Nat Neurosci. Akwụkwọ edemede; dị na PMC Jan 1, 2014.
E bipụtara na ụdị edetu ikpeazụ:
Nat Neurosci. Jul 2013; 16 (7): 10.1038 / nn.3420.
E bipụtara online Jun 2, 2013. doi: 10.1038 / nn.3420
PMCID: PMC3703824
NIHMSID: NIHMS477621
Hui Wang,*,1,2,3 Florian Duclot,*,1,2 Yan Liu,2,3 Zuoxin Wang,2,3 na Mohamed Kabbaj1,2
Mbubre ọhụụ nke onye bipụtara edemede a dị na Nat Neurosci
Hụ ihe ndị ọzọ na PMC na ebe isiokwu e bipụtara.
nkịtị
Na mmadụ na-atụghị anya otu nwanyị praleie vole (Microtus ochrogaster), ntinye ọnụ na-eduga na njikọta ejikọtara ọnụ nke njikọta nke onye mmekọ ma guzozie ya site na ọtụtụ neurotransmitters gụnyere oxygentocin, vasopressin, na dopamine. N'ebe a, anyị nyochara ihe nwere ike ịbụ ihe ọgbụgba nke na-ejikọ ọnụ na njikọta njikọta nke ejikọtara ọnụ. Anyị na-egosi na histone deacetylase inhibitors sodium butyrate na TrichoStatin A (TSA) na-eme ka mmasị nke mmekọ dị na praịmarị nwanyị na-eme ihe n'emeghi ya. Ejikọtara ya na usoro nchịkwa nke oxygentocin (OTR) na vasopressin V1a receiptors (V1aR) na ngụkọta akụrụngwa, site na mmụba na aconelation histone na onye na-akwado ha. Ọzọkwa, a na-egbochi ntụgharị nke akwadoro nke TSA site na mgbochi OTR ma ọ bụ V1aR na njikwa akwara. Ihe di nkpa, ihe ndi mmadu choro ka ndi mmadu meghariri maka mmekorita emeela ka otu ihe megharia ihe omimi nke OTR na V1aR bu ndi n’eme ka ndi ozo mara ihe bu TSA. Nkwupụta ndị a na-egosi na TSA na mating na-eme ka mmasị onye ibe site na ihe omume epigenetic, na-enye ihe akaebe mbụ maka ụkpụrụ iwu nke njikọ nke njikọ ọnụ.
Okwu Mmalite
Njikọ mmekọrịta bụ akparamagwa dị mkpa nke akparamagwa mmadụ na akparamaagwa nke mmekọrịta mmadụ na ibe ya bụ ihe ndị a na-ahụ n'ọtụtụ nsogbu ọria neurochichiatric gụnyere ọrịa ihu mkparịta ụka na nsogbu akpịrịkpa, tinyere ihe riri ahụ na ịda mba.1. The otu na-alụbeghị di na nwunye preles (Microtus ochrogaster) pụta dị ka ihe atụ na-atọ ụtọ maka nyocha nke ntọala neurobiological nke njikọta mmekọrịta, dị ka ụlọ nyocha abụọ na ndị nwere onwe ha na-eguzobe nkekọ ogologo oge.2-4, nke ebidobere site na ịkpa agwa dị mma na -emekọrịta onye mmekọ, nke a na-akpọ mmasị mmekọ5. Ozo a choro onye choro ya nwere otutu neurotransmitters na sistem homonu, tinyere neuropeptides oxytocin na vasopressin (AVP) na mesolimbic dopamine.5.
IN'ozuzu, ntụrụndụ nke onye mmekọ na-ebugharị site na AVP neurotransmission na ventral pallidum na mpụta septum (LS) na ụmụ nwoke, na oxygentocin neurotransmission na nucleus accumbens (NAcc) na prelimbic cortex na prairie nke ụmụ nwanyị6-8.
A na-ejikọkarị ya na ụgwọ ọrụ eke dị ka mating, dopamine na-arụkwa ọrụ dị ka onye ogbugbo dị oke mkpa nke onye ibe na preir prales. Mgba nke dopamine D2 ụdị nnata (D2R) na NAcc na - eme ka arụ ọrụ dopamine D1 ụdị nnabata (D1R) na - egbochi ụdị mmekọ nwoke na nwanyị na - eto nwoke na nwanyị.9-11.
