphakamisa indawo ezinzileyo (> iiyure ezingama-24) echibini elingaphakathi le-GluA1, kunye ne-sucrose eyodwa ekhuthazayo kwi-sucrose eqeqeshiweyo (iintsuku ze-6) i-rat yanele ngokukhawuleza (i-5 min) ukuphakamisa i-GluA1 kwi-membrane ye-plasma ye-extrasynaptic, enokubakho Ukuzoba ii-receptors ezivela echibini elingena ngaphakathi. Sicebisa ukuba isahlulo see-extrasynaptic receptors zibandakanyiwe kwi-PSD, ekhokelela kwisalathiso sokujonga kunye notshintsho lwePSD GluA1, ngaphambi kokuba ichibi le-extrasynaptic libuyele kwisiseko kwi-24 h emva kokukhuthaza. Ezi ziphumo zibonisa ukuba umvuzo wendalo unokunyanzelisa ukurhweba ngabantu ngokurhabaxa kwizilwanyana eziqeqeshiweyo.

Umsebenzi we-CPAR uyimfuneko ukulungiselela indawo ephakanyisiweyo emva kokufakwa kwe-sucrose

I-neurons ephakathi ephakathi ifumana zombini ukufakwa kwe-dopaminergic kunye ne-glutamatergic (ICalabresi, et al., 1992). Ukuvavanya ukubandakanyeka kokungena kwe-glutamate signaling kwindawo ephakanyisiweyo yendawo esiyiqapheleyo emva kokufakwa kwe-sucrose yokufakwa kwiigundane eziqeqeshelweyo, sibeka i-cannulas kwisiseko se-resebens samagundane, kunye nezilwanyana eziqeqeshiweyo kumagumbi okulinganisa enomotoror njengoko kuchaziwe ngasentla. Ngomhla we-8 yoqeqesho lwe-sucrose, siye sangena kwi-Naspm kwi-core ngaphambi kokufakwa kwigumbi lovavanyo lwe-locomotor. Inaliti inciphise umgama ohanjiweyo wezilwanyana zeSurrose kwaye yasusa umohluko phakathi kweSucrose kunye nezilwanyana zamanzi ezibonwe kwangoko emva kokususwa kwebhotile (Umzobo 6A). Ukuqinisekisa ukuba uxinzelelo olubangelwa ukuphatha izilwanyana aluchaphazelekanga ukuphendula kwe-sucrose, safaka injine kwi-saline kwisiseko ngosuku olulandelayo (ngosuku lwe-9 loqeqesho lwe-sucrose); kwakhona uxinzelelo olubalulekileyo lwabonwa kwizilwanyana zeSucrose kwangoko emva kokukhutshwa kwebhotile (Umzobo 6B). Oku kubonisa ukuba i-Naspm yayithintele ngokukodwa ukuphakanyiswa kwe-sucrose-induction of locomotion. Ukufakwa kwi-antikonist ye-NMDAR, APV, embindini ngeentsuku ezilandelayo nako kususe umohluko phakathi kweSucrose kunye nezilwanyana zamanzi (Umzobo 6C), ebonisa ukuba ii-NMDARs ziyafuneka ukuphakanyiswa kwethambeka elizenzekelayo emva kokufakwa kwe-sucrose. Ukujonga ukuba impendulo ekuyo imeko kwigumbi lovavanyo yadlala indima ekufumaneni i-hyperacuction, izilwanyana zabekwa kwigumbi le-35 min ngaphandle kokuqaliswa kwebhotile; akukho mehluko kumgama ohanjiweyo wabonwa phakathi kweSucrose kunye nezilwanyana zamanzi (Umzobo 6D). I-Naspm kunye ne-APV azichaphazelekanga ukusetyenziswa kwe-sucrose (Umzobo 6E), ukubonisa ukuba ii-CPARs eziphambili kunye nee-NMDARs azidingeki ekusebenzeni ngamandla kwe-sucrose. Izilwanyana apho i-cannulas ibingafakwanga kwisiseko se-accumbens (i-2 ngaphandle kwezilwanyana ze-15), njengoko kuvavanywe ukubingelela kwangoko (Umzobo 6F), bezingafakwanga kuphando. Ukuqukumbela, ezi datha ngokudibeneyo zibonisa ukuba ukusetyenziswa kwe-sucrose ngumlinganiso oqeqeshekileyo we-sucrose yenza ukuba ukurhwebeswa kwe-GluA1 kwimizuzu ye-5, kunye nokuvalwa kweendlela zokubonisa okurhweba ngokurhweba ngokungekho mthethweni kuthintele ukuphakama kwemisebenzi ye-locomotor yokuzimela emva kokuhamba kweesuti.

Umzobo 6  

Indawo ekuphakanyiswa kuyo ngokuzenzekelayo emva kokufakwa kwe-sucrose ifuna ii-CPAR nee-NMDAR.

