Impembelelo ye-ΔFosB ngokugqithiseleyo kwi-opioid kunye ne-cannabinoid receptor-signal signaling in the nucleus accumbens (2011)

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

Sim-Selley LJ, Cassidy MP, I-Sparta A, UZachariou V, Nestler EJ, USelley DE.

imvelaphi

ISebe lePhysacology kunye neTochnology kunye neZiko leZifundo ngeziyobisi kunye noTywala, iVenkile yeDyunivesithi yaseMelika eVenayo, eRichmond, eVA 23298, e-USA.

Abstract

Umba ozinzileyo wokukhuphela ΔFosB ubhekiswa kwi-nucleus accumbens (NAc) ngokuchazwa okungapheliyo kweziyobisi ezininzi zokuxhatshazwa, kunye nokubonisa kwe-ΔFosB kwi-striatum kuphucula iipropathi ezinomvuzo ze-morphine kunye necocaine. Nangona kunjalo, isiseko sezobuchwephesha koku kuqatshelweyo asiqondakali ngokupheleleyo. Sisebenzise imodeli yegundane ye-bitransgenic ngentetho engachanekanga ye-osFosB ngaphakathi dopamine I-D (1) i-receptor / dynorphin ene-striatal neurons ukumisela isiphumo sokubonisa kwe-ΔFosB kwi-opioid kunye ne-cannabinoid receptor signature kwi-NAc. Iziphumo zabonisa ukuba umsebenzi we-mu opioid-Mediated G-protein kunye nokuthintela ukuhamba nge-adenylyl cyclase kuphuculwe kwi-NAc yeempuku eziveze ΔFosB. Kwangokunjalo, uthintelo lwe-kappa opioid inhibition ye-adenylyl cyclase yaphuculwa kwi-ΔFosB eveza iimpuku. Ngokwahlukileyo, ukutywinwa kwe-cannabinoid receptor-mediated ahlukanga phakathi kweempuku overexpressing ΔFosB kunye neempuku zokulawula. TIziphumo ze-hese zibonisa ukuba i-opioid kunye ne-cannabinoid receptor signaling ziguqulwe ngokungafaniyo ngokubonakaliswa kwe-ΔFosB, kwaye ibonisa ukuba ibinzana le-ΔFosB linokuvelisa ezinye zeziphumo zalo ngokuphucula i-mu kunye ne-kappa opioid receptor isayina kwi-NAc.

Internet: I-G-protein, i-adenylyl cyclase, i-striatum
Yiya e:

1. intshayelelo

Ii-receptors ze-Opioid kunye ne-cannabinoid CB1 ii-receptors (i-CB1I-R) ziithagethi ze-neurobiological zeeklasi ezimbini ezisetyenziswa ngokubanzi kweziyobisi ezibandakanya i-morphine, i-heroin kunye ne-opioids egunyazisiweyo, kunye nentsangu (Δ9-tetrahydrocannabinol (THC), ngokulandelanayo. Iziphumo ezibi kakhulu ze-opioids kunye nee-cannabinoids zidityaniswa nee-G-protein-coupled receptors ezenza ngokuyintloko G.mna / o Iiproteni kwaye uvelise iimpendulo zehle ezantsi ezinjengethintelo le-adenylyl cyclase (Abasebenzi, 1991, Abasebenzi, et al., 1992, I-Howlett, et al., 2002). Imoto, ukungasebenzi kakuhle kwememori kunye neziphumo zengqondo ze Δ9-THC ziveliswa yi-CB1R (Huestis, et al., 2001, I-Zimmer, et al., 1999), ezisasazwa ngokubanzi kwingqondo, ngamanqanaba aphezulu kwi-basal ganglia, hippocampus kunye ne-cerebellum (I-Herkenham, et al., 1991). Iziphumo zokuhlaziya kunye nezivuzayo zeziyobisi ezifanelekileyo zeklinikhi nezihlukumezayo zixolelaniswe ikakhulu zii-op opioid receptors (MOR) (Matthes, et al., 1996), ezityetyisiweyo kwinkqubo yemilenze kunye nobuchopho (I-Mansour, et al., 1994). Inkqubo ye-mesolimbic, equlathe uqikelelo lwe-dopaminergic ukusuka kwindawo yecarral tegmental tegmental (VTA) ukuya kwi-nucleus accumbens (NAc), idlala indima ebalulekileyo kwiziphumo ezinomvuzo ze-opioids kunye ne-cannabinoids (U-Bozarth kunye noWise, 1984, Vaccarino, et al., 1985, I-Zangen, et al., 2006), kunye nezinye iziyobisi zokuphathwa gadalala (Koob noVolkow, i-2010). Ngaphezu koko, iinkqubo ze-opioid opioid kunye ne-cannabinoid zibandakanyeka kwiziphumo ezivuzayo zezigaba ezininzi zamachiza e-psychoactive (Maldonado, et al., 2006, I-Trigo, et al., 2010). Ke ngoko, kubalulekile ukuba kucaciswe iindlela ngeendlela apho opioid kunye ne-CB1Ukubonisa uphawu kulawulwa kwi-NAc.

Umbuzo ophambili kwicandelo lokusetyenziswa kweziyobisi ngaba kukuchonga iiproteni eziguqula ukutshintsha ukusuka kwisiphumo esibi ukuya kwezamayeza ixesha elide. Into eprintiweyo ye-AP-1 ΔFosB inomdla ngokukhethekileyo kuba iyimveliso eguqukayo yes splice eyahlukileyo yomthi fosb Uhlobo olufunyanwa rhoqo ekuvezweni okuphindaphindiweyo kweziyobisi zokuxhatshazwa okanye imbuyekezo yendalo (McClung, et al., 2004, Nestler, 2008, I-Nestler, et al., 1999). Sifumanise ukuba i-ΔFosB ibethelelwa kwingqondo emva kokubhenqa okuphindaphindiweyo kwi-morphine, Δ9-THC, icocaine okanye i-ethanol, ichiza ngalinye livelisa imodeli yommandla expressionFosBI-Perrotti, et al., 2008). Iziphumo ezifumanekayo kumachiza onke yayikukuba i-ΔFosB ibethelelwa kakhulu apho bekukho khona iziyobisi ezine zibekwa indFosB kumbindi we-NAc kwaye zonke ngaphandle kwa Δ9-THC intetho ebanzi kakhulu kwiqokobhe le-NAc kunye ne-caudate-putamen.