Dịka mkpa, mgbanwe dị iche iche na nkwupụta site na ọmịiko nke oxygentocin na ndị na-anabata AVP V1a, OTR na V1aR, n'otu n'otu, nwere ike imetụta mmasị ibe. N’ụbọchị ụmụ nwanyị na-eme njem, dịka ọmụmaatụ, oke nke OTR na NAcc na-eme ka mmasị onye mmekọ n’onwe ya bụrụ nke nwoke na nwanyị matingiri12-14.
N'azụ usoro nke njikọkọ njikọta abụọ, oxytocin na AVP na-emetụtakwa n'ọtụtụ akparamagwa ọha mmadụ, gụnyere njiko ọha, nnabata, na nlekọta nne.15, 16. Dika odi, ihe ndi mmadu gha eme ihe nwere ike ime ka ogwara ha ihe di ogologo site na usoro ihe omimi, tinyere DNA methylation nke estrogen receptor alfa.17 na mkpụrụ ndụ ihe nketa AVP18, yana histone acetylation nke ndị na-akwalite glucocorticoid receptor na-akwalite19. Ọzọkwa, histone deacetylase (HDAC) na-egbochi, na-eme ka nkwupụta mkpụrụ ndụ pụta site na ịba ụba histone acetylation na ụbụrụ nwere20, nwere ike ịgbanwe mgbanwe ndị a19, ma na-emetụta omume mmekọrịta mmadụ na ibe ya dị ka inwe mmekọahụ21. Dị mkpa, na eriri mkpụrụ ndụ akwara akwara, HDAC inhibitor trichostatin A (TSA) na-akwalite transcript OTR ozugbo site na ịkwalite aconelation mpaghara.22.
Yabụ, enwere ike ịtụle usoro ọmụmụ nọ n'ụdị mmasị onye ibe na preir prair. Iji nwalee hypothesis a, anyị buru ụzọ nyochaa mmetụta nke mmadụ abụọ HDAC inhibitors, sodium butyrate (NaB) na TSA, na ntụnye mmasị mmekọ nwoke na nwanyị dị ka ụmụ nwanyị toro eto. Mgbe nke ahu gasiri, anyi nyochawara udiri ihe nke na eme ka ihe ndi TSA na ewebata udiri onye obula na iyi nwanyi. N'ikpeazụ, anyị chọrọ ịchọpụta ma mgbanwe nke epigenetic nke TSA kpatara na-ebikọ site na mbikọ ọnụ na-abụkwa ihe mgbakasị.
Results
Ọgwụ TSA na-eme ka mmasị onye mmekọ dị
Mmekọahụ naïve nwanyị praịls voles gbara arụ intracerebroventricularly (icv.) na CSF ma ọ bụ CSF nwere 0.08, 0.4 ma ọ bụ 4 ng nke TSA ozugbo n’ihu nwoke na nwoke 6 na-ebikọ n’emebighị mmekọrịta, wee nwalee mmasị ibe ha. Oge awa isii nwoke na nwoke na-ebubigaghị na ya anaghị eme ka nwoke bụrụ nwoke ma ọ bụ nwanyị na-emekọ ihe4 ma otu osila mee ihe omume a n'ọtụtụ ebe iji chọpụta nsonaazụ ọgwụ dị iche iche na-eme ka njikọta nke mmekọ ibe si dị5.
Anụmanụ na-elekọta CSF gosipụtara mmekọrịta na-abụghị nke nhọrọ n'etiti ndị mmekọ ma ọ bụ onye bịara abịa na-eso awa 6 nke ibi n'enweghị nsogbu.t15= 0.76, P = 0.46; 1a fig). Agbanyeghị, anụmanụ jiri ọgwụ mee ihe na usoro ọgwụgwọ niile were onye ya na ya na-emekọrịta ihe karịa oge ọ ga-anọ karịa onye ala ọzọ (t8= 4.35, P = 0.002 maka TSA 0.08 nndi g; t15= 3.63, P = 0.002 maka TSA 0.4 nndi g; na t8= 2.58, P = 0.03 maka TSA 4 nndi g). Ọ dị mkpa, achọtaghị ihe dị iche n'otu otu na arụmọrụ sitos (F3,46 = 1.25, P= 0.30, 1b) na enweghị omume iwe site nwale nwale chere ma onye a na-amabughị ma ọ bụ onye ya na ya hụrụ, na-egosi na nsonaazụ nke TSA bụ nke masịrị ọha na eze, kama ịbụ nke abụọ na mgbanwe nke mkpọda ma ọ bụ nke na-abụghị nke onye ọbịa ahụ.
