I-Proc Natl Acad Sci US A. Jan 19, 2010; I-107 (3): 1217-1222.
Ipapashwe kwi-Intanethi ye-Dec 29, 2009. doi: 10.1073 / pnas.0911998107
PMCID: PMC2824263
Neuroscience
Yan Liu,a,b UBrandon J. Aragona,c Kimberly A. Mncinci,a,b UDavid M. Dietz,b,d,e U-Mohamed Kabbaj,b,d UMichelle Mazei-Robison,e Eric J. Nestler,e kwaye Zuoxin Wanga,b,1
Eli nqaku liye khankanywe ngu amanye amanqaku kwi-PMC.
Abstract
Indawo yokudumisa (Microtus ochrogaster) Luphawu loluntu olugqibeleleyo loluntu olusenza ubumbano emva kokuncancisa, indlela yokuziphatha apho dopamine ephakathi (DA) ifakwe khona. Apha, sisebenzise ii-voes zodonga olungamadoda ukujonga iziphumo zokuvezwa kweziyobisi kukudibana kunye nokujikeleza okune-neural. Kwindlela yethu yokuqala yokuvavanya, i-amphetamine (i-AMPH) indlela yokuziphatha ekhuthaziweyo yavavanywa kusetyenziswa indawo ekhethiweyo (CPP) paradigm kwaye yaboniswa ukuba idluliselwe ngokusebenza kwe-D1-efana ne-receptors ye-DA. Emva koko, sivavanye iimpembelelo zokuboniswa okuphindaphindiweyo kwe-AMPH kwi-bonding. Inkqubo yokulawulwa kwe-Intact kunye ne-saline eyenziwe yenziwa ngabantu abathandayo, ngelixa amadoda enza umlinganiso kunye ne-AMPH kwi-doses esebenzayo yokuphembelela i-CPP isilele ukubonisa ukhetho olukhethiweyo lweqabane. Unyango olunjalo lwe-AMPH lukwaphucule i-D1, kodwa hayi i-D2, i-DA receptor expression in the nucleus accumbens (NAcc). Ngaphaya koko, i-pharmacological blockade ye-D1-efana ne-receptors ye-DA kwi-NAcc yahlangula ukhetho lwayo lomlinganiso okhulayo kumadoda aphethwe yi-AMPH. Ngokudibeneyo, idatha yethu ibonisa ukuba ukubonakaliswa okuphindaphindiweyo kwe-AMPH kunokunciphisa indlela yokuziphatha kwendawo yokudumisa eyamadoda kusetyenziswa indlela ethile ye-receptor ekhethekileyo kwi-NAcc, ekhokelela kukonakala kokonakala kwendibaniselwano.
Yamkelekile ngokubanzi into yokuziphatha ekhuthazayo kunye neemvakalelo ezikhuthaza ukuqina komzimba zilawulwa ngokujikeleza komvuzo wobuchopho kubandakanya inkqubo ye-mesolimbic dopamine (DA) (1, 2). Nangona le nkqubo ihlala ichukunyiswa kukutya kunye nokuziphatha ngokwesondo.3, 4), ikwanyanzeliswa kwezinye izinto ezivele ngokwendalo ezishukumisayo, ezinje ngokudlala phakathi kwabantwana abancinci kunye nolwalamano phakathi kwabazali kunye nabantwana (5-9). Ihlala idityaniswa kuphando ziibhanti zentlalo ezenziwa phakathi kwabantu abadala abatshatileyo, okt. Uphando lwakutsha nje usebenzisa iintlobo zezilwanyana ezinamagqabi ngokwasentlalweni, indalo yeNdawo (Microtus ochrogaster) (10-12), bonisa ukuba umsebenzi omkhulu wommiselo we-neural phantsi kwesiseko sebhondi kunye nokugcinwa kwenzeka ngaphakathi kwe-nucleus accumbens (NAcc) (13-15-I-mesolimbic yengingqi yengqondo ephakathi yokuziphatha1, 2, 16).
Nangona intshukumo yokujikeleza komzimba ivele ukuze ikhuthaze ukuqina komzimba njengokutya, ukhula kunye nomxokelelwano wentlalo (1, 17), isengozini yokusetyenziswa ngobugwenxa ngeziyobisi zokuxhatshazwa (8). Umzekelo, ukuphathwa kweziyobisi zepsypostantant zokuxhatshazwa, ezifana necocaine kunye ne-amphetamine (AMPH), kukhokelela kuguquko oluqhubekayo lomsebenzi we-mesolimbic DA (18, 19). Ifuthe elibi kakhulu kule sekethe ngezi kunye namanye amachiza amachiza acetyiswayo ukunciphisa ixabiso eliqondwayo lezibonelelo zezendalo (20), kubandakanya nezo zasentlalweni (8). Nangona kusaziwa ukuba iziyobisi zinikezela ukungaziphathi kakuhle ekuhlaleni (21), umgaqo wokunxibelelana phakathi kwamava iziyobisi kunye nokuncamathiselwa kwezentlalo akuqondwa kakuhle. Kungenxa yokuba, ngokuyinxalenye, konxibelelwano olunjalo kunzima ukubonisa imodeli kwiintambo zaselabhoratri zemveli ezingabonisi ubumbano lwentlalo phakathi kwabantu abadala.