Izifundo ze-Pharmacological zibonise ukuba ukulawulwa ngokubambisana kwe-dopamine D1 receptor (D1I-R) I-SCH 23390 echasene ne-ΔFosB yokungeniswa kwi-NAc kunye ne-caudate-putamen elandelayo ye-cocaine ye-cocaine okanye ulawulo lwe-morphine, ephakamisa ukubaluleka okunokubakho kwe-D.1Ukubonakalisa i-neurons (Muller no-Unterwald, i-2005, Ewe, et al., 1995). Iziphumo zokungeniswa kwe-ΔFosB kwimikhwa ehambelana neziyobisi kuphandwe kusetyenzwa ngeempuku ze-bitransgenic ezibonisa ΔFosB kubemi abathile be-neuronal ye-NAc kunye ne-dorsal striatum (I-Chen, et al., 1998). Impazamo ezibonisa ΔFosB kwi-dynorphin / D1I-neurons elungileyo kwi-NAc kunye ne-dorsal striatum (umgca 11A) bonisa iimpendulo eziguqulweyo kumachiza okuphathwa gadalala, ngakumbi ukuphucula imvakalelo kwiziphumo ezinomvuzo zecocaine okanye morphine (I-Colby, et al., 2003, UKelz, et al., 1999, UZachariou, et al., 2006). Olutshintsho lwenzekile kungabikho zinguqu kumanqanaba e-MOR okanye kwi-G-protein engaphantsi. Nangona kunjalo, amanqanaba e-dynorphin mRNA ancitshiswa kwi-NAc ye-ΔFosB echaza iimpuku (UZachariou, et al., 2006), ephakamisa ukuba ujoliso lwe-ΔFosB luhlobo lokubhaliweyo lwe-pioide ye-opioid. Ukungeniswa kwe-osFosB kungaphinda kuvelise utshintsho kwindlela yokuziphatha ngokulawula ukwenziwa kwe-receptor kwi-NAc, kodwa oku akunakuphandwa. Ke ngoko, izifundo ezikhoyo zisebenzisa imodeli yegundane ye-bitransgenic ukumisela ukuba ngaba ukugqithisa ΔFosB kwi-dynorphin / D1I-R ene-striatal neurons etshintsha i-MOR-Mediated G-protein yomsebenzi ne-MOR- kunye ne-KOR-Mediated adenylyl cyclase inhibition kwi-NAc. Iziphumo ze-ΔFosB kwi-CB1Umsebenzi we-G-protein-G ophakathi uphinde wavavanywa kuba Δ9Ulawulo lwe -THC lubangela i-ΔFosB kwi-NAc (I-Perrotti, et al., 2008) kunye nenkqubo ye-endocannabinoid iyaziwa ukulawula imijikelezo yomvuzo wobuchopho (UGardner, 2005, Maldonado, et al., 2006), kodwa iziphumo ze-ΔFosB kwinkqubo ye-endocannabinoid azange iphandwe.

Yiya e:

2. Impahla nenkqubo

2.1. Ukuhlaziya

[35S] GTPγS (1250 Ci / mmol), [α-32P] I-ATP (800 Ci / mmol) kunye [3I-H] cAMP (26.4 Ci / mmol) yathengwa kwiPerkinElmer (Shelton, CT). I-ATP, i-GTP, i-GDP, i-cAMP, i-bovine serum albhamuin, i-creatine phosphokinase, ipapaverine, imidazole kunye ne-WIN-55212-2, zithengiwe kwa-Sigma Aldrich (iSt. Louis, MO). I-GTPγS ithengwe kwiRoche Diagnostic Corporation (eChicago, IL). I-DAMGO yanikezelwa yiNkqubo yokuBonelela iziyobisi ngeZiko leSizwe kuZwelonke ngokuSetyenziswa gwenxa kweziyobisi (iR Rockville, MD). I-Econo-1 scintillation fluid yafunyanwa kuFisher Science Science (Norcross, GA). I-ecolite scintillation fluid yafunyanwa kwi-ICN (Costa Mesa, CA). Zonke ezinye iichemicals zifunyenwe kwaSigma Aldrich okanye kwiFisher Science.

2.2. Iimpuku

Iimpuku ze-bitransgenic zeempuku ezivela kwi-NSE-tTA (umgca A) × TetOp-ΔFosB (umgca 11) zaveliswa njengoko zichaziwe kuKelz et al. (UKelz, et al., 1999). Iigundane zeBitransgenic zakhulelwa kwaye zakhulela kwi-doxycycline (100 µg emanzini okusela) ukucinezela ukubonakaliswa kwe-transgene. Kwiiveki ze-8 zeminyaka ubudala, i-doxycycline ishixiwe emanzini kangangesiqingatha seempuku ukuvumela ukugqithisa, ngelixa iimpuku ezisele zigcinwe kwi-doxycycline ukucinezela i-transgene. Iikreyiti zaqokelelwa kwi-8 iiveki emva kwexesha, ixesha apho iziphumo ze-ΔFosB zikhutshelweyo zibukhulu.McClung noNestler, 2003). Umgca wesibini wokudlula kwegundane wasetyenziswa apho u--c-Jun, umchasi ophikisayo ongalunganga we-c-Jun, uboniswa ku-D.1R / dynorphin kunye no-D2R / enkephalin iiseli ze-striatum, hippocampus kunye ne-parietal cortex (I-Peakman, et al., 2003). I-C-Jun kunye neeprotein zosapho ezihambelana ne-Jun ziba mbi nge proteni yosapho ze-Fos kwaye zibophelele kwindawo ye-AP-1 yeentlobo ekujilwe kuzo ukuze zilawule ukukhutshelwa. Nangona kunjalo, ukuthathwa kwe-N-terminus ye-c-Jun (Δc-Jun) kuhambisa ubunzima obuntsokothileyo bokungasebenzi kunye nokukwazi ukuthintela ukubopha kwe-DNA kobume obusebenzayo be-AP-1. Iigundane ze-bitransgenic zeempuku ezivela kwi-NSE-tTA (umgca A) × TetOp-FLAG-Δc-Jun (umgca E) zaveliswa njengoko zichaziwe kwiPeakman et al. (I-Peakman, et al., 2003). Iigundane zeBitransgenic zakhulelwa kwaye zakhulela kwi-doxycycline (100 µg emanzini okusela) ukucinezela ukubonakaliswa kwe-transgene. Ii-Pups zilunyulwe kwiiveki ze-3, zenziwa genotyped, zaza zahlulahlulwa zaba ngamaqela, ngesiqingatha sigcinwe kumanzi aqulethe i-doxycycline kunye nesiqingatha kumanzi okusela aqhelekileyo ukwenza i-FLAG-Δc-Jun. Iikreyiti zaqokelelwa kwiiveki ze-6 emva kwexesha, ixesha apho amanqanaba aphezulu e-FLAG-Δc-Jun ebaliweI-Peakman, et al., 2003). Zonke iinkqubo zezilwanyana beziqhutywa ngokuhambelana neZiko leSizwe leSikhokelo seMpilo kuKhathalelo kunye nokuSetyenziswa kweeLebhu zeLebhu.

