Mgbanwe Na - agbanwe Omimi na Nsogbu Na - eme Ka Efflux Dopamine Na - ebute Mgbe Ọkpụkpụ Coolidge Effect na Rats (1997)

1. Dennis F. Fiorino,
2. Ariane Coury, na
3. Anthony G. Phillips

+Gosi njikọ

1. The Journal of Neuroscience, 15 June 1997, 17 (12): 4849-4855;

nkịtị

Nkọwa nke Coolidge na-akọwa ịmalite inwe mmekọahụ na "anụmanụ" nke nwere mmekọahụ "na-enwe mmekọahụ". N'iburu ọrụ nke usoro mesolimbic dopamine (DA) na ntinye na mmezi nke omume mkpali, a na-eji microdialysis mee ihe iji chọpụta ihe dị njọ (NAC) na-enyefe mgbe a na-enye ya ọzụzụ, ịkwa iko, na ịmalite ịkwa iko. N'akwekọrịta na akụkọ ndị gara aga, e gosipụtara nkwalite nke nwanyị na-enyo enyo n'azụ ákwà ngebichi na mmekorita na mmụba dị ịrịba ama na NAC DA efflux. Nloghachi nke NAC na itinye uche na ụkpụrụ nke ihe jikọrọ ya na oge nke mmekọahụ satiety, ọ bụ ezie na concentrations nke DA metabolites, dihydroxyphenylacetic acid na homovanillic acid, ka dị elu. Ngosipụta nke nwanyi nabatara nwanyi n'azụ ihuenyo mere ka enwewanye mmụba na NAC DA, nke e wulitere elu mgbe ọ na-emegharịghachi ya na nwanyị nwanyi. Ihe ngosi nke oge a na-egosi na ihe mkpali nke nwanyị nwere mmasị na ya nwere ike ịbawanye NAC na nnyefe na oke nwoke na-enwe mmekọahụ, na nke a, nwere ike ịba aka na ịmalite ịkpa mmekọahụ.

Okwu Mmalite

AKỤRỤNGWA NA ỤZỌ

Ndị isi. Ụmụ nwoke Sprague Dawley, bụ ndị e nwetara site na Animal Care Center (na Mahadum nke British Columbia), na ụmụ nwanyị Long-Evans, site na Charles River Canada (St. Constant, Quebec, Canada), nọ na ọdụ ụgbọ mmiri (18 × 25 × 65 cm; onu ogugu ise n'ime ulo ndi ozo. A nọgidere na-edebe ụlọ ndị colony na okpomọkụ nke ~20 Celsius C na ngbanwe 12 hr ìhè / ọchịchịrị. Rats enwere ike inweta nri (Purina Rat Chow) na mmiri.

Ịwa ahụ na akparamàgwà omume n'ihu ụbụrụ microdialysis.A na-eji ụmụ irighiri ụmụ nwanyị na-asọ oyi na-egbuke egbuke n'okpuru halothane gas (Anụsịrị, Ayerst Laboratories) ma ọ dịkarịa ala 4 izu tupu ule. A nabatara ụmụ nwanyị na-akpali akpali site na ntanye nke estradiol benzoate (10 μg) na progesterone (500 μg), 48 na 4 hr, n'otu oge, tupu oge nyocha ọ bụla. A na-ekpuchi oke nwa nwoke maka omume mmekọahụ n'oge abụọ, 4 dịpụrụ iche, na ụlọ Plexiglas (35 × 35 × 40 cm) na nkedo ala. Naanị oke oke nke ruru ogo njirimara, nke gụnyere intromission n'ime 5 min nke ngosi nke nwanyi na ejaculation n'ime 15 min nke mbụ intromission, n'oge nyocha abụọ na-etinye na microdialysis ndu ndu cannulae.