I-neurobiology yolo nxibelelaniso lwasentlalweni, ngakumbi isibini esidityanisayo phakathi kwabantu abadala, ifundwe ngokubanzi kwigumbi lendalo (10-12), kwaye kutsha nje, le ntlobo isekwe njengemodeli esebenzayo yokuvavanya ixabiso lesishukumiso se-AMPH (22). Ngapha koko, zombini ukudalwa kobumbano kunye nokuqiniswa kwe-AMPH kudityanisiwe, ubuncinci ngenxalenye, ngothumelo lwe-DA ngaphakathi kwi-NAcc (14, 15, 23). Ke ngoko, isifundo esikhoyo sisebenzise umfuziselo we-prairie vole ukuseka isimilo sokuziphatha ukuze kufundwe iziphumo zokuvezwa kweziyobisi kwi-bonding yentlalo kwaye kugxilwe kwinkqubo ye-NAcc ye-DA yokubonisa inkqubo ye-neural phantsi kwezi mpembelelo zokuziphatha.
iziphumo
I-AMPH-Ekhutshelwe uXanduva lweNdawo ekhethiweyo (i-CPP) iPhakathi ku-DA kwi-Receptor-Specific Manner.
Ukwenziwa kwe-CPP kuchazwe kukonyuka okukhulu kwexesha elichithwe kwikheyibhuli ye-AMPH ngexesha lokugqibela, emva kweentsuku ze-3 ze-AMPH zokuma, ezinxulumene nesimilo. Nokuba ngaba kutofwe ialinine okanye i-saline eneedosi ezimbini ezisezantsi ze-AMPH ezivavanyiweyo (i-0.1 kunye ne-0.5 mg / kg) izinto ezikhethiweyo zekheji (Ikhiwane. 1A). Nangona kunjalo, abesilisa banemilinganiselo yedosi ephezulu ye-AMPH, kubandakanya i-1.0 (t = 2.87, P <0.01), 3.0 (t = 3.63, P <0.01), okanye 5.0 mg / kg (t = 3.03, P <0.01), ibonakalise i-CPP (Ikhiwane. 1A).
Kuba i-AMPH iyonyusa kakhulu i-neurotransuction ye-DA kwii-voes prairie (24), kwaye i-DA ilamula ukuqiniswa kwe-AMPH kwezinye iintlobo (23), ngoku sivavanye ukumiselwa kwe-receptor ye-DA (i-DAR) ye-AMP-ingene kwi-CPP kwii-voes prairie zamadoda. Izihloko zagqitywa kwi-paradigm ye-CPP, yanyangwa ngesaline okanye i-saline equlathe imithamo eyahlukeneyo ye-nonselective DAR antagonist (haloperidol) ngaphambi kwe-AMPH (1.0 mg / kg) inaliti ngexesha le-3 yeentsuku zokuma kwemeko, yaze yavavanywa i-CPP kwithuba lokugqibela. Izifundo eziphathwe nge-saline (t = 2.69, P <0.01) okanye i-saline equkethe iidosi ezimbini ezisezantsi ze-haloperidol (0.1 mg / kg; t = 3.62, P <0.01; 1.0 mg / kg; t = 3.89, P <0.01) phambi kwemeko ye-AMPH ibonakalise i-CPP eyenziwe nge-AMPH, ngelixa i-haloperidol kwi-5.0 mg / kg ivimbe i-CPP eyenziwe nge-AMPH, ibonisa ukubandakanyeka kwe-DARs kwiziphumo zokuziphatha kwe-AMPH (Ikhiwane. 1B). Ukufumanisa ukuba yeyiphi i-DAR subtype mediates i-AMPH-indened CPP, silawula ngokulandelayo njenge-D1-efana ne-antagonist ethile (SCH23390) okanye i-D2-efana ne-antxonist ethile (i-eticlopride) phambi kwe-injezelo ye-AMPH ngexesha lokulungisa. I-D2-efana ne-antagonism ayivimbanga i-CPP-indened ye-AMPH (t = 3.15, P <0.01 ye-0.5 mg / kg kunye t = 2.60, P <0.05 ye-5.0 mg / kg eticlopride) kodwa i-blockade ye-D1-efana ne-receptors isuse i-CPP eyenziwe nge-AMPH (Ikhiwane. 1B), ukubonisa ukuba i-CPP eyenzelwe i-AMPH ichukunyiswe ngokusebenza kwe-D1-efana, kodwa hayi njenge-D2-efana, ii-receptors kwii-voes zodumo zesilisa.
