Tsela e fokolang ea neuronal inhibition e senola mesebetsi e khahlanong le tsela e sa tobang le e tobileng ea ho matlafatsa (2011)

Nat Neurosci. 2011 Pherekhong; 14 (1): 22-24.

E phatlalalitsoe marang-rang 2010 December 5. Doi:  10.1038 / nn.2703

PMCID: PMC3058296
NIHMSID: NIHMS245934
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Dorsal striatum e bapala karolo ea bohlokoa ho nts'etsopele ea bokhoba ba lithethefatsi; leha ho le joalo, kutlwisiso e nepahetseng ea likarolo tsa striatopallidal (eseng ka kotloloho) le striatonigral (Direct) Tsela li-neurons mabapi le ho laola boits'oaro e lula e le thata.

Ho sebelisa mokhoa oa ho ngola o itšetleha ka polelo e mahareng ea vaerase ea GPCR e nang le enjine4D), re bonts'itse ts'ebetso ea hM4Li-receptors tse nang le clozapine-N-oxygen (CNO) e fokotsitse thabo ea striatal neuron. Ha hM4Li-receptors tsa D li ile tsa hlahisoa ka mokhoa o khethiloeng kapa ka tsela e sa tobang kapa e sa otlolohileng ea methapo ea kutlo litsing, CNO ha ea ka ea fetola likarabo tse mpe tsa locomotor ho amphetamine empa liphetoho tsa boitšoaro tsa polasetiki tse fetotsoeng tse amanang le kalafo e sa khaotseng ea lithethefatsi. Ka ho khetheha, ho senya ts'ebetso ea nakoana ea "striatopallidal neuronal" e nolofalletsang boits'oaro ba boits'oaro nakong eo ho fokotsang thabo ea "striatonigral neurons" ho fokotseng phehello ea eona. Liphumano tsena li fana ka maikutlo a hore litlamorao tse matla tsa lithethefatsi li ka fokotsoa ho tsoa ho boits'oaro bo amanang le tšebeliso ea lithethefatsi khafetsa 'me ba totobatsa ts'ebeliso ea mokhoa ona oa ho fana ka litlatsetso tsa methapo ea methapo methapong ea boitšoaro bo kang maikutlong.

Leha ho na le litlamorao tse mpe tse tlisoang ke ho lemalla lithethefatsi, tšebeliso ea maikutlo le tlhekefetso e ntse e atile. Keketseho ho tloha ho pepesehang lithethefatsi ho tloha tšebelisong ea mehla ea lithethefatsi le ts'ebeliso ea eona ea ho qobelloa, ho tloaela tloaelo le tahlehelo ea taolo e thibelang maikutlo ho kenyelletsa letoto la liphetoho tsa limolek'hule liseleng tsa methapo ea kutlo1,2,3. Striatum e khethiloe e le sebaka sa bohlokoa bakeng sa boits'oaro bo bongata ba boitšoaro le ba neurobiological bo nahanoang e le lits'ebetso tsa mantlha tse tlohelang bokhoba ba tahi1,2,3. Boholo ba li-neuron ka har'a striatum (~ 95%) ke li-neurons tsa "" "" methane ea methapo ea methapo ea methapo ea methapo ea methapo ea mmele (GN). tHat e fapana ho polelo ea bona ea neuropeptide mme ba theha litsela tse peli tse kholo tsa katleho4.

  • Li-MSN tsa Striatopallidal li na enkephalin (ENK) mme li theha tsela e sa tsejoeng
  • athe striatonigral MSNs li na le dynorphin (DYN) le ntho P mme li theha tsela e otlolohileng.

Mefuta e mengata ea khopolo-taba e kholo ea hore lipalo tsena tsa li-MSN li hanyetsana ka mokhoa o sebetsang le ka tšebetso5,6. Leha ho le joalo, ha ho na bopaki bo matla ba ho tšehetsa karolo ea bona e fapaneng taolong ea boits'oaro hobane lipalo tsena tsa sele li kopantsoe 'meleng ebile li sa hlophisehe ho etsa hore ho be bonolo ho khetha ka bolotsana.