Iji nyochaa ma TSA na-eme ka akịkị akwara akwara sonye na njikọta ụbụrụ so na njikọ mmekọ nwoke na nwanyị, ọgwụ ọgwụ dị iche iche batara na TSA (0.4) ng) ma bikọtara nwoke dịka nwoke na - anọghị na mgbakwunye n’oge elekere 30, elekere 2 ma ọ bụ 9. Dị ka o siri dị, enwere ike ịchọpụta mgbanwe mgbanwe dị ukwuu na ọkwa histone H3 acetylation (Lys14) na oge ọ bụla-na NAcc na caudate putamen (P > 0.05 maka otu niile, .Dị 1c & d). Ihe a gosiputara na TSA na-eme ka nhazi nke mmekorita nke onye choro ya di omimi, n’agbanyeghi otutu ihe metutara H3 acetylation na NAcc ma obu caudate putamen.
Ihe di nkpa, sodium butyrate mekwara ka mmekorita nwoke na nwanyi nwee ike dika ihe nlere nke ndi nwanyi na-eso ibikọ ya na nwoke maka awa 6 na-enweghi mmekorita, jikọtara ya na mmụba na histone H3 acetylation (Lys14) na NAcc (Ihe atụ 1). Enwere ike imepụtaghachi nsonaazụ nke TSA na ntinye mmekọrịta na onye ọzọ na-egbochi HDAC, na-atụ aro itinye aka na inhibition HDAC, kama ọ bụghị mmetụta TSA na-agbado na nkwado nke mmekọ. Thattụle na TSA bụ kọmpụta kachasị nke akpọrọ / I HD II inhibitor23, 24, na na agwa omume nke TSA ka ekwuputara karịa nke NaB, anyị họọrọ iji TSA karịa NaB maka nyocha kpọmkwem ụyọkọ akwara dị na akụkụ ndị a.
Molecular mmekọrịta nke TSA na-eme ka mmekọrịta mmadụ na ibe ya bụrụ ọkacha mma
Dika ihe di iche na uzo ihe onyinye ndi mmadu na ndi ozo bu NAcc ka ejikọtara ya na atumatu di iche di iche n'etiti ihe di na nwunye na ndi n’ebughi agha, yana igbanwe mmekorita nke ndi mmadu na ndi ozo.12, 13, 25, 26, anyị enyochala ma usoro nchọta nke akwadoro mmekọrịta nke TSA nwere njikọ yana mgbanwe dị iche iche sitere na usoro ọmụmụ nọ na NAcc.
Usoro ọgwụgwọ TSA (0.4 ng, icv.) webatara mmụba nke ọkwa OTR mRNA na NAcc mgbe oge 2 nke ibi na-ebikọ dịka ejiri ya na njikwa ndị CSF mesoro (t10 = 2.38, P = 0.038, 2a fig), nke ahụ gbara mbọ ịkwagide mgbe oge 9 nke ibi (t9 = 2.17, P = 0.058, 2b). Ọ bụ ezie na enwere ike ịhụ ntakịrị mmụba ma ọ bụrụ na ọ bụghị ihe dị mkpa na V1aR mRNA na oge awa NAcc 2 na-eso ngbanye TSA, ọchọghị ndị otu ndị ọzọ achọpụtara n'otu oge-oge maka mRNA ndị ọzọ tụrụ, gụnyere D1R ma ọ bụ D2R (P > 0.05, Fig. 2a, b). Di nkpa, onweghi ndiiche ndi otu n’otu ihe nlere anya n’eme ka oge obula di otua na atule mRNA obula.P > 0.05 maka otu niile, 2c, d), na-atụ aro na mmụba nke OTR mRNA hụrụ na ụmụ anụmanụ ndị a na-elekọta TSA bụ maka NAcc. Ọzọkwa, ụdị iwu dị ugbu a bụ naanị iso nwoke na nwoke na-ebikọ, dị ka OTR na V1aR mRNA na NAcc nọgidere na-agbanwe agbanwe 2 awa mgbe ọgwụ TSA na-enweghị ibikọ ọnụ (OTR: 100.0% ± 11.70 maka CSF otu, 86.7% ± 12.11 maka TSA otu, t12 = 0.79, P = 0.444; V1aR: 100.0% ± 26.24 maka CSF otu, 92.3% ± 13.75 maka otu TSA, t9 = 0.27, P = 0.791).
