Phello ea ΔFosB ho tlōla tekano tabeng ea pontšo ea li-opioid le ea kannete e amohelehang ho nucleus accumbens (2011)

Neuropharmacology. 2011 Dec;61(8):1470-6. doi: 10.1016/j.neuropharm.2011.08.046.

Sim-Selley LJ, Cassidy MP, Sparta A, Zachariou V, Nestler EJ, Selley DE.

mohloli o moholo

Lefapha la Pharmacology le Toxicology le Institute for Drug and Alcohol Studies, Virginia Commonwealth University Sekolo sa Bongaka, Richmond, VA 23298, USA.

inahaneloang

Ntho e tsitsitseng ea transcription ΔFosB e kenyelletsoa nucleus accumbens (NAc) ka ho ipeha kotsing ea lithethefatsi tse ngata tsa tlhekefetso, 'me polelo e sa tloaelehang ea ΔFosB ka striatum e eketsa maruo a monate a morphine le k'hok'heine. Leha ho le joalo, motheo oa mekhoa ea lipatlisiso tsena o utloisisoa ka ho feletseng. Re ne re sebelisa mohlala oa mouse oa bitransgenic ka polelo e sa hlalosoang ea DFBB ho dopamine D (1) li-neurons tse hlahisang li-receptor / dynorphin ho fumana hore na phello ea polelo ea ΔFosB mabapi le pontšo ea li-opioid le ea cannabinoid ho amohela NAc. Lipatlisiso li bontšitse hore ts'ebetsong ea G-protein e entsoeng ka opioid le thibelo ea adenylyl cyclase e ile ea ntlafatsoa ho NAc ea litoeba tse hlalositseng DFFB. Ka mokhoa o ts'oanang, thibelo ea kappa ea opioid ea adenylyl cyclase e ile ea ntlafatsoa ho ΔFosB e hlahisang litoeba. Ka lehlakoreng le leng, pontšo ea mekhoa ea ho amohela li-cannabinoid e ne e sa tšoane pakeng tsa litoeba tse fokolang maikutlo ΔFosB le litoeba tsa ho laola. TLiphuputso tse entsoeng ka mahlakoreng a mang li bontša hore pontšo ea opioid le cannabinoid receptor e khetholloa ka mokhoa o fapaneng ka ho bonts'a ΔFosB, mme e bontša hore polelo ea ΔFosB e ka hlahisa liphello tsa eona ka tumello ea ho amohela le ea kappa opioid receptor ka NAc.

Keywords: G-protheine, adenylyl cyclase, striatum
Eya ho:

1. Selelekela

Li-receptors tsa opioid le CB ea cannabinoid1 li-receptors (CB1R) ke sepheo sa neurobiological bakeng sa lihlopha tse peli tse sebelisoang haholo tsa lithethefatsi tse akarelletsang morphine, heroin le opioids ea ngaka, le matekoane (Δ9-Tetrahydrocannabinol (THC)), ka ho latellana. Liphello tse bohloko tsa li-opioids le li-cannabinoids li nahanngoa ke li-G-protein-coupled receptors tse sebelisang haholo-holo Gi / o liprotheine le ho hlahisa likarabo tse ka tlaase ho metsoako e kang ho thibela adenylyl cyclase (Bana ba bang, 1991, Childers, le al., 1992, Howlett, et al., 2002). Mokhoa oa ho khanna, oa ho hopola lintho le liphello tsa kelello tsa Δ9-THC e hlahisoa ke CB1R (Huestis, et al., 2001, Zimmer, et al., 1999), tse ajoang haholo bokong, tse nang le litekanyetso tse phahameng ho basal ganglia, hippocampus le cerebellum (Herkenham, et al., 1991). Liphello tse ts'oanang le tse khotsofatsang tsa meriana e mengata e sebetsang le e hlekefetsoang ea li-opioid li nahanngoa haholo-holo ka li-receptors tsa opioid (MOR) (Matthes, et al., 1996), tse ntlafalitsoeng tsamaisong ea limbic le mokokotlo oa bokooa (Mansour, et al., 1994). Mokhoa oa mahlolimbic, o entsoeng ka dopaminergic sepheo sa ventral tegmental (VTA) ho nucleus accumbens (NAc), se phetha karolo ea bohlokoa liphellong tse khotsofatsang tsa li-opioid le li-cannabinoids (Bozarth le Bohlale, 1984, Vaccarino, et al., 1985, Zangen, le al., 2006), hammoho le lithethefatsi tse ling tsa tlhekefetso (Koob le Volkow, 2010). Ho feta moo, mekhoa e metle ea opioid le cannabinoid e ameha liphellong tse putsang tsa lihlopha tse ngata tsa lithethefatsi tsa psychoactive (Maldonado, et al., 2006, Trigo, et al., 2010). Kahoo, ke habohlokoa ho hlakola mekhoa ea opioid le CB1R ho bonahatsa ho laoloa ho NAc.

Potso e ka sehloohong tšimong ea tšebeliso e mpe ea lithethefatsi e bile ho khetholla liprotheine tse lumellanang le phetoho e tlohang ho tloha boemong bo matla ho isa nakong e telele ea liphello tsa lithethefatsi tsa kelello. Ntho ea transcription ea AP-1 ΔFosB e thahasellisa ka ho khetheha hobane ke sefate se tsitsitseng se entsoeng ka truncated sehlahisoa sa fosbe liphatsa tsa lefutso tse hlahelang hangata ha li pepesehela lithethefatsi tsa tlhekefetso kapa meputso ea tlhaho (McClung, et al., 2004, Nestler, 2008, Nestler, et al., 1999). Re fumane hore ΔFosB e kenyelletsoa bokong bo latelang ho pepesehela morphine, Δ9-THC, k'hok'heine kapa ethanol, e nang le lithethefatsi ka 'ngoe e hlahisang mokhoa o ikhethang oa sebaka sa DFFB (Perrotti, et al., 2008). Ho fumana lithethefatsi tse sa khaotseng e ne e le hore ΔFosB e ne e kentse letsoho haholo ho statum, moo lithethefatsi tsohle tse 'nè li ileng tsa susumelletsa ΔFosB kahare ea NAc le bohle ntle le Δ9-THC e susumelitsoe haholo ho NAc shell le caudate-putamen.

Liphuputso tsa bongaka li bontšitse hore li-co-administration tsa dopamine D1 amohela (D1R) mohanyetsi SCH 23390 o thibetse ΔFosB ho kenya letsoho ho NAc le caudate-putamen ka mor'a hore ho be le li-cocaine kapa tsamaiso ea morphine, e leng se bontšang bohlokoa ba D1R-ho hlalosa li-neurons (Muller le Unterwald, 2005, Nye, et al., 1995). Phello ea ΔFosB ho qobelloa ho itšoara ka lithethefatsi e entsoe lipatlisiso ho sebelisa litoeba tsa bitransgenic tse hlalosang DFB ho batho ba itseng ba neuronal ba NAc le dorsal striatum (Chen, et al., 1998). Litoeba tse hlalosang DFBB ka dynorphin / D1R positive neurons ka NAc le dorsal striatum (line 11A) ho bonts'a likarabo tse fetotsoeng lithethefatsi tsa tlhekefetso, haholo-holo kutloisiso e matlafatsang liphello tse khotsofatsang tsa cocaine kapa morphine (Colby, et al., 2003, Kelz, et al., 1999, Zachariou, et al., 2006). Liphetoho tsena li etsahetse ka ho ba sieo ha liphetoho maemong a MOR kapa mekhoa e mengata ea li-G-protheine. Leha ho le joalo, litekanyetso tsa dynorphin mRNA li fokotsoe ka NAc ea ΔFosB e hlahisang litoeba (Zachariou, et al., 2006), ho fana ka maikutlo a hore sepheo se seng sa ΔFosB ke liphatsa tsa lefutso tse nang le peptide e nang le opioid e sa feleng. ΔFosB ho kenya letsoho ho ka 'na ha hlahisa liphetoho tsa boitšoaro ka ho laola pontšo ea receptor ka NAc, empa ho ka etsahala hore ebe ha hoa phenyekolloa. Ka hona, lipatlisiso tsa hona joale li sebelisitse mohlala oa mouse oa bitransgenic ho bona hore na ho tlōla tekano ha DFFB ho dynorphin / D1R e nang le li-neurons tse hlaselang li fetola ketsahalo ea MOR-mediated G-protein e sebetsang le NAc. Phello ea ΔFosB ka CB1Ketsahalo ea G-protein e kopanetsoeng R e boetse e hlahlojoa hobane Δ9-THC tsamaiso e kenyelletsa DFFB ho NAc (Perrotti, et al., 2008) 'me tsamaiso ea endocannabinoid e tsebahala ho laola lipotoloho tsa boko ba boko (Gardner, 2005, Maldonado, et al., 2006), empa phello ea ΔFosB tsamaisong ea endocannabinoid ha ea etsoa lipatlisiso.