Ho hlahloba karolo ea lipalo tsa lisele tsena tse nang le bokhoni ho ntlafatseng mekhoa e latelang ho pepesetsoa lithethefatsi tsa tlhekefetso, re kopantse merero e 'meli ea li-virus: Barekisi ba mofuta oa vaerase ba sebelisang mofuta oa ENK kapa DYN ho lebisa polelo ea transgene ho "striatopallidal kapa striatonigral neurons" , ka ho latellana, le GPCR ea enjine (Gi / o-motsoana oa batho oa mokokotlo oa M4 QETELLO; Moqapi Receptor O Khethisitsoe ka Tsela e Ikhethileng ke Moqapi oa Lithethefatsi; hM4D)7 e hlahisoang ke motsoako o mong oa litheko tsa merianaN-seli8,9 (CNO; Feiga. 1a, h). Ka mor'a polelo ho li-neuron tse tlotsitsoeng, tsamaiso ea CNO e tsosa Gi / o-motsoana oa hM4Li-receptors tsa D kahoo li kenya tšebetsong mekhoa ea kahare ea potasiamo 3 (Kir3), e bakang hyperpolarization le membrane ea nakoana ea neuronal9.

Setšoantšo sa 1  

Thomello ea nakoana le e lebisitsoeng ea ho emisa lisele tsa striatal. (a, h) Limmapa tsa Amplicon tsa leqENK-hM4D/pENK-GFP (a) le leqDYN-hM4D/pDYN-GFP (h) tse shebileng. (b, ke) Microscopy ea "confocal" e bonts'itse hore pENK-hM4D li-receptors li ile tsa hlahisoa ka mokhoa o ikhethileng ho striatopallidal ...

Ho etsa liteko bakeng sa tlhahlobo ea seleng ea ve viral, re sebelisitse li-microscopy tse peli tse hlahang kamora ho kenella ka likokoana-hloko tsa dorsal striatum.Tlhahiso e eketsehileng Fig. 1) HS ea hemagglutinin-tagged hM4Li-receptors tse tlas'a taolo ea "ENK Promoter" (leqENK-hM4D) kapa morekisi oa DYN (leqDYN-hM4D). Re fumane hore leqENK-hM4D polelo e ne e le haholo ka har'a lisele tse nang le li-ENN (90% ea lisele tsa hemagglutinin e ne e le ENK +, 85 kantle ho 94; 6% ea lisele tsa hemagglutinin e ne e le ntho ea P +, 4 liseleng tsa 70; Feiga. 1b) leqDYN-hM4D polelo e ne e le ntho ea mantlha P-e nang le li-MSNs (95% ea lisele tsa hemagglutinin e ne e le ntho P +, 109 tsoa lisele tsa 115; 5% ea lisele tsa hemagglutinin e ne e le ENK +, 5 tsoa lisele tsa 97; Feiga. 1i). Liphetho tse tšoanang li fumanoe kamora ho kenngoa ha livaerase tse ikhethang tse hlalosang protheine e tala ea fluorescent (leqENK-GFP leqDYN-GFP; Lisebelisoa tsa tlatsetso. 2a le 3a).

Ka lebaka la hore striatopallidal MSNs e qala morero oa li-globus pallidus kantle (GPe) le li-striatonigral MSNs ka ho khetheha morerong oa proantia nigra pars reticulata (SNpr), re sebelisitse li-injection tsa "troger tracer" Fluoro-Khauta libakeng tsena tsa boko tse lateloang ke habeli-label fluorescent immunohistochem. ho tiisa hore leqSINGLE leqDYN vaerase e hlahisitse tšoaetso e khethehileng ea tseleng. Re hlokometse hore leqENK-GFP lisele li hokahantsoe le sebaka sa poleloana e khutsitseng ea Fluoro-Khauta kamora ho kenella ho GPe empa eseng SNpr (Tlatsetso ea feiga. 2b) leqDYN-GFP lisele li hokahanngoa le sebaka sa poleloana e khutsitseng ea Fluoro-Khauta e latelang infusions ho SNpr empa eseng GPe (Tlatsetso ea feiga. 3b). Ponaletso ea vaerase ea vaerase ha e fetole palo ea ENK + kapa ntho ea P + neurons sebakeng sa ts'oaetso ea vaerase, ho fana ka maikutlo a hore tšebeliso ea bahlahisi bana ba phetisetso ea bongaka e sa sebetseng e sa kena-kenane le maemo a "neuropeptide" a emeng. Liphetho tsena tsohle li bonts'a hore leqSINGLE leqDYN livaerase tsa vaerase li hlahisa mefuta ea tsona e arohaneng ka tsela e nepahetseng ea lisele.