N'ikwekọ n'ọkwa dị elu OTR mRNA dị elu, ụmụ anụmanụ ndị a na-elekọta TSA gosipụtara ọkwa protein protein OTR dị elu na oge abụọ ahụ na NAcc (awa 2: t10 = 2.34, P = 0.041; Oge 9: t10 = 3.16, P = 0.01, Fig. 2e, f), mana obughi caudate putamen (t10 = 0.41, P = 0.69, Fig. 2g, h). N'ụzọ na-akpali mmasị, ọ bụ ezie na enweghị mgbanwe mgbanwe dị mkpa nke V1aR mRNA nwere ike ịchọpụta na NAcc n'oge 2 ma ọ bụ 9 awa mgbe ọgwụ TSA (Fig. 2a, b), ọkwa protein nke V1aR mụbara nke ukwuu na awa 9, ma e jiri ya tụnyere anụmanụ ndị CSF mesoro, na NAcc (t9 = 3.46, P = 0.007, 2f) ma obughi caudate putamen (t10 = 0.98, P = 0.35, 2h). Agbanyeghị na enwere ụfọdụ mgbanwe, ọkwa nchịkwa D1R na D2R na NAcc na caudate putamen emetụtaghị TSA emetụta nke ukwuu (P > 0.05, Fig. 2e-h).
TSA na-eme ka anakpo histone achota nke oxtr na avpr1a
Mmụba nke ma mRNA na ọkwa protein maka OTR na-eso ibikọ ọnụ mgbe ọgwụgwọ TSA kwupụtara na o nwere ike bụrụ na TSA mụbara ntụgharị nke oxtr, mkpụrụ maka mkpụrụ ndụ maka OTR, karịa ịgbanwe ntụgharị ma ọ bụ ntụgharị nke protein. Ọzọkwa, ọkwa protein protein V1aR dị elu na NAcc, jikọtara ya na ntakịrị ihe ma ọ bụghị mmụba dị ukwuu na ọkwa mRNA na-esochi usoro ọgwụgwọ TSA (Fig. 2). N'iburu n'uche na TSA bụ klas dị ike m na II HDAC inhibitor23, 24, 27, anyị hypothesized na TSA mụbara histone acetylation na oxtr na avpr1a ndị na-akwalite na NAcc, na-emesikwu ike ntụgharị asụsụ ha. Anụ ụmụ anụmanụ ọhụrụ nwetara icv. ogwu ogbugba nke TSA (0.4 ng) ma soro nwoke biri ozugbo n’enweghị nsogbu maka nkeji 30 tupu ịchụ àjà. A họrọ windo 30-min dabere na ọrụ gara aga na-akọ banyere mmụba kachasị nke nariole histone mgbe ọgwụ mpaghara TSA dị na oke na ụmụ oke.28, 29. H3K14 acetylation na oxtr na avpr1a Mgbe ahụ nyochachara ọgwụ mgbochi chromatin.
N'ikwekọ na mmụba nke OTR mRNA na ọkwa protein e gosipụtara na mbụ, anụmanụ ndị a na-elekọta TSA gosipụtara mmụba dị elu (+ 460%) na aconelation histone H3 na oxtr onye na - akwalite gene, dika atụnyere ihe njikwa CSF, na NAcc (t10 = 5.88, P = 0.0002), mana obughi caudate putamen (t9= 0.31, P = 0.76, 3a fig). Ọzọkwa, histone H3 acetylation na avpr1a onye na - akwalite akwagoro elu nke 30 min elu site na njikwa TSA (+ 196%) na NAcc (t10 = 3.12, P = 0.01) ma obughi caudate putamen (t9= 0.38, P = 0.71), dịka e jiri ya tụnyere njikwa ndị CSF mesoro (3b). Ya mere, TSA mụbara saịtị histone acetylation kpọmkwem na NAcc ka oge nkeji 30 mgbe mmalite nwoke na nwoke malitere.