Ama-AMPH Amava Alter Mating-Inended Partner Preference Fuction.
Nangona i-CPP eyenzelwe i-AMPH ifuneka isebenze kwii-D1-receptors (Ikhiwane. 1B), ngaphambili sibonisile ukuba ukuvulwa kwe-receptors ye-D1-efana nokuthintela ukwakheka kwesixokelelwano somtshato (14). Ke ngoko, siye saqonda ukuba ukuvavanywa kwe-AMPH kunokuphazamisa ukuhlangana kwabantu ababhinqileyo kwi-voes prairie voles. Amadoda ahluleke baba ngamaqela amane angafumaniinalaliso (intact), inaliti ye-saline, okanye inaliti ye-1.0 okanye i-5.0 mg / kg AMPH kwiintsuku ze-3 (inaliti paradigm eyaneleyo yokuphelisa i-CPP). Ngosuku lwesine, yonke into eyindoda ibhinqileyo kunye nokwabelana ngesondookwabelana ngesondo kwe-24 h kwaye kuvavanywa kukhetho lweqabane. Iyahambelana nezifundo zangaphambili (14, 25-27), abesilisa abangamadoda nabesilisa abafumene inaliti ye-saline kwiintsuku ze-3 ngaphambi kokuzalela babonisa ukhetho olukhethayo lokulinganisa (amadoda angaphandle; t = 3.05, P <0.01, ityuwa engena ngamadoda; t = 3.21, P <0.01; Ikhiwane. 2A). Nangona kunjalo, amadoda awayexutywe nayiphi na idosi ye-AMPH kwiintsuku ze-3 ngaphambi kokuqhawula umtshato angakhange abonise ukukhetha kwiqabane (Ikhiwane. 2A). Ngokubalulekileyo, ubugqolo be-AMPH abuzange buphazamise ukukhula komzimba ngexesha lokutya (F(3, 26) = 0.26, P = 0.85; Ikhiwane. 2B) okanye umsebenzi wengxoxo ngexesha lokuvavanywa kukhetho lweqabane (F(3, 26)= 2.34, P = 0.10; Ikhiwane. 2C), ebonisa ukuba i-AMPH iphazamisane ngqo nokukhethwa kweqabane lokutshatisa.
Amava e-AMPH aphakamisa ii-Receptors ze-D1 kwi-NAcc.
Ngenxa yokuba i-AMPH i-preexpxpxpxpxpxpender ukhetho lomlingane kunye zombini i-AMP-ingene kwi-CPP (jonga ngentla) kunye nokubhangqa kokubini (14) zilawulwa yi-NAcc DA, saqonda ukuba i-AMPH iza kutshintsha kakhulu ukujikeleza kwe-mesolimbic kwi-voles yamadoda. Iingqondo zezifundo ezivela kolu vavanyo lokuziphatha zikhankanywe apha ngasentla zenziwa ukulungiselela ukufakwa iilebhile ze-mRNAs uphawu lwe-DA. Amadoda aphathwa nge-AMPH (1.0 mg / kg) abonise ukwanda okukhulu kwi-D1 receptor (D1R; t = 3.06, P <0.01), kodwa hayi i-D2 receptor (D2R), i-mRNA ilebheli ngaphakathi kwe-NAcc, xa kuthelekiswa namadoda afumana unyango lwangaphambi kwetyuwa (Ikhiwane. 3 A-C). Nangona kunjalo, akukho mahluko weqela wafunyanwa kubuninzi belebheli ye-mRNA ye-tyrosine hydroxylase (TH), umthuthi we-DA (i-DAT), okanye i-D2Rs ngaphakathi kwendawo yecandelo le-ventral tegmental (VTA) -ingingqi yengqondo ebonelela ngegalelo eliphambili ledopaminergic kwi-NAcc (Ikhiwane. 3 D-G). Ukunyuka kokuthetha kwe-D1R kwi-NAcc kwaqinisekiswa ngakumbi kukucinywa kweNtshona (t = 1.90, P <0.05; Ikhiwane. 3 H kwaye I). Ngokudibeneyo, ezi datha zibonisa ukuba ukubonakaliswa kwe-AMPH kune-receptor- kunye nefuthe lendawo ethile kwinkqubo ye-mesolimbic ye-DA ye-voereades eyindoda-ikhulisa inqanaba le-D1R kwi-NAcc.
I-D1 Receptors kwi-NAcc Mediate AMPH Ukunyanzeliswa kokukhetha kwiQabane.