2.3. Ukulungiselela kweMembrane

Iibhuki zigcinwe kwi-−80 ° C kude kube ngumhla we-assay. Phambi kokuba uthathwe, ingqondo nganye yayiyekisiwe, kwaye i-NAc yahluliwe kwi-ice. Isampulu nganye yayinokungasebenzi kakuhle kwi-50 mM Tris-HCl, 3 mM MgCl2, I-1 mM EGTA, pH 7.4 (i-membrane buffer) ngemivimbo ye-20 esuka kwiglasi homogenizer kwi-4 ° C. I-homogenate yayisisikhulu kwi48,000 × g kwi-4 ° C ye-10 min, iphinde yasebenza kwi-membrane buffer, isentrifuged kwakhona kwi48,000 × g kwi-4 ° C ye-10 min kwaye iphinde yasebenza kwi-50 mM Tris-HCl, 3 mM MgCl2, I-0.2 mM EGTA, i-100 mM NaCl, pH 7.4 (i-assay buffer). Amanqanaba eprotheyini amiselwa yindlela yeBradford (Bradford, 1976) ukusebenzisa i-bovine serum albhamuin (BSA) njengomgangatho.

2.4. Kukhuthazwe u-Agonist [35S] Ukudityaniswa kwe-GTPγS

I-Membranes ifakwe ngaphambili kwimizuzu ye-10 kwi-30 ° C nge-adenosine deaminase (3 mU / ml) kwi-assay buffer. I-Membranes (i-5-10 µg iproteyini) yafakwa incanca kwi-2 hr kwi-30 ° C kwisibambisi esine-0.1% (w / v) BSA, 0.1 nM [35S] GTPγS, 30 µM ​​GDP kunye ne-adenosine deaminase (3 mU / ml) kwaye ngaphandle koqwalaselo olufanelekileyo lwe-DAMGO okanye iWIN55,212-2. Ukubopha okungenakulinganiswa kwalinganiswa nge-20 µM ​​GTPγS. I-incubation iphelisiwe ngokuhlunga ngeefilitha zeglasi ye-GF / B, elandelwa yi-3 yokuhlamba nge-3 ml ye-ice-ebandayo ye-50 mM Tris-HCl, pH 7.4. I-radioacuction yomda yamiselwa yi-fluid scintillation visrophotometry emva kokukhuphela ebusuku iifilitha kwi-Econo-1 fluid fluid.

2.5. I-Adenylyl Cyclase Assay

I-Membranes (i-5-25 µg iprotein) yaqalwa yenziwa nge-adenosine deaminase njengoko kuchaziwe apha ngasentla, emva koko yafakwa kwi-15 min kwi-30 ° C phambi kobukho okanye ukungabikho kwe-1µM forskolin, okanye ngaphandle kwe-DAMGO, U50,488H okanye i-WIN55,212-2. I-50 µM ​​ATP, [α-32P] I-ATP (1.5 µCi), 0.2 mM DTT, 0.1% (w / v) BSA, 50 µM ​​cyclic AMP, 50 µM ​​GTP, 0.2 mM papaverine, 5 mM phosphocreatine, 20 unitsMinine deamin de mamine de mamine de sineh. / ml) kwisixa sokugqibela se3 µl. Phantsi kwale meko, iyonke [α-32I-P] i-cAMP efunyenwe yayihlala ingaphantsi kwe-1% yexabiso elipheleleyo elongeziweyo [α-32P] I-ATP kwisampulu nganye. Ukuphendula kupheliswe ngokubilisa i-3 min kunye [32P] I-cyclic AMP yahlulwa yindlela emibini (Dowex kunye alumina) indlela kaSalomon (USalomon, 1979). [3I-H] cAMP (10,000 dpm) yongezwa kwityhubhu nganye ngaphambi kwekholam ye-chromatography njengomgangatho wangaphakathi. Umsebenzi werediyo wamiselwa yi-fluid scintillation spectrophotometry (ukusebenza ngempumelelo kwe-45% ye 3H) emva kwe-4.5 ml ye-eluate ichithwe kwi-14.5 ml ye-ecolite scintillation fluid.

2.6. Uhlalutyo lwedatha

Ngaphandle kokuba kuboniswe ngenye indlela, idatha ixelwa njengamaxabiso aqinisekileyo ± SE ye4-8 iimvavanyo ezahlukeneyo, nganye nganye yenziwa ngophindaphindo. IshukumiweNet [35S] Ukudibana kwe-GTPγS kubalwa njengokubopha okungaphezulu kwe-agonist. Intshukumo ye-forskolin ekhuthazwayo ye-adenylyl cyclase ichazwa njengomsebenzi we-forskolin ovuseleliweyo-umsebenzi osisiseko (pmol / mg / min). Ipesenti yokuthintela into ye-forskolin-evuswe ye-adenylyl cyclase ichazwa njengesiqhelo (isenzo se-forskolin eshukunyiswayo ukungabikho kwe-agonist - umsebenzi owenziwe nge-forskolin phambi komcimbi we-agonist / net ye-forskolin ekhuthazayo xa kungekho agonist) x 100. Lonke ucazululo lwe-curve-fit kunye nohlalutyo lwamanani lwenziwa kusetyenziswa i-Prism 4.0c (isoftware ye-GraphPad, Inc., iSan Diego, CA). I-curve-isiphumo sempembelelo zahlaziywa ngohlobo lwe-iterative non-linear regression ukufumana i-EC50 kunye Emax iinqobo ezisemgangathweni. Ukubaluleka kwenkcazo yedatha yeziphumo zoxinzelelo kumiselwe ngohlalutyo lweendlela ezimbini (ANOVA), kusetyenziswa idosi ye-agonist kunye ne-gene induction (on okanye off) njengezinto eziphambili. Ukubaluleka kobalo lwamaxabiso afanelekileyo ijikajika (Emax okanye EC50) yagqitywa ngovavanyo lwe-Student ta-two-tailed's test, kusetyenziswa ukulungiswa kwe-Welch okanye inguqu yesikwere sedatha apho kufanelekileyo ukulungisa ukungalingani okungafaniyo (okufunyenwe luvavanyo lwe-F) kwi-EC50 iinqobo ezisemgangathweni.