Leha hM4Mekhoa ea D receptor-based e bontšitsoe ho fetolela tšebetso ea mefuta e meng ea neuronal9, bokhoni ba tsona ba ho ama li-neuron tsa striatal ha li so hlahlojoe. Ka hona, re tšoaelitse methapo ea kutlo ea "" striatal medium spiny neuron "e nang le hM4Li-receptors tsa D tse laoloang ke morekisi oa herpes simplex (HSV), o ile a lokisa likhechana tsa coronal tsa dorsal striatum matsatsi a mabeli hamorao, mme a hlahloba hore na hM4Li-neurons tse pakeng tsa D le e hlalosang lipakeng tsa "spinyatri" li arabela CNO. Re hlokometse hore ts'ebeliso ea lehae ea CNO (10 μM) e ile ea baka hyperpolarization ea bokhoni ba membrane (~ 7 mV, bokhoni ba membrane ea motheo bo fetotsoe ho −80 mV; Feiga. 1c) le ho fokotsa ho hanyetsa ha li-neurons ka mor'a kopo ea CNO (Feiga. 1d, e), ho fana ka tlhahiso ea hore boitšoaro ba potasiamo (ke hore, Kir3-mediated hona joale) e qaliloe ke CNO ho hM4D methapo e hlalosang li-receptor. Ntle le moo, ho fokotsoa ha CNO haholo ho fokolitse palo ea menyetla ea ts'ebetso ea hoke4Li-neuron tse hlahisoang ke D, empa eseng ho lisele tse laolang, ka tsela eo li thibela ka nepo ts'ebetso ea li-neuron tse tšoaelitsoeng ke vaerase Tlhaloso ea hM4D receptors feela ha e a ka ea fetola khanyetsano ea karohano (P = 0.84) kapa ho bolaea motho ka mokhoa o ka u thusangP = 0.64). ((Feiga. 1f, g). Ha e kopantsoe hammoho, lintlha tsena li fana ka maikutlo a hore, e ts'oanang le li-neurons tsa hippocampal9, hM4Mokhoa o thehiloeng ho D / CNO o ka fokotsa thabo ea methapo ea kutlo ea "rat striatal neurons".