Iindlela ezimbini zendlela yokutywinwa kweescrose zinokucingelwa. Inye, ngokunyanzela i-chemosensory okanye i-orosensory, iqalwa yi-sucrose ebopha i-receptor yokunambitheka okumnandi, ehambelana neprotein ye-heteromeric G protein edibeneyo ye-receptor, T1R2 / T2R3 (Kitagawa et al., 2001; UMax et al., 2001; UNelson et al., 2001; USainz et al., 2001). Izakhamzimba ezityebileyo zeekalori zinokukwazi nokulawula ukusebenza kwengqondo ngemigudu ye-metabolic ezimele ngokungcamla, nangona iindlela zingaqondwa kakuhle (de Araujo et al., 2008). Ukwahlula phakathi kwezi ndlela zimbini zeendlela ze-GluA1-zokurhweba ezibanjiswa yi-sucrose, siphindaphinde inkqubo yokuqeqesha kunye namaqela amathathu erhasi (i-4 rats / group) enikwe ukufikelela kwimizuzu ye-5 kwiibhotile eziqukethe amanzi, isisombululo se-25% , okanye i-3% saccharin (Imnandi kwaye iphantsi). Iibhotile zazisuswe kwaye iigundana zihlala imizuzu engama-15 ixesha elide kwiklasi yoqeqesho. Uqeqesho lwaphindwaphindwa ngeentsuku ze7. Iivolumu zelwelo eziselwe yi-sucrose kunye namaqela e-saccharin ayengafani ngokwahlukileyo komnye kwaye zombini zazinkulu kunokusetyenziswa liqela lamanzi, ngokuhambelana nomvuzo ngazo zombini izinto ezimnandi (Umzobo 7A). Ngomhla we-7th wokusela, kwangoko emva kokususwa kwebhotile, kwenziwa iigundane, izithsaba ezingqokelelweyo zavunwa zaza zalungiswa kwiqela ngalinye lovavanyo, iqhezu lePSD lodwa kunye namanqanaba e-GluA1 afunyanwa yi-blot WesternUmzobo 7B). Njengangaphambili, izilwanyana ezitya i-sucrose zibonisa i-GluA1 ephezulu kwiqhezu lePSD elihambelana neqela lamanzi (Umzobo 7C). Ngokuphawulekileyo, i-GluA1 yaphakanyiswa kwakhona kwinxalenye yePSD yezilwanyana ezitya i-saccharin. Akukho mahluko ubalulekileyo kumanqanaba e-GluA1 kuwo onke amaqhezu eseli asuka kwi-accumbens isiseko samanzi, i-sucrose kunye nezilwanyana ze-saccharin, ephakamisa ukuba ukunyuka kwe-GluA1 kwakukhethekileyo kwiqhezu le-synaptic (Umzobo 7D). Kungenxa yokuba i-saccharin ivuselela i-protein efanayo ye-heteromeric G edibeneyo ye-receptor ye-receptor njenge-sucrose (Masuda et al., 2012; UNelson et al., 2001), kodwa ayinalo ixabiso le-caloric, sigqiba kwelokuba ukukhuthaza i-receptor yokunambitheka okumnandi kwanele ukuqalisa ukutyikitya okuphakamisa amanqanaba e-GluA1 kwinqanaba le-nucleus accumbens core synapses.

Umzobo 7  

Uqeqesho lwe-Saccharin lubangela ukwanda kwe-Synaptic GluA1 Efana noQeqeshi lweSunrose.

ingxoxo

Sibonisile ukuba umvuzo we-orosensory, ukusetyenziswa okuphindaphindiweyo kwe-sucrose, okunokubangela ukufakwa kwe-GluA1 i-synaptic ngokufaka kumanyathelo okurhweba ngamanyathelo amaninzi achazwe ngaphambili in vitro. Ukusetyenziswa okuphindaphindiweyo kwe-sucrose ngaphezulu kweentsuku ze-6-7 ezinokubakho zi-encordes ze-electrophysiologically ngokufakelwa kwee-CPAR. Esi siphumo sihamba kunye nokuqokelelwa kweGluA1 kodwa hayi iGluA2 kwi-PSD yesiseko, kwaye yayingumda kwaye ngokwethutyana, njengoko kungekho lutshintsho lwabonwa ngaphambi komhla woqeqesho kwi-7 embindini, kwaye akukho lutshintsho luye lwabonwa kwiqoksi ye-ascobs okanye somatosensory cortex. Uhlalutyo lwe-microncopic luveze ukuba ukuphinda kugxilwe kuphakamise into ezinzileyo (t1/2 > Iiyure ezingama-24) kubemi be-receptors ezingenayo i-GluA1. I-Sucrose ikhawuleze (i-5 min) kwaye okwethutyana (t1/2 <24 hrs) amanqanaba aphakamileyo e-GluA1 equlathe ii-receptors kwiindawo ze-extrasynaptic kwizilwanyana eziqeqeshwe nge-sucrose, ekonyusa inani labemi be-AMPAR abakwaziyo ukusasaza kamva kwi-synapse. I-Synaptic GluA1, zombini emelwe liqhekeza lePSD kwaye ifunyenwe yi-PEG-EM, inyuswe kakhulu kwi-Sucrose ngokuthelekiswa nezilwanyana zamanzi. Ukusuka kwezi ziphumo sicebisa ukuba amanyathelo ngamanyathelo amabini e-extrasynaptic exocytosis elandelwa kukurhweba ngabantu ngokungekho mthethweni ngokufakwa kwe-AMPAR synaptic ngaphambili in vitro (Boehm et al., 2006; IMakino neMalinow, 2009; Ewe et al., 2006; USerulle et al., 2007; I-al et al., I-2005) inokuqaliswa ngokukhawuleza kwi vivo ngumvuzo wendalo.