Ụmụ oke (n = 5) ka anesthetized na ketamine hydrochloride (100 mg / kg, ip) na xylazine (10 mg / kg, ip) tupu ịwa ahụ. A na-etinye ntanye nduzi nyocha microdialysis (ntinye 19) bilataa NAC (nhazi site na bregma: oghere, + 1.7 mm; medial, ± 1.1 mm; ventral, -1.0 mm; okpokoro okpokoro) ma chekwaa na okpokoro isi na eze achan jeweler si kposara. A na-eji ndị na-eduzi ndị na-eduzi ndị na-eduzi ndị mmadụ na-eme ka ndị mmadụ nwee ohere iji mee ka a chọpụta ihe ịga nke ọma. N'ụzọ dị mma, na nnwale ahụ ugbu a, ọ bụ naanị otu cannula dị mkpa maka oke oke ọ bụla. A na-ejide kịtịkpa ụmụ nwoke n'otu ebe na akwa akwa plastic na bedding nke ọkacob maka ihe fọdụrụ n'ime nnwale ahụ. Otu izu mgbe a gwọchara ya, a nwalere oke maka omume mmekọahụ. N'ime akụkụ ọzụzụ a, ụlọ nyocha ahụ nwere ihe ngosi Plexiglas na-agba agba nke kewara ụlọ ahụ n'ime nnukwu ụlọ na obere. A kpọbatara oke ụmụ nwoke n'ime nnukwu ụlọ ahụ na 15 min mgbe e mesịrị, a na-etinye nwanyị n'azụ ákwà ngebichi. Mgbe oge nhazi 15 min, ewepụre ihuenyo ahụ, a na-ahapụkwa oke ndị ahụ iji dozie maka 30 min. A na-eduzi ọzụzụ ọzụzụ atọ, otu n'ime 4 d. Mkpuru oke niile ruru nyocha njirimara n'oge nnọkọ ọ bụla.

Nkọwapụta mmetụta dị na Coolidge. Ejikọtara okporo osisi na-achọpụta 12-18 hr na -adịghị ihe ọ bụla na-emepụta microdialysis tupu enwee nchọpụta nke Coolidge ma tinye ya na nnukwu ogige nke ụlọ nyocha ahụ na-enweghị ohere na nri na mmiri. N'ụtụtụ nke nnwale ahụ, a na-anakọta ihe ntinye microdialysis na 15 min. Nnwale ahụ gụnyere usoro usoro asaa ndị a: (1) ntọala (ma ọ dịkarịa ala 60 min); (2) nwanyị 1 nwanyị n'azụ ákwà ngebichi (15 min); (3) ị na - ejikọta nwanyi 1 ruo oge oge 30 tinyere ugwu; (4) ntinyeghari nke nwanyi 1 n'azụ ihuenyo (15 min); (5) ịnweta 1 15 maka oge minit 3 nke a na-enyeghị ọbịbịa (ọ bụrụ na ịbịakwute emee, a na-ewere usoro a dị ka oge 6); (2) iwebata nwanyi 15 n'azụ ákwà ngebichi (7 min); 2) ikoghari na nwanyi 60 maka XNUMX min.

A na-esere omume na obere ọkụ site n'iji usoro vidio JVC ma hụ na nyocha vidiyo nke dị n'èzí ụlọ ule. Edere usoro omume nke nwoke na nwanyị na-eji kọmputa na software kwesịrị ekwesị (Holmes et al., 1987).

Mgbe nyocha nke microdialysis, e nyere ụmụ anụmanụ nnụnụ nke chloral hydrate ma jiri saline na formalin (4%) mee ihe na nkwanye. A na-esi na skep na-egbuke egbuke, na, mgbe e mesịrị, ngalaba nke coronal jupụtara na cresyl violet iji chọpụta ntinye nke nyocha microdialysis. Naanị nkedo na nyocha nyocha n'ime NAC ka e ji mee ihe maka nyocha nke omume na nyocha.

Microdialysis na HPLC-nchọpụta electrochemical. Ihe nyocha microdialysis bụ ihe mgbatị na-emepụta akpụkpọ anụ eriri (2 mm akpụkpọ ahụ kpughere, 340 μm nke dị na mpụta, 65000 nke na-ahụ maka ịdị arọ molecular, Filtral 12, Hospal) na njedebe distal. A na-emegharị nyocha na 1.0 μl / min na ngwọta nke Ringer (0.01 m sodium phosphate bu, pH 7.4, 1.3 mmCaCl₂, 3.0 mm KCl, 1.0 mmMgCl₂, 147 mm NaCl) site na iji sringe (Hamilton, Reno, NV) na mgbapụta sirinji (ihe ngosi 22, Harvard Apparatus, South Natick, MA). A na-eji nchịkọta nyocha nke nyocha microedialysis iji nweta nyocha microdialysis n'ime ndu ndu. A na-eji ígwè mgbanaka, nke a na-etinye na swentl (Instech 375s) nke dị n'elu ụlọ nyocha ahụ, iji kpuchie ntaneti nyocha (Fiorino et al., 1993).