Yiya e:

3. Iziphumo

3.1. Iziphumo zokubonisa kwe-ΔFosB kwi-opioid kunye ne-cannabinoid receptor-Mediated G-protein activation.

Ukufumanisa ukuba ngaba-MOR- okanye CB1Ukwenza ukuba iprotein ye-R-mediated isebenze yatshintshwa ngu-ΔFosB ongenakuthelekiswa nanto kwi-NAc, evuselelwa yi-agonist-35S] Ukuzibophelela kwe-GTPγS kwavavanywa kulungelelwaniso olulodwa olulungiselelwe kulo mmandla weempuku ze-bitransgenic ziveza imeko (ΔFosB ivuliwe) okanye ngaphandle kokuchaza (ΔFosB off) the ΔFosB transgene. I-MOR ekhethiweyo ye-enkephalin analog DAMGO yayisetyenziselwa ukwenza i-MOR kwaye i-cannabinoid aminoalkylindole WIN55,212-2 yayisetyenziselwa ukwenza i-CB isebenze.1R. Ezi ligands ngaphambili zaboniswa zingama-agonists apheleleyo kwi-MOR nase-CB1R, ngokulandelanayo (UBreivogel, et al., 1998, USelley, et al., 1997). Akuzange kube nokwenzeka ukuvavanya umsebenzi we-G-protein ophakathi we-GOR kuba uphawu luphantsi kakhulu kwingqondo yeentent (Abasebenzi, et al., 1998). Iziphumo zibonise ukukhuthazeka okuxhomekeka kuxinzelelo lwe-G-protein eyenziwa ngababini be-DAMGO kunye ne-WIN55,122-2 kwi-NAc ukusuka kwi-ΔFosB ishiye kunye ne-osFosB kumagundane (Umzobo 1). Ngomsebenzi owenziwa yi-DAMGO (Umzobo 1A), iindlela ezimbini ze-ANOVA zedatha yoxinzelelo-zichaze iziphumo eziphambili ze-osBFosB status (p <0.0001, F = 22.12, df = 1) kunye noxinaniso lwe-DAMGO (p <0.0001, F = 29.65, df = 5) ngaphandle Unxibelelwano olubalulekileyo (p = 0.857, F = 0.387, df = 5). Uhlalutyo olungahambelaniyo nolungelelwaniso lwee-curves-effect curves lubonakalise i-DAMGO E enkulu kakhulumax Ixabiso kwi ΔFosB kwiimpuku (Emax = 73 ± 5.2% ukukhuthaza) malunga ne-ΔFosB off yeempuku (Emax = 56 ± 4.1% yokukhuthaza; p <0.05 eyahlukileyo kwi-ΔFosB kwiimpuku ngovavanyo lomfundi t). IDAMGO EC50 amaxabiso ayengafani phakathi kwe-ΔFosB kwi-BFosB off kunye ne-302 ± 72 nM ngokuthelekisa i-212 ± 56 nM, ngokulandelanayo, p = 0.346).

Umzobo 1 

Iziphumo ze-ΔFosB expression kwi-agonist-ivuswe35S] I-GTPγS ibophelela kwi-NAc. I-Membranes evela kwi-ΔFosB-expression (ΔFosB on) or control (ΔFosB off) amagundane atyiwa njengoko kuchaziwe kwiindlela zokusebenza kusetyenziswa ukugxila okungafaniyo ...

Ngokuchasene neziphumo ezifunyenwe kwi-MOR agonist DAMGO, akukho mehluko dependentFosB oxhomekeke kwisimo se-G-protein wenziwe waqwalaselwa nge-cannabinoid agonist WIN55,212-2 (Umzobo 1B). Iindlela ezimbini ze-ANOVA zedatha ye-WIN55,212-2 yedatha yoxinzelelo ibonakalise eyona nto iphambili kuxinzelelo lwe-WIN55,212-2 (p <0.0001, F = 112.4, df = 7), kodwa hayi ngobume be-osBFosB (p = 0.172 , F = 1.90, df = 1) kwaye kwakungekho nxibelelwano (p = 0.930, F = 0.346, df = 7). Ngokufanayo, kwakungekho siphumo senqanaba le-osBFosB kwi-WIN55,212-2 Emax amaxabiso (103 ± 6% xa kuthelekiswa ne-108 ± 8% ukukhuthaza kwi-ΔFosB phambili kunye nangaphandle kweempuku, ngokulandelelana, p = 0.813 ngovavanyo lwe-T) lomfundi okanye nge-EC.50 amaxabiso (i-103 ± 20 nM xa kuthelekiswa ne-170 N 23 nM kwi-onFosB icinywe nangaphandle kweempuku, ngokwahlukeneyo, p = 0.123).

Ngokusekwe kubume bee-curves kunye nenyaniso yokuba izifundo zethu zangaphambili zibonise i-biphasic WIN55,212-2-curves-effect curves kwi-brain (UBreivogel, et al., 1999, UBreivogel, et al., 1998), ii-curls ze-WIN55,212-2 ziye zahlalutywa kusetyenziswa imodeli yendawo ezimbini. Uhlalutyo lwedatha ethengiweyo lubonise ukuphucuka okuncinci kokulunga kokusebenzisa imodeli yendawo ezimbini (R2 = I-0.933 kunye ne-0.914, isixa sezikwere = 3644 kunye 5463 kwi-ΔFosB kwi-off and off mbeva, ngokwahlukeneyo) ngokuthelekiswa nemodeli yendawo enye (R2 = I-0.891 kunye ne-0.879, isixa sezikwere = 6561 kunye ne-6628 kwi-ΔFosB kwi-off and off mbezi, ngokulandelanayo). Nangona kunjalo, akukho mahluko ubalulekileyo wafunyanwa phakathi kwe-ΔFosB kwiimpazamo nangaphandle kweempuku nokuba yeyiphi Emax okanye EC50 amaxabiso eziza eziphezulu okanye ezisezantsiItafile ezongezelelweyo 1), nangona bekukho indlela eya kwiEC esezantsi50 Ixabiso kwindawo ephezulu ye-potency kwindawo yeempuku nge-ΔFosB kwi-EC50phezulu = 28.0 ± 10.6 nM) ngokuthelekiswa nezo ΔFosB zivaliwe (EC50phezulu = 71.5 ± 20.2 nM; p = 0.094). Ngaphezu koko, akubangakho mpembelelo yesimo se-ΔFosB kwisiseko [35S] I-GTPγS ibopha kwi-membcane ye-NAc (i-253 N 14 xa ithelekiswa ne-226 ± I-14 fmol / mg kwi-ΔFosB kwi-off and off mbezi, ngokulandelanayo, p = 0.188). Ezi datha zibonisa ukuba i-genFosB kwi-NAc yeempuku inyusa ukungasebenzi kwe-MOR ngaphandle kokuchaphazela kakhulu i-CB.1Umsebenzi we-protein ophakathi okanye osisiseko se-G-protein.