Utshintsho kumanqanaba e-synaptic GluA1 lujongwe kuphela emva kweeseshini zoqeqesho ze-7, ucebisa ukuba inkqubo yosuku olunye iyafuneka. Uvavanyo lwebhayoloji kwi vivo, Asizange siqwalasele ukwanda okukhulu kwamanqanaba e-PSD GluA1 engundoqo ngeentsuku ze-1, i-3, kunye ne-5 yoqeqesho lwe-sucrose; kuphela emva kweentsuku ze-7 zoqeqesho lwe-sucrose yayiyi-GluA1 kwi-PSD iphakanyiswe kakhulu. Kuvavanyo lwe-microncopy yovavanyo lwe-elektroni, sabona ukuba i-Sucrose / izilwanyana zamanzi, ezazikhe zaqeqeshwa kwiintsuku ze-6 kodwa zaza zangafumani khuthazo lwe-sucrose ngeyure ze-24, zabonisa imeko yokunyuka kwePSD GluA1. Ezi zilwanyana zikwabonisa ukuphakama okunomdla kwe-GluA1 xa kuthelekiswa nezilwanyana zamanzi, kodwa akukho lutshintsho kwi-membrane ye-extrasynaptic GluA1 yaqwalaselwa. Kwezi ziphumo sifikelela kwisigqibo esithathu. Okokuqala, ii-receptors ze-GluA1-equlathe i-AMPA ziqokelelene ngamandla kunye nokukhuthaza okulandelayo kwe-sucrose. Ngenxa yokuba izifundo zangaphambili zibonakalise ukuba ukukhutshwa kwe-sucrose ye-induces ye-dopamine kukhutshwa kwi-accumbens (Cacciapaglia et al., 2012; McCutcheon et al., 2012; URada et al., 2005), kunye nokuba ii-D1Rs zinokuqhuba uguqulelo lwasekhaya lweGluA1 kwi-dendrites (Smith et al., 2005), ukukhutshwa kwe-dopamine emva kokungena kwe-sucrose kunokucaphukisa i-GluA1 synthesis yokhokelela ekuqokeleleni kwe-GrasA1 ye-intraspinous. Ngenye indlela ukunyuka okunyanzelisayo kunokubonakalisa ukurhweba nge-GluA1 ukusuka kumasayithi asemaphandleni. Kusenokwenzeka ukuba urhwebo olugqithisileyo oluvela kolu hlobo oluphakamileyo lwe-intraspinous inegalelo kwichibi le-extrasynaptic kwi-membrane ye-plasma. Okwesibini, ukujongwa kokonyuka kwe-membrane ye-extrasynaptic GluA1 e-Sucrose, kodwa ingeyiyo kwi-Sucrose / Amanzi okanye izilwanyana zamanzi zicebisa ukuba ii-receptors ze-extrasynaptic mhlawumbi zihamba ngenyathelo lesibini kwi-synapse okanye zibekiwe ngaphakathi kwe-24 hr kulandela ukusetyenziswa kwe-sucrose, extrasynaptic echibini ixesha elifutshane. Okwesithathu, ukuphakama kweSuprose yezilwanyana i-PSD GluA1 xa kuthelekiswa nezilwanyana eziManzi, kodwa hayi iSuprose / Izilwanyana zamanzi zikwacebisa ukuba emva kwesikhuselo ngasinye se-sucrose, ii-receptors zihamba emva kwexesha kwisiphambuka ukusuka echibini lama-receptors ashenxiswa ngokukhawuleza ziye kwi-membrane ye-plasma ye-extrasynaptic. Asinakucacisa ukuba ukurhweba ngabantu nge-GluA1 ngqo ukusuka echibini elingene ngaphakathi ukuya kwisirinji. Indlela enjalo nangona kunjalo ibonakala ingathandeki ukuba inikwe izifundo ezibonakalisa iGluA1 ifakwe kwi-extrasynaptic (Boehm et al., 2006; IMakino neMalinow, 2009; Ewe et al., 2006; USerulle et al., 2007; I-al et al., I-2005). Ezi ziphumo zibonisa umboniso wokuqala wokuba ikhosi ye-GluA1 yokurhweba ngabantu ngokungekho mthethweni (<5 min) kunye nendlela eqatshelweyo in vitro ziyajongwa kwi vivo. Ukongeza, iziphumo zethu zicebisa ukuba ukuphindaphinda okunomvuzo okonwabisayo kuguqula amandla okuphinda kusebenze ngokuphakamisa echibini lama-receptors athambileyo anakho ukurhweba.

Ngenxa yokuba i-saccharin yaphembelela ukurhweba nge-GluA1 ngokufanayo ne-sucrose, umxholo we-caloric we-sucrose awufuneki. I-Saccharin ivuselela i-receptor yencasa emnandi efanayo, i-T1R2 / T2R3, njengoko sucrose (Masuda et al., 2012; UNelson et al., 2001), sukusebenza kakubi kwe-receptor kungenzeka ukuba kuqale ukufakwa kwe-GluA1 kwii-NTN synapses. I-Sucrose iphakamisa ukukhutshwa kwe-dopamine kwii-accumbens ezivela kwi-VTA neurons (Cacciapaglia et al., 2012; McCutcheon et al., 2012; URada et al., 2005lukuya e-GluA1 ukurhweba ngomhlaba. Yindlela ke enxibelelanisa i-receptor enambitheka emnandi kwiVTA kusenokwenzeka ukuba ingumbindi weplastiki efundwe apha.