TSA na-eme ka mmasị ibe dị n'etiti OTR na V1aR
Site na usoro nyocha ndị gara aga, usoro ihe eji arụ ọrụ na-apụta, ebe n'oge ibikọ ọnụ, TSA na-eme ka anakwere histone acetylation na oxtr na avpr1a ndị na-akwalite, mechaa meziwanye ntụgharị okwu ha ma rụpụta ogo protein OTR na V1aR dị elu ruo awa 9 mgbe mmalite nke oge ibikọ ọnụ. Ihe dị mkpa bụ na mmetụta TSA a bụ nke akọwapụtara na ntanetị ka caudate putamen anaghị enwe mmetụta. N'ebe a, anyị nwalere ma mmụba a na-eweta TSA na OTR na V1aR nwere njikọta na ntinye nke mmasị mmekọ ibe. Pramụ nwanyị prairie voles nwetara ntụtụ intra-NAcc nke TSA (0.04)ng n'akụkụ ọ bụla) na ma ọ bụ na-enweghị ntụtụ tupu (30minutes tupu ntụtụ TSA, 0.5ng kwa akụkụ) nke CSF ma ọ bụ CSF nwere otu n'ime ụzọ abụọ dị iche iche OTR, OTA (B) na OTA (T), ma ọ bụ V1aR antagonist (V1aRA). Ngwa ngwa ogwu ogbugba TSA, ụmụ nwanyị ebikọtara ọnụ na nwoke maka awa 6 na-enweghị mating, ule nyocha nke ibe.
Anụmanụ na-emeso CSF egosighi mmasị onye ibe (t5 = 0.17, P = 0.87, 4a fig). Agbanyeghị, anụmanụ ndị a na-elekọta TSA nọrọ oge dị ukwuu na ndị ha na ha na-emekọrịta ihe karịa ka ha na onye “ọbịa” na-ekwu, na-atụ aro na otu mkpụrụ ọgwụ TSA na-abanye na NAcc zuru ezu iji kwado njikọ ibe ya na-enweghị mating (t5 = 7.04, P = 0.0009). N'ụzọ na-akpali mmasị, mgbochi nke ma ọ bụrụ na OTR ma ọ bụ V1aR site na ọgwụgwọ tupu ya na OTA (B), OTA (T), ma ọ bụ V1aRA gbochiri mmetụta nke TSA (P > 0.05 maka otu niile). Ebe ọ bụ na enweghị otu esemokwu dị na ọrụ locomotor (F4,32 = 1.89, P = 0.14, 4b), data ndị a na-atụ aro na TSA na NAcc na-eme ka usoro njikọ nke mmekọ dị na ntanetị sitere na OTR- na V1aR.
Atinggwakọta akụ na-akpata neuroadaptations yiri TSA
Site na nlele ndị anyị mere, anyị chọpụtara na ikike ịmụba nke oxygentocin na vasopressin neurotransmission na nwanyị nke NAcc ezuru iji kwalite ụdị mmekọ ibe na enweghị mating. Iji nyocha ma neuroadaptations ndị a na-emekwa n'oge ebilite nke eke nke mmasị ibe, a na - ebikọ nwoke iyi praịmarị na nwoke n'oge 24 awa na ọnụnọ, nke na - akpali mmasị onye ibe4, wee chụọ àjà. Anyị hụrụ mmụba na ma OTR na V1aR mRNA na ọkwa protein na NAcc, dịka e jiri ya tụnyere ụmụ nwanyị naïve mmekọahụ (OTR: + 38%, t10 = 2.68, P = 0.02 maka mRNA, na + 58%, t8 = 3.05, P = 0.01 maka protein; V1aR: + 89%, t14= 2.53, P = 0.02 maka mRNA, na + 26%, t20 = 2.23, P = 0.037 maka protein, Fig. 5a, b).