3.2. Iziphumo ze-ΔFosB kwi-opioid kunye ne-cannabinoid receptor-mediated inhibition ye-adenylyl cyclase

Ukuvavanya isiphumo sentetho engafezekisiyo yeNgeniso ye-ΔFosB ekuguqulweni kwemisebenzi esetyenziswayo ye-MOR kunye ne-CB.1R, inhibition ye1 µM ​​forskolin-ekhuthazwayo ye-adenylyl cyclase yomsebenzi yahlolwa kwi-membrane ye-NAc. Ukongeza kwi-MOR- kunye ne-CB1Isithintelo se-R-Mediated inhibition yomsebenzi we-adenylyl cyclase, iziphumo zomsebenzi we-KOR zavavanywa kusetyenziswa i-UORNUMX ye-KOR ekhethiweyoUZhu, et al., 1997), kuba iziphumo zangaphambili zibonise ukuba i-dynorphin mRNA yayijolise kwi-ΔFosB kwimodeli ye-bitransgenic (UZachariou, et al., 2006). Iziphumo zabonisa ukuba i-DAMGO, U50,488 kunye ne-WIN55,212-2 nganye ivelise inhibition yokuxhomekeka koxinzelelo lomsebenzi we-adenylyl cyclase kuzo zombini ΔFosB off kunye ne-ΔFosB kumagundane (Umzobo 2). Iindlela ezimbini ze-ANOVA yedatha yeziphumo ze-DAMGO (Umzobo 2A) ibonakalise iziphumo eziphambili ze-osBFosB imeko (p = 0.0012, F = 11.34, df = 1) kunye noxinaniso lwe-DAMGO (p <0.0001, F = 29.61, df = 6), kodwa akukho nxibelelwano lubalulekileyo (p = 0.441, F = 0.986 , df = 6). Uhlalutyo olungahambelaniyo nolungelelwaniso lwee-curves ze-DAMGO ziveze i-DAMGO EC esezantsi kakhulu50 Ixabiso kwi-osBFosB kwiimpuku (101 ± 11 nM) xa kuthelekiswa ne-osBFosB kwiimpuku (510 ± 182 nM, p <0.05 ngovavanyo lomfundi t). Nangona kunjalo, kwakungekho mahluko ubalulekileyo kwi-DAMGO Emax amaxabiso (20.9 ± 1.26% kuthelekiswa ne19.8 ± 1.27% inhibition in ΔFosB on and off gice, ngokulandelanayo, p = 0.534).

Umzobo 2 

Iziphumo zokubonisa kwe-ΔFosB kwithintelo lomsebenzi we-adenylyl cyclase kwi-NAc. I-Membranes evela kwi-ΔFosB-expression (ΔFosB on) or control (ΔFosB off) iimpuku zazicingelwa njengoko kuchaziwe kwiindlela kubukho be1 µM ...

I-KOR-Mediated adenylyl cyclase inhibition nayo yahlukile njengomsebenzi wentetho engenakuphikiswa ye-genFosB (Umzobo 2B). Iindlela ezimbini ze-ANOVA ze-U50,488 yedatha yesiphumo soxinzelelo ibonisa iziphumo eziphambili ze-osBFosB status (p = 0.0006, F = 14.53, df = 1) kunye noxinaniso lwe-U50,488 (p <0.0001, F = 26.48, df = 3) , ngaphandle konxibelelwano olubalulekileyo (p = 0.833, F = 0.289, df = 3). Uhlalutyo olungahambelaniyo nolungelelwaniso lwee-curve-effect curves ziveze i-U50,488 enkulu ye-Emax Ixabiso kwi-osBFosB kwiimpuku (18.3 ± 1.14% inhibition) xa kuthelekiswa ne-osBFosB kwiimpuku (12.5 ± 2.03% inhibition; p <0.05 eyahlukileyo kwi-ΔFosB ngovavanyo lwe-Student t), kungekho mahluko ubalulekileyo kwi-U50,488 EC50 amaxabiso (i-310 ± 172 nM xa kuthelekiswa ne-225 N 48 nM kwi-onFosB icinywe nangaphandle kweempuku, ngokwahlukeneyo, p = 0.324).

Ngokuchasene neempembelelo ezibonwe nge-MOR ne-KOR, kwakungekho siphumo sibalulekileyo sokuthetha okungafunekiyo kwe-genFosB kwisithintelo se-adenylyl cyclase yi-cannabinoid agonist WIN55212-2 (Umzobo 2C). Iindlela ezimbini ze-ANOVA ze-WIN55,212-2 idatha yesiphumo soxinzelelo ibonakalisa isiphumo sokuxinzezeleka kweziyobisi (p <0.0001, F = 23.6, df = 2), kodwa hayi ngobume be-osBFosB (p = 0.735, F = 0.118, df = 1) kwaye kwakungekho nxibelelwano lubalulekileyo (p = 0.714, F = 0.343, df = 2). Ngaphaya koko, kwakungekho siphumo senqanaba le-osBFosB kwisiseko okanye kwimisebenzi ye-forskolin evuselelekayo ye-adenylyl cyclase ngokungabikho kwayo nayiphi na i-agonist. Umsebenzi osisiseko we-adenylyl cyclase yayiyi-491 ± 35 pmol / mg / min kwi-ΔFosB kwiimpuku xa kuthelekiswa ne-546 ± 44 kwi-osBFosB kwiimpuku (p = 0.346 ngovavanyo lwabafundi). Kwangokunjalo, umsebenzi we-adenylyl cyclase phambi kwe-1 forM forskolin yayiyi-2244 ± 163 pmol / mg / min kwi-ΔFosB kwiimpuku xa kuthelekiswa ne-2372 ± 138 pmol / mg / min kwi-osBFosB kwiimpuku (p = 0.555).