Kusenokwenzeka ukuba ukurhweba nge-GluA1 ngokukhawuleza emva kokungena kwi-sucrose kudlala indima kulawulo lwe-locomotion engazenzisiyo. Ewe, kwizilwanyana eziqeqeshelwe ukuzimela, ukuthintelwa kwee-CPAR kuthintele ukuphakama okuzenzekelayo kwemisebenzi ye-locomotor kwangoko emva kokungena kwe-sucrose. Umgama ewonke ohanjiweyo ziigundane emva kokusetyenziswa kwe-sucrose ukulinganiswa ngeentsuku ezilandelelanayo zaphakanyiswa kakhulu kwixesha le-3 min ngokukhawuleza emva kokusetyenziswa kwe-sucrose ngosuku lwesixhenxe loqeqesho. Umsebenzi owandisiweyo ngokukhawuleza emva kokuba i-sucrose ibonwe iqala ngemini ye-3 yoqeqesho, kodwa ayizange ihluke kakhulu kude kube yimini ye-7. Eli xesha lekhondo lomsebenzi lihambelana nexesha lexesha lokuqokelelwa kweGluA1 kwii-dendrites ezingumxholo wokuqala.. Isongezo esandisiweyo sisiphumo esisebenzayo sokurhweba ngabantu nge-CPAR kwii-synapses ze-MSN kwisiseko se-accumbens ukusukela kwi-Naspm yokufaka kwisithinteli kuthintele ukwanda komsebenzi. Ukuthintelwa kwendawo leyo yaphakanyiswa yi-NMDA receptor inhibitor kubonise ukuba ukubhengezwa kwe glutamate kusetyenziswa ii-NMDA receptors kunye neeCPAR kwakuyimfuneko ukuphakamisa ukusebenza kwendawo. Ukufakwa kwenqanawa, nangona kunjalo, akuzange kuchatshazelwe luphawu lwe-glutamate signaling, ukugcina ngezifundo zangaphambili ezibonisa ukuba isiseko se-ascobs sibandakanyeka kwi-orchestration yeempendulo zemoto ezinxulumene nomvuzo we-orosensory kodwa ungazisebenzisi ngokwakho (USmith, 2004). Ikhosi yexesha elifanayo lokuphuhliswa kwe-hyperlocomotion iye yanikwa ingxelo yokuphuhliswa koxinzelelo lweemeko ezincomekayo kwizilwanyana ezondliwe ngokutya kwazo kwemihla ngemihla kwindawo ekhethekileyo (UMateyu et al., 1996). Ukuba impendulo yangoku ibingumboniso we-hyperactivity ovela kukubhangqa umxholo kunye ne-sucrose, nangona kunjalo, ngekwandulela ukuhanjiswa kwe-sucrose, engakhange ibonwe. Kuyenzeka ukuba izifundo zibonise umboniso wokuhlaziya. Olunye uvavanyo luya kuba yimfuneko ukwahlulahlula ukuba ingaba ukuphakanyiswa okuphezulu emva kokufakwa kwe-sucrose kwakumnandi wokuphononongwa ngokuchasene nefomathi yokuziva ngemoto okanye enye into. Ngayiphi na imeko, ukuphakanyiswa kwe-locomotion okuzenzekelayo kufuna ukutyhilwa kwe-glutamate, kwaye kwaphumela, ubuncinci kwinxalenye, ukusuka kufakwa kwee-CPAR kwisiseko se-accumbens.

Umsebenzi owandisiweyo we-locomotor emva kokungena kwe-sucrose unokubangela ngqo ngokuthe ngqo kubukho obuqaphelekayo be-synobes basic synapses, njengoko ukonyuka kokuphuma kwindlela ngqo ye-basal ganglia kukhuthaza i-locomotion ngokusebenzisa i-disthhibition ye-motor thalamus (I-Sesack kunye neGrace, 2010). TI-synenties ene-potentiated enokwenzeka kakhulu ukuba ihlale kumgaqo othe ngqo we-bokella ii-neurons eziphambili, ezibonisa ii-D1R. Ukuchaphazeleka kwendlela ehambelana ngqo ne-neurons ye-neurons kuya kubangela ukuba umsebenzi we-D1R ubangele ukurhweba nge-GluA1-ene-AMPARs kwii-synapses kwezi zi-neurons ezilandela ukukhutshwa kwe-dopamine. Isiphumo esinokubakho sinokunyusa umsebenzi kwi-inhibitory ukuqikelelwa kwe-neurons yendlela ngqo kwi-basal ganglia kwimveliso yokukhupha, ngaloo ndlela ukubulala i-motor thalamus kunye nokukhuthaza imisebenzi ye-motor cortex (UGerfen noSurmeier, 2011; Kravitz et al., 2010; I-Sesack kunye neGrace, 2010). I-synaptic potentiation eqaphelekileyo emva kokungena okuphindaphindiweyo kwe-sucrose inokwenzeka ngokuthe ngqo kwindlela ethe ngqo yomgudu ngenxa yokuba i-dopamine esebenza nge-D1 receptor inokubangela iphosphorylation ye-GluA1 S845, ekhokelela ekurhwebeni ngomphezulu.

Uninzi lophononongo luvavanye iimpembelelo zokuvuselela okuphindaphindiweyo nge-cocaine elandelwa ukurhoxa, unyango olunikezela iziphumo ezinamandla kwimisebenzi yenkqubo yomvuzo kwaye ekugqibeleni kukhokelela ekunyanzeliseni i-cocaine, ebonakaliswa ngokuphendula okuphezulu kwemoto kwi-cocaine, ukuthanda iziyobisi kunye nokubuyela umva (Kalivas et al., 1998). Inaliti ephindaphindiweyo ye-IP kunye necocaine yeentsuku ze-5-10 elandelwa kukurhoxa kukhokelela kukonyuka okuthe ngcembe kwiintsuku ze-14 ebusweni beGluA2-equkethe ii-receptors ze-AMPA (Boudreau et al., 2007; UKourrich et al., 2007). Nangona kunjalo, kwiintsuku ze-45 zokurhoxiswa kulandela i-10 d yokulawula ngokwakho, ukunyuka okukhulu kwesalathiso sokulungiswa kwaqwalaselwa kwii-rat zeMNN (McCutcheon et al., 2011b) Ukubonisa ukwanda kwee-CPAR. Ke ngoko, ukurhweba nge-CPAR kuqapheleke emva kokungena kwi-sucrose, emsebenzini wangoku, kunye nolawulo lwe-cocaine, nangona kunjalo phantsi kweemeko ezahlukeneyo zonyango.. Kungenxa yokuba iziphumo zangoko zokuzilawula ze-cocaine okanye inaliti (umzekelo, kwimizuzu ye-5) ayaziwa, isenzo se-cocaine asinakuthelekiswa ngqo nomsebenzi wangoku we-sucrose. Ngokunjalo, ayaziwa ukuba ii-CPARs ziyaqhubeka kwi-synNeses ze-MSN zezilwanyana eziqeqeshelwe ukuzimela emva kokuyeka uqeqesho lwe-sucrose okanye ukuba ezo zilwanyana zibonakalisa ubuntu bokuziva emva kokurhoxa ixesha elide.