Eya ho:

2. Lisebelisoa le mekhoa

2.1. Reagents

[35S] GTPγS (1250 Ci / mmol), [α-32P] ATP (800 Ci / mmol) le [3H] CAMP (26.4 Ci / mmol) li rekiloe ho PerkinElmer (Shelton, CT). ATP, GTP, GDP, CAMP, serum albumin, creatine phosphokinase, papaverine, imidazole le WIN-55212-2, li ile tsa rekoa ho Sigma Aldrich (St. Louis, MO). GTPγS e rekiloe ho tloha Roche Diagnostic Corporation (Chicago, IL). DAMGO e fanoe ke Lenane la Phepelo ea Lithethefatsi la Setsi sa Sechaba sa Tlhekefetso ea Meriana (Rockville, MD). Econo-1 scintillation fluid e fumanoe ho Fisher Scientific (Norcross, GA). Ecolite scintillation fluid e fumanoe ho ICN (Costa Mesa, CA). Lik'hemik'hale tse ling tsohle li fumanoe ho Sigma Aldrich kapa Fisher Scientific.

2.2. Litoeba

Litoeba tsa bitransgenic tsa banna tse tsoang NSE-tTA (line A) × TetOp-ΔFosB (line 11) li ile tsa hlahisoa joalokaha ho hlalositsoe ho Kelz et al. (Kelz, et al., 1999). Litoeba tsa Bitransgenic li ile tsa emoloa 'me tsa tsosoa ka doxycycline (100 μg ka metsi a nooang) ho thibela polelo ea litsela. Nakong ea libeke tsa 8, doxycycline e ile ea tlosoa metsing bakeng sa halofo ea litoeba ho lumella polelo ea litloaelo, athe litoeba tse setseng li ne li bolokiloe ka doxycycline ho thibela litlōlo tsa molao. Likhopolo li ile tsa bokelloa libeke tsa 8 hamorao, nako eo liphello tsa transcription tsa ΔFosB li fokolang haholo (McClung le Nestler, 2003). Sehlopha sa bobeli sa lintlheng tsa phetoho se ne se sebelisoa moo Dc-Jun, e leng mohanyetsi ea hanyetsang oa C-Jun, o hlalositsoeng ka D1R / dynorphin le D2R / enkephalin lisele tsa striatum, hippocampus le parietal cortex (Peakman, et al., 2003). C-Jun le liprotheine tsa lelapa la Jun li fokotseha le liprotheine tsa lelapa la Fos ebe li tlama ho sebaka sa AP-1 sepheo sa liphatsa tsa lefutso ho laola phetolelo. Leha ho le joalo, truncation ea N-terminus ea c-Jun (Δc-Jun) e etsa hore mokhoa o rarahaneng oa ho ngoloa ka molao o se o sebetse ebile o khona ho thibela DNA ho tlamisa mekhoa e sebetsang ea AP-1. Litoeba tsa bitransgenic tsa banna tse tsoang NSE-tTA (line A) × TetOp-FLAG-Δc-Jun (line E) li hlahisitsoe joalokaha ho hlalositsoe Peakman et al. (Peakman, et al., 2003). Litoeba tsa Bitransgenic li ile tsa emoloa 'me tsa tsosoa ka doxycycline (100 μg ka metsi a nooang) ho thibela polelo ea litsela. Li-pups li ne li longoa libeke tsa 3, li-genotyped, 'me li arohane ka lihlopha, li na le halofo li bolokiloe metsing a nang le doxycycline le halofo metsi a nooang a kamehla bakeng sa ho hlahisa maikutlo a FLAG-Δc-Jun. Likopi li bokelitsoe libeke tsa 6 hamorao, nako eo maemong a phahameng a FLAG-Δc-Jun a lekantsoeng (Peakman, et al., 2003). Lisebelisoa tsohle tsa liphoofolo li ne li khannoa ho ea ka Mekhatlo ea Sechaba ea Tataiso ea Bophelo bakeng sa tlhokomelo le tšebeliso ea liphoofolo tsa liphoofolo.

2.3. Ho itokisa sekheo

Bokooa bo ne bo bolokoa ho-80 ° C ho fihlela letsatsi la teko. Pele ho leka, boko bo bong le bo bong bo ne bo qhibilihisoa, 'me NAC e ne e aroloa ka leqhoa. Sampula e 'ngoe le e' ngoe e ne e tšoantšoa le 50 mM Tris-HCl, 3 mM MgCl2, 1 mM EGTA, pH 7.4 (buffer membrane) le likhetho tsa 20 tse tsoang ho homogenizer ea khalase ho 4 ° C. Homogenate e ne e le centrifuged 48,000 × g ka 4 ° C bakeng sa 10 min, e tsosolositsoe ka mochine oa membrane, e khutliselitsoe hape ho 48,000 × g ka 4 ° C bakeng sa minara ea 10 mme e tsosolosoa ka XMUMX mM Tris-HCl, 50 mM MgCl2, 0.2 mM EGTA, Naxl XMUMX mM, pH 100 (testay buffer). Maemo a protheine a ne a khethiloe ke mokhoa oa Bradford (Bradford, 1976) ho sebelisa serum albumin (BSA) e le tekanyetso.

2.4. Agonist-Stimulated [35S] GTPXS Ho koala

Litobele li ne li e-s'o qobelloa bakeng sa metsotso ea 10 ka 30 ° C le adenosine deaminase (3 mU / ml) ka tekanyo ea liteko. Li-membrane (5-10 μg liprotheine) li ne li kenngoa ka 2 hr ka 30 ° C ka tape e nang le 0.1% (w / v) BSA, 0.1 nM [35S] GTPγS, 30 μM GDP le adenosine deaminase (3 mU / ml) le ntle le likhahla tse nepahetseng tsa DAMGO kapa WIN55,212-2. Ho tlamahala ho sa hlokahaleng ho ne ho lekantsoe ka 20 μM GTPγS. Tsubollo e ile ea felisoa ka ho hloekisoa ka lihlahisoa tsa galase tsa galase tsa GF / B, tse lateloang ke ho hlatsoa ha 3 le 3 ml ea metsi a batang 50 mM Tris-HCl, pH 7.4. Motsoako oa radioactivity o ne o khetholloa ke spectrophotometry ea scintillation ea metsi ka mor'a hore ho sebelisoe bosiu bo lekaneng ho li-filter ho Econo-1 scintillation fluid.

2.5. Adenylyl Cyclase Assay

Li-membrane (5-25 μg protein) li ne li e-na le li-adenosine deaminase tse hlalositsoeng ka holimo, ebe li kenngoa bakeng sa 15 min ka 30 ° C ka ho ba teng kapa ho ba sieo ha 1μM forskolin, ntle le DAMGO, U50,488H kapa WIN55,212-2. 50 μM ATP, [α-32P] ATP (1.5 μCi), 0.2 mM DTT, 0.1% (w / v) BSA, 50 μM AMP, 50 μM GTP, 0.2 mM papaverine, 5 mM phosphocreatine, 20 diyuniti / ml creatine phosphokinase le adenosine deaminase (3 mU / ml) karolong ea ho qetela ea 100 μl. Tlas'a maemo ana, kakaretso [α-32P] cAMP e fumanoeng e fumaneha ka kakaretso e ka tlaase ho 1% ea kakaretso ea kakaretso [α-32P] ATP ka mohlala o mong le o mong. Karabo e ile ea felisoa ka ho pheha bakeng sa 3 min le [32P] Mokhoa oa ho etsa li-cyclic AMP o ne o arotsoe ke mekhoa e mabeli ea khoeli (Dowex le alumina) ea Salomon (Salomon, 1979). [3H] CAMP (10,000 dpm) e ne e kenngoa thupeng e 'ngoe le e' ngoe pele ho khalase ea chromatography e le tekanyo ea ka hare. Tlhaselo ea motsoako e ne e khethoa ke spectrophotometry ea scintillation (45% e sebetsang hantle bakeng sa 3H) ka mor'a hore 4.5 ml ea ho qhibiliha e qhibilihe ka 14.5 ml ea Ecolite scintillation fluid.