Ngaphambili sibonisa ukuba, kwi-voes prairie voles, kusebenze ii-D1Rs ngaphakathi kwe-NAcc kuthintele ukwakheka kokukhetha kweqabane (14), kwaye isifundo esikhoyo sibonisa ukuba ukubonakaliswa kwe-AMPH-kulawula ii-D1Rs kwi-NAcc (Ikhiwane. 3). Ke ngoko, siye savavanya i-hypothesis ethi ukunganyanzeliswa kwe-AMPH kunyanzeliso lwezinto ezikhethwayo liqabane kulawulwa zii-D1R ngaphakathi kwe-NAcc. Amaduna afumene i-cannulation ye-stereotaxic ejolise kumazwe amabini kwi-NAcc shell (Ikhiwane. 4A). I-Artificial cerebrospinal fluid (CSF) yodwa okanye i-CSF equlathe imithamo eyahlukileyo ye-D1-efana ne-receptor antagonist SCH23390 yafakwa kwi-NAcc ngaphambi kokulimala kwe-AMPH (1.0 mg / kg) ngeentsuku ze3. Emva koko, izifundo zabhangqwa kunye nebhinqa kwi-24 h kwaye zavavanywa kukhetho lweqabane. Njengokulinga apha ngasentla (Ikhiwane. 2), Ukuvezwa kwe-AMPH kuthintele ukukhethwa kweqabane lokutshatisa okukhuthazayo kumadoda afumane inaliti ye-intra-NAcc ye-CSF okanye idosi ephantsi ye-SCH23390 (Ikhiwane. 4B). Nangona kunjalo, abesilisa abatofwe ngedosi ephezulu ye-SCH23390 (100 ng / kg) ukhetho olukhethiweyo kwiqabane (t = 2.55, P <0.05), ebonisa ukuba i-D1R blockade kwi-NAcc isuse ukungahambi kakuhle kwe-AMPH yokwakheka kokukhethwa kweqabane (Ikhiwane. 4B). Akukho mahluko weqela ufunyenwe kwisantya sokumanyanisa ngexesha lokuhamba okanye imisebenzi ye-locomotor ngexesha lovavanyo lokhetho lweqabane.
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Kolu phando, siphindaphinda ukufumanisa kwethu kwangaphambili ukuba ukubonakaliswa kwe-AMPH kuzisa i-CPP kwii-voes prairie zamadoda (22) kwaye ubonise ukuba ukwenziwa kwe-D1R kwi-NAcc kuyadingeka koku kuziphatha, iziphumo ezingqinelanayo nezifundo kolunye uhlobo lweentaka (28). Olu fundo, kunye nezifundo zangaphambili, licebisa ukuba iindlela ezizimeleyo ze-DAergic kwi-NAcc ilawule i-AMPH- kunye neendlela zokuziphatha ezishukunyiswa liqabane: Ukuziphatha okukhuthaziweyo kwe-AMPH (CPP) kugqaliselwe yi-D1R, ngelixa indlela yokuziphatha ekhuthazwayo yeqabane (ukuthanda iqabane) kuqhutywa yi-D2R Ukwenza kusebenze kwaye kuthintelwe ukusebenza kwe-D1R ngaphakathi kwe-NAcc (13-15). [Kubalulekile ukuqaphela ukuba, kwi-NAcc, kwi-D2R isebenze i-CPP ebangelwa zezi ziyobisi phantsi kweemeko ezithile (29) kwaye ukusebenza kwe-D1R kubandakanyeka kwezinye iindlela zokuziphatha ezixhaswe eluntwini, ezifana nezo zibhekiswe ngqo kwinzala (7, 30)].
Umahluko wokumiselwa kwe-DAergic kummiselo we-CPP kunye nokwakheka kokukhethwa kweqabane ikakhulu kufezekiswa ngumahluko kwinqanaba lokuvuselela-elenza inkqubo yokuxinana ye-DA isebenze kwi-DAR subtypes ezahlukeneyo. Ngenxa yomahluko wokudibana kobumbano, ukuphakanyiswa okumandla kwi-DA kuxinekile ukuze kusebenze ii-D1Rs eziphantsi, ngelixa ukunyuka okuthe tye kojoliso lwe-DA kukhetha ukusebenza ngobuninzi kwii-D2Rs (31). Kwiindawo zokudumisa, i-AMPH ikhupha unyuso olukhulu kuninzi lwe-DA (24) ngokuthelekisa oko kukhutshiwe15, 25). Ezi datha zibonisa ukuba inyuke ngokulingeneyo kuxinzelelo lwe-DA ngexesha lokusebenzisana (15, 25) Vumela ukwenza kusebenze okukhethekileyo kokuzibandakanya okuphezulu kwe-D2Rs kwaye ke ngoko kuququzelele ukwakhiwa kweqhina. Kwelinye icala, ukwanda okungqingqwa kukonyuka kwiDA emva kolawulo lwe-AMPH kunokwenzeka ngokwaneleyo ukuba kusebenze ii-D1Rs ezizezinye, ngalo ndlela kuququzelele i-CPP eyenzelwe i-AMPH.