Ukuqonda ukuba ivuselela kangakanani inkuthazo yokulawula i-plastiki kunye nokuziphatha kubaluleke kakhulu ekujonganeni nokulutha, i-hyperphagia, ukungcakaza kwe-pathological, kunye nokunye ukuphazamiseka kokuziphatha (Basar et al., 2010; EBerridge, 2009; ILischer neMalenka, 2011). Ukusetyenziswa kakhulu kweswekile kufaka isandla kwisifo sokutyeba kakhulu (Hu kwaye Malik, 2010), kwaye nangona zinokubakho ngokusetyenziswa gwenxa kweziyobisi (Avena et al., 2008), indlela yokusebenza kwayo ayikhange ihlolwe ngokubanzi. Iziphumo zangoku zisekela izinto ezisisiseko somvuzo ongena mbuyekezo apho izifundo zamva zinokujongana nolawulo lwezinto ezintsokothileyo, ngokunokwenzeka zibonelela ngeendlela zokuphinda zijongane ne-pathologies ezinxulumene nomvuzo.

Imibulelo

Sibulela amalungu eLebhu yeZiff, yakudala neyangoku, ngoncedo lobuchwephesha kunye neengxoxo eziluncedo, kubandakanya uH. B. Fernholz, B. Jordan, W. Lu, G. Rameau, S. Restituito kunye no-Y. Serulle. Lo msebenzi uxhaswe liZiko leSizwe lezeMpilo yeNgqondo iPredoctoral Fsoci F31MH76617-01 kunye ne-NIH yoQeqesho iSibonelelo 5T32DC000063 kwiNkqubo yoQeqesho kwiYunivesithi yaseNew York kwi-Neurosciences (DST), R01NS061920 evela kwiZiko leSizwe lokuPhazamiseka kweNeurological and Stroke (EBZ), 1R21MH091445- I-01 evela kwiZiko leSizwe lezeMpilo yeNgqondo kunye neOfisi yoPhando ngezeMpilo yaBasetyhini, iNkqubo yeKharman Family Foundation Grants kwiNkqubo yoPhando lweeNdlela zokutya, iNgxowa-mali yoMngeni woPhando kunye ne-P30EY13079 (CA), iZiko leLizwe kuXhaso lokuSetyenziswa gwenxa kweziyobisi i-DA003956 kunye neBhaso elizimeleyo loMphandi ovela kwi-NARSAD (KDC), iZiko leLizwe le-Deafness kunye nezinye iingxaki zokuNxibelelana zinika i-DC009635 kwi-RCF, nangesibonelelo sembewu kwiZiko lokuGqwesa kwiZiyobisi kwiZiko lezeMpilo laseNew York iLangone.

Imihlathi

Ingxabano yomdla: Ababhali baxela ukuba akukho mdla wokukhuphisana kwezemali.

