Chirwere chinonzi opioid-induced neuroplasticity ye dopaminergic neurons mu ventral tegmental nzvimbo inokonzera mhuka uye opiate mubayiro (2014)

COMMENTS: The DeltaFosB inoongorora isu tinowanzotaura zvese zvakanyanyisa nucleus accumbens . Kubva pane yakakosha yekudzidza neyeyu mumwe muongorori (Mitemo yemasikirwo emitemo uye yemishonga Mutemo we Common Common Neural Plasticity Maitiro neAFosB seMurevereri Mukuru (2013)):

Nekudaro, mibairo yechisikigo neyezvinodhaka haingoshanduke munzira imwechete yeiyo neural, inosangana pane mamwe mamorekuru mamoreketa, uye pamwe mune mauroni akafanana muNucleus Accumbens, kukurudzira kushingairira kwekukurudzira uye "kuda" kwemhando mbiri dzemubairo.

Izvo zvinotorwa: meth, cocaine, uye zvepabonde vose vanoita zvinhu zvakafanana zvinokosha kune mamwe masero emagetsi mumugwagwa wemubairo (nucleus accumbens), chero zvingaita zvakasiyana mune imwe nzvimbo muuropi. Izvi zvakabvisa chirevo chinowanzotaurwa chekuti mibairo yezvisikwa nemishonga inopesana nenzira nemigumisiro.

Ichi chidzidzo chitsva chakaongorora zvinoitwa pabonde neVTA. Iyo VTA ndipo panotangira iyo dopamine-inogadzira maseru etsinga anotanga - uye iwo anoenda kune iyo nucleus accumbens, kumberi cortex, uye amygdala. Chaizvoizvo iyo VTA ndiyo sosi (tsime) yeazhinji eedu dopamine. Ona iyi 2 mifananidzo yedunhu remubairo: Pic1, Pic2

Vatsvakurudzi vakaona kuti zvepabonde (kunopera) zvinokonzera kuti masero eVTA muchekerere (mune varume). Mapoka emagetsi uye avo vanonzi dendrites inosanganisira gorozi dambudziko reuropi. Izvi ndizvo chaizvo zvinonzi heroin zvinodhaka kune VTA (Kwete nguva imwe chete ye heroin, asi heroin isingaperi). Cherechedza kuti iyi shrinkage yeVTA cell bodies inowanikwa mu heroin yevanhu.

Kuita zvepabonde-induced cell shrinkage kunowedzera kwemazuva 7. Kuchinja kunokonzerwa nekuita zvepabonde kwakadzokera kune imwe nguva mumazuva e30, asi vatsvakurudzi vaongorore mazuva 1, 7, uye 30.

Iyo shrinkage yemasero miviri mu heroin kupindwa muropa kunoguma yakadzika dopamine mune iyo nucleus accumbens - kana izvo zvatinodaidza desensitization. Vatsvakurudzi vakapa morphine kumakonzo kuti vaongorore mhinduro yavo (mushure mebonde), asi hapana chakaitika. Kazhinji makonzo anofarira morphine zvakanyanya, asi apa ivo vaishushikana kwenguva. Muchidimbu, iyo post-ejaculatory rats 'mubairo wedunhu yakanga isiri-yekuteerera kune yakaderera nhanho ye heroin. Vatsvakurudzi vakafungidzira kuti mizinga yepamusoro yaizodiwa kuti iite "yakajairika" rat reaction.

Kupfupisa - Bonde (kwechinguva) rinoita chaizvo zvakafanana chinhu muVTA sezvinoita kupindwa muropa neheroin: shrinkage yedopamine inogadzira tsinga maseru masero. Izvi zvinotungamira kudzikisa dopamine munzvimbo yemubairo, uye kusapindura kushoma kune zvinodhaka - uye zvinotora mazuva manomwe mazuva ekurapa makonzo kuti awanezve.

 

J Neurosci. 2014 Jun 25;34(26):8825-36. doi: 10.1523 / JNEUROSCI.0133-14.2014.

Pitchers KK1, Coppens CM2, Beloate LN3, Fuller J2, Van S2, Frohmader KS2, Laviolette SR2, Lehman MN4, Coolen LM5.

ngovepo

Mubairo wepanyama uye zvinodhaka zvekushungurudzika zvinoshandura pazisolambic nzira uye kuita shanduro inowanzo ya neural plasticity mu nucleus accumbens. Kugara kwenguva refu kune maopiates kunoita kuti mapurasitiki ave ne dopaminergic neurons we ventral tegmental area (VTA), iyo inodzora morphine mubayiro wokushivirira.

Pano, tinoedza maonero ekuti kusununguka kunokonzera kusununguka kweapioid inogadzikana muVTA kunokonzera marphological change yeVTA dopamine masero muvarume vese, iyo in-inodzora kurarama kwenguva refu kwechiitiko-yakakonzerwa kusimbisa unhu hwepabonde.

Kutanga, ruzivo rwepabonde rwakaderedza VTA dopamine soma ukuru 1 nemazuva 7, asi kwete mazuva 30 mushure mekupedzisira kwekusangana. Izvi zvakavharidzirwa neapoxone mushure mega rega sangano rekugadzirisa; saka, VTA dopamine cell plasticity yakanga yakatarisana nezviito zveapioid isingagumi.

Zvakare, VTA plastiki yakabatanidzwa nehutano hwakashandurwa opiate, sezvo varume vane hutano hwepabonde havana kuumba nzvimbo yakagadzirirwa ye 0.5 mg / kg morphine.

Zvadaro, zvakagadziriswa kana chiito chinogumira cheipioid chinosangana nemubairo wepabonde nekuyeuka mumakumbo evarume akashandiswa neapoxone munguva yekusangana, kana sangano kana intra-VTA. Naloxone haina kudzivisa kutanga kwekuita-kukonzerwa nekufambisa kwehutano hwepabonde pamusoro pekudzokorora maitiro ekugadzirisa, kana nzvimbo yakagadziriswa yekuda kubata. Asi zvakadaro, naloxone kurapwa yakaratidza nguva yakareba-yekutaura kwechiitiko-induced facilitation zvekuita zvepabonde uye neural activation mu mesolimbic nzvimbo dzinokonzerwa nekubatana-kunokonzerwa nemamiriro ezvinhu.

Pamwe chete, idzi data dzinoratidza kuti maipioid anochengetedza panguva yekubatana inoita neural plasticity muVTA dopamine neurons inoonekwa yakakosha kumufaro we morphine uye yekuyeuka kwenguva refu yehutano hwemararamiro.

 

ziviso

Makomborero emhando yemafambisirwo anopindirana nemesocorticolimbic system (Meisel uye Mullins, 2006; Hoebel et al., 2009; Frohmader et al., 2010a; Pitchers et al., 2010a; Young et al., 2011; Blum et al., 2012). Zvinodhaka zvekushungurudza zvinokonzera kushanduka kweveurural muhurongwa uhwu, iyo in-inobatsira pakukura nekuratidza kushandiswa kwezvinodhaka (Hyman et al., 2006; Nestler, 2012). Isu takagara takatsunga kuti ruzivo nemararamiro emararamiro echisikigo, kureva, ruzivo rwepabonde mumakumbo evarume, zvinokonzera neural plasticity mu nucleus accumbens (NAc), kusanganisira kuwedzera dendritic spines (Pitchers et al., 2010a) uye deltaFosB (Pitchers et al., 2013). Mukati-kutendeuka, ipurasitiki inoita zvepabonde inonyanya kukosha pamigumisiro yehutano hwepabonde pamushure pekubatana, inoratidzirwa sekufambisa kwekutanga nekuita kwehutano hwepabonde (Pitchers et al., 2010b, 2012, 2013). Uyezve, ruzivo rwepabonde rinoshandura kuterera kune psychostimulants, kusanganisira kukurudzirwa kwekuita basa uye kuwedzerwa mubayiro (Frohmader et al., 2010a; Pitchers et al., 2010a, 2013).

IAc ndeimwe iyo inowanikwa pasi ye dopaminergic neurons mu ventral tegmental area (VTA). VTA dopamine neurons inoshandiswa panguva yekubatana uye shure kwekutsvaga kune zvakagadziriswa cues predictive zvepabonde zvepabonde (Balfour et al., 2004; Frohmader et al., 2010a), kuburikidza nechigumo che opioid peptide (EOP) -kuchengetedza μ-opioid receptors (MORS; Matthews uye German, 1984; Johnson neNorth, 1992; Klitenick et al., 1992; Ikemoto et al., 1997; Balfour et al., 2004). Nokudaro, kutarisana nemamiriro ezvinhu ehutano hwekuita zvepabonde kunokonzera kusunungurwa kweEOP neVTA dopamine-cell activation, iyo inobatsira kuita zvepabonde (Mitchell naStewart, 1990; van Furth et al., 1995; van Furth naVan Ree, 1996) uye dopamine kusunungurwa muNac (Fiorino et al., 1997).

Kudzokororwa kunowanikwa kune maopiates anonyanyisa kunokonzera marphological change muVTA (Mazei-Robison et al., 2011; Mazei-Robison naNestler, 2012), kuderedza soma ukuru hweVTA dopamine neurons (Sklair-Tavron et al., 1996; Spiga et al., 2003; Chu et al., 2007; Russo et al., 2007; Mazei-Robison et al., 2011), zvakaderedza mapurogiramu emapuroteni ekushaya ropa (Beitner-Johnson et al., 1992), kuwedzerwa kwekudzivirirwa kwedopamine masero, uye kuderedzwa kwekutsvaga axoplasmic uye dopamine yakawanikwa ku NAc (Beitner-Johnson et al., 1992; Mazei-Robison et al., 2011). Izvozvi VTA dopamine neuron zvinokonzera kuti morphine mupiro wekushushikana uye inopera sezvavanoparadzanisa mukati memwedzi wekupindwa muropa nezvinodhaka (Russo et al., 2007). Ikozvino haisi pachena kana ipurasitiki muVTA dopamine neurons yakasiyana nezviito zveopiates kana kana iyo yakabudiswa nekusunungurwa kweEOP panguva yemafambiro ekugutsikana.

Pano, tinoedza pfungwa yekuti mupiro wemupiro wehupenyu hunokonzera kushaya kweuroplasticity yakafanana neyoyo inokonzerwa neopiates, uye naizvozvo, iyo maopiates anotendeuka pahutuputi hwemapurisa hunokosha pamararamiro emararamiro ezvokwadi uye mubayiro wekuyeuka. Isu tinoedza kana hutano hwepabonde mumakumbo evarume hunoderedza hukuru hwe soma hweVTA dopamine neurons kuburikidza nehutano hunoenderana neEOP action muVTA. Uyezve, tinoongorora kana kuchinja kweEOP-kunokonzerwa kushandiswa muVTA dopamine neurons kunobatanidzwa nekusimbiswa kwekuita kwekuita zvipo zvekupa mubayiro uye kupiwa kwekukurudzira kune zvinyorwa zvinobatanidzwa nemubairo wechisikigo, nekukonzera kubvumira mutemo kune mubayiro we morphine.

Zvinhu uye Nzira

Animals

Mukuru wevarume Sprague-Dawley rats (200-225 g) akawanikwa kubva kuCharles River uye akagara muzvikamu zviviri mumakamuri akanyatsogadziriswa pa 12 h chiedza / rima mumamiriro ose ekuedza (chiedza chinobva pa 10: 00 AM kunze kwekuedzwa kwe morphine , mwenje pa 5: 00 PM). Zvokudya nemvura zvakanga zviripo ad libitum kunze kwekuedza kuenzanisa. Stimulus females yaive yakasvibiswa uye yakasimwa zvishoma nezvishoma ne 5% 17-β-estradiol benzoate SILASTIC capsules (1.98 mm mukati mukati, 0.5 cm cm, Dow-Corning). Injections ye progesterone (subcutaneous, 500 μg mu 0.1 ml yemafuta esame) yakashandiswa 3-6 h isati yaedzwa kuti iite zvekuita zvepabonde. Mitemo yose yakagamuchirwa neYunivhesiti yeWestern Ontario neYunivhesiti yeMichigan Animal Care Committees, uye yakaganhurwa neCanada Council pamusoro pePil Care uye National Institutes of Health mirayiridzo inosanganisira mhuka dzemhuka dziri mukutsvakurudza.

Nguva yeVTA dopamine soma size shanduko

Zuva roga roga kutora.

Kudzidza nguva yenguva yekuchinja mune dopamine neuron soma size muVTA, zviitiko zvepabonde uye zvisizvo zvakaurayiwa pa 1, 7, kana 31 d (n = 5-8 neboka) mushure mekupedzisira kwekubatana (ruzivo) kana kutonga (noive). Mapoka ane ruzivo rwepabonde akafananidzirwa nekuita zvepabonde panguva yekupedzisira yekugadzirisa, pamwe chete nenhamba yakazara ye ejaculations pamusoro pemisangano shanu (muyero we5 yeboka rimwe nerimwe), uye haina kusiyana pane imwe nzira yepabonde.

Mating sessions.

Vanhurume vasina zvekuita zvepabonde vakagoverwa kune chimwe chezviitiko zviviri zvekuongorora: zvepabonde kana zvepabonde. Mhuka dzepabonde dzakagamuchirwa dzakabvumirwa kuti dzibatanidze kashanu pamasikati anoenderana nevakadzi vanogamuchira mumakamuri emayero evhareta (60 × 45 × 50 cm) kusvikira kuratidzirwa kwekushongedza kana kusvika ku 1 h (chero yakasvika pakutanga). Makomba akacheneswa zvakakwana ne 70% ethanol solution uye matanda matsva akawedzerwa pakati pekugadzirisa maitiro. Kuita zvepabonde kwakaitwa munguva yerima (2-6 h mushure mekutanga kwemazuva). Mhuka chete dzakasvibiswa panguva inenge yechina yezvikamu zvishanu zvekukwirirana dzakaonekwa sepabonde uye dzakabatanidzwa mukuedza. Zvose zvikamu zvekubatana zvakaonekwa uye zvepabonde zvakanyorwa. Nhamba yezvikwata (M) inogadziriswa (IMs), gomo latency (ML; nguva kubva pakusunungurwa kwevakadzi kusvika kunekutanga kukwira), intromission latency (IL; nguva kubva pakusunungurwa kwevakadzi kusvika pakutanga kuisa mukati), uye ejaculation latency (EL; nguva kubva pakutanga kusanganisa kusvika pakuiswa) yakanyorwa (Agmo, 1997). Mhuka dzisingadi dzakagadziriswa muchengeti wakachena wechirwere ye 1 h panguva imwe chete nevanhurume vane ruzivo rwepabonde vanobatana muimba imwechete, zvakadai kuti vakananga kune zvinonhuwira zvevakadzi, uye maitiro akafanana ekuvhiringidza uye nharaunda yakasikwa sevarume vane ruzivo.