Ukuphathwa ngqo komgaqo wokuziphatha okwenziwa yi-DA kuhambelana nezifundo zangaphandle ze-electrophysiological kwimigudu yokuhamba ngokukhululekileyo ebonisa ukuba iziyobisi kunye nokukhuthaza kwendalo kusonjululwa ngabantu abahlukileyo be-neuronal ngaphakathi kwi-NAcc (32). Nangona izifundo ezinje nge-elektriksiyali zingakwaziyo ukubona inkqutyana ye-receptors ye-DA evezwe kwii-neurons ezizodwa, izifundo zenzululwazi zibonakalisa ukuba i-NAcc yenziwe ngee-neuron ze-proion ezibonisa nokuba zii-D1R okanye ii-D2R ezine-coexpression encinci.33). Ii-Psychostimulants zikhetha ngokukhethekileyo kusebenze indlela yentsingiselo ye-intracellular esezantsi kwi-D1Rs (njengoko kumiselweyo ngokwanda kwephosphorylation yeemolekyuli zokubonisa) (34) kwaye ngaphambili sibonisile ukuba ukwanda kokusebenza kwezi ndlela zotyikityo kuthintela ukwakheka kukhetho lweqabane (35). Ke ngoko, nangona ukuhanjiswa kwe-DA ngaphakathi kwi-NAcc kudlala indima ebalulekileyo ekuphenduleni kwe-AMPH kunye nokwenziwa kokukhethwa kweqabane, ezi ndlela zokuziphatha zinokuthi zinqamlezwe kusetyenziswa i-microcircuitries eyahlukileyo ebandakanya iinkqubo zokuqikelela zestrakta (33, 36). Oku kubangela umdla kuba ukwakhiwa kwamabhondi kufakwa phakathi ngokwenza i-neuron ye-D2R-eveze iprojekthi yokuqala kwi-porral pallidum (33), omnye ummandla wobuchopho obalulekileyo ngokudibanisa izibini (26).
Kolu phononongo, sinika ubungqina bokuba amava e-AMPH athintela ukuqhawula umtshato kunye. Njengoko uvavanyo lokukhetha iqabane kolu vavanyo lwenziwa kwi-48 h emva kokuvezwa kwe-AMPH yokugqibela (okt, emva kokuba ichiza lenziwe umlo ngokupheleleyo), ezi datha zibonisa isiphumo esiqhubekayo se-AMPH kwibhondi yentlalo. Umatshini omnye onokubakho, ohambelana nefuthe le-AMPH kolunye uhlobo (37, 38), apho i-AMPH inokuthintela is bonding ngokwanda kwentetho ye-D1R ngaphakathi kwi-NAcc. Olu luvo luxhaswa yindima echasayo ye-NAcc D1Rs ekwakheni iqabane lokukhetha (14, 24) kwaye ngedatha yangoku ebonisa ukuhlangulwa kokukhethwa kweqabane elikhuselayo ne-intra-NAcc D1R blockade kwizilwanyana eziphathwe yi-AMPH (Ikhiwane. 4). Into enomdla kukuba, ngaphambili sibonisile ukuba, kwi-voes prairie voles, up-regulation of NAcc D1Rs emva kweeveki ze-2 ze-bonding zenza ukuba bakhethe uburhabaxa kubantu abangabaziyo, kubandakanywa nabasetyhini abamkeleyo ngokwesini, ukucebisa ukuba le nkqubo ye-neural neendlela ezivele ngayo Gcina izibini esele zisekiwe (14). Isifundo esikhoyo sicebisa ukuba i-AMPH yenziwe ngobuchule le neuroplasticity, ekhokelela ekunciphiseni kwe-AMPH ukungasebenzi kokubini. Kuyenzeka ukuba abesilisa abaphathwa nge-AMPH banokudibanisa obhinqileyo kunye nokuzikhupha kwi-AMPH ukurhoxiswa, kwaye ke lo mbutho ungathandekiyo unokubangela ukuthothisa kwe-AMPH. Uphononongo lwangaphambili lubonise ukuba ukurhoxa kwishedyuli eyandayo ye-d-amphetamine ukungaziphathi kakuhle ngokwesini kwizidudu zamadoda (39). Kwisifundo sethu, nangona kunjalo, ii-voes ezivela kumaqela awahlukeneyo onyango zibonise iindlela ezifanayo zokundala (Ikhiwane. 2B), ebonisa ukuba mhlawumbi babengekho kwimeko yokurhoxa kwi-amphetamine. Nangona kunjalo, oku kunokwenzeka kuqinisekisa ukuqhubeka kophando.