Ihe nyocha nke Microdialysate, nke gụnyere DA na metabolites dihyroxyphenylacetic acid (DOPAC) na homovanillic acid (HVA), dị iche iche site na chromatography nke na-agbanwe agbanwe (nke dị na Beckman, Fullerton, CA, 5 μm, 15 cm, 4.6 mm, ) site na iji nchekwa 0.083m sodium acetate, pH 3.5 (5% methanol). Achọpụtara nchọpụta electrochemical (EC) site na nchọpụta nyocha. Ngwa ahụ nwere mgbapụta Bio-Rad (Richmond, CA), ihe eji eme ihe na Valco Instruments (Houston, TX) EC10W abụọ, na ESA (Bedford, MA) Coulochem II EC detector, na onye na-edekọ ihe nkiri abụọ (Kipp na Zonen, Bohemia, NY). Nchọpụta electrochemical nchọpụta bụ ndị a: 1 electrode, + 450 mV; 2 electrode, -300 mV; na nchekwa nche, -450 mV. Ụdị nyochaghachi ihe nyocha, eduzi na vitro na n'ime ụlọ okpomọkụ, bụ 22% maka DA, 18% maka DOPAC, na 18% maka HVA.

IHE

NKWASỊTA

N'akwekọrịta na akụkọ ndị mbụ, ihe ndị na-apụta ugbu a gosipụtara mmelite na-ebuwanye ibu nke na-emetụta ụbụrụ nwoke na nwoke na-enwe mmekọahụ ma na-atụle ya.na vivo microdialysis (Mas et al., 1990; Pfaus et al., 1990;Pleim et al., 1990; Damsma et al., 1992; Wenkstern et al., 1993; Fumero et al., 1994; Mas et al., 1995a,b,d). Tụkwasị na nke a, nsonaazụ ndị a na-enye ihe mgbakụta na-adịghị mma maka mmekorita nke mmekọahụ na mmeghasị nkwado nke ịkpụzi mmiri na nzaghachi nye nwanyị na-anabata akwụkwọ (mmetụta Coolidge). Ihe ngosi nke oge a na-egosi na ihe mkpali nke nwanyi na-anabata nwanyị nwere ike ịmalite iwelie NAC na nnyefe na oke oke nwoke nwere mmekọahụ, nke nwere ike, n'aka nke ya, jikọọ aka na mmekorita nke mmekọahụ. Nke a bụ nke mbụ pụta ìhè na mmụba ntakịrị na NAC na n'oge ngosi nke nwanyi di iche iche n'azụ ihuenyo ma mee ka o doo anya dị ka mmụba ọzọ a na-akpọ mgbe a na-emegharịghachi ya na nwanyị 2 (Fig.1).

Ọnụnọ nke ndị inyom nabatara na azụ na ihuenyo rụpụtara mmụba siri ike na NAC na efflux (44% site na akara ngosi) yiri nke dị na ihe edere na nyocha ndị gara aga site na iji nhazi ahụ (30%, Pfaus et al., 1990; 35%,Damsma et al., 1992). Ọzọkwa na nkwekọrịta na ọmụmụ ihe ndị a bụ nchọpụta na NAC DA efflux na-eme ka ọ dịwanye mma n'oge a na-emegharị (na> 95% n'elu ntọala na nnwale ugbu a). Ọ bụ ezie na anyị nwere ike ịlele akụrụngwa na-eri mmụọ dị ka ihe jikọrọ ya na nkwalite NAC DA (Wenkstern et al., 1993; Wilson et al., 1995), ọ dị mkpa iji nyochaa okwu ahụ bụ "mmetụ" na "ihe eji eme ihe" n'ihe gbasara mmekọahụ. Ọ bụ ezie na oge nwata nwanyị nọ n'azụ ihuenyo ahụ bụ nanị ihe na-agụ agụụ ma ọ bụ ịkwadebe, omume a na-eme n'oge a na-eme ka a ghara ịsị na ọ bụ ihe na-adịghị mma. Ebe ọ bụ na "agụụ" nwere ike iji kọwaa àgwà niile na-eduga n'inweta àgwà nke mkpali (mmechi), àgwà kachasị mma nke nwoke na-egosipụta mgbe ọ na-arụsi ọrụ ike n'oge "consummatory" kachasị mma dị ka mkpali; nwoke na-etinye oge ya na mgbalị ya n'ịchụso nwanyị ahụ ka ọ na-elekọta ya. Na nke a, anyị nwere ike ịmekọrịta NAC na nnyefe ya yana akparamagwa nke oke nwoke na nwanyi.