2.6. Tlhahlobo ea lintlha

Ntle le ha ho bontšitsoe ka tsela e 'ngoe, data e tlalehoa e le litekanyetso tsa bohlokoa ± SE tsa liteko tse fapaneng tsa 4-8, e' ngoe le e 'ngoe e neng e etsoa ka katleho. Net-stimulated [35S] GTPyS ho tlama e baloa e le agonist-susumelitsoeng tlama tlaase tlaase tlaal binding. Kakaretso ea motsoako oa forskolin e susumelitsoeng ke adenylyl cyclase e hlalosoa e le mosebetsi oa forskolin-o tsosolositsoeng - ts'ebeliso ea basal (pmol / mg / min). Tšebetso ea li-percentage ea mosebetsi oa forskolin-o susumelitsoeng ke adenylyl cyclase o hlalosoa e le (ts'ebetso ea toskolin-e tsosolositsoeng ka ho ba sieo ha agonist-net forskolin-ts'ebetso e tsosolositsoeng ho ba teng ha agonist / net forskolin-ts'ebetso e tsosolositsoeng ka ho ba sieo ha agonist) × 100. Lipheputso tsohle tse loketseng le tsa lipalo li ne li etsoa ka Prism 4.0c (GraphPad Software, Inc., San Diego, CA). Likhahla tsa maqakabetsing li ile tsa hlahlojoa ke khatello ea litsela e sa tsitsitseng ho fumana EC50 le E, max litekanyetso. Tlhaloso ea lintlha tsa data-concentration e entsoeng ka mahloriso e ne e khethoa ka ho hlahloba ka tsela e le 'ngoe ea li-variance (ANOVA), ka ho sebelisa tekanyo ea bo-agonist le ho kenngoa ha liphatsa tsa lefutso (ho tloha kapa ho tima) e le lintho tse ka sehloohong. Ntho ea bohlokoa ea lintlha tsa mekhoa e metle ea sekhahla (E, max kapa EC50T-teko ea Seithuti se se nang mehatla e 'meli, e sebelisa khalemelo ea Welch kapa phetoho ea methapo ea data moo ho hlokahalang ho lokisa liphapang tse sa lekanang (tse fumanoeng ke F-test) ho EC50 litekanyetso.

Eya ho:

3. Liphetho

3.1. Phello ea polelo ea ΔFosB mabapi le ts'ebeliso ea protheine ea G-opioid le ea cannabinoid

Ho fumana hore na MOR kapa CB1Mokhoa o sebetsang oa G-protheine o kopantsoeng ke R o fetotsoe ke polelo e hlalosang mantsoe a hau ea ΔFosB ka NAc, e susumelitsoeng ke agonist [35S] GTPγS e tlamisa e ne e hlahlojoa ka lisele tse ka thōko tse lokiselitsoeng ho tloha sebakeng sena sa litoeba tsa bitransgenic maemo a hlalosang (ΔFosB ka) kapa ho sa hlahise (ΔFosB holimo) phapang ea FFB. MOR-sele-selective enkephalin analog DAMGO e ne e sebelisetsoa ho etsa MOR le mefuta-futa ea aminoalkylindole WIN55,212-2 e ne e sebelisetsoa ho etsa hore CB1R. Li-ligands li kile tsa bontšoa hore ke li-agonists tse feletseng ho MOR le CB1R, ka ho latellana (Breivogel, et al., 1998, Selley, et al., 1997). E ne e sa khonehe ho hlahloba ketsahalo ea G-protein e tsamaisanang le KOR hobane pontšo e le tlaase haholo bokong ba rodent (Childers, le al., 1998). Liphello li bontšitse ts'ebetsong ea ts'ebetso ea G-protein ka bobeli DAMGO le WIN55,122-2 ho NAc ho tloha ΔFosB ho tloha le ΔFosB ho litoeba (Setšoantšo sa 1). Bakeng sa mosebetsi oa stimulated DAMGO (Setšoantšo sa 1A), ANOVA ea litsela tse peli ea tlhaiso-leseling e hlahisitse litlamorao tse kholo tsa boemo ba osBFosB (p <0.0001, F = 22.12, df = 1) le mahloriso a DAMGO (p <0.0001, F = 29.65, df = 5) e se nang Tšebelisano ea bohlokoa (p = 0.857, F = 0.387, df = 5). Tlhahlobo e sa sebetseng ea khatello ea maikutlo ea li-curve-effect curves e senotse DAMGO E e kholo haholo, max e bohlokoa ho DFFB ka litoeba (E, max = 73 ± 5.2% tsusumetso) ho ea ho DFB ho tlosa litoeba (E, max = Tsosoloso ea 56 ± 4.1%; p <0.05 e fapaneng le ΔFosB ho litoeba ka teko ea Seithuti). TLHOKOMELISO EC50 litekanyetso li ne li sa tšoane pakeng tsa ΔFosB le DFFB ho tloha litoeba (302 ± 72 nM ho latela 212 ± 56 nM, ka ho latellana, p = 0.346).

Setšoantšo sa 1 

Phello ea maikutlo a ΔFosB ka agonist-a susumelitse [35S] GTPXS e tlama ka NAC. Litokelo tse tsoang ho ΔFosB-ho hlalosa (ΔFosB ka) kapa ho laola (ΔFosB ho tloha) litoeba li ile tsa lekoa joalokaha li hlalositsoe Mekhoa e sebelisang libaka tse sa tšoaneng ...

Ho fapana le liphetho tse fumanoeng le MOR agonist DAMGO, ha ho maemo a DFFB a itšetlehileng ka boemo ba G-protheine e sebelisoang ka ho hlahisa li-agonist agonist WIN55,212-2 (Setšoantšo sa 1B). ANOVA ea litsela tse peli ea data ea WIN55,212-2 ea litlamorao e hlahisitse sephetho sa mantlha sa mahloriso a WIN55,212-2 (p <0.0001, F = 112.4, df = 7), empa eseng ea boemo ba osBFosB (p = 0.172 , F = 1.90, df = 1) 'me ho ne ho se tšebelisano (p = 0.930, F = 0.346, df = 7). Ka mokhoa o ts'oanang, ho ne ho se na tšusumetso ea boemo ba ΔFosB ho WIN55,212-2 E, max litekanyetso (103 ± 6% ho latela 108 ± 8% tsusumetso ho DFB ho tloha le ho tlohela litoeba, ka ho latellana, p = 0.813 ke T-test ea T) kapa EC50 litekanyetso (103 ± 20 nM ho feta 170 ± 23 nM ka DFBB le ho tlohela litoeba, ka ho latellana, p = 0.123).