3.3. Iziphumo ze-ΔcJun kwi-opioid kunye ne-cannabinoid receptor-mediated inhibition ye-adenylyl cyclase

Kungenxa yokuba ukungafikeleli kokubonisa kwe-ΔFosB yothutho lwesandisi-mqondiso sokuthintela ukusuka ku-MOR nakwi-KOR ukuya kwi-adenylyl cyclase kwi-NAc, kwakungumdla ukumisela ukuba into ethintelayo engalunganga ye-ΔFosB-ephakathi ekhutshelweyo inokuguqula isigama se-opioid receptor ngendlela eyahlukileyo. Ukuphendula lo mbuzo, inhibition ye-forskolin-evuswe ngumsebenzi we-adenylyl cyclase ngu-DAMGO kunye ne-U50,488 yahlolwa kwimisebenzi yolungiso evela kwi-NAc yeempuku ze-bitransgenic ziveza imeko ΔcJun. Iziphumo aziboniswanga ntshukumo ibalulekileyo expressioncJun expression ekuthinteleni umsebenzi we-adenylyl cyclase yi-MOR okanye KOR (Umzobo 3). Iindlela ezimbini ze-ANOVA zee-curves ze-DAMGO zibonisa isiphumo esiphambili soxinaniso lwe-DAMGO (p <0.0001, F = 20.26, df = 6), kodwa hayi imeko ye-ΔcJun (p = 0.840, F = 0.041, df = 1) kwaye kwakungekho nxibelelwano lubalulekileyo (p = 0.982, F = 0.176, df = 6). Ngokufanayo, kwakungekho mahluko ubalulekileyo ku-Emax okanye EC50 amaxabiso phakathi kweempuku kunye ΔcJun kwi (Emax = 23.6 ± 2.6%; EC50 = 304 ± 43 nM) okanye ΔcJun off (Emax = 26.1 ± 2.5%, p = 0.508; EC50 = 611 ± 176 nM, p = 0.129). Iziphumo ezifanayo zabonwa nge-U50,488, ezinje ngeendlela ezimbini ze-ANOVA zempembelelo zoxinzelelo ezibonisa isiphumo soxinzelelo (p <0.0001, F = 11.94, df = 6), kodwa hayi imeko ye-JcJun (p = 0.127 , F = 2.391, df = 1) kwaye kwakungekho nxibelelwano lubalulekileyo (p = 0.978, F = 0.190, df = 6). Ngokunjalo, kwakungekho mahluko ubalulekileyo ku-Emax okanye EC50 amaxabiso phakathi kweempuku kunye ΔcJun kwi (Emax = 14.8 ± 2.9%; EC50 = 211 ± 81 nM) okanye ucime (Emax = 16.7 ± 1.8%, p = 0.597; EC50 = 360 ± 151 nM, p = 0.411).

Umzobo 3 

Iziphumo ze-ΔcJun expression ekuthinteleni umsebenzi we-adenylyl cyclase kwi-NAc. I-Membranes evela kwi-ΔcJun-expression (ΔcJun on) okanye control (ΔcJun off) iingcambu zazifakelwe ubukho be-DAMGO (A), U50,488H (B) okanye WIN55,212-2 ...

I-JcJun expression nayo ayichaphazelekanga kuthintelo lwe-adenylyl cyclase kwi-NAc yi-cannabinoid agonist. Iindlela ezimbini ze-ANOVA ze-WIN55,212-2 curveffe curves zibonise eyona nto iphambili kuxinzelelo lwe-WIN55,212-2 (p <0.0001, F = 15.53, df = 6), kodwa hayi uhlobo lwe-genotype (p = 0.066, F = 3.472, df = 1) kwaye kwakungekho nxibelelwano lubalulekileyo (p = 0.973, F = 0.208, df = 6). Ngokunjalo, kwakungekho mahluko ubalulekileyo kwi-WIN55,212-2 Emax amaxabiso (13.0 ± 2.3% kunye ne-13.6 ± 0.9% inhibition in ΔcJun ngokuchasene neempuku, ngokwahlukeneyo, p = 0.821) kunye okanye kwi-EC.50 amaxabiso (208 ± 120 nM kunye 417 ± 130 nM kwi-ΔcJun ngokuchasene neempuku, ngokwahlukeneyo, p = 0.270). Ke, nangona bekukho imeko encinci yokunciphisa i-potency ye-WIN55,212-2 kwimigundwane echaza ΔcJun, i-transgene ayitshintshanga kakhulu ekutshintsheni i-cannabinoid inhibition ye-adenylyl cyclase. Ngapha koko, akubangakho mpembelelo yesimo se-ΔcJun kwisiseko sebhasiki okanye ye-forskolin evuselelweyo ye-adenylyl cyclase. Umsebenzi we-basal adenylyl cyclase was 1095 ± 71 pmol / mg / min kunye ne-1007 ± 77 pmol / mg / min (p = 0.403) kwiimpuku nge-ΔcJun okanye ngaphandle, ngokwahlukeneyo. Umsebenzi we-adenylyl cyclase uchukunyiswe yi-1 µM ​​forskolin yayingu-4185 ± 293 pmol / mg / min ngokuchasene ne-4032 ± 273 pmol / mg / min (p = 0.706) kwiimpuku nge-ΔcJun okanye ngaphandle, ngokwahlukeneyo.

3.4. Ingxoxo

Iziphumo zolu phononongo zivelise ukusebenza kwe-MOR-mediated G-protein activation kunye nokuthintela ukuhamba nge-adenylyl cyclase kwi-NAc yeempuku nge-transgenic transgenic expression of ΔFosB in dynorphin / D1R inee-neurons. Uthintelo lwe-KOR-mediated inhibition ye-adenylyl cyclase yaphuculwa kwi-NAc ye-ΔFosB echaza amagundane, ecetyiswa ukuba i-ΔFosB ilawula inkqubo ye-opioid ye-endo native kwi-NAc. I-DAMGO Emax ixabiso lalinkulu lokuvuselelwa kwe-MOR [35S] Ukudibana kwe-GTPingS, kunye ne-EC yayo50 Ixabiso lalingaphantsi kwe-adenylyl cyclase inhibition, kwi-ΔFosB ngaphezulu kokubonakalisa iigundane xa kuthelekiswa nolawulo lweempuku. Ezi ziphumo zibonisa ukuba kunokwenzeka ukuba ugcino lwe-receptor lokuguquguqulwa kwe-endor kodwa hayi i-G-protein yokuqhutywa phantsi kweemeko zokuvavanywa. Iziphumo zokufumanisa ukuba ubukhulu be-inhibition ye-adenylyl cyclase yi-KOR agonist yachaphazeleka yi-ΔFosB expression icacisa indawo yogcino ephantsi ye-KOR yokulamla, ehambelana namazinga asezantsi e-KOR yokubopha kwingqondo yegundane (I-Unterwald, et al., 1991). Ngokwahlukileyo, i-CB1Umsebenzi we-G-protein-G ophakathi kunye nokuthintelwa kwe-cymbase ye-adenylyl ayichaphazwanga yizwi le-ΔFosB, Ukucebisa ukuba iinkqubo ze-opioid kunye ne-cannabinoid zahlukile kwimpendulo yazo kwi-ΔFosB kwezi Nons neurons.