Nweta ohere nke abụọ, akwụkwọ nwanyi mere ka a gbanwee nkwenye na isiokwu ọ bụla. Nnyocha ndị gara aga egosiwo na ọnụ ọgụgụ ka ukwuu nke oke na-ekwe ka ha na-edozi iji satiety, na-eji usoro omume omume yiri nke ahụ mee ihe na nnwale ahụ ugbu a, ọ maliteghị ịme nkata mgbe a nwalere 24 hr mgbe e mesịrị (Beach na Jordan, 1956). O yikarịrị ka ọnụnọ nke ihe mkpali nke nwanyi 2, nke nwere ike ịgụnye ihe ndị na-eme ka ahụ dị mma yana nke anya na ntinye anya, mere ka e nwee nkwenye. Ajụjụ na-adọrọ mmasị, bụ nke a ka ga-aza, ọ bụ site na usoro usoro nwoke oke na-ama ọdịiche nke nwanyị nwanyị si n'aka nwanyị nke o jikọtara n'oge na-adịbeghị anya. Ebe maka usoro ahụ nwere ike ịdọrọ na usoro kachasị mma. A kọwawo na iguzosi ike n'ezi ihe nke usoro ihe a dị oké mkpa maka mmetụta nchịkwa Coolidge na hamsters (Johnston na Rasmussen, 1984). Otú ọ dị, usoro ntụrụndụ nke vomeronasal-accessory, Otú ọ dị, a kọwara usoro nchekwa pheromonal n'oge na-adịbeghị anya na ụmụ oke (Kaba et al., 1994), bụkwa onye na-eto eto. Na nke a, ọ dị ịrịba ama na enwere mmụba na NAC na nnyefe na-eji na vivo agba ume na oke nwoke na-eji ihe ndina na-ekpughere oke ụmụ nwanyị na estrus (Louillot et al., 1991; Mitchell na Gratton, 1992). Ọzọkwa, ngwa nke K⁺ kpọmkwem na nkpuchi akwara vomeronasal nke ngwa ngwa olfactory nke ngwa ngwa, tinyere nkwụnye olfactory nke ngwa ngwa n'onwe ya, zuru ezu iji mewanye NAC na nnyefe (Mitchell na Gratton, 1992).

Ihe mbụ 15 min nke na-ejikọta ya na nwanyị 2 nwere njikọ na nnukwu mmụba na NAC DA. N'adịghị ka nwanyi 1, mmekọrịta ya na 2 bụ nwanyi enweghi mmụba na NAC DA nke ịdị n'otu n'oge mmemme ahụ (12%) ma ọ bụ na-eme ihe (34%). Otú ọ dị, ntakịrị mmụba ndị a na NAC NA, na-emekọ nke ọma na njedebe nke omume mmekọahụ gosipụtara na 2 nwanyị jiri ya tụnyere nwanyị 1. A nọgidere na-etinye concentrations nke metabolite n'oge oge satiety, na-eme ka e nwee ihe mgbaru ọsọ ọhụrụ (ihe nrịba 10 na 12) nke e buliri elu site na mbido nke mbụ (ihe atụ 1).

Ọdịdị nke anụ ahụ na-arịwanye elu na DOPAC na HVA concentrations n'oge mmechi na-adabere n'usoro ha dị ka metabolites nke nne na nna, DA. E kwuwo na microdialysis metabolite concentrations, ọ dịkarịa ala n'oge a na-akpa àgwà nke na-adịghị na-achụ nta ọgwụ, nye ndepụta bara uru nke neural ọrụ (Damsma et al., 1992; Fumero et al., 1994). Eziokwu ahụ bụ na a nọgidere na-etinye concentrations nke metabolite ọbụna n'oge oge enweghi mmekọahụ na nnwale a, mgbe ụda ihe dị iche iche laghachiri na-eme ka a ghọta ụkpụrụ nke usoro ntọala, na-etinye obi abụọ banyere aro a.