Kumiselwe kakuhle ukuba i-neural plasticised eyanyanzeliswa iziyobisi ibalulekile ekusebenziseni iziyobisi (40). Ii-Psychostimulants zitshintsha ngokumangalisayo ngobume beplastiki kwinkqubo ye-mesolimbic DA (41) kwaye kukho ubungqina bokuphakamisa ukuba utshintsho olunjalo lunyamezele ngakumbi kwi-D1 echaza i-neurons (37). Ngaphaya koko, izifundo ze-electrophysiological zibonise ukuba amava e-cocaine anganciphisa i-neuroplasticity elandelayo kwi-NAcc (42). Ngokwenyani, ukuhlengahlengiswa kwendalo okulungiselelwe iziyobisi kwi-NAcc kubonakalisa ulungelelwaniso lwendalo olwenzeka ngokwendalo emva kokubhencwa kwendalo kwimeko ezintsonkothileyo (43). Ke ngoko, nangona umlutha weziyobisi uchazwa ngokubanzi njengengxaki yokufunda kunye nememori (1, 40, 44,, kuhlala kuthathelwa ingqalelo yokuba ilahleko ebangelwa zezi ziyobisi inokubakho ngenxa yotshintsho olwongeziweyo kwimijikelezo ye-mesolimbic nayo inokuba yimfuneko yokuba likhoboka. Unciphiso olunjalo lweplastiki lunokunciphisa umda wokuziphatha kwakhona ekufuneni iziyobisi. Akumangalisi ke ngoko ukuba ii-voels eziphathwe nge-AMPH zibonise ukunciphisa ukuziphatha kwe-bonding. Iziphumo zangoku zokuvezwa kwe-AMPH kwisibini sokuqhawula umtshato ziyahambelana nezifundo zangaphambili ezibonisa isiphumo sokungasebenzi kakuhle kwengqondo kwezinye iindlela zokuziphatha, kubandakanya nokuziphatha kwabafazi (45-47) kunye nomdlalo wentlalo (48-50). Ngokudibeneyo, umsebenzi onjalo ubonelela ngesithembiso sokuba uvavanyo lwendlela iziyobisi kunye nenkuthazo yentlalo inxibelelana ngayo nengqondo zinokwandisa ukuqonda kwethu konxibelelwano olomeleleyo phakathi kokuziphatha kwezentlalo kunye nokusetyenziswa kweziyobisi ebantwini (21).
Impahla nenkqubo
Umxholo.
Izihloko yayingamabhongo endoda angamadoda okulala ngesondo avela kwigumbi lokuzala lokuzala. Izifundo zilunyulwe kwiintsuku ze-21 zeminyaka kwaye zigcinwa kwisibini sabantwana besini esifanayo kwizisele zeplastiki (i-12 × 28 × 16 cm) apho amanzi kunye nokutya kubonelelwe nge-ad libitum. Zonke iibheji zagcinwa phantsi kwe-14: Umjikelo wobumnyama be-10, kwaye iqondo lobushushu lalimalunga ne-20 ° C. Zonke izifundo zazimalunga neentsuku ze-90 ubudala xa kuvavanywa. Ukuqunjelwa kwe-stereotaxic kunye nokufakwa ngaphakathi kwamachiza e-DA kuchazwe ngokucacileyo kwenye indawo (14).
Uvavanyo lweehambo.
Uvavanyo lweCPP lwenziwa njengoko bekuchaziwe ngaphambili (22) ngaphandle kwezi zilandelayo. Ukukhethwa kwekheji lokuqala kwisifundo ngasinye kumiselwe kwi-30-min preest ngemini ye-1. Izihloko zalungiswa emva koko, ngexesha leeseshini ze-40, nge-AMPH ukuya kwisisele esingakhethwanga kunye ne-saline eya kwikheji ekhethiweyo (zombini i-AMPH kunye nenaliti ye-saline yanikwa ngosuku olufanayo, i-6 h ngaphandle) kwiintsuku ezilandelanayo ze-3 (iintsuku ze-2-4). Emva koko, izifundo zavavanywa (kwi-posttest) zobukho be-CPP ngomhla we-5.
Uvavanyo lokhetho lweqabane lwenziwe njengoko kuchaziwe ngaphambili (14). Ngamafutshane, izixhobo zovavanyo zazine-cage ephakathi (12 × 28 × 16 cm) edityaniswe ziityhubhu ezixineneyo (i-7.5 × 16 cm) ukuya kwizindlu ezibini ezifanayo ezifanayo kwindlu nganye yesilwanyana esishukumisayo. Izilwanyana ezivuselelayo yayililo "iqabane" eliqhelekileyo (iqabane lesetyhini lesi sifundo) kunye "nomntu ongaziwayo" (ibhinqa elingakhange lihlangane naloo mbandela) ezazitsalwa ngokukhuselekileyo ngaphakathi kwizisele zazo ezahlukeneyo ngaphandle kokudibana ngqo omnye komnye. Ekuqalisweni kovavanyo lwe-3-h, izifundo zabekwa kwindawo esembindini kwaye zavunyelwa ukuba zihambe ngokukhululekileyo kuyo yonke izixhobo zokusebenza. Izinto zokuziphatha zarekhodwa kusetyenziswa inkqubo yokurekhoda ividiyo yexesha. Iingcali ezingaboniyo kubuqhetseba zihlolisise ikhasethi kwaye yarekhoda indlela yokuziphatha kwesifundo. Ukukhetha iqabane kuchazwe njengomxholo ochitha ixesha elingakumbi kunxibelelwano ngecala kunye neqabane kunasemzini, njengoko kubonisiwe kwiisampulu ezibini. t vavanya (27).