Immunofluorescence labeling.

Mhuka dzaive dzakanyanyisa anesthetized kuburikidza ne sodium pentobarbital (270 mg / kg, ip) uye yakashandiswa zvisina kumira ne 50 ml ye 0.9% saline, ikatevera 500 ml ye 4% paraformaldehyde mu 0.1 m sodium phosphate buffer (PB). Ubongo hwakabviswa uye hwakasvibiswa nokuda kwe1 h muchitsiko chemukati (RT) mune imwe yakagadziriswa, uye yakanyorerwa mu 20% sucrose uye 0.01% sodium azide mu 0.1 m PB yekuchengetedza pa 4 ° C. Zvikamu zveCoronal zvakagurwa pa 35 μm pane microtome inotonhora (H400R, Microm) uye yakaunganidzwa muzvinyorwa zvina zvakafanana mucyoprotectant solution (30% sucrose, 30% ethylene glycol mu 0.1 m PB) ndokuchengetwa pa 20 ° C. Zvose zvakashongedzwa zvakaitwa paR RT nekudengenyeka zvinyoro uye zvipfeko zvinokonzera ne 0.1 m PBS, pH 7.35, pakati pekudzivirira. Zvikamu zvakaiswa kune 1% H2O2 ne10 min kuparadza peroxidases isingagumi, ndokuvharidzirwa 1 h mubato remubatanidzwa (PBS +: PBS ine 0.4% Triton X-100; Sigma-Aldrich) uye 0.1% bovine albumin (Jackson Immuno Research Laboratories). Zvadaro, zvikamu zvakange zvichinyorwa usiku humwe paR RT mumushonga tyrosine hydroxylase (TH) -antibody (1: 20 000; Millipore). Mushure mekutanga kwechirwere chekuchengetedzwa kwemunhu, zvikamu zvakaiswa mu Alexafluor 555-conjugated mbu anti-mouse antibody (1: 100; Invitrogen, Eugene, OR) ye30 min. Pakupedzisira, zvikamu zvakashambidzwa ne 0.1 m PB, zvakasungirirwa paSirafrost Plus magirazi, akaomeswa, uye akafukidzwa negelvatol ane anti-fading agent 1,4-diazabicyclo (2,2) octane (DABCO; 50 mg / ml, Sigma-Aldrich; Lennette, 1978).

Data analysis: neuron soma size.

Mifananidzo yeT-immunoreactive (IR) neuroni muVTA yakatorwa pa 40 × kukudzwa pamatatu matatu kusvika kumawere caudal (Balfour et al., 2004). Kwakanga kusina kusiyana kwakasiyana kunoonekwa pakati pemasero emazinga akasiyana. Iko soma ukuru hweT-IR neuroni yakarongedzwa uchishandisa ImageJ (National Institutes of Health). Nharaunda, nzvimbo, uye chitenderedzwa zvakayerwa sezvinorondedzerwa Sklair-Tavron et al. (1996). Avhareji emasero e25 nemhuka (akabatanidza ose mazinga 3 VTA) akaongororwa uye masero chete aine nucleus yakajeka akaiswa. Kune mhuka imwe neimwe, nzvimbo inorehwa, mhirizhonga, uye tsvina yakaverengwa. Nokuda kwezvinyorwa kuongororwa nzira mbiri ANOVA yakashandiswa [zvikonzero: ruzivo rwepabonde (zvepabonde rakaonekwa kana zvepabonde pasina) uye nguva (1, 7, kana 31 d)] inotevera post hoc kuenzanisa achishandisa nzira yeHolm-Sidak nechinomiriro chezinga re0.05.

VTA kwete-dopamine kuchinja

Biweekly mating sessions.

Kuti uone kana ruzivo rwepabonde panguva dzekugadzirisa zuva nezuva dzinodiwa kuti udzike muhutano hweT-IR neuron soma, VTA dopamine neurons yemhuka dzakange dzanyura panguva yezvikamu zvishanu zvekugadzirisa zvinyorwa zvakaongororwa. Zvirongwa zvekuenzanisa zvakarondedzerwa pamusoro apa, asi kwenguva yevhiki dze2.5. Ubongo hwakakonzerwa 7 d mushure mokunge uchinyatsopedzana kana kubata.

Immunoperoxidase labeling.

Mukuwedzera, yakaedzwa kana kushandiswa kwehutachiona hunoshandiswa nema immunoperoxidase uye chromogen yekucherechedza, kunobvumirawo kuonekwa kweT-IR soma shanduko kuchinja. Perfusion uye tissue processing yaive mufambiro sezvinotsanangurwa pamusoro apa. Kutevera kurapa ne 1% H2O2 uye PBS +, zvikamu zvakashandiswa usiku humwe paR RT mumushonga polyclonal tyrosine hydroxylase (TH) -antibody (1: 20 000; Millipore). Mushure mekutanga kushandiswa kwevanhu vanorwara, zvikamu zvakange zvakabatanidzwa ne-biotin-conjugated goat anti-rabbit IgG (1 h, 1: 500 muPBS +; Vector Laboratories), avidin-biotin-horseradish peroxidase (1 h, ABC elite; 1: 1 000 muPBS ; Vector Laboratories), uye 3,3'-diaminobenzidine tetrahydrochloride (10 min, 0.02%, DAB; Sigma-Aldrich) yakasimbiswa nickel sulfate mu (0.02% mu 0.1 m PB) nehydrogen peroxide (0.015%). Zvikamu zvakashambidzwa zvakakwana muC0.1 m PB kuti igadzire kupindura uye yakasimudzirwa kune coded Superfrost pamwe nemirasigiramu emirasi (Fisher) ne 0.3% gelatin muDdH2O. Kutevera kubuda kwemvura, mashizha ose akanga akafukidzirwa neDPX yakakura (dibutyl phthalate xylene; Sigma-Aldrich).

Data analysis: neuron soma size.

TH-IR masero akaongororwa enzvimbo, perimeter, uye kutenderera sezvakatsanangurwa pamusoro. Uye zvakare, TH-IR masero mune substantia nigra (SN), muzvikamu zvakafanana zvinoshandiswa pakuongororwa kweVTA TH-IR masero, akaongororwa. Pakupedzisira, zvichitevera kuongorora kweVTA uye SN TH-IR masero, zvikamu zvakabatanidzwa zvichishandisa cresyl violet uye asiri-TH-IR maseru akaongororwa vachishandisa nzira dzakafanana sekutsanangurwa pamusoro apa. Misiyano pakati pemapoka ane ruzivo uye ane ruzivo akafananidzwa achishandisa maviri-akashatisa Mudzidzi t miedzo inokosha inokosha ye0.05.

Migumisiro yeaixone pane zvakaitika-induced dopamine soma kukura kweusimba

Kuti uone kana MASHUMI aifarira basa mune zvepabonde-akashandurwa kuchinja mu dopamine neuron soma size, MOR yakavharwa munguva yekuita zvepabonde. Hafu yemhuka yakawana ruzivo rwepabonde, asi imwe hafu yakabatwa asi yakaramba ichiita zvepabonde. Mhuka dzehupombwe dzakagadziriswa dzakabvumirwa kuti dzibate pa 5 mazuva akatevera. Muchikamu chevanhu vanoziva zvepabonde uye vasina mapoka, mhuka dzakabatwa neNonselective MOR inopikisa naloxone (10 mg / kg, sc; Sigma-Aldrich, yakagadziriswa mu 0.9% saline) kana saline 30 min kusati kwasvika mukadzi (ane ruzivo) kana asati agadzirisa (naive); nokudaro kuumba mapoka mana ekuenzanisa: zvepabonde neveine saline (Naive Sal), vanoita zvepabonde naxone (Naive NLX), vane ruzivo rwepabonde (Exp Sal), uye anex sex experienced (Exp NLX; n = 5-8 neboka). Naloxone kurapwa kwaisava nemigumisiro yakakosha pane chero ipi zvayo yehutano hwepabonde, pane chero ipi zvayo ye5 d, uye maawaxone- uye maitiro ekurapwa ane saline akanga akafanana mukuziva zvepabonde. Mhuka dzose dzakaurayiwa kuburikidza nehupenzi husina kukwana 7 d mushure mekupedzisira kwekugadzirisa. Chikamu, immunohistochemistry, uye kuongorora kwedhiyabhorosi (nzira mbiri ANOVA; zvikonzero: ruzivo rwepabonde uye kurapwa kwemishonga) kwekuita dopamine soma kuitiswa sezvinotsanangurwa pamusoro apa.

Morphine yakagadzirirwa nzvimbo inofarira

Ongororo yekugadzira.

Kare, Russo et al. (2007) yakaratidza kusingaperi morphine kunobatsira kutsungirira kune morphine mubayiro. Sezvo ruzivo rwepabonde uye rurefu rwe morphine runokonzera kuderera kwakadaro mukukura kwechioma che dopamine neurons muVTA, kushandiswa kwebasa rekuita zvepabonde morphological changes kwakaongororwa nemubayiro we morphine. Mhuka dzezvepabonde uye dzisina maturo dzakaparadzaniswa kuva mapoka matanhatu ekuenzanisa (n = 9-13 neboka) maererano nemitemo yepabonde (vanoita zvepabonde kana vanoziva) uye morphine dose (0.5, 5.0, kana 10.0 mg / kg, ip) uye yakaongororwa kuti nzvimbo inogadziriswa nemafungiro e morphine (CPP).

Morphine-CPP.

Kugadzirisa kwakaitika 1 d mushure mekupedzisira kwekugadzirisa masangano uye mapoka akafananidzwa nekuita zvepabonde panguva yekupedzisira yekugadzirira. Shanduro yePPP yakashandiswa yaiva yekare, mazuva ekugadzirisa, uye mushure mokuedzwa, uye zvigadzirwa zvaive zvakavakirwa Tenk et al. (2009). Muchidimbu, zvishandiso zveCPP (MED Associates) zvaive nemakamuri matatu akasarudzika. Pakati pechikamu chega chega, zvishandiso zvakacheneswa zvakaringana ne 70% ethanol mhinduro kudzikisira inonamira zvirevo. Kuti uone zvaunofarira zvega, kwekunyepedzera kwaiitwa panguva iyo mhuka dzakapihwa mukana wekusununguka kune zvese zvishandiso zve15 min. Seboka, mhuka hadzina kuratidza kukosha kwakakosha kune imwe kamuri, asi mhuka yega yega yaive nekadiki kekutanga. Makonzo airatidza kuda kwakanyanya kune imwe yemakamuri (> 200 s mutsauko pakati penguva yakashandiswa mune imwe neimwe yemakamuri; <5% yemhuka) panguva yekufungidzira vakasiyiwa muchidzidzo. Munguva yekugadzirisa mamiriro, mushonga waive wakapetwa kune wekutanga waisarudzika kana isina kufananidzwa mukamuri uchishandisa unbiased paradigm (Tzschentke, 2007) uye mhuka dzaiva mumakamuri e30 min. Mhuka dzakanga dzakakonzerwa ne saline (ip) mangwanani (9: 00 AM kusvika 12: 00 PM) uye yakavharirwa mukamuri ye-saline-paired (control). Mumazuva masikati (1: 00-4: 00 PM), mhuka dzakakonzerwa ne morphine (ip, 0.5 mg / kg, 5.0 mg / kg kana 10.0 mg / kg; morphine sulfate yakagadziriswa mu 0.9% saline, Johnson Matthey) uye akavharirwa kusvika kumakamuri e-morphine-paired. Mhuka dzakanga dzichiiswa pasi pamazuva maviri ekugadzirisa. Zuva rinotevera (3 d mushure mekupedzisira kwekuroorana) mushure mokuedzwa, zvakagadziriswa zvakafanana nezvakaitika kare, zvakaitwa. Kuenzanisa kuongorora, nguva inoshandiswa mumakamuri efafine-paired panguva yekuedzwa yakaenzaniswa nenguva yakashandiswa mumakamuri ane saline-paired mushure mekuongororwa kwevanhu vane zvepabonde kana varume vane ruzivo mukati megaji imwe neimwe vachishandisa pairi t muedzo. p <0.05 yaifungidzirwa kuverengera kukosha. Kuwedzera ekudzora mapoka emhando dzepabonde uye vane ruzivo mhuka vakagamuchira saline mumakamuri maviri akaparadzaniswa uye asina kuvharwa kuti vashande semabatiro asina kunaka. Hapana misiyano munguva yakashandiswa pakati pemakamuri yakawanikwa kune chero boka.

Migumisiro yeaconicone yakagadziriswa pamusana pekugadzirisa-kuwedzerwa kwekuita zvekuita zvepabonde

Ongororo yekugadzira.

Chiitiko chepabonde chinoguma mukufambisa kwehupombwe hunochengeterwa kwekunenge kwe1 mwedzi (Pitchers et al., 2012). Kuti uongorore mhinduro yekuvhara MOR kuburikidza nehutano hwakakonzerwa nehutano hwemhando dzepabonde, mhuka dzakagadzirirwa zvepabonde dzakagamuchira iyoxone kana saline musati mavhiki mashanu anotevera achienzanisa (n = 12 imwe neimwe) sezvinotsanangurwa pamusoro apa. Imwe vhiki mushure mekugadzirisa kwekugadzirisa kwekupedzisira, kuedza kwekupedzisira kwekugadzirisa kwaiitwa panguva iyo mhuka dzose dzakabvumidzwa kuti dzibatane kusvikira imwe yejjulation kana kusvika ku1 h. Hapana naloxone kana kuti chirwere chinonzi saline chaitungamirirwa musati matanho pazuva rokupedzisira. Zviratidzo zvekubatana zvakafananidzwa nekuona kana iyoxone yakagadzirirwa kana chiitiko chepabonde-chakakonzera kushandiswa kwekusangana (zuva 1 vs zuva 5) kana kugadziriswa kwekufambisa uku (zuva 5 vs kuongorora) uchishandisa nzira mbiri ANOVA [zvikonzero: kurapa (saline neyexxone ) uye zuva (zuva 1, zuva 5, kana muedzo)] uye nzira yeHolm-Sidak post hoc kuenzanisa. Pamusoro pevose vanoongorora miedzo, p <0.05 yaifungidzirwa kuve inoverengeka yakakosha.