E le bopaki bo eketsehileng ba mohopolo, re ile ra leka hore na hM4Li-receptors tsa D li ka thibela li-neurotransication potolohong e tsitsitseng eo ho eona tšebetso ea neural e ts'oaroang ke ts'usumetso e amanang le boits'oaro. Ka lebaka leo, re tšoaelitse li-neuron tsa sebakeng sa moea (VTA) tse nang le hM4Li-receptors tsa D tse tlasa taolo ea papatso ea HSV, e bonts'a ka matla li-dopamine neurons10, hape e sebelisitse "cyclic voltammetry" e potlakisang sekhahla ho lekola liphetoho tlatsong ea dopamine ka har'a li-nucleus ka mor'a ho tlisa moputso o sa lebelloang oa moputso oa lijo11. Tsamaiso ea CNO e fumane ka mokhoa o phollotsoeng lijo tsa pellet-evoke dopamine ka har'a li-bokellase tsa nucleus ha li bapisoa le koloi (Tlhahiso e eketsehileng Fig. 4). Kamora nako, re ile ra etsa liteko tsa hore na ho fokotseha hoa mefuta ea lisele tse itseng tsa methapo ea methapo ho striatum Ka vivo e ka fetola bokhoni ba amphetamine ho tsosa polelo ea Fos. Psychostimulants, joalo ka amphetamine, ke li-activator tse matla tsa c-fos striatum12 e tla eketsa c-fos ka bobeli striatonigral le striatopallidal neurons tlasa maemo a rona a liteko13. Ntle le ts'ebeliso ea eona e le lets'oao la tšebetso ea li-neuronal, psychostimulant induction ea c-fos ho nahanoa hore e bapala karolo ea bohlokoa ho qaleng le ho boloka mekhoa ea tlhaho e amanang le psychomotor sensitization1,14. Re fumane hore ts'ebetso ea CNO-mediated activation ea pENK-hM4D li-receptors li fokotsehile haholo palo eohle ea lisele tsa amphetamine-ikiwa tsa c-Fos striatum (Feiga. 1k 'me Tlatsetso ea feiga. 5a). Phokotso ena e etsahetse ka li-neuron tse peli tsa bohlokoa tsa hemagglutinin (ke hore, tse hlalosang hM4D receptors) le hemagglutinin-negative neurons (ke hore, tse sa hlahiseng hM4Li-receptors; Feiga. 1l), ho fana ka maikutlo a phello ea puo ea methapo e pakeng tsa hM4Li-neuron tse hlalosang li-D le li-neuron tse sa bonoeng. Phokotso ea bohlokoa palong eohle ea lisele tsa amphetamine-evoke c-Fos le palong ea lisele tsa hemagglutinin-c-Fos le tsona li ile tsa bonoa ha hM4Li-receptor tsa D li ile tsa kenngwa tšebetsong tse otlolohileng tsa tsela e otlolohileng (Feiga. 1n, o 'me Tlatsetso ea feiga. 5b). Habohlokoa, litlamorao tsena ha li bakoe feela ke polelo ea vaerase ea receptor e ncha ho tloha ka polelo ea e ka ba ea leqENK-hM4D kapa leqDYN-hM4D li-receptors ha ho se na phekolo ea CNO ha li na ho ama palo ea lisele tsa amphetamine-evoked Fos (Likokoana-hloko tse tlatsetsang. 6 le 7). Kahoo, liphumano tsena li bonts'a ts'ebetso ea CNO-mediated activation ea hM4Li-receptors tsa D le tsona li ka lebisa ho fokotseha ha ts'ebetso ea neuronal ka ho fokotsa ho lokolloa ha li-neurotransmitter le ho bonts'a matšoao a kenang ka hare.

Ho pepesetsoa khafetsa ho lithethefatsi tse lemalloang ho ka lebisa ho eketseheng ho tsoelang pele le boiphetetso ba karabelo ea boits'oaro, eo hangata ho thoeng ke tšusumetso ea boitšoaro. Habohlokoa le ho feta, maikutlo a kenyelletsang a mang a lipotoloho tse tšoanang tsa neural tse kentsoeng kholisong ea bokhoba ba lithethefatsi tsa batho3. Mona re sebelisa lisebelisoa tsa rona tsa nalane ho fuputsa phello ea ho nyahamisoa ka ho khetheha ha tšebetso ea methapo mabapi le nts'etsopele ea maikutlo a amphetamine. Re kholoa hore li-neurons tsa tsela e tobileng le e sa tobang li na le likarolo tse hanyetsanang moo li-neurons tsa striatonigral li hatelletsang maikutlo le methapo ea kutlo ea striatopallidal e hatellang maikutlo, e lumellanang le likarolo tsa bona tse reriloeng ka kotlolloho ea ts'ebetso ea ts'ebetso le ts'oaetso ka ho latellana.5,6. Ka lebaka leo, re ile ra leka hore na ebe li-neurons tse fokotsang methapo ea kutlo li ka ba le tšusumetso e matla ho maikutlo a amphetamine-dosing a emisang boemo ba maikutlo le hore na ho fokotseha hoa thabo ea neuronal ea "striatonigral neurons" ho ka thibela sensitization ho protocol eo ka tloaelo e hlahisang maikutlo a matla a matla.