Iziphumo ze-ΔFosB kwisibonakaliso se-opioid receptor-Mediated iyahambelana nengxelo yethu yangaphambili yokuba intetho ye-ΔFosB kwisistriatum iguqulwe yingozi kunye neziphumo ezibi ze-morphine (UZachariou, et al., 2006). Iziphumo ezizezinye zolu phononongo yayikukuba iimpuku zine-transgenic expression of ΔFosB in dynorphin / D1I-striatal neurons yayinovelwano ngakumbi kwi-morphine kwimeko yemeko yendawo ngakumbi kunolawulo. Ngaphaya koko, le miphumo ilingisiwe kukuthetha okungaphakathi kwe-ΔFosB ngenaliti ethile yendawo kwi-NAc. Olu luqwalaselo luhambelana neziphumo zangoku ezibonakalisa ukuphuculwa kwe-MOR kwi-NAc.

Ngaphambili siye saluchonga uvavanyo lwe-gene I-dynorphin njengengjoliso ye-ΔFosB, kwaye iphakamise ukuba i-dynorphin encitshisiweyo ihambelane nepropathi enomvuzo ye morphine kwi ΔFosB bitransgenic mbezi (UZachariou, et al., 2006). Iziphumo ezikhoyo zibonisa ukuba i-KOR-mediated inhibition of adenylyl cyclase kwi-NAc iphuculwe kwiimpuku ze-ΔFosB, ezinokubonisa ukunyuka kokunyanzeliswa kokuziva kwe-KOR emva kokuncitshiswa kwe-dynorphin. Izifundo zangaphambili zibonise ukuba i-KOR iphinde yaphakanyiswa kwimimandla ethile yobuchopho beempuku ze-prodynorphin knockout, kubandakanya ne-NAc (Clarke, et al., 2003).

Ngokuchasene ne-ΔFosB, ukuchaza okungafikelelekiyo kwe-ΔcJun, isenzi esinegalelo elingalunganga lesiphazamiso se-JFosB esibambisene neqabane likaCJun, alizange litshintshe inhibition ye-adenylyl cyclase yi-MOR okanye i-KOR agonists. Ezi ziphumo zibonisa ukuba amanqanaba asiseko entetho ye-ΔFosB, aphantsi kakhulu, awadlali ndima ebalulekileyo ekugcineni isiginali ye-opioid receptor kweli nqanaba lokuhambisa umqondiso kwi-NAc. Inyaniso yokuba imeko enomvuzo enefomphine yancitshiswa ngamazwi ΔcJun kwisifundo sethu esedlule (UZachariou, et al., 2006) Icebisa nokuba ukwenziwa kwe-morphine kwe-ΔFosB ngexesha lokumiswa kwemeko kubalulekile kulawulo lweendlela zokuziphatha kwichiza okanye iziphumo zokukhutshelwa kwe-ΔFosB ngaphandle kwalezo ezichaphazela ukubonakaliswa kwesiginali ngu-opioid receptors kunokuba nefuthe kumvuzo we-opioid. Nakuyiphi na imeko, iziphumo zophando zibonisa ngokucacileyo ukuba, xa intetho ye-ΔFosB iphakanyisiwe ngaphezulu kwamanqanaba asisiseko kwi-drioral dynorphin / D1Ukubonakalisa i-neurons, kukho ukonyuka okumandla kokudityaniswa kwe-MOR kunye ne-KOR ekuthinteleni isiphikiso se-adenylyl kwi-NAc.

Iindlela zokucacisa i-MOR- kunye ne-KOR -edi-mediation ziphuculwe yi-ΔFosB ooverxpression azicacanga, kodwa ngaphambili sibonisa ukuba amanqanaba e-MOR, ahlolwe ngu-3H] i-naloxone ebophayo, ayifani kwi-NAc ye-ΔFosB kwiitayile ezichaseneyo (UZachariou, et al., 2006). Uphononongo olufanayo lwafumanisa ukuba i-GcyiAmanqanaba eprotheni ye-1 kunye ne-2 awachaphazelekanga kulo mmandla ngu-ΔFosB expression. Nangona kunjalo, uhlalutyo lwentambo lwangaphambili lokulungelelanisa lubonisa ukuba i-Gcyo IMRNA yavuselelwa kwi-NAc ye-ΔFosB kwiimpuku (McClung noNestler, 2003). Kuya kuba ngumdla kwizifundo ezizayo ukuvavanya ngokubanzi ifuthe le-transgenic ΔFosB expression on G-protein subunit expression kwinqanaba leprotein kunye nokuchazwa kweeproteni ezininzi zemodyuli ye-G-protein.

Kuyathakazelisa ukuba i-ΔFosB expression ayizange iphucule i-CB1Ukunxibelelana ngomnxibelelanisi kwi-NAc. Kungenzeka ukuba utshintsho kwi-CB1Ukubonakalisa kwenzeka kwi-discrete yabantu be-neurons ecekeceke kulungiselelo lonke lwe-NAc. Umzekelo, ulawulo lwe Δ9-I-THC ibangelwe kakhulu yi-ΔFosB kumbindi, kodwa hayi iqokobhe, le-NAc (I-Perrotti, et al., 2008). Mnaengekho, kubonisiwe ukuba umngeni ngo-Δ9-THC emva kolawulo oluphindiweyo luka Δ9-THC yonyuse ukukhutshwa kwe-dopamine kwisiseko se-NAc, kodwa yanciphisa ukukhutshwa kwembumba (Cadoni, et al., 2008). Kubalulekile ukuba uqaphele ukuba umgca we-11A womgca weempuku ze-bitransgenic ziveza ΔFosB kuphela kwi-dynorphin / D1I-neurons ephakathi elungileyo ye-striatum, kodwa i-CB1R zibonakaliswe zombini i-dynorphin / D1R kunye ne-enkephalin / D2I-striatal neurons elungileyo (UHohmann kunye noHerkenham, 2000), kunye nakwizikhululo zamaqabane athembekileyo (URobbe, et al., 2001). Ukucaciswa komlawuli ombi kakhulu we-ΔFosB ophakathi okhutshelweyo, ΔcJun, akwabikho siphumo sibalulekileyo kwi-cannabinoid receptor signaling, nangona i-ΔcJun ibonakaliswa ngokungafanelekanga kuzo zombini1 kunye noD2-Ukugcina abantu abaninzi bephakathi bephakathi be-spiny neurons kwezi mbezi (I-Peakman, et al., 2003). Kuyenzeka, nangona kunjalo, ukuba i-basal ΔFosB expression is ngokwaneleyo ngokwaneleyo ukuba ΔcJun ayinakuchaphazela ukubonakaliswa kwe-receptor, njengoko kucetyiswayo ziziphumo ze-MOR ne-KOR. Kusenokwenzeka ukuba i-CB1Ukubonakalisa ukuqiniswa ngokuthozamileyo kukuziswa kwe-basal ΔFosB, yokuba ukunyusa ngakumbi ukubonakaliswa kwe-ΔFosB okanye ukuvimba izenzo zayo nge-ΔcJun kuneziphumo ezincinci ezingafikanga kwinqanaba lokubaluleka kobalo. Inkxaso engathanga ngqo kolu toliko inokubonwa ngokuthelekisa i-WIN55,212-2 EC50 amaxabiso phakathi kweempuku ezibonisa ΔcJun kuthelekiswa ΔFosB. Umyinge we-WIN55,212-2 EC50 Ixabiso le-adenylyl cyclase inhibition in mbezi ngokuchazwa kwe-ΔcJun kwi-EC yayo50 Ixabiso lokwenza i-protein-ye-protini kwimigundane ene-ΔFosB yayiyi-4.0, ngelixa umlinganiso ofanayo kwiimpuku ngaphandle kokungeniswa kwenye ye-transgene yayiyi-1.2.