Zvimwe zvekudzidzira kuedza

Systemic naloxone pazuva rekuedza.

Kuratidza kuti maitiro ekuita zvepabonde akashandurwa pakupedzisira kwekuenzanisa zuva rekutsvaga hauna kukonzerwa nekushayikwa kweivaxone, taitora irixone kana saline pazuva rekupedzisira rekuedza kusangana nemhuka dzakagamuchira kubatana dzakabatana neapoxone apo dzikawana ruzivo rwepabonde. Kunyanya, mhuka dzose dzakagamuchira joxone injection (10 mg / kg, sc) 30 min kusati kwasangana kusvika kune imwe yega yega panguva ye 5 mazuva anoitevera. Pamuedzo zuva 7 d gare gare, inenge hafu yezvipfuwo yakagamuchira jekoni yeteroxone (10 mg / kg, n = 7) kana saline (n = 6) 30 min vasati vatanga kupiwa mukadzi anogamuchira. Kuita zvepabonde kwakaonekwa uye kwakanyorwa. Zviratidzo zvekubatana zvakafananidzwa nekuona kana iyoxone yakagadzirirwa kana chiitiko chepabonde-chakakonzera kushandiswa kwekusangana (zuva 1 vs zuva 5) kana kugadziriswa kwekufambisa uku (zuva 5 vs kuongorora) uchishandisa nzira mbiri ANOVA [zvikonzero: kurapa (saline neyexxone ) uye zuva (zuva 1, zuva 5, kana muedzo)] uye nzira yeHolm-Sidak post hoc kuenzanisa. Pamusoro pevose vanoongorora miedzo, p <5% yaifungidzirwa kuve inoverengeka yakakosha.

Migumisiro yeaixone pamusoro pemaonero emhedziso ekufambiswa kwepabonde.

Migumisiro yeaixone (10 mg / kg, sc) kurapwa panguva yekugadzirisa yakaedzwa pazviitiko zvepabonde zvakazotevera panguva yekupedzisira yekuenzanisa zuva zuva, iro rakaitwa chete 1 d mushure mekusangana kwekupedzisira (saline, n = 5; naloxone, n = 4).

Systemic naloxone pretreatment.

Kuti uone kana ichidzokorora kurapwa kwejaxone chete inokonzera kuora mwoyo kwehupombwe 7 d mushure mekurapa kwekupedzisira, mhuka dzepabonde dzisingabatsiri dzakagamuchira mashanu zuvazone zuva nezuva (10 mg / kg, sc) kana kuti injini ye saline mumazuva anotevera mushure mokuedzwa kwekusangana 7 d mushure mekupedzisira naxone kana injini yerineni. Pachidzidzo chekupedzisira chezuva, mhuka dzisina kugamuchira jekiseni. Kuita zvepabonde kwakaonekwa uye kwakanyorwa sezvinotsanangurwa pamusoro apa. Zviratidzo zvekubatana zvakafananidzwa pakati pemapoka achishanda asina kubhadhara t miedzo. Pamusoro pevose vanoongorora miedzo, p <5% yaifungidzirwa kuve inoverengeka yakakosha.

Systemic naloxone uye mubayiro wepabonde.

Imwe mukana wekuti naloxone inowana mhedzisiro pakuratidzira kwekuchengetedzwa kwekufambiswa kwehunhu hwepabonde ndeyekuti naloxone inovhara mhedzisiro inobatsira yehunhu hwepabonde. Kuti uedze izvi, CPP paradigm yakaitwa yebonde nekukurumidza ichitevera naloxone kana jekiseni re saline mune vanhurume vasina ruzivo rwepabonde. Maitiro eCPP akafanana neakatsanangurwa pamusoro pe morphine-CPP, kusanganisira kunyepedzera, mazuva ekugadzirisa, uye post-bvunzo.

Kuita zvepabonde kwakaparadzana nekambani isina kufanirwa. Mune imwe nzira yakasiyana, mhuka imwe neimwe yakagamuchira jekiseni yeteroxone (n = 12) kana saline (n = 11) 30 min vasati vatambira kugona kumudzimai anogamuchira. Ivhareji yenguva yekugadzirisa chikwata chaiva ~13 min. Maminitsi masere mushure mekujaira, mhuka yacho yakaiswa muimba yepamusoro ye30 min. Pane rimwe zuva raifungidzirwa, mhuka dzakagamuchira jekisi yejaixone kana saline (chero ipi yavakagamuchira vasati vakura), uye yakaiswa muimba isina kubhadhara ye 30 min. Zvadaro, mushure mokuedzwa wakaitwa, zvakaenzana nehupenzi. Kuti aone kusarudzwa kwekamuri, nguva inoshandiswa muimba yepamusoro panguva yekufungidzira uye mushure mokuedzwa yakaenzaniswa. Nekuverenga nhamba, kuwirirana t miedzo yakashandiswa kuenzanisa kudiwa uye kusiyana kwezvikamu, uye nguva muimba yepakati paine chikwata chekutanga uye chekuongorora-chekuita kuti aone kana chirongwa chikuru chePCP chakaumbwa nokuda kwekuita zvepabonde. p <0.05 yaionekwa seinokosha.

Migumisiro ye-intra-VTA naloxone pamusana pekuita-kutungamirirwa nekuita zvekuita zvepabonde

Ongororo yekugadzira.

Kuti uone kana chirongwa cheEOP chaiwo muVTA, chaiva nemhosva yehutano hwekuita zvepabonde-hunoita kuchinja kwehutano hwemhando dzepabonde, mhuka dzakasvibiswa mukati memunharaunda yeaixone kana saline muVTA musati mavhiki mashanu ekugadzirisa zuva nezuva. Chimiro chemuitiro wechimiro chakange chakafanana neyese ye systemic naloxone. Mhuka dzezvepabonde dzakagamuchirwa kuti dzisangana panguva ye 5 mushure memazuva kusvikira imwe ejaculation kana kusvika ku 1 h. Maminitsi gumi nemashanu asati atanga mukadzi anogamuchira, mhombwe idzi dzinogamuchirwa zvakashata zveextone (10 μg / μl pamusana wehemisphere; 0.5 μl volume; yakagadziriswa mu 0.9% saline) kana saline (0.5 μl per hemisphere). Bilateral microinjections yakashandiswa pakuyerera kwe0.5 μl / min pane 1 min interval yakateverwa nehuwandu 1 min nejaya rejojo ​​rakasiiwa munzvimbo kuti riparidzire. Iyo jena cannula yakabva yatsiviwa ne dummy cannula uye guruva. Imwe vhiki mushure mekupedzisira kwekugadzirisa (zuva rekuedza), mhuka dzose dzakasungirirwa zvakare zvakare kuchinjwa pasina kuvharirwa kweteroxone kana saline. Mufananidzo 3A inotsanangura chimiro chekuedza. Kuongorora kwedhiyabhorosi kwakaitwa sezvinotsanangurwa mu systemic naloxone kuedza.

Kuvhiyiwa kunogona.

Makuve evanhu akange anestitizedeki neine intraperitoneal injection (0.1 ml / kg) yeketamine (0.87 mg / ml) uye xylazine (0.13 mg / ml), ndokuiswa mune zvigadzirwa zve stereotaxic (Kopf Instruments). Bilateral 21-gauge guide cannulas (Plastics One) yakaderedzwa kuburikidza nemakomba maduku mumharukoni muuropi kuenda kuVTA pa -4.8 mm AP, ± 0.75 mm ML kubva kuBregma uye -7.8 mm DV kubva kumusoro kwehenyare maererano Paxinos uye Watson (2013). Cannulas dzaive dzakachengetedzwa nemazino acrylic akatevedzana nemapundu matatu akaiswa muhenya. Mhuka dzakapiwa 2-vhiki yekudzikinura nguva, uye dzakatarirwa zuva rega rega kuti dzigadzirise kushandiswa nekujekesa nzira dzinoshandiswa munguva yekuedza kuzvibata.

Cannula kuiswa kugadziriswa.

Kuiswa kwe cannulas kwakaongororwa uchishandisa TH-immunostaining kusimbisa kuti VTA yakanyatsorongedzwa. Zvisikwa zvipenyu chete zvakakonzerwa zvakananga mukutsanangurira (boka rekupedzisira boka: ruzivo rwune ruzivo n = 8; anexpected naxone n = 6). Zvimwe nhatu mhuka dzakagamuchira intra-VTA iyoxone injections inotungamirirwa kunze kweVTA yakanga yakabatanidzwa pamwe chete mu "boka" rakakanganiswa. Boka rakarasika rakarongedzwa pachavo kuti rive sekutonga kwemaatomical uye Mann-Whitney U muedzo wakashandiswa kuenzanisa maitiro pazuva rekupedzisira rekuedza neapoxone uye varume vane hutano hwakagadziriswa.

Mating-associated contextual cue-induced pERK kutaura

Ongororo yekugadzira.

Kuvepo kune cage umo varume vanowana kuwirirana ruzivo rwakaratidzwa kuita kuti MOR kushandiswe muVTA uye mabasa emagetsi eVTA neAcc (Balfour et al., 2004). Nokudaro, nzvimbo inobatanidza inoshanda sechikonzero chekuchengetedza chirevo chepabonde. Izvo zvidzidzo zvemazuva ano zvakaongororwa kana MOR kushandiswa panguva yehutano hunodiwa kunotevera hurumende yakagadzirirwa-induced neural activation. Naloxone kana kuti saline yakashandurwa nenzira yepamutemo (ip) 30 min isati yaiswa panzvimbo yakatarisana uye kuiswa kwevakadzi vanogamuchira kubata (vane ruzivo), kana kuti kusati kwaitwa kushandiswa kwekutonga kwakave nekuiswa mugadziriro yekugadzirisa kunze kwekutaurirwa kwevakadzi (kusatenderera mamiriro ezvinhu; naive). Nokudaro, mapoka mana ekuenzanisa akasika: Naive Sal, Naive NLX, Exp Sal, uye Exp NLX. Imwe vhiki mushure mekupedzisira yekugadzirisa chirongwa, hafu yezvipfuwo mumapoka ega ega akange akazarurirwa gorosi rinokurudzirwa (Exp varume: zvekuita zvepabonde zvakagadziriswa cues) kana kutakura cage (varume vasingatendi: vasingatendi / vasina kutora cues), asi imwe hafu haina kuzarurirwa chero zvinyorwa uye panzvimbo yazvo yakaramba iri mumakamuri emisha (kuti aone zvinyorwa zvitsanangudzo pERK). Iyi paradigm inoongorora yakabudisa mapoka e8: Naive Sal-No Cue, Naive Sal + Cue, Naive NLX-No Cue, Naive NLX + Cue, Exp Sal-No Cue, Ex NLX + Cue, Exp NLX + Cue (n = 4 imwe neimwe kunze kweNaive NLX-No Cue, n = 3). Mhuka dzakashandurwa 10-15 min mushure mokunge zvinyorwa. Kudzora mhuka zvakabviswa pamakamuri avo emisha uye zvakashandiswa panguva imwe chete.

Immunohistochemistry.

Chikamu uye immunohistochemistry zvakaitwa sezvinotsanangurwa pamusoro apa. Pano, taishandisa sarbit polyclonal antibody p42 uye p44 MAP Kinases ERK1 uye ERK2 (pERK; 1: 4 000; Cell Signaling Technology). The primary antibody has been widely identified in the literature (Roux uye Blenis, 2004; Murphy uye Blenis, 2006; Frohmader et al., 2010b). Uyezve, kusaregererwa kwechirwere chekutanga chinorambidza muviri wose uye kushandiswa kweWestern blot yehutachi uhwu hunobudisa zviratidzo zviviri pane zvakakodzera maserosi masero.

Data analysis.

pERK-immunoreactive (-IR) masero akaverengwa munzvimbo dzinoverengeka dzeuropi achishandisa kamera yeruchiramu lucida yakaiswa kune Leica DMRD microscope: NAc [nheyo (C) uye shell (S); 400 × 600 μm; medial prefrontal cortex; mPFC; anterior cingulate area (ACA); prelimbic cortex (PL); infralimbic cortex (IL); 600 × 800 μm mumwe nomumwe], caudate-putamen (CP; 800 × 800 μm), uye anolateral amygdala (BLA; 900 × 1200 μm; Balfour et al., 2004; Frohmader et al., 2010b; Pitchers et al., 2010b). Zvikamu zviviri zvakaverengwa munharaunda yeuropi uye nhamba dzemasero munzvimbo dzakaenzana dzekuongororwa dzaive dzakaverengwa senhamba dzemasero pa mm2. Izvo zviyero zviviri zvakaenzaniswa nemhuka yega yega kuverengwa kweboka rinoreva. Itai mapoka maviri mukati mevanhu vane ruzivo rwepabonde kana mapoka asina kufanana akaenzaniswa achishandisa nzira mbiri ANOVA [zvikonzero: kurapa kwezvinodhaka (NLX kana Sal) uye cue (cue kana cue cue)] inotevera post hoc kufananidza kushandisa Holm-Sidak kana Mann-Whitney kuongororwa kwepamusoro apo zvakakodzera nehuwandu hunokosha hwe p <0.05. MuNac shell yemhuka dzine ruzivo rwepabonde, pakanga paine chimiro chakasimba chekukosha kwezvinhu uye nekudaro, kuenzanisa kwepaviri kwakaitwa kuenzanisa saline (Sal-No Cue) uye saline cue (Sal + Cue) mapoka chete.

Mifananidzo.

Dhidhijimu mifananidzo yakashandiswa uchishandisa CCD kamera (Macrofire, Optronics) yakabatanidzwa a Leica microscope (DM5000B) nekatsva kamera zvirongwa. Mifananidzo yakatengeswa kuAdobe Photoshop 9.0 software. Mifananidzo haina kuchinjwa nenzira ipi zvayo kunze kwekugadziriswa kwekubwinya uye kusiyana.