Ucaphulo

  1. IAvena NM, Rada P, Hoebel BG. Ubungqina bokulutha iswekile: indlela oziphethe ngayo kunye nemithambo-luvo yokuphazamiseka, ukutyeba kakhulu iswekile. I-Neurosci Biobehav Rev. 2008; 32: 20-39. [Inkcazelo yamahhala ye-PMC] [PubMed]
  2. Basar K, Sesia T, Groenewegen H, Steinbusch HW, Visser-Vandewalle V, Temel Y. I-Nucleus ibutha kunye nokuxhonywa. Iprog Neurobiol. I-2010; 92: 533-557. [PubMed]
  3. IBerridge KC. 'Ukuthanda' kunye 'nokufuna' umvuzo wokutya: iziqwengana zobuchopho kunye nendima yokuphazamiseka kwindlela otya ngayo. I-physiology kunye nokuziphatha. Ngo-2009; 97: 537-550. [Inkcazelo yamahhala ye-PMC] [PubMed]
  4. I-Boehm J, Kang MG, Johnson RC, Esteban J, Huganir RL, Malinow R. Synaptic yokufakwa kwe-AMPA receptors ngexesha le-PMP lilawulwa yindawo ye-phosphorylation ye-PKC kwi-GluR1. Neuron. I-2006; 51: 213-225. [PubMed]
  5. IBoudreau AC, Reimers JM, Milovanovic M, Wolf ME. I-receptors ye-AMPA yeseli kwiseli ye-ratus ye-ratus inyuka ngexesha lokurhoxiswa kwe-cocaine kodwa ingene ngaphakathi emva komngeni wecocaine ngokunxulunyaniswa nokutshintsha okuguqulweyo kwe-mitogen-activated kinases protein. J Neurosci. I-2007; 27: 10621-10635. [Inkcazelo yamahhala ye-PMC] [PubMed]
  6. UBrebner K, Wong TP, uLiu L, uLiu Y, uCampsall P, uGrey S, uPhelps L, uPhillips AG, u-Wang YT. I-Nucleus iqokelela uxinzelelo lwexesha elide kunye nokuvakaliswa kokuziphatha. Inzululwazi. I-2005; 310: 1340-1343. [PubMed]
  7. I-Cacciapaglia F, iLungu leSaddoris, i-Wightman RM, i-Carelli RM. Ukwahlula okhutshelweyo kwe-dopamine ekhupha amandla kwi-nucleus eqokelele isiseko kunye neqokobhe lokulandelela okwahlukileyo kokuziphatha-okujolise ekuziphatheni kwe-sucrose. I-Neuropharmacology 2012 [Inkcazelo yamahhala ye-PMC] [PubMed]
  8. UCalabresi P, Maj R, Pisani A, uMercuri NB, uBernardi G. Uxinzelelo lwexesha elide lwe-synaptic kwi-striatum: ubume bomzimba kunye ne-pharmacological. J Neurosci. I-1992; 12: 4224-4233. [PubMed]
  9. Carr KD, Chau LS, Cabeza de Vaca S, Gustafson K, Stouffer M, Tukey DS, restituito S, Ziff EB. I-AMPA receptor subunit GluR1 isezantsi kwe-D-1 dopamine receptor stimulation in nucleus accumbens shell mediates yandisa ubungakanani bomvuzo weziyobisi kwiigundane ezithintelwe kukutya. I-Neuroscience. I-2010; 165: 1074-1086. [Inkcazelo yamahhala ye-PMC] [PubMed]
  10. UConrad KL, Tseng KY, Uejima JL, Reimers JM, Heng LJ, Shaham Y, Marinelli M, Wolf ME. Uyilo lwe-accumbens I-GluR2-engekho i-AMPA receptors i-mediates incubation yokunqwenela kwe-cocaine. Indalo. I-2008; 454: 118-121. [Inkcazelo yamahhala ye-PMC] [PubMed]
  11. Usuku JJ, Carelli RM. I-nucleus accumbens kunye nokufunda ngomvuzo wasePavlovian. Neuroscientist. I-2007; 13: 148-159. [Inkcazelo yamahhala ye-PMC] [PubMed]
  12. de Araujo IE, Oliveira-Maia AJ, Sotnikova TD, Gainetdinov RR, Caron MG, Nicolelis MA, Simon SA. Umvuzo wokutya ngokungabikho kokubonisa i-receptor signating. Neuron. I-2008; 57: 930-941. [PubMed]
  13. I-Ehlers MD, Heine M, Groc L, Lee MC, Choquet D. Ubunzima bokutsalwa kwe-GluR1 AMPA receptors ngomsebenzi we-synaptic. Neuron. I-2007; 54: 447-460. [Inkcazelo yamahhala ye-PMC] [PubMed]
  14. I-Esteban JA, Shi Shi, Wilson C, Nuriya M, Huganir RL, Malinow R. PKA phosphorylation ye-AMPA receptor subunits ilawula ukurhweba ngabantu ngokungekho mthethweni. Nat Neurosci. I-2003; 6: 136-143. [PubMed]
  15. IGerfen CR, uSurmeier DJ. Ukuguqulwa kweenkqubo ze-striatal proion nge-dopamine. Uvavanyo lonyaka lwe-neuroscience. I-2011; 34: 441-466. [Inkcazelo yamahhala ye-PMC] [PubMed]
  16. IGrueter BA, Brasnjo G, Malenka RC. I-Postynaptic TRPV1 ibangela uxinzelelo lweeseli oluchaziweyo lwexesha elide kwi-nucleus accumbens. I-neuroscience yendalo. I-2010; 13: 1519-1525. [Inkcazelo yamahhala ye-PMC] [PubMed]
  17. U-K, Ingoma L, i-Cummings ye-LW, uGoldman J, uHuganir RL, uLee HK. Ukuzinziswa kwe-Ca2 + -permeable receptors AMPA receptors in perisynaptic site by GluR1-S845 phosphorylation. I-Proc Natl Acad Sci US A. 2009; 106: 20033-20038. [Inkcazelo yamahhala ye-PMC] [PubMed]
  18. I-Hu FB, iMalik VS. Iziselo ezineswekile kunye nomngcipheko wokutyeba kakhulu kunye nohlobo lwe-2 yeswekile: ubungqina besifo. I-physiology kunye nokuziphatha. Ngo-2010; 100: 47-54. [Inkcazelo yamahhala ye-PMC] [PubMed]