Kwimo ye-Hybridization kunye ne-Western Immunoblotting.
Amagqabantshintshi athile achaphazelekayo1 Table) zazisetyenziselwa i-D1R, D2R, TH, kunye ne-DAT kwi-lm mRNA yokulebhelisha. Ukwenza iilebheli kwenziwa kunye 35Iziphumo zelebheli kunye nolawulo lwe-mRNA yesiphawu ngasinye se-DA njengoko kuchaziwe ngaphambili (51). Uhlalutyo lwe-Western blotting, iprotheyini ye-DAR yakhutshwa kwi-supernatant ye-NAcc tishu ze-punches zaze zathwaxwa njengoko kuchaziwe ngaphambili (52).
Ukulinganiswa kwedatha kunye nohlalutyo.
I-CPP kunye nezinto ezikhethwa liqabane zichazwe ziisampulu ezibini t iimvavanyo. Umahluko weqela kwimidlalo yokuphelisa ukhula ngexesha le-6 yokuqala yokubhangqa nomntu obhinqileyo kunye nohlobo lwamakheji ngexesha lovavanyo lokhetho lweqabane luye lwahlalutywa yi-ANOVA. Ukuxinana kwamehlo kwe-D1R kunye ne-D2R mRNA ilebheli kwi-NAcc kunye ne-TH, i-DAT, kunye ne-D2R mRNA yokulebhelisha kwi-VTA yagqitywa kwi-autoradiograms kusetyenziswa inkqubo yomfanekiso wecompyuter (NIH IMAGE 1.64). Idatha yathunyelwa njengepesenti yeentsingiselo zeqela lokulawula i-saline kunye neqela umahluko lahlaziywa t iimvavanyo. Okokugqibela, uxinano lokujonga lwe-D1R kunye ne-D2R kwelebheli ye-x-ray kwifilimu yovavanyo lwaseNtshona. t iimvavanyo.
Uyilo lovavanyo.
I-1a yesilingo iseke ijika yedosi-yokuphendula ye-AMP ebangelwa yi-AMPH. Izihloko zahonjiswa kwizixhobo ze-CPP ngemini ye-1, yabelwa ngokulandelelana kwelinye lamaqela amathandathu okuvavanya afumana i-ip (ip) yealine enezinto ezahlukeneyo zoxinzelelo lwe-AMPH [0 (n = 12), 0.1 (n = 8), 0.5 (n = 9), 1.0 (n = 12), 3.0 (n = 12), okanye 5.0 mg / kg (n = 13)] ngeentsuku ze-3 (usuku lwe-2-4) lweemeko, kwaye emva koko kuvavanyelwa i-CPP kwindawo yokuqala ngomhla we-5.
I-1b yesilingo ityhile iindima ze-receptors ze-DA kwi-AMPH eyenzelwe i-AMP. Izihloko zahonjiswa kwizixhobo ze-CPP zaza zabelwa ngokulandelelana kwelinye lamaqela asibhozo okuvavanywa afumene inaliti (sc) yesaline (n = 10) okanye i-saline equlathe ukugxila okungafaniyo kwe-nonselective DA receptor antagonist [haloperidol; 0.1 (n = 8), 1.0 (n = 8), okanye 5.0mg / kg (n = 8)], okanye i-D1-efana ne-SCH23390; 0.5 (n = 7) okanye 5.0 mg / kg (n = 7)] okanye D2-njenge-DA receptor antagonist ethile [eticlopride; 0.5 (n = 8) okanye 5.0 mg / kg (n = 8)]. Imizuzu engamashumi amathathu kamva, idosi yomkhawulo ye-AMPH (1.0mg / kg), eyanyanzela i-CPP kwi-Testion 1a, yasetyenziselwa i-AMPH imeko yokuphanda. Emva kweentsuku ze-3 zemeko ye-AMPH, zonke izifundo zifumene i-CPP posttest.