Results

Zvepabonde ruzivo-rwakakonzera kuchinja muVTA dopamine masero

Chiitiko chepabonde chakaguma nekuderera kweVTA dopamine soma size (Fig. 1A-C). Chiitiko chepabonde chakanyanya kuderedza nzvimbo uye kuperera kwe soma yeVTA TH-IR seli (nzvimbo: F(1,31) = 23.068, p <0.001; mukomberedzo, F(1,31) = 18.225, p <0.001). Paive zvakare neakakosha akakosha mhedzisiro yenguva (nzvimbo: F(2,31) = 6.377, p = 0.005; perimeter, F(2,31) = 4.389, p = 0.021) uye kubatana kukuru pakati pezviitiko uye nguva (nzvimbo: F(2,31) = 5.284, p = 0.011; perimeter, F(2,31) = 4.347, p = 0.022). Pairwise kuenzanisa kwakaratidza kuti nzvimbo uye mhirizhonga yemasero e-TH-IR akanyanya kuderedzwa 1 uye 7 d mushure mekupedzisira zuva rekuita zvepabonde mumhuka dzine ruzivo rwepabonde kana zvichienzaniswa nekudzivirira zvepabonde [Fig. 1B, nzvimbo: p = 0.002 (1 d), p <0.001 (7 d); C, parimeter: p = 0.009 (1 d), p <0.001 (7 d)]. Mhedzisiro yehunhu hwepabonde yakaparara painoteverwa nenguva yemubairo kuzvidzora sosi saizi yeT-IR neurons yakadzoserwa kune yekutanga basara 31 d mushure mekupedzisira musangano wekusangana (Fig. 1B, nzvimbo: p = 0.798; C, parimeter: p = 0.785). Kuita zvepabonde-kukonzerwa kushandura kwakange kusina kuonekwa mu circular pane chero nguva yenguva (Fig. 1D). VTA dopamine soma ukuru hwekukura kwakanga isingabatsiri pamazuva ose ekugadzirisa, sezvo ruzivo rwekukwirirana mukati mezvishanu zvinyorwa zvekudzivirira kwake zvakagadzirawo zvakaderera VTA dopamine soma size (Fig. 2A,B, E-H, nzvimbo: p = 0.004; perimeter: p <0.001). Mukupesana, ruzivo rwepabonde haruna kukanganisa saizi yeT-IR soma mune substantia nigra (Fig. 2C, I-J, nzvimbo: p = 0.13; perimeter: p = 0.16) kana kushandura ukuru hwema soma pedyo neVTA isina-TH-IR neurons (Fig. 2D, E-H, nzvimbo: p = 0.46; perimeter: p = 0.45).

Mufananidzo 1. 

Endogenous opioid-induced soma size shanduko dzeVTA dopamine neurons. A, Mumiririri mifananidzo yeVTA dopamine neurons kubva kune zvepabonde nevechiitiko chinoratidzira kuratidza kuderedza soma ukuru 7 d mushure mokunge mushure mokugadzirisa. Nhamba yezinga, 5 μm. Nhamba yezvakawanda inoratidza kuti hutano hwepabonde (Exp, mazamu madema) hwakakonzera kuderedza kunzvimbo munzvimbo (B; mu μm2) uye chitarisiko (C; mune μm) yeVTA dopamine masero, 1 d (Naive, Exp; n = 6) uye 7 d (Naive, n = 5; Exp, n = 6), asi kwete 31 d (Naive, n = 6; Exp, n = 8) mushure mekusangana kwekupedzisira, zvichienzaniswa nekutonga kwepabonde (Naive, white bars). Nharaunda yakaderedzwa kusvika ku84% mune varume vane ruzivo kana vachienzanisa neveveid control pa 1 kana 7 d. Perimeter yakaderedzwa kusvika ku91.6 uye 90% mumapoka ane ruzivo kuenzaniswa nekutonga pa1 uye 7 d resp. Kwakanga kusina kuitika pa circularity (D). Iyi dopamine cell plasticity munzvimbo (E) uye chitarisiko (F) yakadziviswa na naloxone (NLX, n = 8), asi kwete saline (Sal, n = 7) kurapwa panguva yekubatana, 7 d mushure mekupedza kusangana kwekupedzisira pakuenzanisa nekuenzanisa nekudzivirira zvepabonde (Sal, n = 5; NLX, n = 6). Data inomiririra zvinoreva ± SEM; * inoratidza kusiyana kukuru kunoenzaniswa nekudzivirira zvepabonde kwezuva rimwechete (B, C) kana kuti kuenzaniswa nesarine-inorambidzwa zvekuita zvepabonde uye maitiro anonzi naxxone akaitirwa zvepabonde (E, F).

Mufananidzo 2. 

Chiitiko chepabonde hachina kutapudza soma ukuru mu substantia nigra dopamine neurons kana VTA nondopamine neurons. VTA TH-IR neuron soma nzvimbo (A; mu μm2) uye chitarisiko (B; mu μm) mune zvepabonde (mutsvuku) uye vane ruzivo (madema) mhuka dzakawana ruzivo kuburikidza nekukwirirana kaviri pavhiki panzvimbo pane nguva dzinotevera. Substantia nigra TH-IR soma area (C) uye VTA kwete-TH-IR soma nzvimbo (D) mune zvepabonde (chichena) uye vane ruzivo (ruvara) mhuka. Data inomiririra zvinoreva ± SEM; * inoratidza kusiyana kwakakura kuenzaniswa nekudzivirira zvepabonde kwepabonde. Mumiririri mifananidzo inoratidza TH-IR (maronta) neuroni muVTA yekuita zvepabonde (E), uye ruzivo (F) varume. G, H, Mufananidzo wakakwirira-wekodhi unoratidzwa nemuseve mukati E uye F respectively. Nissl-stained neurons inoratidzwa mubhuruu mumifananidzo iyi. Mumiririri mifananidzo inoratidza TH-IR neuroni muSN yevanhu vanoita zvepabonde (I) uye ruzivo (J) varume. Mizariro mizinga: E-J, 20 μm.

Zvepabonde ruzivo-rwakakonzera soma kukura kwekukura kweVTA dopamine neurons inobva pane opioid receptor activation

Kuderedzwa kweVTA dopamine neuron soma chiyero chakakonzerwa nehutano hwepabonde hwakadzivirirwa neNonselective MOR anoragonist naloxone, inotungamirirwa mushure mega rega sarudzo. Naloxone kurapwa mushure mekugadzirisa maitiro kunobatsira zvikuru munzvimbo (F(1,22) = 4.738, p = 0.041) uye yakaenderera mberi kune zvakakosha pamiganhu (F(1,22) = 2.892, p = 0.052). Kukurukurirana kunokosha pakati pezviitiko uye nerakisixone kurapwa kwakawanikwa panzvimbo (F(1,22) = 5.578, p = 0.027) uye mumiririri (F(1,22) = 8.167, p = 0.009). Pairwise kuenzanisa kwakaratidza kuti ruzivo rwepabonde mumhuka dzakarapwa nesarine zvakaderedzwa nzvimbo uye mhirizhonga yeVTA dopamine neurons 7 d mushure mekupedzisira kwekugadzirisa mukomana uchienzaniswa nevarume vanoita zvepabonde vanoita zvepabonde (Fig. 1E, nzvimbo: p = 0.018; F, parimeter: p = 0.007). Kusiyana neizvi, mhuka dzakabatwa neaixone dzakabatwa nepabonde dzakanga dzisingatendi nevarumexone-vaibata vasiri varume (Fig. 1E, nzvimbo: p = 0.483; F, parimeter: p = 0.330). Mukuwedzera, soma ukuru hwechizivo hwemhuka yakakonzerwa nehaini yakanyanya kuderera kana ichienzaniswa nemafungiro aneeviloni (Fig. 1E, nzvimbo: p = 0.002; F, parimeter: p = 0.002). Izvi zvakakonzerwa neaixone yaiva yakananga nokuda kwehutano hwepabonde, sezvo naxxone kurapwa chete haina kukanganisa ukuru hweT-IR soma soma mune zvepabonde vainexx-vakatapwa varume kana vachienzaniswa nesarine-treated controls (Fig. 1E,F). Uyezve, izvi zvakakonzerwa naivaxone pamusana pezvakaitika-induced soma kupera kwemaitiro hazvina kukonzerwa nemigumisiro yaivaxone pamusoro pekuita zvepabonde, sezvo mufambiro wekukwirirana wakanga usina kusiyanisa zvakanyanya pakati naxaxone- uye varume vanorapwa nevarine, kunze kwenguva yakareba yekutanga kubata mushure mokunge (mushure mekudzivirira nguva) mumaloxone-akabatwa nevarume panguva yekutanga uye yechishanu sangano rekusangana (p = 0.03 uye p = 0.004, zvichienderana). Zvose saline- uye neapoxone-vakatapwa varume vakagadzirirwa kudijulisa panguva imwe neimwe yezvikamu zvishanu zvokugadzirisa.

Kuita zvepabonde-kunokonzerwa morphine mubayiro kushivirira

Migumisiro yehutano hwezvepabonde paVTA dopamine soma yakakonzerwa nechiitiko cheEOP muVTA yakafanana neyo inonzi yakanyanyisa opiates (Sklair-Tavron et al., 1996; Russo et al., 2007). Nokudaro, yakaedzwa kana mupiro wemubairo-wakakonzerwa neVTA dopamine cell plasticity inobata mubayiro we opiate morphine. Zvechokwadi, ruzivo rwepabonde rwakakonzera morphine kubhadhara kushivirira, zvakafanana nemigumisiro yenguva refu opiates (Russo et al., 2007). Vanhurume vanoita zvepabonde vakakundikana kuumba CPP ye 0.5 mg / kg morphine dose; asi, varume vane zvepabonde vanoita zvepabonde vakaita chirongwa chePPP nokuda kwechiyero ichi, sezvinoratidzwa nekupedza nguva yakawanda munguva yekamuri yefafine-paired yakaenzaniswa nekamuri ye saline-paired panguva yekuongorora (Fig. 3; p = 0.039). Vose vanoita zvepabonde vasina ruzivo uye vane ruzivo vanopedza nguva yakawanda kwazvo munguva yakawanda yekamphine-paired yakaenzaniswa neka saline-paired chamber nepamusoro maitiro e morphine: 5.0 mg / kg (Fig. 3; Naïve: p = 0.029; Exp: p = 0.012) uye 10.0 mg / kg (Fig. 3; Naïve: p <0.001; Exp: p = 0.002).

Mufananidzo 3. 

Migumisiro yehutano hwepabonde pamufaro we morphine. Nguva inoshandiswa mu saline- (Sal) kana kuti morphine- paired (Mor; 0.5, 5 kana 10 mg / kg bodyweight) makamuri munguva yechipupuriro mune zvepabonde (Naive, n = 10-13) kana ruzivo (Exp, n = 9-13) varume. Dhiyabhorosi inoratidzirwa seanoreva ± SEM; * inoratidza kusiyana kwakakura kuenzaniswa neSal-paired chamber mumhuka imwechete. NS, Kwete kukosha.

Zvepabonde ruzivo-rwakakonzerwa nekufambisa kwehutano hunoenderana neopioid receptor activation

Zvakawanikwa kusvika ikozvino zvinoratidza kuti EOP inobata muVTA panguva ye 5 zuva rimwe nerimwe maitiro ekugadzirisa mashoma anokonzera plastiki yeVTA dopamine neurons yakafanana nemigumisiro yekusingaperi morphine kana heroin self-administration (Russo et al., 2007; Mazei-Robison et al., 2011). Takafungidzira kuti VTA dopamine soma-size kuderedza kunokosha pamakadziro ekudzidza uye zvakananga nekuda kwepabonde-ruzivo rwekuita zvekuita zvepabonde maererano nekukurudzira nekushanda. Iyi pfungwa yakaedzwa kuburikidza nekudzivirira MOR kuburikidza naroxone panguva yekugadzirisa uye kuongorora migumisiro pakuita zvekuita zvepabonde kunokonzerwa nehutano hwepabonde panguva yezvikamu zvishanu zvekurara kwezuva nezuva. Dhiyabhorosi inowanikwa Mufananidzo 4 nokuda kwekutanga uye yechishanu kugovera zvidzidzo chete, sezvo aya ari data inonyatsoratidza kusangana-kwakagadziridzwa kushandiswa kwehutano hwekuita. Uyezve, nguva yakareba-inoguma yeaconxone kurapwa panguva yekugadzirisa maitiro inoshandiswa pakuchengetedzwa kwechiitiko-kukonzerwa nekufambisa kwekuita maitiro, panguva yekuedzwa kwekupedzisira kukwidza 1 vhiki imwe gare gare. Mufananidzo 4A inoratidza chimiro chekuedza. Ikoko kwakanga kune chinhu chikuru chinokosha chekugadzirisa chikwata pane zvose zvigadziro zvekuita zvepabonde (mount latency: F(2,55) = 11.286, p <0.001; kupindira latency: F(2,55) = 8.767, p <0.001; ejaculation latency: F(2,55) = 10.368, p <0.001) uye naloxone kurapwa pane latency yekukwira (F(1,55) = 6.585, p = 0.013) uye kutumirwa (F(1,55) = 7.863, p = 0.007). Pairwise kuenzanisa kwakaratidza kuti iyoxone kurapwa kwakagadzirisa tsika dzepabonde panguva yekutanga yekugadzirisa mukono nekuti mhuka yejonixone yakanga yakareba kwenguva refu kusvika kune yakatanga gomo (p = 0.002) uye kutumirwa (p = 0.002) inofananidzwa nehutori hwemasimba pazuva rokutanga rokusangana. Izvi zvinoxone inowanikwa pamaitiro ekutanga zvepabonde zvakaderedzwa nehutano hwepabonde uye kwete kucherechedzwa panguva ipi zvayo yemazuva ekuenzanisa maitiro (Tafura 1). Uyezve, iyoxone inotungamirira mushure mechimwe chezvikamu zvishanu zvekugadzirisa kusina kukwanisa kudzivisa kutanga kuita zvepabonde nehutano hwepabonde. Zvinopindirana nekusimbiswa kwemigumisiro yehutano hwezvepabonde, varume vanorapwa nesarine vakaratidza kuderedzwa kwema latency kuti vaende (Fig. 4B; p = 0.032) kutengesa (Fig. 4C; p = 0.033) uye ejaculation (Fig. 4D; p <0.001) panguva yechikamu chekusangana kwechishanu ichienzaniswa neyekutanga kusangana, iyo yakaratidza kufambiswa kwehunhu hwepabonde. Saizvozvo, nevanxone-vanobatwa vanhurume vanoratidzwa zvakanyanya mapfupi latency kukwira (Fig. 4B; p <0.001), kupinza (Fig. 4C; p <0.001), uye ejaculation (Fig. 4D; p = 0.017) pachishanu achienzaniswa nezuva rokutanga. Uyezve, varume vaikonzerwa neaconxone vakanga vasina kusiyana nemararamiro ekurara mune chero ipi zvayo panguva yechikamu chechishanu chekusangana.

Mufananidzo 4. 