E itšetlehile ka sebōpeho sa li-curves le hore lithuto tsa rona tsa pele li bontšitse biphasic WIN55,212-2-methapo ea mahlaseli a boko ba bokong (Breivogel, et al., 1999, Breivogel, et al., 1998), lithapo tsa WIN55,212-2 li ile tsa boela tsa hlahlojoa ho sebelisa mekhoa e mabeli ea sebaka. Ho hlahlojoa ha boitsebiso bo fapaneng bo bontšitse ntlafatso e fokolang ea botle bo nepahetseng ho sebelisoa setšoantšo sa maqephe a mabeli (R2 = 0.933 le 0.914, likarolo tse ngata = 3644 le 5463 ka DFFB le ho khaola litoeba ka ho latellana) ha li bapisoa le setšoantšo se le seng sa sebaka (R2 = 0.891 le 0.879, likarolo tse ngata = 6561 le 6628 ka DFFB le ho tlohela litoeba ka ho latellana). Leha ho le joalo, ha ho phapang e khōlō e ileng ea fumanoa pakeng tsa ΔFosB le ho tlosa litoeba ho E, max kapa EC50 litekanyetso tsa libaka tse phahameng kapa tse tlaase tsa potency (Letlapa le tlatsetsang 1), le hoja ho ne ho e-na le mokhoa o lebisang ho EC e tlaase50 bohlokoa ho sebaka se phahameng sa potency ho litoeba tse nang le DFFB (EC50phahameng = 28.0 ± 10.6 nM) ho bapisoa le ba nang le DFFB (EC50phahameng = 71.5 ± 20.2 nM; p = 0.094). Ho feta moo, ho ne ho se na phello ea boemo ba DFBB ka basal [35S] GTPγS e tlama ka lisele tsa NAc (253 ± 14 ho latela 226 ± 14 fmol / mg ka ΔFosB ho tloha le ho tlohela litoeba, ka ho latellana, p = 0.188). Litlaleho tsena li bonts'a hore polelo e sa utloahaleng ea mantsoe a DFBB ka NAC ea litoeba e ekelitse ts'ebetso ea protheine ea G-mohopolo oa MOR ntle le ho ama CB haholo1Mesebetsi ea R-mediated kapa ea bas-G ea protheine.

3.2. Phello ea ΔFosB ka thibelo ea li-opioid le ea cannabinoid e amohelang li-adenylyl cyclase

Ho hlahloba phello ea polelo e hlalosang mantsoe ea TFFB e fokolang ka mokhoa oa ho feto-fetoha ha mesebetsi e ka tlaase ho MOR le CB1R, ho thibela ha 1 μM forskolin-e susumelitsoeng ke mosebetsi oa adenylyl cyclase e ile ea hlahlojoa ka li-membrane tsa NAc. Ho phaella ho MOR- le CB1Ho thibela li-acenylyl cyclase ho e-na le mekhoa e mengata, liphello tsa mosebetsi oa KOR le tsona li ile tsa hlahlojoa li sebelisa KOR-selective e feletseng ea agonist U50,488 (Zhu, et al., 1997), hobane liphetho tse fetileng li bontšitse hore dynorphin mRNA e ne e hlasetsoe ke ΔFosB ka mohlala oa bitransgenic (Zachariou, et al., 2006). Lipatlisiso li bontšitse hore DAMGO, U50,488 le WIN55,212-2 li hlahisa tšenyo e itšetlehileng ka mahloriso a li-adenylyl cyclase ka bobeli ΔFosB le DFFB ka litoeba (Setšoantšo sa 2). Tsela e 'meli ea ANOVA ea data-concentration e sebetsang ka DAMGO (Setšoantšo sa 2A) e senotse litlamorao tse kholo tsa boemo ba osBFosB (p = 0.0012, F = 11.34, df = 1) le mahloriso a DAMGO (p <0.0001, F = 29.61, df = 6), empa ha ho na tšebelisano e kholo (p = 0.441, F = 0.986 , df = 6). Tlhahlobo e sa sebetseng ea khatello ea maikutlo ea li-curve tsa litlamorao tsa DAMGO e senotse DAMGO EC e tlase haholo50 boleng ho osBFosB ka litoeba (101 ± 11 nM) ha e bapisoa le ΔFosB litoeba (510 ± 182 nM, p <0.05 ke t-test ea moithuti). Leha ho le joalo, ho ne ho se na phapang e kholo ho DAMGO E, max litekanyetso (20.9 ± 1.26% ho latela 19.8 ± 1.27% ho thibela ho DFB ho tloha le ho tlohela litoeba, ka ho latellana, p = 0.534).

Setšoantšo sa 2 

Phello ea polelo ea ΔFosB mabapi le thibelo ea mosebetsi oa adenylyl cyclase NAC. Litokelo tse tsoang ho ΔFosB-ho hlalosa (ΔFosB ka) kapa ho laola (ΔFosB ho tloha) litoeba li ile tsa lekoa joalokaha li hlalositsoe Mekhoa ka pel'a 1 μM ...

K'homphieutha ea adenylyl cyclase e kopantsoeng le KOR e ne e boetse e fapane le mokhoa oa ho bua ka mantsoe a fetoletsoeng ka mantsoe a hau a ΔFosB (Setšoantšo sa 2B). ANOVA ea litsela tse peli ea data ea litlamorao tsa U50,488 e bonts'itse litlamorao tse kholo tsa boemo ba osBFosB (p = 0.0006, F = 14.53, df = 1) le mahloriso a U50,488 (p <0.0001, F = 26.48, df = 3) , ntle le tšebelisano ea bohlokoa (p = 0.833, F = 0.289, df = 3). Tlhahlobo e sa sebetseng ea khatello ea maikutlo ea likhahla tsa khatello ea maikutlo e senotse U50,488 E e kholo, max boleng ho osBFosB ho litoeba (18.3 ± 1.14% inhibition) ha e bapisoa le ΔFosB litoeba (12.5 ± 2.03% inhibition; p <0.05 e fapaneng le ΔFosB ka t-test ea Seithuti), ho se na phapang e kholo ho U50,488 EC50 litekanyetso (310 ± 172 nM ho feta 225 ± 48 nM ka DFBB le ho tlohela litoeba, ka ho latellana, p = 0.324).

Ho fapana le liphello tse ileng tsa etsoa le MOR le KOR, ho ne ho se na phello e kholo ea mantsoe a transgenic a DFFB ka thibelo ea adenylyl cyclase ke ea agonist ea cannabinoid WIN55212-2 (Setšoantšo sa 2C). ANOVA ea litsela tse peli ea data ea litlamorao tsa WIN55,212-2 e bonts'itse phello e kholo ea khatello ea lithethefatsi (p <0.0001, F = 23.6, df = 2), empa eseng ea boemo ba osBFosB (p = 0.735, F = 0.118, df = 1) hape ho ne ho se na puisano e kholo (p = 0.714, F = 0.343, df = 2). Ntle le moo, ho ne ho se na phello ea boemo ba ΔFosB ho basal kapa forskolin-stimulated adenylyl cyclase ts'ebetso ha ho se na agonist efe kapa efe. Ketsahalo ea basal adenylyl cyclase e ne e le 491 ± 35 pmol / mg / min ho ΔFosB ho litoeba ha e bapisoa le 546 ± 44 ho ΔFosB litoeba (p = 0.346 ke t-teko ea Seithuti). Ka mokhoa o ts'oanang, ts'ebetso ea adenylyl cyclase boteng ba 1 µM forskolin e ne e le 2244 ± 163 pmol / mg / min ho osBFosB ho litoeba khahlanong le 2372 ± 138 pmol / mg / min ho osBFosB litoeba (p = 0.555).