  19. UIsaac JT, Ashby MC, McBain CJ. Indima ye-GluR2 subunit ekusebenzeni kwe-AMPA receptor kunye ne-synaptic plasticity. Neuron. I-2007; 54: 859-871. [PubMed]
  20. I-Jordan BA, i-Fernholz BD, i-Boussac M, i-Xu C, iGrigorean G, iZiff EB, iNeubert TA. Ukuchongwa kunye nokungqinwa kweeprotheyinokuxinana kwenqanaba leepenteni. Iiproteomics zeMol Cell. I-2004; 3: 857-871. [PubMed]
  21. IKalivas PW, uPierce RC, uCornish J, uSorg BA. Indima yokuziva unomdla wokunqwenela kunye nokuphinda ubenakho ukubandakanyeka kwi-cocaine. J. Pharmacol. I-1998; 12: 49-53. [PubMed]
  22. I-Kitagawa M, Kusakabe Y, Miura H, Ninomiya Y, Hino A. Ukuchongwa kofuzo lweMolekyuli yohlobo lomamkeli womntu ongena luthando. Unxibelelwano lwebhayoloji kunye ne-biophysical research. I-2001; 283: 236-242. [PubMed]
  23. UKourrich S, uRothwell PE, uKlug JR, uThomas MJ. Amava cocaine alawula i-bidirectional synaptic plasticity kwi-nucleus accumbens. J Neurosci. I-2007; 27: 7921-7928. [PubMed]
  24. IKravitz AV, iFreeze BS, iParker PR, uKay K, uThwin MT, uDeisseroth K, uKreitzer AC. Ukulawulwa kweendlela zokupaka i-parkinsonian ngokulawulwa kwe-optogenetic yecandelo lesiseko se-basal ganglia. Indalo. I-2010; 466: 622-626. [Inkcazelo yamahhala ye-PMC] [PubMed]
  25. LaPlant Q, Vialou V, Covington HE, 3rd, Dumitriu D, Feng J, Warren BL, Maze I, Dietz DM, Watts EL, Iniguez SD, et al. I-Dnmt3a ilawula indlela yokuziphatha kwemvakalelo kunye ne-spine plastikiity kwi-nucleus accumbens. Nat Neurosci. I-2010; 13: 1137-1143. [Inkcazelo yamahhala ye-PMC] [PubMed]
  26. Liu SJ, Zukin RS. I-Ca2 + -permeable receptors AMPA kwi-synaptic plastikiity kunye nokufa kwe-neuronal. Iindlela kwi-neurosciences. I-2007; 30: 126-134. [PubMed]
  27. Lu W, Isozaki K, Roche KW, Nikoll RA. Ukujolisa kwe-Synaptic ye-AMPA receptors kulawulwa yindawo yeCaMKII kwilokhi yokuqala ye-GluA1. I-Proc Natl Acad Sci US A. 2010; 107: 22266-22271. [Inkcazelo yamahhala ye-PMC] [PubMed]
  28. Lu W, Roche KW. Ummiselo wasemva kokurhweba ngabantu ngokusetyenzwa kwe-AMPA Uluvo lwangoku kwi-neurobiology 2011 [Inkcazelo yamahhala ye-PMC] [PubMed]
  29. I-Luscher C, i-Malenka RC. Ubukho beplastiki ye-synaptic yeziyobisi ezihanjiswe iziyobisi: ukusuka kutshintsho lweemolekyuli ekujikelezeni ngokutsha kwesekethe. Neuron. I-2011; 69: 650-663. [Inkcazelo yamahhala ye-PMC] [PubMed]
  30. Makino H, Malinow R. AMPA receptor ukufakwa kwii-synapses ngexesha le-PDP: indima yokuhamba kwe-lateral kunye ne-exocytosis. Neuron. I-2009; 64: 381-390. [Inkcazelo yamahhala ye-PMC] [PubMed]
  31. U-Mameli M, uHalbout B, uCreton C, u-Engblom D, uPititna JR, uSpangel R, uLuscher C. Cocaine-evoke i-synaptic plasticity: Ukunyanzeliswa kwe-VTA kubangela uhlengahlengiso kwi-NAc. Nat Neurosci. I-2009; 12: 1036-1041. [PubMed]
  32. IMasuda K, Koizumi A, Nakajima K, Tanaka T, Abe K, Misaka T, Ishiguro M. Ukubonakaliswa kweendlela zokubophana phakathi komntu onambithekayo okumnandi okumnandi kunye nobunzima obuphakathi kweesampulu zobunzima. I-PloS enye. I-2012; 7: e35380. [Inkcazelo yamahhala ye-PMC] [PubMed]
  33. UMatthews K, uWilkinson LS, uRobbins TW. Ukwahlulwa okuphindaphindiweyo koomama kwiigundwe ezisaphambi kokuzalwa kuzisa iimpendulo zokuziphatha kwizikhuthazi eziphambili kunye ezinemeko yokungena ebudaleni. IPhysol Behav. I-1996; 59: 99-107. [PubMed]
  34. UMax M, Shanker YG, Huang L, Rong M, Liu Z, Campagne F, Weinstein H, Damak S, Margolskee RF. I-Tas1r3, encodor i-receptor entsha yomkhethe, iyinyani kwisimemo sokuphendula esimnandi. Imfuzo yendalo. I-2001; 28: 58-63. [PubMed]
  35. McCutcheon JE, Beeler JA, Roitman MF. Iimpawu ezixeliweyo zokuxela kwangaphambili zivuselela ukukhutshwa kwe-phasic dopamine enkulu kunokukhupha i-saccharin-predicative cues. Ukuncipha. I-2012; 66: 346-351. [Inkcazelo yamahhala ye-PMC] [PubMed]
  36. UMcCutcheon JE, u-Wang X, u-Tseng KY, u-Wolf ME, u-Marinelli M. I-calcium calcium-evumayo ii-receptors ze-AMPA zikhona kwi-nucleus accumbens synapses emva kokurhoxa ixesha elide kwi-cocaine ye-self-management kodwa hayi i-cocaine elawulwa yi-cocaine. Ijenali ye-neuroscience: Ijenali esemthethweni yoMbutho weNeuroscience. I-2011a; 31: 5737-5743. [Inkcazelo yamahhala ye-PMC] [PubMed]