Opioid endogenous inobata basa rakakosha mukufambisa-kutungamirirwa mukuita zvepabonde. A, Experimental design. B-D, Kuita zvepabonde zvipimo zvevarume vanobatwa ne saline (Sal, white bars, n = 11) kana naloxone (NLX; mabheti madema, n = 12) ne systemic administration. Dhiyabhorosi inoratidzirwa ndeye latency kukwira (B; miviri), intromission (C; masekondi), uye ejaculation (D; miviri) pamazuva 1 uye 5 yemashanu ekutevera mazuva ekuberekwa. Mukuwedzera, deta inoratidzirwa pakuedzwa kwekupedzisira kusangana, 7 d mushure mechikamu chechishanu chekugadzirira. Dhiyabhorosi inowanikwa seanoreva ± SEM; + inoratidza kusiyana kukuru pakati pezuva 1 ne5 mukati mekurapa; * inoratidza kusiyana kukuru pakati pekuedza zuva nezuva zuva 5 mukati mekurapa; # inoratidza kusiyana kwakakura pakati peaconxone uye saline mapoka mukati mezuva.

Ona tafura iyi: 

Tafura 1. 

Naloxone hutungamiri musati matanho yakawedzerwa latency kuwedzera uye kupindira chete pazuva rokutanga rokusangana

Kusiyana neizvi, naxone kurapwa munguva yekuita zvepabonde zvidzidzo zvakakanganisa kuchengetedza kwekuziva-kunokonzerwa nekuita zvepabonde pamusi wokupedzisira wekuenzanisa kwezuva. Zuva rekuedzwa rakaitwa 7 d mushure mekupedzisira kwekugadzirisa chirongwa mukushayikwa kwejoxone injection. Kurapa-kurapwa kwakarongwa nevarume kwakaratidza kutarisira-kunokonzerwa nekufambisa kwepabonde. Kunyanya, latency yakasimuka, inopission, uye ejaculation haina kusiyanisa pakati pechikamu chechishanu chekusangana uye zuva rokupedzisira rekuongorora (Fig. 4B-D). Izvo, iyoxx-yakarapwa varume yakaratidza kuwedzera kukuru kwe latencies kuisa (Fig. 4B; p = 0.033), kutengesa (Fig. 4C; p = 0.036) uye ejaculation (Fig. 4D; p = 0.049) pazuva rekuedza richienzaniswa nechishanu chekugadzirisa chirongwa. Uyewo, pazuva rekuedza mhuka ye-naloxone yakawanikwa ichireruka zvakanyanya pane varume vanorapwa nevarini sezvinoratidzwa neverefu latencies kukwira (Fig. 4B; p = 0.017) uye kutumirwa (Fig. 4C; p = 0.043). Nokudaro, lexaxone kurapwa kwakadzivisa kugadziriswa, asi kwete kutanga, kukura kwekuziva-kukonzera kushandiswa kwehutano hwepabonde. Izvi zvitsva zvinoratidza chikamu chakakosha cheEOP-inokonzerwa neVTA dopamine neuron plasticity kwenguva refu yekuratidzwa kwekusimbiswa kwemararamiro emararamiro emhuka.

Mamwe matsva ekuwedzera kuongororwa akaitwa kuti aone kuti migumisiro yeopioid receptor blockade pakurasikirwa kwenguva refu kwekusimbisa zvepabonde yakaitika yakasununguka kubva pakushayikwa kweaconxone kutungamira pazuva rokupedzisira rekuenzanisa zuva (Fig. 5A,B), zvakanyatsorasikirwa nekurasikirwa kwenguva refu, asi kwete kugadzirisa nguva pfupi yekufambisa kufambidzana (Fig. 5E,F), hazvina kukonzerwa nokuratidzwa zuva nezuva neapoxone oga (Fig. 5C,D), uye havana kubatwa nekurasikirwa kwepabonde mune zvepabonde mumurume anexx-akabatwa (Fig. 5G,H). Kutanga, kuratidzira kuti kuchinja kwepabonde pakuchinja kwekupedzisira kwekusangana hakuna kukonzerwa nekushayikwa kwaivaxone, iyoxone kana saline yaitungamirirwa nezuva rokupedzisira rekuedza kusangana kune mhuka dzakagamuchira kubatana dzakabatana neapoxone pavakawana ruzivo rwepabonde (Fig. 5A). Ikoko kwakanga kune chinhu chinonyanya kukosha chekutsvaga zuva pane latencies kukwira (Fig. 5B; F(2,27) = 30.031, p = 0.038) uye kutumirwa (Tafura 2; F(2,27) = 10.686, p = 0.048). Kwakanga kusine chinhu chikuru chekugadzirisa zuva pane latency kusvika kujjulation (Tafura 2; F(2,27) = 2.388, p = 0.109) Sezvinorondedzerwa pamusoro apa, naxaxone kurapwa panguva yekubatana hakubvumiri nekufambisa kwehutano hwepabonde panguva yekutanga yezvidzidzo zvepabonde zvishanu. Mapoka ose maviri (saline- uye akabatanidzwa neapoxone-akabatwa sezvakaitwa nemishonga yakagamuchirwa panguva yekuedzwa kwekugadzirisa kwekupedzisira; vose vakagamuchira naroxone panguva yekugadzirisa) vakaratidza hutano hwekuita zvepabonde pazuva 5 yakaenzaniswa nezuva 1 uye vakaratidza zvakadzama latency later to first mount (Fig. 5B; saline: p = 0.033; naloxone: p = 0.014) uye kutumirwa (Tafura 2; saline: p = 0.034; naloxone: p = 0.026). Mhuka dzakagamuchira iyoxone kana saline pamusana wekupedzisira mating test day yakanga yakareba latencies kukwira (Fig. 5B; saline: p = 0.018; naloxone: p = 0.029) uye kutumirwa (Tafura 2; saline: p = 0.019; naloxone: p = 0.020) yakaenzaniswa nezuva rechishanu rekugadzirisa ruzivo. Nokudaro, kutungamirirwa kwejaxone kana saline pazuva rekuedzwa nguva pfupi kusati kwasangana kusina kukanganisa kukonzerwa neaconxone kurapwa munguva dzekuita zvepabonde uye kuregererwa kwenguva refu kwekuita zvepabonde kwakafanana kune izvo zvakaratidzwa mhuka dzisina kugamuchira jekisi pamusangano wokupedzisira wekuenzanisa zuva (Fig. 4).

Mufananidzo 5. 

Majoi endogenous anokosha mukuratidza kwenguva refu kwekuziva-kukonzerwa kushandiswa kwehutano hwepabonde. A, Ongororo yekugadzirira kuedza kuitika kweNLX mishonga pazuva rekuedza. B, Gomo latency mumazuva 1 uye 5 kwemazuva mashanu anotevera mazuva ekugadzirisa uye zuva rokupedzisira rekuedza kusangana (Test) mushure me saline (grey) kana kutijoxone (yakasviba) injection. Data inomirira zvinoreva ± SEM. * inoratidza kusiyana kukuru pakati pezuva 1 uye zuva 5 mukati mekurapa. # inoratidza kusiyana kwakakura pakati pekuedzwa zuva uye Zuva 5 mukati mekurapa. C, Kuongororwa kwekuedza kwekuedza kuedza kushanda kweandroxone pre-treatment chete pasina ruzivo rwepabonde pamabatiro ekubata. D, Gomo latency pakupedzisira kwekutsvaga zuva rekuedza, mazuva e7 achitevera mazuva e5 eine saline kana kuti injection yaaloxone mukushayikwa kwekubatana. Data inomirira zvinoreva ± SEM. E, Kuongororwa kwekuedza kuedza kana chirwere ichoxone chakakanganisa kuratidzwa kwenguva pfupi kwemaitiro ekuita zvepabonde mumhuka dzezvepabonde. F, Gomo latency masikati 1 uye zuva 5 yemazuva mashanu ekukwirirana uye zuva rokupedzisira rekuenzanisa zuva, 1 zuva rimwe zuva 5 pamberi pe-saline (grey) kana kuti injini yexidon (yakasviba). Data inomirira zvinoreva ± SEM. * inoratidza kusiyana kukuru pakati pezuva 1 uye zuva 5 mukati mekurapa. G, Kuongororwa kwekuedza kwekuedza kuongororwa kuti ikoxone kurapwa yakavhara migumisiro inobatsira yekuita zvepabonde. H, Nguva inoshandiswa mukukwiridzira kamuri yakavharidzirwa (mumasekondi mashoma) panguva yekutanga (yakachena) uye mushure mokuedzwa (mutsvuku) wemhuka inogamuchira iyoxone kana saline musati matanho. Data inomiririra zvinoreva ± SEM; * inoratidza kusiyana kwakakura kuenzaniswa ne pretest.

Ona tafura iyi: 

Tafura 2. 

Dhiyabhorosi inoratidzirwa ndeyekudzivirira kune kuisa mukati uye kushandiswa kwemashure (masekondi) kubva kuhutano hwekuedza kunoitwa kuti uone kuti migumisiro yeBL blockade pakurasikirwa kwenguva refu kwekusimbiswa kwepabonde yakaitika yakasununguka kubva pakushayikwa kweaconxone kutungamira pazuva rokupedzisira rekuedza kusangana

Kuti uone kana yaiva nexaxone kurapwa pawakabatana nehutano hwepabonde uye usati wadzokororwa kunexxone neyo iyo yakakonzera kuora mwoyo kwehutano hwepabonde 7 d mushure mekurapa kwekupedzisira, mhuka dzepabonde dzakagamuchira zvirwere zvishanu zvezuva nezuva zvejaixone kana kuti injini ye saline mushure mokunge mushure mokunge uchitarisana 7 d gare (Fig. 5C). Hapana kusiyanisa kwakakosha kwaionekwa kune chero ipi zvayo yakasarudzwa pakati pe saline neapoxone pretreated groups (Fig. 5D; gomo latency; intromission latency: saline 139.7 ± 40.3 vs naloxone XUMUM ± 121.83; ejaculation latency: saline 42.55 ± 887.9 vs naloxone 70.0 ± 1050.8). Izvi zvinoratidza kuti naloxone chete haikwanisi kuchinja maitiro ekuita zvepabonde anotevera, akafanana nekushaikwa kwemigumisiro yejaxone oga paVTA dopamine neuron plasticity.

Tinofunga kuti iyoxone kurapwa panguva yekuwana chiitiko chepabonde kunokanganisa kurarama kwenguva refu kwekuita zvepabonde-kukonzerwa nekuita zvekuita zvepabonde. Kuti uedze izvi zvakare, migumisiro yeaixone kurapwa panguva yekugadzirisa yakaedzwa pamushure mekuita zvepabonde munguva yekupedzisira yekuenzanisa, izvo zvakaitwa chete 1 d mushure mekusangana kwekupedzisira (kuongorora kwekuedza; Fig. 5E). Ikoko kwakanga kune chinhu chikuru chakakosha chekugadzirira zuva pagomo (Fig. 5F; F(2,20) = 19.780, p <0.001) uye kutenderera kwemukati (Tafura 2; F(2,20) = 19.041, p <0.001). Pakange pasina chakakosha chiitiko chezuva pane ejaculation latency (Tafura 2; F(2,20) = 3.042, p = 0.070). Zvakafanana nechakurondedzerwa pamusoro apa, varume vose (pasinei nemaronda e saline kana kutixxx) vakaratidzira kufambisa tsika dzepabonde munguva dzezvikamu zvishanu zvekuita zvepabonde zviratidzo zvinoratidzirwa nechepfupi latency latten (Fig. 5F; saline: p = 0.002; naloxone: p = 0.018) uye kutumirwa (Tafura 2; saline: p = 0.006; naloxone: p = 0.009) pazuva 5 yakaenzaniswa nezuva 1. Saizvozvowo, nzira dzakagadziriswa dzepabonde dzakaratidzwa nezuva rekuenzanisa kana richienzaniswa nezuva 1 rinoratidzwa nechepfupi latency lating (Fig. 5F; saline: p = 0.001; naloxone: p = 0.020) uye kutumirwa (Tafura 2; saline: p = 0.004; naloxone: p = 0.009). Zvinotonyanya kukosha ndezvekuti, iyoxone kurapwa panguva yekubatana haina kuchinja zvakanyanya kugadzirisa-kukonzerwa nekufambisa kwehutano hwepabonde paakaedzwa 1 d mushure mekuita zvepabonde, pasina kuzvipira kwaivaxone pamuzuva uyu wokupedzisira wekutsvaga.

Chekupedzisira, isu takaedza mukana wekuti naloxone inowana mhedzisiro yekutaura kwenguva refu kwekufambiswa kwetsika yekuita zvepabonde nekuda kwekuvhara mhedzisiro ye naloxone pazvinhu zvakanaka zvinobatsira zvebonde. Nekudaro, naloxone yakatumirwa pakarepo isati yamera haina kushandura kuumbwa kweCPP yekukweva (Fig. 5G), zvichiratidza kuti nanxone kurapa haina kuchinja mupabonde mubayiro. Zvose saline- uye mapoka ezvikambiso akaitwa neapoxone akaumba huwandu hwePPP hwekuita zvepabonde sekuratidzwa nekuwedzerwa nguva yakapedzerwa muimba yepabonde-paired chamber (Fig. 5H; saline: p = 0.038; naloxone: p = 0.002) panguva yechipupuriro chekuenzaniswa nechokutanga. Nokudaro, naloxone haibatsiri kukanganisa kwayo pakuchengetedzwa kwehutano hwepabonde nekudzivisa mubayiro unobatanidza nehutano hwepabonde.