3.3. Phello ea ΔcJun ka thibelo ea li-opioid le ea cannabinoid e amohelehang ea li-adenylyl cyclase

Kahobane polelo e fetisisang ea mantsoe a hau ea ΔFosB e fetisetsoang ho fetisetsoa ha signal ho tloha MOR le KOR ho adenylyl cyclase ho NAc, e ne e thahasella ho bona hore na inhibitor e kholo ea inthanete ea ΔFosB e fetisitsoeng e ne e tla fetola pontšo ea ho amohela li-opioid ka tsela e fapaneng. E le ho rarolla potso ena, ho thibela tšebetso ea forskolin-stimulated acenylyl cyclase e entsoeng ke DAMGO le U50,488 e ile ea hlahlojoa ka likhechana tse lokiselitsoeng ho tloha NAc ea litoeba tsa bitransgenic ka tsela e hlalosang DNA. Liphello ha li bontše tšusumetso e matla ea polelo ea ΔcJun ho thibela mosebetsi oa adenylyl cyclase ke MOR kapa KOR (Setšoantšo sa 3). ANOVA ea litsela tse peli ea li-curve tsa litlamorao tsa DAMGO e bonts'itse phello ea mantlha ea mahloriso a DAMGO (p <0.0001, F = 20.26, df = 6), empa eseng ea boemo ba JcJun (p = 0.840, F = 0.041, df = 1) mme ho ne ho se na tšebelisano ea bohlokoa (p = 0.982, F = 0.176, df = 6). Ka mokhoa o ts'oanang, ho ne ho se na phapang e kholo ho E, max kapa EC50 litekanyetso pakeng tsa litoeba le Dccun (E, max = 23.6 ± 2.6%; EC50 = 304 ± 43 nM) kapa ΔcJun off (E, max = 26.1 ± 2.5%, p = 0.508; EC50 = 611 ± 176 nM, p = 0.129). Liphetho tse ts'oanang li bonoe ka U50,488, joalo ka hore ANOVA ea litsela tse peli ea li-curve-effect curves e bonts'itse phello e kholo ea mahloriso (p <0.0001, F = 11.94, df = 6), empa eseng ea boemo ba JcJun (p = 0.127 , F = 2.391, df = 1) mme ho ne ho se na puisano e bohlokoa (p = 0.978, F = 0.190, df = 6). Ka mokhoa o ts'oanang, ho ne ho se na phapang e kholo ho E, max kapa EC50 litekanyetso pakeng tsa litoeba le Dccun (E, max = 14.8 ± 2.9%; EC50 = 211 ± 81 nM) kapa tima (E, max = 16.7 ± 1.8%, p = 0.597; EC50 = 360 ± 151 nM, p = 0.411).

Setšoantšo sa 3 

Phello ea polelo ea ΔcJun e thibelang mosebetsi oa adenylyl cyclase ka NAc. Lisele tsa ho tloha ka DcJun-expressing (ΔcJun on) kapa taolo (ΔcJun off) litoeba li ne li koaletsoe ka pel'a DAMGO (A), U50,488H (B) kapa WIN55,212-2 ...

Polelo ea JcJun le eona ha ea ka ea ama haholo thibelo ea adenylyl cyclase ho NAc ke agonist ea cannabinoid. ANOVA ea litsela tse peli ea maqhubu a sephetho sa WIN55,212-2 a bonts'itse sephetho sa mantlha sa mahloriso a WIN55,212-2 (p <0.0001, F = 15.53, df = 6), empa eseng ea genotype (p = 0.066, F = 3.472, df = 1) mme ho ne ho se na puisano e bohlokoa (p = 0.973, F = 0.208, df = 6). Ka mokhoa o ts'oanang, ho ne ho se na phapang e kholo ho WIN55,212-2 E, max mekhoa ea bohlokoa (13.0 ± 2.3% le 13.6 ± 0.9% ho thibela ho se hokae ka litoeba, ka ho latellana, p = 0.821) le kapa EC50 mekhoa ea bohlokoa (208 ± 120 nM le 417 ± 130 nM ka PAC ho ea ka litoeba, ka ho latellana, p = 0.270). Ka hona, le hoja ho ne ho e-na le mokhoa o fokolang oa ho fokotseha ha potso ea WIN55,212-2 ka litoeba tse hlalosang ΔcJun, litlōlo tsena ha lia ka tsa fetola tsela e matla ea ho thibela li-adenylyl cyclase. Ho feta moo, ho ne ho se na phello ea boemo ba ΔcJ ka mosebetsi oa basal kapa oa forskolin o susumelitsoeng ke acenylyl cyclase. Mosebetsi oa basallyl cyclase e ne e le 1095 ± 71 pmol / mg / min le 1007 ± 77 pmol / mg / min (p = 0.403) ka litoeba tse nang le DcJun kapa ka ntle ho tsona. Ketsahalo ea Adenylyl cyclase e tsosolositsoeng ke 1 μM forskolin e ne e le 4185 ± 293 pmol / mg / min ho feta XUMUMX ± 4032 pmol / mg / min (p = 273) ka litoeba tse nang le DcJun kapa ka ntle ho tsona.

3.4. Puisano

Liphello tsa phuputso ena li senoletse ts'ebetso e matlafatsang ea pro-G-protein e entsoeng ka MOR le ho thibela li-acenylyl cyclase ho NAc ea litoeba tse nang le polelo e hlalosang ea DFFB ka dynorphin / D1R e nang le neurons. K'homphieutha e entsoeng ka KOR ea mosebetsi oa adenylyl cyclase e ile ea boela ea ntlafatsoa NAC ea ΔFosB e hlahisang litoeba, e fana ka maikutlo a hore ΔFosB e laola tsamaiso ea opioid e mamellang ka NAc. DAMGO E, max boleng bo ne bo le boholo bakeng sa MOR-e susumelitsoeng [35S] GTPγS e tlama, le EC ea eona50 boleng bo ne bo le tlaase bakeng sa thibelo ea adenylyl cyclase, ho DFFB ho fetisa litoeba ho bapisoa le litoeba tsa taolo. Liphuputso tsena li fana ka maikutlo a monyetla oa ho fumana sebaka sa polokelo bakeng sa ho feto-fetoha ha motlakase empa eseng G-protein e sebetsang tlas'a maemo a tlhahlobo a hlahlojoang. Ho fumanoa hore thibelo e kholo ea adenylyl cyclase ke KOR agonist e ile ea angoa ke polelo ea ΔFosB e fana ka sebaka se tlase sa polokelo bakeng sa karabo ea KOR, e lumellanang le maemo a tlase a KOR a ho kopanya libaka ka boko ba mouse (Unterwald, et al., 1991). Ka lehlakoreng le leng, CB1Mesebetsi ea G-protein e tsamaisanang le R le ho thibela li-acenylyl cyclase ha lia ameha ke polelo ea DFFB, e fana ka maikutlo a hore tsamaiso ea opioid le ea cannabinoid e fapana ka karabelo ea bona ho ΔFosB li-NAc neurons tsena.

Phello ea ΔFosB mabapi le pontšo ea li-opioid receptor-mediated e lumellana le tlaleho ea rona ea pele e bontšang hore ΔFosB e hlalositsoeng ka lebalatum e fetotse liphello tse matla le tse sa foleng tsa morphine (Zachariou, et al., 2006). E 'ngoe ea liphuputso tseo e ne e le litoeba tse nang le maikutlo a tloaelehileng a DFFB ka dynorphin / D1Litšoelesa tse hlaselang li ne li e-na le kamano e ntle le morphine sebakeng sa maemo a fapaneng ho feta taolo. Ho phaella moo, phello ena e ne e etsisoa ke polelo e nang le mefuta e mengata ea ΔFosB ka ente e khethehileng ea sebaka ka NAC. Liphuputso tsena li lumellana le liphetho tsa morao-rao tse bontšang hore ponts'o ea MOR e ntlafalitsoe ho NAc.

Re kile ra tsebahatsa liphatsa tsa lefutso dynorphin e le sepheo sa DFFB, 'me e hlalositsoe hore e fokotsoe ke dynorphin e tla ba e lumellanang le thepa e ntlafatsang ea morphine ho DFFBB litransgenic litoeba (Zachariou, et al., 2006). Liphello tsa hona joale li bontša hore thibelo ea KOR ea cyenylase ea adenylyl ka NAc e ntlafatsoa ho ΔFosB e hlahisang litoeba, tse ka 'nang tsa bontša ho eketseha ha phoso ho senoloa KOR ka mor'a ho fokotsa dynorphin. Liphuputso tse fetileng li bontšitse hore KOR e ne e fokotsehile libakeng tse itseng tsa boko ba litoeba tsa prodynorphin tsa ho kokota, ho kenyeletsa le NAc (Clarke, le al., 2003).