Thutong ea pele, re sebelisitse mofuta oa kalafo o kenyelletsang tekanyo ea maikutlo a locomotor taolong ea GFP (litlatsetso tse 'ne tsa lithethefatsi). Kamora nako ea ho tlohela, ho ile ha fanoa ka phephetso e leka-lekaneng ea amphetamine (2 mg / kg) kamora ho ba sieo ha CNO e kentsoeng ts'ebetso ea ts'ebetso ea methapo ea methapo ho bona hore na sensitization e ne e phehella. CNO-mediated ts'ebetso ea pENK-hM4D li-receptors nakong ea kalafo ea amphetamine ha lia ka tsa fetola karabelo e matla ea locomotor ho amphetamine (Setšoantšo sa 2a). Leha ho le joalo, ts'ebetso ea CNO e kopantsoeng le ts'ebetso ea methapo ea methapo ea methapo methapong e sa tsamaelaneng e ile ea nolofalletsa nts'etsopele ea matla a matla a matla ha a bapisoa le taolo ea GFP (Feiga. 2b). Ntlafatso ena ea maikutlo e ile ea bolokoa ho phephetso ea amphetamine, e ileng ea etsoa beke e le 'ngoe hamorao kamora ho hloka kalafo ea CNO (Feiga. 2c, d). Litlamorao tsena li ka hlahisoa ke phokotso e itšetlehileng ka CNO ea tšebetso ea li-neuron tsa striatopallidal ka hM4D receptors hobane hM4Polelo ea D receptor ntle le kalafo ea CNO ha e hlahise maikutlo a locomotor ho regimen ena e bonolo ea amphetamine (Tlhahiso e eketsehileng Fig. 6).

Setšoantšo sa 2  

Ho fokotsa butle-butle thabo ea "striatopallidal kapa striatonigral neurons" ho ne ho e-na le litlamorao tse khahlanong le maikutlo a amphetamine. (a, e) Likarabo tse sebetsang tsa locomotor ho amphetamine ho latela ts'ebetso ea ts'ebetso ea CNO e hlahisitsoeng ke pENK-hM4D (a) le leqDYN-hM4D (e) li-receptors. ...

Bakeng sa ho tseba hore na li-neuron tsa striatonigral li ka laola ho arabela ha lithethefatsi ka mokhoa o hanyetsanang, ka mor'a moo re ile ra leka phello ea biochemically fokotsa thabo ea striatonigral neurons ho dorsomedial striatum nakong ea phekolo ea lithethefatsi tsa kalafo ea amphetamine e hlahisang maikutlo a matla ho taolo ea GFP (litheko tse ts'eletseng tsa lithethefatsi ) hammoho le nakong ea phephetso e tlase ea amphetamine (0.5 mg / kg) moo ho se nang pherekano ea tšebetso ea methapo ea methapo. Joalo ka ho fokotseha ha tsela e sa sebetseng, CNO-Mediated e fokotseha ka thabo ea methapo ea kutlo e otlolohileng nakong ea kalafo ea amphetamine ha e fetole karabelo e matla ea locomotor ho amphetamine (Feiga. 2e). Leha nts'etsopele ea sensitization e hlahile e ts'oanang le lits'ebetso tsa GFP tse latelang ts'ebetso ea ts'usumetso ea CNO ea pDYN-hM4D li-receptors nakong ea karolo ea kalafo (Feiga. 2f), sensitization ha ea ka ea phehella ho pDYN-hM4D sehlopha empa se ntse se bolokiloe ka matla taolong ea GFP (Feiga. 2g, h). Litlamorao tsena li ka hlahisoa ke ho fokotseha ho itšetlehileng ka CNO ha ts'ebetso ea li-neuron tsa striatonigral ke hM4D receptors hobane hM4Polelo ea "receptor" ea ho se sebetse ha kalafo ea CNO ha e a ka ea thibela nts'etsopele ea sensomization ea sensomotor kaha sensitization e ne e bonoa nakong ea karolo ea kalafo le phephetsong ea lithethefatsi (Tlhahiso e eketsehileng Fig. 7). Lintlha tsena li fana ka maikutlo a hore li-neuron tsa striatonigral li ka ba tsa bohlokoa haholo ho laoleng liphetoho tse amanang le boitšoaro tsa nako e telele e le litholoana tsa tšebeliso e mpe ea lithethefatsi.