Kukurudzira kuita zvepabonde kunoenderana neEOP chiito muVTA

Kusimbisa kuti EOP nharaunda chaiyo muVTA yekuita kuti kushandiswa kwenguva refu kwekuita zvepabonde kuve kwenguva yakareba, chirongwa chekuedza chinorondedzerwa mu Mufananidzo 3A yakadzokororwa ne intra-VTA naloxone infusions panzvimbo ye systemic injections. Zvigumisiro zvaive zvakafanana ne systemic administration inotsanangurwa pamusoro apa. Ikoko kwaive nechinonyanya kukosha chekugadzirisa zuva pamatambudziko ose ekuita zvepabonde (Fig. 6A, gomo latency: F(2,33) = 4.494, p = 0.019; B, intromission latency: F(2,33) = 4.042, p = 0.027; C, ejaculation latency: F(2,33) = 5.309, p = 0.010) uye intra-VTA naloxone kurapa pa latencies kuisa (F(1,33) = 7.345, p = 0.011) uye kutumirwa (F(1,33) = 6.126, p = 0.019). Intra-VTA naloxone haina kudzivirira kutanga kwepfungwa-kukonzerwa nekufambisa kwehutano hwepabonde panguva ye 5 d yekukwirirana, sezvo varume vanonzi naxaxone-vakatariswa vakaratidza kuderedzwa kwema latency kuisa (Fig. 6A; p = 0.001), kutengesa (Fig. 6B; p <0.001), uye ejaculation (Fig. 6C; p = 0.001) pazuva 5 yakaenzaniswa nezuva 1. Varume vanonzi Naloxone-vakatorwa havana kusiyana nevarume vanorapwa nevarine pazuva rechishanu rokukwirirana mune chero ipi zvayo. Intra-VTA naloxone kurapwa, sekushandiswa kwehutungamiri, kwakakonzera zvikuru kuwedzera gomo (Fig. 6A; p <0.001) uye kutenderera kwemukati (Fig. 6B; p <0.001) pazuva rekutanga rekukwikwidza kana richienzaniswa nevarume vaibatwa nesaline, iyo isina kucherechedzwa panguva dzekuteedzana dzinotevera (panguva iyo naloxone- nevarume vaibatwa nesaline vasina kusiyana). Chimwe chiitiko chisingatarisirwe ndechekuti mune kuyedza uku, varume vakarapwa munyu havana kuratidza kuverengera kwakakosha kufambisa kwegomo kana kusvetuka kwemashure (sezvakaratidzwa mune zvese zviedzo zvinotsanangurwa pamusoro), uye chete ejaculation latency yakapfupiswa pazuva rechishanu zvichienzaniswa nezuva rekutanga. (munyu: p = 0.001).

Mufananidzo 6. 

Zviyo zvinogadziriswa muAVTA zvinopindira ruzivo rwakaitwa-ruzhinji rwekuita zvepabonde nekuchengetedza kwenguva refu. Kuita zvepabonde zvirongwa zvevarume vanobatwa ne saline (Sal, white bars, n = 8) kana NLX (mabheji matsvuku, n = 6) ine intra-VTA utungamiri. Dhiyabhorosi inoratidzirwa ndeye latency kukwira (A), intromission (B), uye ejaculation (C) mumazuva 1 uye 5 kwemazuva mashanu akatevedzana ekuberekwa. Mukuwedzera, deta inoratidzwa nokuda kwekupedzisira kwekutsvaga zuva rekuedza, 7 d inotevera zuva 5 pakusina kwe saline kana kuti injection yaaloxone. Data inomiririra zvinoreva ± SEM; + inoratidza kusiyana kukuru pakati pezuva 1 ne5 mukati mekurapa; * inoratidza kusiyana kukuru pakati pekuedza zuva nezuva zuva 5 mukati mekurapa; # inoratidza kusiyana kwakakura pakati peapoxone neSal mapoka mukati mezuva. Zvirongwa zveCononic VTA zvikamu (H, -4.60; I, -5.00; J, -5.25 kubva kuBregma) inoratidza intra-VTA nzvimbo yejojo ​​yezvipfuwo zvose Muedzo 5 (saline; white; uchishandisa template mifananidzo kubva ku Swanson Brain Maps (Swanson, 2004). Cannulas yakanga iri pamwe chete, asi injection nzvimbo dzinomiririrwa unilaterally kuitira nyore kutaura. fr, Fasciculus retroflexus; ML, Medial lemniscus; SN.

Intra-VTA naxolone kurapwa kwakadzivirira kugadziriswa kwehutano hwehutano hwakarongeka hunoonekwa mune varume vane hutano hwepabonde, zvakafanana nemigumisiro ye systemic naroxone. Kunyanya, pamusi wokupedzisira wekuedzwa zuva renaxx-treated men had longer latencies to mount (Fig. 6A; p = 0.011), kutengesa (Fig. 6B; p = 0.010), uye ejaculation (Fig. 6C; p = 0.015) inofananidzwa nechishanu chekugadzirisa chirongwa uye ichienzaniswa nevarume vane chirwere chesaini pazuva rekuedzwa kwekupedzisira (Fig. 6A, p = 0.006; B, p = 0.008). Kusiyana neizvi, mhuka dzakabatwa nesaini dzakanga dzisingatendi mu latencies kuti dzisimudze uye dzipindire pakati pekupedzisira kwekuedza zuva nezuva zuva 5 yekubatana. Izvo zviitiko zvaive zvakananga kubudiswa kwaNaxone kuVTA, sevarume vane nzvimbo dzinoparadza nzvimbo dziri pedyo asi kwete kutarisa VTA (Fig. 6D; n = 3) akaratidzirwa kwenguva refu yekuita zvepabonde zvakafanana nekurapa-kurapwa kwekutarisa (ML, IL = 53 ± 6.245, EL = XUMUMI ± 389 uye zvakasiyana zvikuru nekuenzaniswa ne-intra-VTA mhuka yexxoni pazuva rokupedzisira rokubata pamuviri (ML, IL: p = 0.029; EL: p = 0.0395).

EOP tsanangudzo inotarisirwa kugadziriswa-inogadziriswa inogadziriswa cue-induced neural activation

Kubva pane zvakawanikwa kusvika ikozvino, takafungidzira kuti EOP kushandiswa muVTA panguva yekuwana ruzivo uye kuenderera mberi kweVTA dopamine soma kuderedza ukuru kwakakosha kupiwa kwekukurudzira kwekukwanisa kufambisa zvinokonzerwa nemubairo uye nekudaro, kuchengetedzwa kwechikonzero chekuita zvepabonde. Kuti uedze iyi hypothesis, migumisiro yekuvhara maopioid receptors munguva yekubata ruzivo pamusoro pezvakaitwa neural's induced by subsequent exposure to contextual cues predictive of sexually rewarded (sex-associated contextual cues) yakarongedzwa. Mhuka dzepabonde dzisiri dzepabonde dzakanga dzakataridzirwa kune zvakatipoteredza, asi idzi dzakanga dzisingabatanidzwi nehutano hwepakutanga, naizvozvo vakanga vasingatongogumiriri. Pakupedzisira, nheyo dzepamusoro dzePERK dzakagadzirirwa mune zvekuita zvepabonde uye mapoka ekudzivirira maitiro akasara mumakamuri epamba uye asingatauri kune chero cue (-No Cue). Kusimbisa uye kuwedzera zviwanikwa zvekare (Balfour et al., 2004), kuiswa kune zvinyorwa zvemashoko zvakabatanidzwa nemubhadharo wepabonde wepakutanga zvakanyanya kuwedzera kutaura kwemashoko muvarume vane zvepabonde mune NAC (Fig. 7) uye mPFC (Fig. 8A-C), asi hazvina kukonzera neuronal activation muBLA (Fig. 8D) kana CPU (data isingaratidzi). Paiva nemigumisiro mikuru ye cue exposure mu NAC core (F(1,12) = 12.1941, p = 0.004), ACA (F(1,12) = 5.541, p = 0.038), uye PL (F(1,12) = 5.241, p = 0.041), uye naloxone kurapwa mu NAC core (F(1,12) = 6.511, p = 0.025), ACA (F(1,12) = 15.242, p = 0.002), uye PL (F(1,12) = 7.336, p = 0.019). Paiva nekubatana kwakakosha muNac core (F(1,12) = 10.107, p = 0.008), ACA (F(1,12) = 16.060, p = 0.002), PL (F(1,12) = 8.235, p = 0.014), uye IL ((F(1,12) = 6.965, p = 0.022). Kutanga, kuve nekubatanidza kuwirirana kweCue exposure kunowedzera kuwedzera muzvinyorwa zvepabonde zvinoshandiswa mune zvehutano (Exp Sal ​​+ Cue) zvichienzaniswa nemitemo yakanga isingatauri kune chero ipi zvayo uye yakatorwa kubva muimba yeimba (Exp Sal-No Cue) mu NAC core (Fig. 7A; p <0.001), uye mPFC nharaunda ACA (Fig. 8A; p = 0.001), PL (Fig. 8B; p = 0.003), uye IL (Fig. 8C; p = 0.029). Kusiyana neizvi, mumhuka dzakasvibiswa zvepabonde, kusangana nemamiriro ezvinhu, izvo zvisingabatanidzwi nemubairo wepabonde, hazvina kukonzera chero nzvimbo dzepfungwa dzese (Naive Sal + Cue yakaenzaniswa naveve Sal-No Cue; Maonde. 7, 8), kuratidza kuti kuiswa kwepERK kunonyanya kujekesa kwehutano hwehutano-hunoenderana. Uyezve, ruzivo rwepabonde chete haruna kuchinja hutano hwemashoko huri munharaunda dzeuropi, sezvo pakanga pasina kupesana pakati pemapoka akatorwa kubva kumakamuri emba, kana kuti zvepabonde kana chiitiko, uye anobatwa ne saline kana naloxone.

Mufananidzo 7. 

Kugadziriswa kwemaopioid action inotarisirwa neural activation mu NAc inokonzerwa nepabonde-inosanganiswa nemamiriro ezvinhu. Numeri yePERK-IR masero pamam2 mu nucleus accumbens core (A) uye shell (B) mune zvepabonde (chichena) uye vane ruzivo (Exp; nyeredzi) mhuka dzakagara dzichitungamirirwa ne systemic NLX kana saline (Sal) panguva yekugadzirisa (Exp male) kana kugadzirisa zvirongwa (Naive varume). Mapoka akange akazarurirwa kune zvinyorwa (Cue), izvo zvaive zvakabatana-zvekubatanidza mune zveExp males uye zvematongerwe enyika muNaive mhuka, kana kutorwa kubva kumakamuri emisha (No Cue; inoratidzwa nokusasha kweCue-label). Dhiyabhorosi inowanikwa seanoreva ± SEM; * inoratidza kusiyana kwakasiyana nekuenzaniswa nesarine-yakatangwa kwete chekudzivirira-kuonekwa pachena (Naive Sal-No Cue uye Exp Sal-No Cue); # inoratidza kusiyana kwakasiyana nekuenzaniswa neSal-treated Cue-yakajeka Exp boka (Exp Sal ​​+ Cue). Mumiririri mifananidzo yePERK-IR masero pamam2 muAc core yevarume vane zvepabonde neSar (C, D) kana NLX (E, F) iyo inogona kutorwa kubva pamba yemusha (No Cue, C, E), kana kuti yakazarurirwa kune zvinyorwa zvinowirirana (Cue; D, F). N = 4 boka rimwe chete kunze kweNaive NLX (No Cue), n = 3. ac, anterior commissure. Nhamba yezinga, 100 μm.

Mufananidzo 8. 

Migumisiro yeaixone pamating cue-induced pERK kutaura mune dzimwe VTA chinangwa nzvimbo. Numeri yePERK-IR masero pamam2 mune zvepabonde (chichena) uye vane ruzivo (Exp; nyeredzi) mhuka dzaive dzakarongedzwa ne systemic NLX kana saline (Sal) panguva yekugadzirisa nekugadzirisa uye dzakasununguka kune zvinyorwa (Cue) kana imba yepamusoro (hapana cues) mu ACA (A), PL (B), IL (C), uye BLA (D). Dhidhiyo inomiririra kureva ± SEM; * inoratidza kusiyana kwakasiyana nekuenzaniswa nesarine-yakatangwa kwete chekudzivirira-kuonekwa pachena (Naive Sal-No Cue uye Exp Sal-No Cue); # inoratidza kusiyana kwakasiyana nekuenzaniswa neSal-treated cue-pachena yakarongedza zvekuita zvepabonde (Naive Sal + Cue).

Mukusimbisa kwehana yedu, ikoxone kurapwa panguva yekuita zvepabonde zvakanyanyisa kuregererwa kwepERK kuburitswa nehutano hwehutano hwakabatanidzwa. PREK kutaura mumadzinza aya anexone-akaonekwa akawanikwa akashandurwa (Exp NLX + Cue) haana kusiyana nemashoko ekutanga emashoko ekureva mune zvevamwe vevanhu vasina zvekuita zvepabonde kana kuti vane ruzivo rwakatorwa kubva kumakamuri emisha (Naive Sal-No Cue kana Naive NLX- Kwete Cue). Uyezve, kutaura kwemanzwiro mumadzinza anexx-akaonekwa akamboonekwa (Exp NLX + Cue) akanga akadzika zvakaderera kana achienzaniswa nemhuka dzakagadziriswa dzakagadziriswa (salon cue) muNac core (Fig. 7A; p = 0.002) uye mPFC subregions (Fig. 8A, ACA: p <0.001; B, PL: p = 0.002; C, IL: p = 0.015).

MuAc shell, nzira mbiri ANOVA kuongororwa haina kubereka maitiro akawandisa emigumisiro yezvikonzero cue exposure uye nalxone kurapwa. Kunyange zvakadaro, kuenzanisa kwechipiri kwakaratidza kuti cue exposure yakaita kuti pERK iri muboka rinotarisana nevarume vanoita zvepabonde (Exp SAL + Cue) yakaenzaniswa nesarudzo isina kuonekwa yaSaline naive control group (Fig. 7B; Naive SAL-Hapana Cue: p = 0.0163).

hurukuro

Chidzidzo chezvino chinoratidza kuti EOP inoshanda muVTA panguva yekuita zvepabonde, mararamiro emhando yepanyama, yakakonzera kuderedzwa kwakasimba asi kwenguva pfupi mukukura kwe soma yeVTA dopamine masero. Kupera muhukuru hwema soma hauna kuonekwa muVTA isina-dopamine neurons, kana mu dopamine neurons mune pedo substantia nigra, zvichiratidza kuti shanduko iyi yaiva yakananga kuVTA dopamine masero. Iyi VTA dopamine plasticity inowanikwa yakafanana neyo inokonzerwa nekusingaperi opiate exposure (Sklair-Tavron et al., 1996; Russo et al., 2007; Mazei-Robison et al., 2011) uye zvakakonzera kushivirira kwakafanana kune opiate yakawanda (morphine) mubayiro. Takaratidza kuti VTA dopamine plastiki inokosha kune nguva refu (kuchengetedza) asi kwete nguva pfupi (kukurudzira), kusimbiswa kwehutano hwepabonde uye mubayiro-inobatanidza kuwanikwa-neural mabasa (pERK) muVTA nzvimbo dzinotarisirwa: NAc uye mPFC. Izvi zvakawanikwa zvinoratidzira basa reVTA dopamine plasticity mukutaura kwenguva refu kwekukurudzirwa kwechikonzero chekuwana mubairo wezvakaitika kare predictive cues kana mubairo yekuyeuka.