Ho fapana le ΔFosB, polelo e fokolang ea tlhaho ea ΔcJun, mofuthu o ka sehloohong o se nang mofuthu oa molekane oa ΔFosB ea kopanetsoeng molekane, ha oa ka oa fetola thibelo ea adenylyl cyclase ea MOR kapa KOR. Liphetho tsena li bontša hore litekanyetso tsa basal tsa polelo ea ΔFosB, e batlang e le tlaase, ha e bapale karolo ea bohlokoa ho boloka pontšo ea li-opioid receptor ho lekaneng lena la ho fetisetsoa ha signal ho NAc. Taba ea hore phello e thabisang ea morphine e fokotsehile ke polelo ea ΔcJun ho ithutong ea rona e fetileng (Zachariou, et al., 2006) e fana ka maikutlo a hore tlhahiso ea morphine ea ΔFosB nakong ea ts'ebetso ea boemo ba bohlokwa ke ea bohlokoa ho laola likarabo tsa boitšoaro ho lithethefatsi kapa hore liphello tsa phetolo ea ΔFosB tse ling ntle le tse amang palo e kholo ea li-receptors li ka 'na tsa ama moputso oa opioid. Leha ho le joalo, liphello tsa thuto ea hona joale li bontša ka ho hlaka hore, ha polelo ea DFFB e phahame ka holimo ho maemo a basal ka ho hlasela dynorphin / D1R-ho hlalosa li-neurons, ho na le keketseho e matla ho kopanngoa ha MOR le KOR ho thibela li-adenylyl cyclase ho NAc.

Mokhoa oo MOR-le KOR-pontšo e fetiselitsoeng ka oona e ntlafatsoa ka ho hlaka ha ΔFosB ho sa hlaka, empa re kile ra bontša hore maemo a MOR, a hlahlojoa ke [3H] naloxone e tlama, ha e fapane le NAc ea ΔFosB ha e bapisoa le litoeba (Zachariou, et al., 2006). Thuto e tšoanang e fumane hore GaiLi-protein tsa 1 le 2 ha lia ameha sebakeng sena ka polelo ea DFFB. Leha ho le joalo, lipatlisiso tsa liphatsa tsa lefutso tse fetileng li bontšitse hore Gao MRNA e ne e hlophisitsoe ka NAC ea DFFB ka litoeba (McClung le Nestler, 2003). Ho tla ba le thahasello liphuputsong tsa nakong e tlang ho hlahloba ka ho tebileng phello ea polelo ea DFFB ea G-protein e nang le poleloana ea liprotheine hammoho le polelo ea li-protein tse ngata tsa G-protheine.

Hoa thahasellisa hore polelo ea ΔFosB ha ea ntlafatsa CB1Pontšo ea R-mediated NAC. Ho ka etsahala hore liphetoho CB1R ho bontša hore ho na le libaka tse sa tšoaneng tsa li-neurone tse patehileng hohle ho hlophisoa ha NAc. Ka mohlala, tsamaiso ea Δ9- THC e hlohlellelitse ΔFosB haholo-holo, empa eseng khetla, ea NAc (Perrotti, et al., 2008). Kee le mahlomola, ho bontšitsoe hore bothata le D9-THC e latela tsamaiso ea khafetsa ea Δ9-THC e ekelitse ho lokolloa ha dopamine ka NAC ea motheo, empa ho fokotsoa ho lokolloa sekhethong (Cadoni, et al., 2008). Hape ke habohlokoa ho hlokomela hore line ea 11A ea litoeba tsa bitransgenic e hlalosa DFFB feela ka dynorphin / D1R positive spiny neurons ea striatum, empa CB1R li hlalosoa ka bobeli dynorphin / D1R le enkephalin / D2R tse ntle tse hlaselang (Hohmann le Herkenham, 2000), hammoho le li-terminals tsa afferentts tsa cortical (Robbe, et al., 2001). Tlhaloso ea molaoli o ka sehloohong oa tsamaiso ea ΔFosB-e ngotsoeng lipakeng, ΔcJun, le eona e ne e se na phello e kholo tabeng ea pontšo ea kankere ea li-cannabinoid, le hoja ΔcJun e hlalositsoe ka bobeli D1 le D2-e nang le baahi ba li-neurone tse bohareng ba li-mouse (Peakman, et al., 2003). Leha ho le joalo, hoa khoneha hore polelo ea basing ea ΔFosB e lekane ka ho lekaneng hore ΔcJun e ke ke ea ama pontšo ea ho amohela, joalokaha e hlahisoa ke liphetho le MOR le KOR. Hape ho ka etsahala hore CB1R ho bontša hore ka boikokobetso bo matlafatsoa ke polelo e ka sehloohong ea DFFB, e leng ho eketsa ka ho eketsehileng polelo ea DFFB kapa ho thibela liketso tsa eona le ΔcJun ho ne ho e-na le liphello tse fokolang feela tse sa kang tsa fihla tekanyo ea lintlha tsa bohlokoa. Tšehetso e sa tobang bakeng sa tlhaloso ena e ka bonoa ka ho bapisa WIN55,212-2 EC50 litekanyetso pakeng tsa litoeba tse hlalosang ΔcJun le DFFB. Karolelano ea WIN55,212-2 EC50 bohlokoa ba ho thibela li-adenylyl cyclase ho litoeba tse nang le polelo e susumelitsoeng ea ΔcJun ho EC ea eona50 bohlokoa ba tlhahiso ea G-protheine litulong tse nang le polelo e susumelitsoeng ea ΔFosB e ne e le 4.0, empa karolelano e lekanang le litoeba tse se nang tlhahiso ea hore e ne e le transgene e ne e le 1.2.

Ntle le moo, li-cannabinoids li ka 'na tsa hlahisa maikutlo a DFFB ntle le phello e tobileng ho CB1R ho bontša. Tabeng ena, li-cannabinoids li ka khona ho ikarabella ho liphello tsa kelello tsa lithethefatsi tse ling ka taelo ea "transposal" e tsamaisanang le litšoantšo. KeHa e le hantle, tsamaiso ea Δ9-THC e hlahisa tšusumetso e kholo ea li-opioid le amphetamine (Cadoni, et al., 2001, Lamarque, et al., 2001), e lumellanang le khopolo-taba ena. Ho feta moo, tsamaiso ea khafetsa ea kannete ea cannabinoid CP55,940 e ile ea tlalehoa hore e tla eketsa ts'ebetso ea protheine ea G-mediated ho NAc, le eona e le ho bua ka mokhoa o hlakileng oa ho hlalosa ΔFosB sebokeng sa hona joale (Vigano, et al., 2005). Phello ea polelo ea ΔFosB ka Δ9Mekhoa e ikemetseng -THC e sa hlahlojoa, empa liphello tsa hona joale ha li thibele ho sebelisana. Liphello tsa sena le thuto ea rona ea pele (Zachariou, et al., 2006) bonts'a liphetoho tse bakoang ke FosB ho MOR le KOR / dynorphin ho striatum. Liphello tse thabisang tsa D9-THC, e lekantsoeng ke khetho ea sebaka, e felisoa ka litoeba tsa MOR, ha ho tlosoa KOR ho hlokomoloha D9-THC e thibela ho hanyetsa 'me e senola D9-THC sebaka se khethollang (Ghozland, et al., 2002). Ka mokhoa o ts'oanang, sebaka sa boemo bo tsitsitseng ho Δ9-THC ha e fumanehe ka ho kokota ha pro-dynorphin ha e bapisoa le litoeba tsa mofuta o hlaha (Zimmer, et al., 2001). Lintlha tsena li bontša hore Δ9-THC e ka ba e khotsofatsang haholo ka mor'a hore ΔFosB e kenyelletsoe le ho kenngoa ha moralo oa MOR ho bontša ka ho fokotseha ha polelo ea dynorphin.

Ka kakaretsoy, liphello tsa thuto ena li bontšitse hore polelo ea ΔFosB ka D1R / dynorphin methapo e ts'oanang e hlaselang ea MOR-le KOR e bonts'oa mofuteng karolong ea G-protein e thibelang ho thibela mosebetsi oa adenylyl cyclase ho NAc. Tlhaloso ena e lumellana le lithuto tse bontšitseng karolo ea tsamaiso ea opioid e tšoarellang ka moputso (Trigo, et al., 2010), le ho fana ka mochine o ka sebelisoang ke DFF-mediated effects on the reward. Ka lehlakoreng le leng, CB1Pontšo ea R-mediated ka NAc ha ea ka ea ameha haholo ka polelo e hlaselang ea ΔFosB tlas'a maemo a hlahlojoang, le hoja lithuto tse eketsehileng li hlokehile ho fumana phello ea DFB ho kenya mokhoa oa ho qetela oa ho qetela.