Ka ho phethela, datha tsena li fana ka bopaki ba pele bakeng sa li-neurons tse mahlonoko le tse hanyetsanang taolong ea ts'ebetso ea ts'ebeliso ea ts'ebeliso ea lithethefatsi mabapi le ts'ebeliso ea lithethefatsi.. Ntle le moo, ho haella ha phello ea phetisetso ea neuronal ho karabelo e matla ea locomotor ho amphetamine ho fana ka bopaki bo bong ba hore methapo e laolang karabelo e matla ea lithethefatsi e fapane le e fetolang litekanyetso tse mamellang tse etsahalang ka ho pepeseha hoa lithethefatsi khafetsa. Kamora nako, ho roala li-virar tse khethehileng tsa virot tse ikhethang tse nang le li-receptor tse nang le bokhoni ba ho fetola tšebetso ea methapo ntle le ho senya ts'ebetso ea sele e fana ka mokhoa o mocha le o matla oa ho theha motheo oa ts'ebetso ea molek'hule.

Boitsebiso bo Eketsehileng

lumela hore baa fokola

Mosebetsi ona o ne o tšehelitsoe ke lithuso tsa NIH K99 DA024762 (SMF), T32 GM07266 le T32 GM07108 (DE), T32 AA009455 le F32 DA026273 (MJW), R21 DA021793 (PBPX) NXXUMX (X01) ), le R023206 DA19 (JFN), Meputso ea Katleho bakeng sa Bo-rasaense ba Boithuto (DE) le Moputso oa Boipheliso oa NarsAD oa NARSAD

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Phatlalatso ea lithahasello

Bangoli ba phatlalatsa hore ha ba na thahasello ea tlholisano ea lichelete.

References

1. Berke JD, Hyman SE. Neuron. 2000; 25: 515-532. [E fetotsoe]
2. Neslter EJ. Nat Rev Neurosci. 2001; 2: 119-128. [E fetotsoe]
3. Robinson TE, Berridge KC. Ho lemalla. 2001; 96: 103-114. [E fetotsoe]
4. Smith Y, Bevan MD, Shink E, Bolam JP. Neuroscience. 1998; 86: 353-387. [E fetotsoe]
5. Shen W, Flajolet M, Greengard P, Surmeier DJ. Mahlale. 2008; 321: 848-851. [Tlhahiso ea mahala ea PMC] [E fetotsoe]
6. Durieux PF, et al. Nat Neurosci. 2009; 12: 393-395. [E fetotsoe]
7. Conklin BR, et al. Mekhoa ea Nat. 2008; 5: 673-678. [Tlhahiso ea mahala ea PMC] [E fetotsoe]
8. Alexander GM, et al. Neuron. 2009; 63: 27-39. [Tlhahiso ea mahala ea PMC] [E fetotsoe]
9. Armbruster BN, Li X, Pausch MJ, Herlitze S, Roth BL. Proc Natl Acad Sci USA. 2007; 104: 5163-5168. [Tlhahiso ea mahala ea PMC] [E fetotsoe]
10. Choi KH, et al. Ann NY Acad Sci. 2003; 1003: 372-374. [E fetotsoe]
11. Clark JJ, et al. Mekhoa ea Nat. 2010; 7: 126-129. [Tlhahiso ea mahala ea PMC] [E fetotsoe]
12. Harlan RE, Garcia MM. Mol Neurobiol. 1998; 16: 221-267. [E fetotsoe]
13. Badiani A, Oates MM, Day He, Watson SJ, Akil H, Robinson TE. Behav Brain Res 1999; 103: 203-209. [E fetotsoe]
14. Hyman SE, Malenka RC. Nat Rev Neurosci. 2001; 2: 695-703. [E fetotsoe]