  37. McCutcheon JE, Loweth JA, Ford KA, Marinelli M, Wolf ME, Tseng KY. Iqela I mGluR isebenze ibuyisa ingqokelela ye-calcium esetyenziswayo ye-AMPA kwii-nucleus accumbenssynapses ngenqubo yokuxhomekeka kweprotein kinase C. J Neurosci. I-2011b; 31: 14536-14541. [Inkcazelo yamahhala ye-PMC] [PubMed]
  38. I-Nedelescu H, iKelso CM, iLazaro-Munoz G, i-phenura M, i-CK, i-Ledoux JE, i-Aoki C. I-GeoR1-ene-receptors ye-AMPA idlulisela kwii-asymmetric synapses kwi-amygdala yangemva kwesigaba sokuqala sokwakheka kwememori: i-elektrikhi ye-elektrikhi. funda. Ijenali ye-neurology yokuthelekisa. I-2010; 518: 4723-4739. [Inkcazelo yamahhala ye-PMC] [PubMed]
  39. UNelson G, Hoon MA, Chandrashekar J, Zhang Y, Ryba NJ, Zuker CS. Ii-receptors ezimnandi zaseMammalia. Iseli. I-2001; 106: 381-390. [PubMed]
  40. Owu MC, Derkach VA, Guire ES, Soderling TR. Ukurhweba nge-membrane ye-extrasynaptic kulawulwa yi-GluR1 i-serine 845 phosphorylation primes AMPA receptors ye-AMPA yokufumana ithuba elide. J Biol Chem. I-2006; 281: 752-758. [PubMed]
  41. I-Pascoli V, iTuriault M, uLuscher C. Ukujika kwe-cocaine-evoked synaptic potentiation ye-cocaine. Indalo. I-2012; 481: 71-75. [PubMed]
  42. Tyala K, Pelkey ​​KA, Bortolotto ZA, Morita D, Terashima A, McBain CJ, Collingridge GL, Isaac JT. Ukudityaniswa okwethutyana kwe-GluR2-ye-AMPA yokwamkela i-receptors ngexesha le-hippocampal yexesha elide. Nat Neurosci. I-2006; 9: 602-604. [PubMed]
  43. URada P, Avena NM, Hoebel BG. Ukubetha yonke imihla iswekile iphinda ikhuphe i-dopamine kwiqokobhe leqokelelo. I-Neuroscience. I-2005; 134: 737-744. [PubMed]
  44. URoche KW, O'Brien RJ, Mammen AL, Bernhardt J, Huganir RL. Ukubonakaliswa kweendawo ezininzi zephosphorylation kwi-AMPA receptor GluR1 subunit. Neuron. I-1996; 16: 1179-1188. [PubMed]
  45. I-Rumpel S, i-LeDoux J, iZador A, iMalinow R. I-Postynaptic receptor yokurhweba ngabantu ngokungekho mthethweni kwindlela yokufunda ngokudibeneyo. Inzululwazi. I-2005; 308: 83-88. [PubMed]
  46. USainz E, Korley JN, Battey JF, Sullivan SL. Ukuchongwa kwelungu lelinye losapho le-T1R yosapho lwe-receptors Ijenali ye-neurochemistry. I-2001; 77: 896-903. [PubMed]
  47. USerulle Y, uZhang S, uNinan I, uPuzzo D, uMcCarthy M, uKhatri L, u-Arancio O, uZiff EB. Ukusebenzisana kwe-GluR1-cGKII kulawula ukurhweba nge-AMPA receptor. Neuron. I-2007; 56: 670-688. [Inkcazelo yamahhala ye-PMC] [PubMed]
  48. I-Sesack SR, iGrace AA. ICortico-Basal Ganglia inethiwekhi yomvuzo: i-microcircuitry. I-Neuropsychopharmacology: ushicilelo olusemthethweni lweKholeji yaseMelika yeNeuropsychopharmacology. I-2010; 35: 27-47. [Inkcazelo yamahhala ye-PMC] [PubMed]
  49. USmith GP. I-Accumbens dopamine iguqula into enefuthe lokuvuselela i-orosensory by sucrose. Umdla. I-2004; 43: 11-13. [PubMed]
  50. Smith WB, Starck SR, Roberts RW, Schuman EM. Ukunyusa amandla e-dopaminergic yokufakelwa kweprotein yendawo yongeza ukucaciswa komhlaba kwe-GluR1 kunye nokuhanjiswa kwe-synaptic kwi-hippocampal neurons. Neuron. I-2005; 45: 765-779. [PubMed]
  51. Ilanga X, Milovanovic M, Zhao Y, Wolf ME. I-dopamine receptor ye-acute kunye engapheliyo yokunyanzeliswa kwemodyuli ye-receptor ye-AMPA kwi-nucleus iqokelela i-neurons ecociweyo nge-preonalal cortex neurons. Ijenali ye-neuroscience: Ijenali esemthethweni yoMbutho weNeuroscience. I-2008; 28: 4216-4230. [Inkcazelo yamahhala ye-PMC] [PubMed]
  52. Ilanga X, Zhao Y, Wolf ME. I-dopamine receptor stimulation modulates i-AMPA receptor synaptic yokufakelwa kwi-neurons yangaphambili ye-cortex. J Neurosci. I-2005; 25: 7342-7351. [PubMed]
  53. UThomas MJ, uBeurrier C, uBonci A, uMalenka RC. Uxinzelelo lwexesha elide kwi-nucleus accumbens: ulungelelwaniso lwe-neural yokuvuselela isimilo kwi-cocaine. Nat Neurosci. I-2001; 4: 1217-1223. [PubMed]
  54. I-Ungless MA, uWhistler JL, uMalenka RC, uBonci A. Ukuvezwa kwecocaine yodwa kwi-vivo induces yexesha elide kwi-dopamine neurons. Indalo. I-2001; 411: 583-587. [PubMed]
  55. UWhitlock JR, uHeynen AJ, uSwingr MG, uBhere MF. Ukufunda kufaka ithuba elide kwi-hippocampus. Inzululwazi. I-2006; 313: 1093-1097. [PubMed]