Lintlha-khōlō tsa Patlisiso

  • Pontšo ea MOR e ntlafatsoa mokokotlong oa litoeba tse hlalosang DFF
  • KOR ho thibela li-adenylyl cyclase e boetse e ntlafatsoa ka litoeba tse hlalosang ΔFosB
  • Tlhaloso ea DFF ha e fetole CB1R ho bonahatsa mokokotlong oa accumbens
Eya ho:

Boitsebiso bo Eketsehileng

Eya ho:

Liteboho

Bangoli ba leboha Hengjun He, Jordan Cox le Aaron Tomarchio bakeng sa thuso ea theknoloji le [35S] GTPγS e tlama litekanyetso. Phuputso ena e ne e tšehetsoa ke USPHS Grants DA014277 (LJS), DA10770 (DES) le P01 DA08227 (EJN).

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Mongolo o botlaaseng ba leqephe

Boitlhotlhollo ba Mohoeletsi: Ena ke faele ea PDF ea mongolo o ngotsoeng ka letsoho o sa amoheloang o amoheletsoeng bakeng sa ho hatisoa. Joaloka tšebeletso ho bareki ba rona re fana ka phetolelo ena ea pele ea mongolo o ngotsoeng ka letsoho. Mongolo o ngotsoeng ka letsoho o tla ba le ho kopitsa, ho kenya lihlopha le ho hlahloba bopaki bo hlahang pele o hatisoa ka mokhoa oa oona oa ho qetela. Ka kopo hlokomela hore nakong ea mekhoa ea tlhahiso ea lihlahisoa e ka fumanoa e ka amang litaba, le litsebiso tsohle tsa molao tse sebetsang koranteng e amanang le eona.