Izvo zvakanyatsorongedzwa kuti ruzivo rwepabonde runoguma mukufambisa kwehutano hwepabonde hunotevera, kusanganisira kukurumidza kukurumidza kutanga kwekusangana nekuwedzera kwekuita (Balfour et al., 2004; Pitchers et al., 2010a,b, 2012). Iyi yekufambisa kana kusimbiswa kwepabonde kunochengetwa kwekuda kwe28 d mushure mokugadzikana (Pitchers et al., 2012). Mukuwedzera, zvakaratidzwa kuti hutano hwepabonde uye hunoenderana nehuwandu hwehutano hwehupombwe hunokonzera MOR kudzidzira mukati muVTA uye kuita neuronal activation mukati mezisolimbic system, kusanganisira muVTA (dopamine uye nondopamine neurons), NAc, PFC, uye BLA (Balfour et al., 2004, 2006). Iko yakanyatsosimbiswa VTA dopamine neurons inobata basa rinokosha mukudzidza uye kupiwa kwekukurudzira kwekupa mubayiro wekupa mubayiro (Berridge naRobinson, 1998; Berridge et al., 2009; Flagel et al., 2011) uye vari kutsvaga kuti mubairo utaure (Schultz, 2010). Ikozvino kuwanikwa kunowedzera pane ruzivo rwedu rwezvino kuburikidza kuratidza kuti mubairo-wakakonzerwa neVTA neuroplasticity inokosha pamabasa aya, uye anobva pane MOR kushandiswa neEOP muVTA. Ikozvino hazvizivikanwe kuti EOP ndiyo MOR ligand inoshanda muVTA munguva yehutano hwemurume. Kunyange zvazvo zvose β endorphin uye enkephalin zvakabatanidzwa mukukurudzirwa kwekurudziro yezvokudya zvekudya (Hayward et al., 2002), izvi zvinoramba zvichisimbiswa pakuita zvepabonde zvevarume. Takave takazivisa kuti β-endorphin neurons haisati yaitwa panguva yekubatana, uye hapana kuwedzera muPOMC mRNA; saka, zvichiratidza kuti β endorphin inogona kunge isiri iyo EOP inokosha inobata muVTA panguva yekukwirirana (Davis et al., 2007). Iyi VTA dopamine plastiki yakanga yakakosha pamabasa ekushandiswa kweveural mu mPFC, NAc, uye VTA mushure mekunge vatarisana nemigumisiro yekuita zvepabonde-kufanotaura nezvezvakatipoteredza. Uyezve, VTA dopamine plasticity yakanga yakakosha nokuda kwekugara kwenguva refu kuratidzwa kwekutanga uye kushanda kwepabonde. Kusiyana neizvi, VTA neuroplasticity inokonzerwa nehutano hwepabonde haina kudiwa nokuda kwehutano hwehutano sehupombwe (yakagadzirirwa neCPP) uye nguva pfupi yekufambisa zvekuita zvepabonde nekushanda (panguva yekuita zvepabonde kana 1 d gare gare) yakaramba yakasimba pasinei naMOG kudzivirirwa panguva yekubatana (Mehrara naBaum, 1990). Pane kudaro, deta inoratidza kuti VTA dopamine neuroplasticity inopindirana kwenguva refu (7 d mushure mekuita zvepabonde pakupedzisira; Pitchers et al., 2012) kuratidzwa kwe "kuda" zvepabonde mubayiro uye kukwidziridzwa kwakagadziriswa kune mating cues (Miller naBaum, 1987; Berridge naRobinson, 1998).

Mhuka dzinotarisana nepabonde dzakaratidza kubvumira kutsvaga kumufaro we morphine, zvakafanana nemigumisiro yegumbo rinoshanda mumakonzo, imwe yomuitiro wekupa mubairo, chiitiko chakavharwa neaconxone kurapwa (Lett et al., 2001, 2002) uye vakatsunga kuva vanovimba neVTA dopamine cell plasticity (ikozvino kuwanikwa). Kufanana nemubairo wepanyama, kudzokorora kune maopiates morphine kana heroin zvinoguma nekuderera kwechinguva kweVTA dopamine soma size (Sklair-Tavron et al., 1996; Spiga et al., 2003; Russo et al., 2007; Mazei-Robison et al., 2011). Uyezve, kubvuma kuvhara kwenguva pfupi kunokonzera mubayiro kushivirira, sezvinotsanangurwa nepamusoro pemishonga inodhaka inotarisirwa kuita masangano ekubhadhara (Shippenberg et al., 1987; Russo et al., 2007), uye zvinokonzera kuzvipa mhuka kuti kuwedzere kudhakwa kwezvinodhaka (Ahmed et al., 2000; Walker et al., 2003). Nokudaro, EOP uye maopiates vanoita zvinowanzoitwa neural substrates kuti vashandise mukana wokushivirira pavanotanga kubviswa, izvo zvinogona kuratidzira mvumo yekugadzirisa homeostatic nzira yekudzivisa kukurudzira nekudzokorora zvakare (Koob naLe Moal, 2005). Kusiyana neizvi, panguva yakareba kwenguva refu opiate zvinodhaka, kushivirira kunoderedzwa kunzwisisika kune zvipo zvinokonzera kumushonga (Harris naAston-Jones, 2003; Aston-Jones naHarris, 2004; Harris naGewirtz, 2004). Zvinonakidza, ruzivo rwepabonde rwakakonzerwa nekurara kwepabonde nguva ye7-28 d yakawanikwa ichiita kuti kuve nekukurudzirwa kwepfungwa yepistimululant (Pitchers et al., 2010a), iyo inobva pane kubata-induced deltaFosB kutaura uye dopamine receptor 1 kushandiswa mu NAc (Pitchers et al., 2013). Nokudaro, ruzivo rwepabonde rwekuita zvepabonde hunokonzera panguva imwechete opiate mubayiro wekushushikana uye psychostimulant mubayiro wekusimudzira, kunyange zvazvo nguva refu yekurega zvepabonde panguva ye morphine mukana wokushivirira kunoramba kuri kuedzwa. Tinoratidza kuti izvi zvinopesana nemigumisiro yemishonga inogona kunge yakarongedzwa nemhando dzakasiyana dzemapuranga epurasitiki munzvimbo dzakasiyana dzemasolimbic system: VTA EOP chiito uye dopamine plasticity inobatanidza opiate mubayiro wokushivirira (ikozvino kuongorora), asi NAc deltaFosB kutaura kutonga psychostimulant kukudzivirira (Pitchers et al., 2013). Zvose izvi zviitiko zvinogona kuita kuti kuwedzera kwemishonga inotora (Ahmed naKoob, 1998, 1999; Ahmed et al., 2000, 2002, 2003; Walker et al., 2003).

Maitiro emakemikari ayo EOP anokonzera VTA dopamine neurons panguva yekuzvipira maitiro evhudziri haazivikanwi. I IRS2-Akt-mTORC2 mugwagwa ndechemurevereri mukuru wekurasika soma ukuru muVTA yakakonzerwa ne morphine yakadzokororwa (Jaworski et al., 2005; Russo et al., 2007; Mazei-Robison et al., 2011). Kudzokorora morphine kutungamirirwa kwakakonzera kuchinja muhukuru hwe dopamine neurons muVTA kunogona kudziviswa ne-intra-VTA kuputika kweuropi-yakagadzirirwa neurotrophic factor (BDNF; Sklair-Tavron naNestler, 1995). BDNF inoshandisa nzira ino kuburikidza neTrBB kuisa (Russo et al., 2007), receior kinase nehupamhi hwepamusoro neBDNF uye chikamu che IRS2-Akt nzira (Seroogy uye Gall, 1993; Numan uye Seroogy, 1999), uye inoratidzwa pane dopamine uye GABA neurons muVTA. Kuderedzwa kwezvikamu zvakasiyana-siyana zve IRS2-Akt nzira yekushandisa viral vector gene transfer technology inoshandisa migumisiro yekusingaperi opiate exposure. Uyezve, migumisiro yeipiate kufungidzirwa inogona kununurwa kuburikidza nekudzorera iyi nzira yekucherechedza (Russo et al., 2007) uye kunyanya kufungidzira kwechikamu cheMTORC2 kunodzivirira morphine-inokonzerwa neVTA dopamine soma kuderedza (Mazei-Robison et al., 2011). Naizvozvo, basa rekare rekutsvaga migumisiro yekusingaperi maopiates paVTA dopamine soma size inoratidza kuti morphine-induced downregulation yemugwagwa we IRS2-Akt-mTOR zvose zvakakwana uye zvakakosha kuitira izvi (Mazei-Robison naNestler, 2012). Nokudaro, iri kuedza kufungidzira kuti migumisiro yehutano hwepabonde paVTA dopamine neuroplasticity yakafanana yakabatana neBDNF uye IRS2-Akt-mTORC2 mugwagwa.

Mukupedzisa, kuongorora ikozvino kwakaratidza kuti VTA neuroplasticity inokonzerwa nechiitiko nemararamiro emararamiro evanhu, kunyanya kuburikidza nekudzokorora kwehutano hwevanhu hwepabonde. Kunyanya, EOP chiito muVTA kuderedza dopamine soma size, iyo inofungidzirwa kuti ibatanidzwe nekuwedzera kweeural neyakagadzikana uye kudarika dopamine yakaguma inoguma ne hypodopaminergic system, uye inoshandura mesolimbic system inoshandiswa mukuita kune zvinyorwa izvo zvinofananidzira zvepabonde. Uyezve, VTA neuroplasticity inokosha pakukurudzira nekukurudzira chiyeuchidzo, asi kwete nekuda kwehutachiona hwekuita zvepabonde. Pakupedzisira, VTA neuroplasticity inokonzerwa nemararamiro emararamiro evanhu akatevera nguva shomanana yekubhadhara kusagadzikana kunokonzera opiate mubairo uye zvingakanganisa kudzvinyirirwa pakukura kwezvinodhaka.

Mashoko Omuzasi

  • Yakagamuchira January 12, 2014.
  • Revision yakagamuchirwa May 17, 2014.
  • Yakagamuchirwa May 20, 2014.

  • Tsvakurudzo iyi yakatsigirwa nezvipo kubva kuCanada Institute of Health Research kusvika kuLMC uye Natural Sciences uye Engineering Research Council kuKKP

  • Vanyori vanoti hakuna kupikisana mune zvemari.

  • Mharidzo inofanira kutaurirwa kuna Dr Lique M. Coolen, University of Mississippi Medical Centre, Dhipatimendi rePhysiology uye Biophysics, 2500 North State Street, Jackson, MS 39216-4505. [email inodzivirirwa]

References

  1. ↵
    1. Agmo A

    (1997) Chirwere chevarume chepabonde. Ubongo Res Brain Res Protoc 1: 203-209.

  2. ↵
    1. Ahmed SH,
    2. Koob GF

    (1998) Kuchinja kubva pakuenzanisa kusvika pakuwedzerwa kwezvinodhaka zvinodhaka: shandura mu hedonic set point. sainzi 282: 298-300.

  3. ↵
    1. Ahmed SH,
    2. Koob GF

    (1999) Kuwedzerwa kwenguva refu mune zvakatemwa zve cocaine self-administration shure kwekuwedzera mumakonzo. Psychopharmacology 146: 303-312.

  4. ↵
    1. Ahmed SH,
    2. Walker JR,
    3. Koob GF

    (2000) Kuramba kuchiwedzera mukukurudzirwa kutora heroin mumakonzo ane nhoroondo yezvinodhaka. Neuropsychopharmacology 22: 413-421.

  5. ↵
    1. Ahmed SH,
    2. Kenny PJ,
    3. Koob GF,
    4. Markou A

    (2002) Neurobiological uchapupu hwe hedonic allostasis inobatanidza nekuwedzera kwekushandisa cocaine. Nat Neurosci 5: 625-626.

  6. ↵
    1. Ahmed SH,
    2. Lin D,
    3. Koob GF,
    4. Parsons LH

    (2003) Kuwedzera kwekodineine self-administration hazvibvi pane zvakagadziriswa nekasekasi-induced nucleus accumbens dopamine levels. J Neurochem 86: 102-113.

  7. ↵
    1. Aston-Jones G,
    2. Harris GC

    (2004) Nhengo dzebhondi dzekubata kutsvaga zvinodhaka munguva yekupera kwekusiya. Neuropharmacology 47: 167-179.

  8. ↵
    1. Balfour ME,
    2. Yu L,
    3. Coolen LM

    (2004) Kuita zvepabonde uye zvepabonde zvinosangana nezvimwe zvinhu zvinokonzerwa nekuita zvepabonde zvinoita kuti mazisolimbic system muvarume varove. Neuropsychopharmacology 29: 718-730.

  9. ↵
    1. Balfour ME,
    2. Brown JL,
    3. Yu L,
    4. Coolen LM

    (2006) Mipiro inogona kuitika yehutachiona kubva kune imwe nzvimbo yepamusoro yekodhi neyeural activation maererano nekuita zvepabonde munhurume yemurume. Neuroscience 137: 1259-1276.

  10. ↵
    1. Beitner-Johnson D,
    2. Guitart X,
    3. Nestler EJ

    (1992) Neurofilament mapuroteni uye mazisolimbic dopamine system: inowanzogadziriswa nehupenyu husingaperi hunopera morphine uye chine cocaine chisinganzwisisiki mu rat rat ventral tegmental. J Neurosci 12: 2165-2176.

  11. ↵
    1. Berridge KC,
    2. Robinson TE

    (1998) Chii chinonzi dopamine mubhadharo: hedonic zvinokonzera, mubayiro wekudzidza, kana kuti kukurudzirwa? Ubongo Res Brain Rev Rev 28: 309-369.

  12. ↵
    1. Berridge KC,
    2. Robinson TE,
    3. Aldridge JW

    (2009) Kuparadzanisa zvikamu zvemubayiro: "kufarira", "kuda", nekudzidza. Curr Opin Pharmacol 9: 65-73.

  13. ↵
    1. Blum K,
    2. Werner T,
    3. Makwara S,
    4. Zvikwata P,
    5. Bowirrat A,
    6. Giordano J,
    7. Oscar-Berman M,
    8. Ndarama M

    (2012) Kuita zvepabonde, zvinodhaka, uye dombo 'n' roll: kufungidzira kufanana kwemaziso anombic kushandiswa sekushanda kwemubairo wezvinyorwa zvepamuviri. J Psychoactive Drugs 44: 38-55.