Ọganihu dị elu nke DA metabolite concentrations hụrụ na nnwale a gosipụtara na profatic medial preoptic mpaghara (mPOA) nke DA metabolites hụrụ na oke n'ụbọchị nke mbụ mgbe ha jedebere iji satiation (Mas et al., 1995a,b). A naghị ahụkarị nkwupụta elu nke DOPAC na HVA concentrations na NAC ma ọ bụ mPOA mgbe oge oge na-agakọ bụ oge a kara aka, nke dị mkpirikpi karịa oge achọrọ iji ruo satiation. Dịka ọmụmaatụ, ọtụtụ nchọpụta egosiwo na ọnụ ọgụgụ DOPAC nọ na-amụba ma nọgide na-ebuli elu mgbe a na-ejikọta ya ma na-ajụ ụkpụrụ dị elu n'oge na-adịghị anya mgbe ewepụrụ nwanyị (Pfaus et al., 1990; Pleim et al., 1990; Damsma et al., 1992;Hull et al., 1993; Wenkstern et al., 1993; Hull et al., 1995). Na ọmụmụ ihe site na Mas et al. (1995b), basal extracellular concentrations nke DOPAC na HVA na mPOA nọgidere na-elu n'elu 4 ụbọchị kwekọrọ na oge nke mmekorita mmekọahụ. Site na ụbọchị nke anọ, tupu ụmụ anụmanụ amaliteghachi ịkpụkpụ, nkwonkwo basal nke ndị metabolites nọ nso na ụkpụrụ omume. Ndị na-ede akwụkwọ jiri usoro mgbanwe nke neurochemical mee ka ndị ahụ hụrụ mgbe ha nyesịrị nke DA ndị na -Zetterström et al., 1984; Imperato na DiChiara, 1985) ma kwadoro na enweghi ike ime ka ọnọdụ nke mmekorita nke nwoke na nwanyi ghara ikpuchi ya site na mkpofu prolactin, nke nwere ike ime dika "neuroleptic"Mas et al., 1995a,b,d). O doro anya na ochichi neuroleptic na-esonyere na mmụba na extracellular metabolite concentrations na DA efflux (Zetterström et al., 1984; Imperato na DiChiara, 1985). O di nwute, Mas et al. (1995a,b) enweghi ike ịchọpụta mPOA na iche. N'ọmụmụ ihe ugbu a, ihe dị iche iche na NAC laghachiri na-emetụta ụkpụrụ omume, ma ọnụnọ DOPAC na HVA ka dị elu. Ụkpụrụ a ekwekọghị na ọrụ maka neuroleptic na-adọrọ adọrọ na-arụ ọrụ na NAC iji mee ka agụụ mmekọahụ nwee mmekọahụ.

Nyere ntinye aka nke mesolimbic DA neurons na omume mkpali (Fibiger na Phillips, 1986; Blackburn et al., 1992; Kalivas et al., 1993; LeMoal, 1995) na uche ha maka ihe omumu nke gburugburu ebe obibi (Fabre et al., 1983; Schultz, 1992; Mirenowicz na Schultz, 1994), mmụba ndị a na-arịwanye elu na nchoputa nke NAC na nzaghachi nye ndị inyom nwanyị ahụ na-ekwenye n'echiche bụ na ọrụ na usoro DA a dị mkpa maka ịmalite ịkpaghasị mmekọahụ. Na mgbakwunye, akụkọ nke mmetụ na ọkụ na-abawanye na DA nnyefe (Hull et al., 1993, 1995;Mas et al., 1995b; Sato et al., 1995) na ọrụ neuronal (Shimura et al., 1994) na mPOA nke oke oke nwoke na nwanyi na-egosi na nke a nwere ike inye aka mee ka e nwee mmelite nke a na-enwe mmetụta nke Coolidge.

N'ikwekọ n'ozuzu ọrụ maka usoro mesolimbic DA na mkpali, ọ bụ nke ọma kwenyere na ihe ndị ọzọ gbasara DA na-ebuli elu n'oge, n'oge, na ozugbo o richara nri, ya na nlọghachi na njirimara ala ~30 min mgbe e mesịrị (Wilson et al., 1995). A maara nke ọma na ihe oriri na-emetụta ihe oriri na-edozi ahụ site na nri. Ụmụ mmadụ na ụmụ anụmanụ na-ajụ nri nke e nyere ha ka ha nọrọ na satiety na ingest ihe oriri ndị ọzọ a na-eribeghị (Na-apụta, 1986). Nke a na-ebuli ajụjụ a banyere ma ọ bụ mmepụta mmiri na NAC ga-amụba nhọrọ site na ngosi nke ụdị akwụkwọ nri, ma ọ bụghị site na nri e riri n'oge na-adịbeghị anya iji nọrọ jụụ n'ụzọ yiri nke ahụ a kọrọ na ọmụmụ ihe ugbu a n'ihe gbasara ya. akpali agụụ mmekọahụ. Ọ bụrụ na a kwadoro, mmekọrịta a dị n'etiti ihe mgbagwoju anya nke nkwụghachi ụgwọ, satiety, na mesolimbic DA nnyefe ga-apụta ọrụ dị oke mkpa maka usoro nchịkwa nke usoro a nke usoro mkpali na-akpali, nke nwere ike iduga ajọ nsogbu nke iri nri na mmekọahụ .

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