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References

  • Bozarth MA, RA ea Bohlale. Makala a amohelehang a opiate a amohelehang a arolelana moputso le ho itšetleha ka 'mele. Saense. 1984;224: 516-517. [E fetotsoe]
  • Bradford MM. Tsela e potlakileng le e thata bakeng sa quantification ea microgram tse ngata tsa protheine e sebelisang molao-motheo oa ho kopanya li-protein. Anal. Likokoana-hloko. 1976;72: 248-254. [E fetotsoe]
  • Breivogel CS, Childers SR, Deadwyler SA, Hampson RE, Vogt LJ, Sim-Selley LJ. Nōka ea khale ea delta9-Tetrahydrocannabinol kalafo e hlahisa ho lahleheloa ha nako ea li-G-protheine tse amohelitsoeng ka kannete ka bokong. J. Neurochem. 1999;73: 2447-2459. [E fetotsoe]
  • Breivogel CS, Selley DE, Childers SR. Cannabinoid receptor agonist e sebetsang bakeng sa ho susumetsa [35S] GTPyS e tlama ho ratana le cerebellar membrane e tsamaeang le ho fokotseha ha agonist ho fokolang ha GDP. J. Biol. Chem. 1998;273: 16865-16873. [E fetotsoe]
  • Cadoni C, Pisanu A, Solinas M, Acquas E, Di Chiara G. Ts'ebetso ea ts'ebeliso ea tlhaho ka mor'a hore li phethoe ka ho hlaka ho Delta 9-tetrahydrocannabinol le ho fokolisa morphine. Psychopharmacology (Berl) 2001;158: 259-266. [E fetotsoe]
  • Cadoni C, Valentini V, Di Chiara G. Ho khothaletsoa ho delta 9-tetrahydrocannabinol le cross-sensitization le morphine: liphetoho tse fapaneng ho khetholla sekhetla le phetoho ea mantlha ea dopamine. J. Neurochem. 2008;106: 1586-1593. [E fetotsoe]
  • Chen J, Kelz MB, Zeng G, Sakai N, Steffen C, Shockett PE, Picciotto MR, Duman RS, Nestler EJ. Liphoofolo tsa li-transgenic tse nang le inducible, tse lebisitsoeng ho liphatsa tsa lefutso bopoong. Mol. Pharmacol. 1998;54: 495-503. [E fetotsoe]
  • Bana ba SR. Liemeli tsa bobeli tse kopanetsoeng ho opioid. Bophelo Sci. 1991;48: 1991-2003. [E fetotsoe]
  • Childers SR, Fleming L, Konkoy C, Marckel D, Pacheco M, Sexton T, Ward S. Opioid le ho amohela li-cannabinoid ho thibela li-adenylyl cyclase bokong. Ann. NY Acad. Sci. 1992;654: 33-51. [E fetotsoe]
  • Bana ba nang le SR, Xiao R, Vogt LJ, Sim-Selley LJ. Kappa e fumanoang ke li-opioid receptor [35S] GTPγS e tlama ka boko ba likolobe: Ho hloka bopaki ba kappa2- ho thibela li-G-li-protein. Likokoana-hloko. Pharmacol. 1998;56: 113-120. [E fetotsoe]
  • Clarke S, Zimmer A, Zimmer AM, Hill RG, Kitchen I. Karolo ea libaka tse khethollang li-micro-, li-receptor tse kang marta le kappa-opioid empa li sa amohele li-receptor tse kang li-1 tse nang le likopi tsa enkephalin le litoeba tsa ho kokota dynorphin. Khopolo-taba. 2003;122: 479-489. [E fetotsoe]
  • Colby CR, Whisler K, Steffen C, Nestler EJ, Boiketsetso ba DW. Tlhahlobo e hlakileng ea sele ea striatal mofuta oa DeltaFosB e eketsa khothatso ea cocaine. J. Neurosci. 2003;23: 2488-2493. [E fetotsoe]
  • Gardner EL. Endocannabinoid pontšo ea tsamaiso le moputso oa boko: ho hatisoa ka dopamine. Pharmacol. Likokoana-hloko. Behav. 2005;81: 263-284. [E fetotsoe]
  • Ghozland S, Matthes HW, Simonin F, Filliol D, Kieffer BL, Maldonado R. Motivational liphello tsa li-cannabinoids li kopanngoa ke li-receptor tsa ma-opioid le kappa-opioid. J. Neurosci. 2002;22: 1146-1154. [E fetotsoe]
  • Herkenham M, Lynn AB, Johnson MR, Melvin LS, de Costa BR, Rice KC. Boikarabello le ho tseba sebaka sa li-receptor tsa kannete ea bokhoni ba bokhoni: boithuto bo bongata ba in vitro ea boithuto. J. Neurosci. 1991;11: 563-583. [E fetotsoe]
  • Hohmann AG, Herkenham M. Sebaka sa meriana ea receptor CBR (1) e fumanoang ka methapo ea rat striatum: phuputso e entsoeng ka bobeli boemong ba hybridization. Synapse. 2000;37: 71-80. [E fetotsoe]
  • Howlett AC, Barth F, Bonner TI, Cabral G, Casellas P, Devane WA, CC Felder, Herkenham M, Mackie K, Martin BR, Mechoulam R, Pertwee RG. International Union of Pharmacology. XXVII. Mekhabiso ea li-receptor tsa kannete. Tlhahlobo ea Litlhapi. 2002;54: 161-202.
  • Huestis MA, Gorelick DA, Heishman SJ, Preston KL, Nelson RA, Moolchan ET, Frank RA. Thibelo ea liphello tsa matekoane a tsuba ka CB1-e khethollang motho ea amohelang molaetsa SR141716. Sethapo. Gen. Psychiatry. 2001;58: 322-328. [E fetotsoe]
  • Kell MB, Chen J, Karl J, Karl K Wheller, Kelli Keller, Kelli MB, Karl Keller, Nestler EJ. Tlhaloso ea motsoako oa deltaFosB oa khatiso boko bo laola ho utloahala ha cocaine. Tlhaho. 1999;401: 272-276. [E fetotsoe]
  • Koob GF, Volkow ND. Meriana ea methapo ea meriana. Neuropsychopharmacology. 2010;35: 217-238. [Tlhahiso ea mahala ea PMC] [E fetotsoe]
  • Lamarque S, Taghzouti K, Simon H. Phekolo ea meriana e nang le Delta (9) -tetrahydrocannabinol e eketsa karabo ea mofuthu ho amphetamine le heroin. Liphello tsa ts'oaetso ea ho lemalla lithethefatsi. Neuropharmacology. 2001;41: 118-129. [E fetotsoe]
  • Maldonado R, Valverde O, Berrendero F. Ho kenngoa ha tsamaiso ea endocannabinoid ho lemalla lithethefatsi. Trends Neurosci. 2006;29: 225-232. [E fetotsoe]
  • Mansour A, Fox CA, Thompson RC, Akil H, Watson SJ. polelo ea mRNA ea ho-opioid receptor rateng ea CNS: papiso le ho kenya mo-receptor. Resin ea Boko. 1994;643: 245-265. [E fetotsoe]
  • Matthews HWD, Maldonado R, Simonin F, Valverde O, Slowe S, Kitchen I, Befort K, Dierich A, LeMeur M, Dolle P, Tzavara E, Hanoune J, Roques BP, Kieffer BL. Ho lahleheloa ke analgesia e entsoeng ka morphine, phello ea moputso le matšoao a ho tlohela litoefong tse hlokang liphatsa tsa lefutso tsa μ-opioid receptor. Tlhaho. 1996;383: 819-823. [E fetotsoe]
  • McClung CA, Nestler EJ. Tlhophiso ea liphatsa tsa lefutso le moputso oa cocaine ke CREB le DeltaFosB. Nat. Neurosci. 2003;6: 1208-1215. [E fetotsoe]
  • McClung CA, Ulery PG, Perrotti LI, Zachariou V, Berton O, Nestler EJ. DeltaFosB: molek'hule oa limolek'hule bakeng sa ho fetola nako e telele bokong. Resin ea Boko. Mol. Resin ea Boko. 2004;132: 146-154. [E fetotsoe]
  • Muller DL, Unterwald EM. D1 dopamine receptors e ikamahanya le marang-rangFOSB ho kenngoa ho rat ratatum ka mor'a tsamaiso ea morphine e bohareng. J. Pharmacol. Exp. Ther. 2005;314: 148-154. [E fetotsoe]
  • Nestler EJ. Tlhahlobo. Mekhoa ea phetoho ea ho lemala: karolo ea DeltaFosB. Philos. Trans. R. Soc. London. B. Biol. Sci. 2008;363: 3245-3255. [Tlhahiso ea mahala ea PMC] [E fetotsoe]
  • Nestler EJ, Kelz MB, Chen J. DeltaFosB: mokena-lipakeng oa molek'hule oa nako e telele oa neural le plasticity ea boitšoaro. Resin ea Boko. 1999;835: 10-17. [E fetotsoe]
  • Nye HE, Hope BT, Kelz MB, Iadarola M, Nestler EJ. Liphuputso tsa litsebi tsa thuto ea lefapha la tlhophiso ea tlhahiso ea antigen e sa foleng e amanang le li-FOS ke k'hok'heinine ea striatum le nucleus accumbens. J. Pharmacol. Exp. Ther. 1995;275: 1671-1680. [E fetotsoe]
  • Peakman MC, Colby C, Perrotti LI, Tekutalla P, Carle T, Ully P, Chao J, Duman C, Steffen C, Monteggia L, Allen MR, Stock JL, Duman RS, McNeish JD, Barrot M, Self DW, Nestler EJ , Schaeffer E. Ha e khonehe, polelo e boletsoeng ke sebaka sa boko bo bobe ba c-Jun ka litoeba tsa tlhaho e fokotsa ts'oaetso ea cocaine. Resin ea Boko. 2003;970: 73-86. [E fetotsoe]
  • Rowland RI, Rowland RI, Rowland R Row, Rowland Row, Rowland RI, Rowland RI, Rowland RI, Rowland RJ, Rowland RJ, Rowland EJ. Mekhoa e fapaneng ea DeltaFosB ho kenngoa ka bokooa bokong ka lithethefatsi tsa tlhekefetso. Synapse. 2008;62: 358-369. [Tlhahiso ea mahala ea PMC] [E fetotsoe]
  • Robbe D, Alonso G, Duchamp F, Bockaert J, Manzoni OJ. Sebaka sa sebakeng sa heno le mekhoa ea liketso tsa li-receptor tse nang le li-cannabinoid li-synapses tsa glutamaterg tsa nucleus ea mouse. J. Neurosci. 2001;21: 109-116. [E fetotsoe]
  • Salomon Y. Adenylate cyclase test. Keletso. Cyclic Nucleotide Res. 1979;10: 35-55. [E fetotsoe]
  • Selley DE, Sim LJ, Xiao R, Liu Q, Childers SR. Ha ho amoheloa opioid-stimulated [35S] GTPγS e tlamang ka thalamus ea rat le lisele tse hlahisitsoeng: Litsela tsa ho fetisetsa litšoantšo tse tsamaisang matla a agonist. Mol. Pharmacol. 1997;51: 87-96. [E fetotsoe]
  • Trigo JM, Martin-Garcia E, Berrendero F, Robledo P, Maldonado R. Ts'ebetso ea opioid e sa feleng: karolo e tloaelehileng ea ho lemalla lithethefatsi. Tšebeliso ea Lithethefatsi e Tšepahala. 2010;108: 183-194. [E fetotsoe]
  • Unterwald EM, Knapp C, Zukin RS. Ho fumanoa ha methapo ea metsoako ea κ1 le κ2 li-receptor tse nang le boko ba likhopo le likolobe. Resin ea Boko. 1991;562: 57-65. [E fetotsoe]
  • Helena FJ, Helena FE, Koob GF. Ho thibeloa ha nucleus accumbens opiate receptors ho thibela heroin e kenyang chelete moputsong. Psychopharmacology (Berl) 1985;86: 37-42. [E fetotsoe]
  • Vigano D, Rubino T, Vaccani A, Bianchessi S, Marmorato P, Castiglioni C, Parolaro D. Mekhoa ea likokoana-hloko e amanang le ho se lumellane ha lipakane pakeng tsa tsamaiso ea li-cannabinoid le opioid. Psychopharmacology (Berl) 2005;182: 527-536. [E fetotsoe]
  • Zachariou V, Bolanos CA, Selley DE, Theobald D, Cassidy MP, Kelz MB, Shaw-Lutchman T, Berton O, Sim-Selley LJ, Dileone RJ, Kumar A, Nestler EJ. Karolo ea bohlokoa bakeng sa DeltaFosB nucleus accumbens morphine khato. Nat. Neurosci. 2006;9: 205-211. [E fetotsoe]
  • Zangen A, Solinas M, Ikemoto S, Goldberg SR, RA ea bohlale. Libaka tse peli tsa boko bakeng sa moputso oa cannabinoid. J. Neurosci. 2006;26: 4901-4907. [E fetotsoe]
  • Zhu J, Luo LY, Li JG, Chen C, Liu-Chen LY. Ho ts'ebetsoa ha motho ea bitsoang kappa opioid receptor ka li-agonists ho ntlafatsa [35S] GTPγS e tlamella likarolo: ho iketsetsa matla le matla a li-ligands. J. Pharmacol. Exp. Ther. 1997;282: 676-684. [E fetotsoe]
  • Zimmer A, Valjent E, Konig M, Zimmer AM, Robledo P, Hahn H, Valverde O, Maldonado R. Ha ho letho la delta -9-tetrahydrocannabinol e bakoang ke litoeba tsa litoeba tse senyehileng tsa dynorphin. J. Neurosci. 2001;21: 9499-9505. [E fetotsoe]
  • Zimmer A, Zimmer AM, Hohmann AG, Herkenham M, Bonner TI. Ho shoela ho eketsehileng, boikaketsi, le hypoalgesia mokhoeng oa kokotlo oa CB1 mokuli oa ho kokota. Proc. Natl. Acad. Sci. USA 1999;96: 5780-5785. [Tlhahiso ea mahala ea PMC] [E fetotsoe]