Uvavanyo lwe-2 luhlolisise ukuba amava e-AMPH aphazamisekile kwi-bonding. Izifundo zabelwa ngokulandelelana kwelinye lamaqela amathathu ovavanyo afumene i-ip sindano ye-saline (n = 8) okanye i-saline equkethe i-1.0 mg / kg (n = 8) okanye 5.0 mg / kg (n = 7) i-AMPH kube kanye ngemini ngeentsuku ze-3 ngokulandelelana-iparadigm ebangele i-CPP kwi-voes yediries eyindoda. Ngosuku lwesine, izifundo zadityaniswa zibhinqile ibhanti ye-estrogen ye-24 h (14), kwaye ke kuvavanywa kuvavanyo lweqabane le-3-h. Ukulawula iimpembelelo ezinokubakho zezitofu kukubhanti, iqela lesine lovavanyo lwenyani lwamaduna angakhange afumane nanye inaliti (n = I-6) yahanjwa ngabasetyhini abane-estrogen-primed ye-24 h kwaye yavavanywa kukhetho lweqabane. Zonke iimvavanyo zokuziphatha zazenziwe zaza zangqinwa ividiyo ukuze kuqinisekwe ukuba iyamkelwa. Ixesha lokufundwa kwamacandelo kunye neqabane kunye nomphambukeli lagqitywa. Ukongeza, imisinga yokuqhawuka komtshato ngexesha le-6 yokuqala yokubhanqa kunye nomsebenzi we-locomotor (eboniswe kukuwela ikheyi) ngexesha lovavanyo lwe-3-h lomlinganiso okhethwayo wagqitywa. Emva kovavanyo lokukhetha iqabane, izifundo zabulawa kwangoko. Zonke iingqondo zivuniwe, zomkhenkce emkhenceni owomileyo, kwaye zigcinwe kwi-−80 ° C ukulungiselela ukufakwa iileyibheli ye-mRNA ye-DA.
Uvavanyo lwe-3 luvavanye ukuba ukungqinisisa isibini se-AMPH kunxulunyaniswa notshintsho kwimisebenzi ye-DA ye-mesolimbic. Iingqondo ezivela kwizifundo ezifumene i-saline (n = 8) okanye 1.0 mg / kg AMPH (n = 8) kwi-Testion 2 yanqunyulwa kwi-cryostat yaba ngamacandelo e-coronal (ubukhulu be-14 μm) eyayithwelwe kwi-Superfrost / plus slides (iFisher Science Science). Amacandelo obuchopho kwizithuba ze-98-werem kwenziwa ukuba kufakwe iileyibheli ze-D1R, D2R, TH, kunye ne-DAT mRNA. Ngenxa yokuba ii-voles ezinamava nge-1.0 mg / kg AMPH zibonisa ukonyuka kwe-D1R, kodwa ingeyiyo i-D2R, ukulebhelisha kwe-mRNA kwi-NAcc, kuthelekiswa nolawulo oluchaphazelekileyo nge-saline, amaqela amabini ongezelelweyo ezifundo ezifunyenweyo atofwe iinaliti zesaline (n = 6) okanye 1.0 mg / kg AMPH (n = 6), 24 h yokuzalela, kunye novavanyo lokukhetha iqabane, njengoko kuchaziwe ngasentla. Izihloko zachazwa kwaye iibhutsi zazicwecwa kwi-cryostat ebukhulu be-300 μm. Izithintelo zethishu zathatyathwa kabini kwi-NAcc zalungiswa nge-D1R kunye ne-D2R Western blotting.
Uvavanyo lwe-4 luhlolisise ukuba ingaba i-activation ye-D1-receptors kwi-NAcc inoxanduva lokuthothisa i-AMPH. Izihloko zafakelwa kwisikhokelo se-cannulae ngokujikelezayo ijolise kwibhola ye-NAcc. Emva kweentsuku ze-3 zokufumana kwakhona, babelwa ngokulandelelana kwelinye lamaqela amathathu olingo apho bafumana inaliti ye-intra-NAcc ye-CSF (200 nL / side, n = 11), okanye i-CSF equkethe i-0.4 (n = 6) okanye 100 ng / icala (n = 7) SCH23390. Imizuzu engamashumi amathathu kamva, bafumana i-ip sindano ye-1.0 mg / kg AMPH. Le nkqubo yaphindaphindeka kwiintsuku ezilandelelanayo ze3. Ngosuku lwesine, izifundo zabhangqwa ngumfazi ovunyiweyo we-estrogen ye-24 h emva koko zavavanywa kukhetho lweqabane.
Imibulelo
Ababhali bayambulela uKyle Gobrogge, uClaudia Lieberwirth, uKelly Lei, noMelissa Martin ngokufunda le ncwadi ebhaliweyo. Lo msebenzi uxhaswe ngamaZiko eSizwe amaZiko ezeMpilo i-MHR01-58616, DAR01-19627, kunye ne-DAK02-23048 ukuya ku-ZW
Ucaphulo
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