  14. ↵
    1. Chu NN,
    2. Zuo YF,
    3. Meng L,
    4. Lee DY,
    5. Han JS,
    6. Cui CL

    (2007) Purogiramu yemagetsi yekusimudzira yakashandura kushandiswa kwesero kwekamuri uye kuwedzerwa kweBDNF muzinga re ventral tegmental munharaunda dzisingagumi dzoropa-morphine. Ubongo Res 1182: 90-98.

  15. ↵
    1. Davis BA,
    2. Fitzgerald ME,
    3. Brown JL,
    4. Amstalden KA,
    5. Coolen LM

    (2007) Kushanda kwePOMC neurons panguva yakasununguka yakawanda asi kwete kuita zvepabonde mumakumbo evarume. Behav Neurosci 121: 1012-1022.

  16. ↵
    1. Fiorino DF,
    2. Coury A,
    3. Phillips AG

    (1997) Dynamic changes in nucleus accumbens dopamine efflux panguva yeCoolidge effect muvarume rats. J Neurosci 17: 4849-4855.

  17. ↵
    1. Flagel SB,
    2. Clark JJ,
    3. Robinson TE,
    4. Mayo L,
    5. Czuj A,
    6. Willuhn I,
    7. Akers CA,
    8. Clinton SM,
    9. Phillips PE,
    10. Akil H

    (2011) Basa rinosarudzira dopamine mukumutsa-mubayiro kudzidza. muzvarirwo 469: 53-57.

  18. ↵
    1. Frohmader KS,
    2. Pitchers KK,
    3. Balfour ME,
    4. Coolen LM

    (2010a) Kusanganisa mafaro: kuongororwa kwemigumisiro yemishonga yekuita zvepabonde kune vanhu nemhuka dzemhuka. Horm Behav 58: 149-162.

  19. ↵
    1. Frohmader KS,
    2. Wiskerke J,
    3. Wane RA,
    4. Lehman MN,
    5. Coolen LM

    (2010b) Methamphetamine inobata pamagetsi evanhu vanoita zvepabonde muvarume. Neuroscience 166: 771-784.

  20. ↵
    1. Harris AC,
    2. Gewirtz JC

    (2004) Nyeredzi yakakwirira panguva yekurega kubva mumuviri wakaoma morphine: muenzaniso weipiate kubvisa uye kuzvidya mwoyo. Psychopharmacology 171: 140-147.

  21. ↵
    1. Harris GC,
    2. Aston-Jones G

    (2003) Kukurudzira uye kuidzidzira kwakapfurikidza nekutevera opiate kurega: uchapupu hwekuwedzerwa kwenguva refu kwemubairo kugadziriswa. Neuropsychopharmacology 28: 865-871.

  22. ↵
    1. Hayward MD,
    2. Pintar JE,
    3. Low MJ

    (2002) Kukanganiswa kwemubairo muzvimiti zvinoshaya beta-endorphin uye enkephalin. J Neurosci 22: 8251-8258.

  23. ↵
    1. Hoebel BG,
    2. Avena NM,
    3. Bocarsly ME,
    4. Rada P

    (2009) Utachiona hwemhuka: chimiro chekuzvibata uye chekufambisa chinokonzerwa nehutano hwekusvuta mumakonzo. J Mhirizhonga Med 3: 33-41.

  24. ↵
    1. Hyman SE,
    2. Malenka RC,
    3. Nestler EJ

    (2006) Neural nzira dzokunyadzisa: basa rekuwana mubayiro-uye chiyeuchidzo. Annu Rev Neurosci 29: 565-598.

  25. ↵
    1. Ikemoto S,
    2. Kohl RR,
    3. McBride WJ

    (1997) GABA (A) gadziriro inogadziriswa mumhepo yekare inonzi ventral tegmental inowedzera extracellular mazinga e dopamine mu nucleus accumbens yemakonzo. J Neurochem 69: 137-143.

  26. ↵
    1. Jaworski J,
    2. Spangler S,
    3. Seeburg DP,
    4. Hoogenraad CC,
    5. Sheng M

    (2005) Kudzora dendritic arborization ne phosphoinositide-3'-kinase-Akt-mammalian chinangwa che rapamycin nzira. J Neurosci 25: 11300-11312.

  27. ↵
    1. Johnson SW,
    2. North RA

    (1992) Opioids inokurudzira dopamine neurons ne hyperpolarization yemainterior emunharaunda. J Neurosci 12: 483-488.

  28. ↵
    1. Klitenick MA,
    2. DeWitte P,
    3. Kalivas PW

    (1992) Mutemo we somatodendritic dopamine kubudiswa mu ventral tegmental nzvimbo ne opioid uye GABA: in vivo microdialysis kudzidza. J Neurosci 12: 2623-2632.

  29. ↵
    1. Koob GF,
    2. Le Moal M

    (2005) Plasticity yemubairo neurocircuitry uye "rutivi rwenyama" rwezvinodhaka zvinodhaka. Nat Neurosci 8: 1442-1444.

  30. ↵
    1. Lennette DA

    (1978) Shanduro yakanyatsosimbiswa yeumunofluorescence microscopy. Am J Clin Pathol 69: 647-648.

  31. ↵
    1. Lett BT,
    2. Ipai VL,
    3. Koh MT

    (2001) Naloxone inogadzirisa nzvimbo inofungidzirwa nzvimbo inokonzerwa nefudu inoshandisa rats. Physiol Behav 72: 355-358.

  32. ↵
    1. Lett BT,
    2. Ipai VL,
    3. Koh MT,
    4. Flynn G

    (2002) Kusati kwambova neruzivo nemhepo inotungira magetsi inobereka kuvhiringidzwa kwepamuchinjiko kumigumisiro inobatsira ye morphine. Pharmacol Biochem Behav 72: 101-105.

  33. ↵
    1. Matthews RT,
    2. German DC

    (1984) Uchapupu hwemagetsi ehupupuriro hwekukurudzirwa kwechikwata chechirwere chinonzi dopamine neurons ne morphine. Neuroscience 11: 617-625.

  34. ↵
    1. Mazei-Robison MS,
    2. Koo JW,
    3. Friedman AK,
    4. Lansink CS,
    5. Robison AJ,
    6. Vinish M,
    7. Krishnan V,
    8. Kim S,
    9. Siuta MA,
    10. Galli A,
    11. Niswender KD,
    12. Appasani R,
    13. Horvath MC,
    14. Neve RL,
    15. Worley PF,
    16. Snyder SH,
    17. Hurd YL,
    18. Cheer JF,
    19. Han MH,
    20. Russo SJ,
    21. and unto.

    (2011) Basa rokutaura kweMTOR uye neuronal mu morphine-induced adapations in ventral tegmental area dopamine neurons. Neuron 72: 977-990.

  35. ↵
    1. Mazei-Robison MS,
    2. Nestler EJ

    (2012) Opiate-induced molecular uye cellular plasticity ye ventral tegmental nzvimbo uye locus coeruleus catecholamine neurons. Cold Spring Harb Perspect Med 2: a012070.

  36. ↵
    1. Mehrara BJ,
    2. Baum MJ

    (1990) Naloxone inokanganisa kutaura asi kwete kuwanikwa nemajuku evarume vane nzvimbo yakagadziriswa nzvimbo inofanirwa kugadziriswa nevakadzi vane hupfumi. Psychopharmacology 101: 118-125.

  37. ↵
    1. Meisel RL,
    2. Mullins AJ

    (2006) Chiitiko chepabonde mumakumbo emadzimai: nzira dzemagetsi nemigumisiro inoshanda. Ubongo Res 1126: 56-65.

  38. ↵
    1. Miller RL,
    2. Baum MJ

    (1987) Naloxone inhibits kukwirirana uye inogadziriswa nzvimbo inofanirwa kune an estrouskadzi muvarume rats munguva pfupi mushure mekusimudzira. Pharmacol Biochem Behav 26: 781-789.

  39. ↵
    1. Mitchell JB,
    2. Stewart J

    (1990) Kukurudzira maitiro ekuita zvepabonde mumakumbo ehurume anokonzerwa nemajoka e-intra-VTA yeapiates. Pharmacol Biochem Behav 35: 643-650.

  40. ↵
    1. Murphy LO,
    2. Blenis J

    (2006) MAPK zviratidzo zvakananga: nzvimbo yakakodzera panguva yakakodzera. Trends Biochem Sci 31: 268-275.

  41. ↵
    1. Nestler EJ

    (2012) Nzira dzekushandura zvinodhaka. Clin Psychopharmacol Neurosci 10: 136-143.

  42. ↵
  • ↵
    1. Paxinos G,
    2. Watson C

    (2013) Uropi hwehutachiona mumakoronga e stereotaxic (Academic, Boston), Ed 7.

  • ↵
    1. Pitchers KK,
    2. Balfour ME,
    3. Lehman MN,
    4. Richtand NM,
    5. Yu L,
    6. Coolen LM

    (2010a) Neuroplasticity muonolimbic system inokonzerwa nemubairo wepanyama uye inotevera mubayiro wekudzivisa. Biol Psychiatry 67: 872-879.

  • ↵
    1. Pitchers KK,
    2. Frohmader KS,
    3. Vialou V,
    4. Mouzon E,
    5. Nestler EJ,
    6. Lehman MN,
    7. Coolen LM

    (2010b) DeltaFosB mu nucleus accumbens inokosha pakusimbisa migumisiro yehupombwe. Genesinese Bongo Behav 9: 831-840.

  • ↵
    1. Pitchers KK,
    2. Schmid S,
    3. Di Sebastiano AR,
    4. Wang X,
    5. Laviolette SR,
    6. Lehman MN,
    7. Coolen LM

    (2012) Mubairo wezvakanaka wezvakanaka unoshandura AMPA uye NMDA kuparidzirwa kwekugamuchira uye basa mumusasa wekuunganidza. PLoS One 7: e34700.

  • ↵
    1. Pitchers KK,
    2. Vialou V,
    3. Nestler EJ,
    4. Laviolette SR,
    5. Lehman MN,
    6. Coolen LM

    (2013) Makomborero ezvakanaka uye emishonga anobata pane zvinowanikwa neural plasticity nzira ne DeltaFosB senhengo inokosha. J Neurosci 33: 3434-3442.

  • ↵
    1. Roux PP,
    2. Blenis J

    (2004) ERK uye p38 MAPK-yakagadzirwa protein kinases: mhuri yemapurotini kinases nemabasa akasiyana-siyana ehupenyu. Microbiol Mol Biol Rev 68: 320-344.

  • ↵
    1. Russo SJ,
    2. Bolanos CA,
    3. Theobald DE,
    4. DeCarolis NA,
    5. Renthal W,
    6. Kumar A,
    7. Winstanley CA,
    8. Renthal NE,
    9. Wiley MD,
    10. Self DW,
    11. Russell DS,
    12. Neve RL,
    13. Eisch AJ,
    14. Nestler EJ

    (2007) IRS2-Akt nzira in midbrain dopamine neurons inoronga maitiro ekuzvibata uye magetsi kune maopiates. Nat Neurosci 10: 93-99.

  • ↵
    1. Schultz W

    (2010) Mitambo yakawanda ye dopamine neurons. F1000 Biol Rep 2: 2.

  • ↵
    1. Seroogy KB,
    2. Gall CM

    (1993) Tsanangudzo yeurourotrophins nekati midzi dopaminergic neurons. Exp Neurol 124: 119-128.

  • ↵
    1. Shippenberg TS,
    2. Bals-Kubik R,
    3. Herz A

    (1987) Kukurudzira zvinhu zveipioids: uchapupu hwokuti kushandiswa kwema delta-receptors kunobatanidza kusimbisa nzira. Ubongo Res 436: 234-239.

  • ↵
    1. Sklair-Tavron L,
    2. Nestler EJ

    (1995) Zvinopesana nezvinokonzerwa ne morphine uye neurotrophins, NT-3, NT-4, uye BDNF, pane locus coeruleus neurons in vitro. Ubongo Res 702: 117-125.

  • ↵
    1. Sklair-Tavron L,
    2. Shi WX,
    3. Lane SB,
    4. Harris HW,
    5. Bunney BS,
    6. Nestler EJ

    (1996) Chronic morphine inobatsira kuchinja kuoneka mu morphology ye mesolimbic dopamine neurons. Proc Natl Acad Sci USA 93: 11202-11207.

  • ↵
    1. Spiga S,
    2. Serra GP,
    3. Puddu MC,
    4. Foddai M,
    5. Diana M

    (2003) Morphine kubvisa-kunokonzera kusagadzikana muVTA: kuvhiringidzika kwema-scanning microscopy. Eur J Neurosci 17: 605-612.

  • ↵
    1. Swanson LW

    (2004) Mamepu ehutano: chimiro chehuropi hwepachi (Chikoro, San Diego), Ed 3.

  • ↵
    1. Tenk CM,
    2. Wilson H,
    3. Zhang Q,
    4. Pitchers KK,
    5. Coolen LM

    (2009) Mubairo wepabonde muvarume vhudzi: migumisiro yehutano hwepabonde pamamiriro ezvinhu anofanirwa kunzvimbo inobatanidza ne ejaculation uye inopromission. Horm Behav 55: 93-97.

  • ↵
    1. Tzschentke TM

    (2007) Muyero wekuenzanisa nemamiriro ezvinhu anofanirwa (CPP) paradigm: update yemakore gumi apfuura. Mupombwe Biol 12: 227-462.

  • ↵
    1. Van Furth WR,
    2. van Ree JM

    (1996) Kukurudzira zvepabonde: kubatanidzwa kweapioid inogumira mu ventral tegmental nzvimbo. Ubongo Res 729: 20-28.

  • ↵
    1. Van Furth WR,
    2. Wolterink G,
    3. van Ree JM

    (1995) Mutemo wehutano hwehupombwe: kubatanidzwa kweuropi opioid uye dopamine. Ubongo Res Brain Rev Rev 21: 162-184.

  • ↵
    1. Walker JR,
    2. Chen SA,
    3. Moffitt H,
    4. Inturrisi CE,
    5. Koob GF

    (2003) Kuratidzwa kweiorioid inogara kwenguva refu inobereka heroin yakawedzera kuzvitonga mumakonzo. Pharmacol Biochem Behav 75: 349-354.

  • ↵
    1. Young KA,
    2. Gobrogge KL,
    3. Liu Y,
    4. Wang Z

    (2011) Inonzi neurobiology yekubatana pamwe chete: mazano kubva kumudzimai wemudzimai mumwe chete. Front Neuroendocrinol 32: 53-69.

    •