Kungenjalo, i-cannabinoids inokubangela i-ΔFosB expression ngaphandle kokuchaphazela ngqo kwi-CB1Ukubonakalisa. Kule meko, i-cannabinoids inokuguqula ukuphendula kwiziphumo zengqondo kunye nezinye iziyobisi nge-ΔFosB-Mediated transcriptal regulation. Mnainyani, ulawulo lwe-Δ9-THC ivelisa imbonakalo enqamlezayo kwi-opioids kunye ne-amphetamine (Cadoni, et al., 2001, I-Lamarque, et al., 2001), ehambelana nale hypothesis. Ngaphaya koko, ukuphindaphindiweyo kolawulo lwe-cannabinoid agonist CP55,940 kwathiwa yonyusa ukusebenza kwe-MOR-mediated G-protein kwi-NAc, ngokufanayo kwiigundane ezichaza ngokungalunganga ΔFosB kwisifundo esikhoyo (UVigano, et al., 2005). Iziphumo ze ΔFosB expression kwi Δ9Iindlela zokuziphatha eziphakathi -THC azivavanywa, kodwa iziphumo ezikhoyo azithinteli ukusebenzisana. Iziphumo zoku kunye nohlolisiso lwethu lwangaphambili (UZachariou, et al., 2006) bonisa utshintsho lwe-ΔFosB-in-MOR kunye ne-KOR / dynorphin kwi-striatum. Iziphumo ezivuzayo ze Δ9-THC, njengoko ilinganiswe kukhetho lwendawo, iyapheliswa kwiimpuku ze-MOR, ngelixa ucinyiwe lwe-KOR echongiweyo Δ9-THC Beka ubuqhophololo kwaye utyhilelwe Δ9-THC indawo ekhethiweyo (Ghozland, et al., 2002). Ngokufanayo, indawo ekhethiweyo yindawo yokuphambukela Δ9-THC ayikho kwi-pro-dynorphin knockout ngokuthelekiswa neempuku zohlobo lwasendle (I-Zimmer, et al., 2001). Ezi datha zibonisa ukuba Δ9-THC inokuba ngumvuzo ngakumbi emva kokungeniswa kwe-ΔFosB kunye nokungeniswa kwesiphumo se-MOR kunye nokuncitshiswa kwentetho ye-dynorphin.

Ngamafutshaney, iziphumo zolu phando zibonise ukubonakaliswa kwe-ΔFosB ku-D1I-R / dynorphin elungileyo ye-striatal neurons iphucule ukuqiniswa kwe-MOR- kunye ne-KOR-Mediated inhibition ye-G-protein Mediated inhibition yomsebenzi we-adenylyl cyclase kwi-NAc. Oku kufumanisa kuyahambelana nezifundo eziye zadlala indima yenkqubo ye-opioid opoid kumvuzo (I-Trigo, et al., 2010), kunye nokubonelela ngesixhobo esinokwenzeka se-ΔFosB-Mediated imiphumela kumvuzo. Ngokwahlukileyo, i-CB1Ukuboniswa kwe-R-Mediated signative kwi-NAc akuchatshazwanga kakhulu yintetho ye-riFosB ye-striatal phantsi kweemeko ezivavanyiweyo, nangona ezinye izifundo ziqinisekisiwe ukumisela isiphumo sokungeniswa kwe-ΔFosB kwinkqubo ye-endocannabinoid.

Amagqabantshintshi ophando

  • Ukuboniswa kwe-MOR kuyaphuculwa kwii-nucleus accumbens zeempuku ezibonisa ΔFosB
  • I-KOR inhibition ye-adenylyl cyclase ikwaphuculiswa kwiimpuku ezibonisa ΔFosB
  • Ukuchazwa kwe ΔFosB ayitshintshi i-CB1Ukutywina kwi-nucleus accumbens
Yiya e:

Izinto ezongezelelweyo

01

Yiya e:

Imibulelo

Ababhali bayambulela uHengjun He, uJohn Cox kunye no-Aaron Tomarchio ngoncedo lobugcisa [35S] I-GTPγS yokubopha iminyanya. Olu phononongo luxhaswe ziZibonelelo ze-USPHS DA014277 (LJS), DA10770 (DES) kunye ne-P01 DA08227 (EJN).

Yiya e:

Imihlathi

Iphepha elichazayo ukuba awusenanto oyifunayo: Le fayili yeFayile yombhalo wesandla ongabhalwanga owamkelwe ukushicilelwa. Njengenkonzo kumakhasimende ethu sinika le ngcaciso yokuqala kwincwadi yesandla. Umbhalo wesandla uza kufumana ukukopishwa, ukufakela, nokuphonononga ubungqina obunokubakho ngaphambi kokuba kukhutshwe kwifomu yayo yokugqibela. Nceda uqaphele ukuba ngexesha lokuveliswa kweeprogram ezinokuthi zifumaneke ezinokuthi ziphazamise umxholo, kunye nazo zonke izisemthethweni ezichasayo ezisetyenziswa kwiphephancwadi.

Yiya e:

Ucaphulo

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