Dịka ma mRNA na ọkwa protein maka OTR na V1aR mụbara site na ibikọ ọnụ na mating, anyị nyochapụrụ ma usoro iwu a nwere nkwalite epigenetic nke oxtr na avpr1a mkpụrụ ndụ ihe nketa. Ndi ohuru nwanyi ji otu nwoke wee rue otutu 6 tinye ya na ntuba onu na H3K14 oxtr na avpr1a ndị na - eme ihe ngosi site na njikwa mgbochi chromatin. N'ikwekọ na OTR na V1aR mRNA na ọkwa protein, praị nwanyị voles na-egosipụta ọkwa dị elu nke H3K14 na oxtr na avpr1a na-akwalite na NAcc, ma e jiri ya tụnyere ndị nwanyị naïve nwanyị (oxtr: t9 = 2.64, P = 0.02; avpr1a: t9 = 2.91, P = 0.017 5c, d). Ihe omuma ndi a n’egosi na udiri obibi na ntukwasiobi ndi mmadu na acho ime ka umu nwanyi mekwaa ihe ngosi nke OTR na V1aR na NAC site na usoro ogwu, dika ahuru mgbe TSA gwachara.
Nkwurịta
N'ime ọmụmụ ihe dị ugbu a, anyị na-akọ akụkọ izizi usoro iwu metụtara ịmụmụ ọnụ ọgụgụ nke njikọ mmekọ ụdị n'ụdị. Nke mbu, anyi gosiputara na inwetakwu akuko histone na NAcc site na nchịkwa nke onye na - egbochi HDAC na - eme ka mmasị onye mmekọ dị na praịmarị nwanyị bụrụ okenye. TAgụ, anyị edepụtarala ihe akaebe doro anya na ime ka mmasị nwoke na nwoke bụrụ nwoke ma ọ bụ nwanyị ka a na-ebikọ, n'ihi na ibikọ ọnụ na nwoke na nwoke na-amụba oxtr na avpr1a ngosipụta mkpụrụ ndụ ihe nketa site na agbakwụnye histone acetylation na NAcc. Nlekọta TSA na NAcc kpaliri mmasị onye mmekọ ma dugara n'ọkwa dị elu nke OTR mRNA na protein n'ime NAcc. Ọzọkwa, ọ bụ ezie na nwanyị X3 acetylation emetụtaghị ọgwụ ụmụ nwanyị na-agwọ ọrịa TSA, mmụba dị ịrịba ama nke acone aconele. oxtr A hụrụ onye kwalitere na NAcc ka oge nkeji 30 na-esote njikwa TSA. N'ikpeazụ, igbochi OTR na NAcc ezuru iji gbochie mmasị ndị mmekọ TSA kwadoro. Ebe ọ bụ na achọpụtara mgbanwe mgbanwe ndị dị otu a na-esochi ibikọ ọnụ na mating, n'okpuru usoro a maara iji kpalie mmasị onye mmekọ, data anyị na-ebute ihe atụ maka ụkpụrụ epigenetic nke omume mmekọrịta. Dna -akpata ime nwoke, TSA, ma ọ bụ mating, na - ebute ngwa ngwa H3 histone na oxtr onye na - akwalite ya na NAcc nke na - eme ka ntụgharị ya dịkwuo elu, nke na - ebute OTR mRNA dị elu na ọkwa protein, na - emesịa kwalite njikọ ibe.
N’ebe ụmụ nwanyị na-eme ihe nkiri, awa 6 nwoke na nwoke n’ebikọghị ọnụ na-eme ka mmekorita nwoke na ibe ya ghara ike4, ejiriwo omume a mee ihe iji nyochaa nsonaazụ ọgwụ aghụghọ na-akpata mmasị nke mmekọ ibe5. Na omumu anyi, ebe saline- ma obu CSF ejiri aka mee ihe emeputaghi ihe ndi mmadu choro, preir nke nwanyi dika ndi NeB ma obu TSA mebere. Dịka NaB na TSA emetụtaghị mkpọmkpọ ebe ahụ dum, mmetụta ha na mmasị ibe ha yiri ka akparamagwa akparamagwa kama inwe mmetụta na locomotion. Nleba anya nke ihe omimi gosiputara uto a nke TSA. N'ezie, ọ bụ ezie na a na-elekọta ya icv,, anyị mechara chọpụta mgbanwe mgbanwe nke mkpụrụ ndụ na NAcc mana ọ bụghị n'usoro dị n'akụkụ ahụ, caudate putamen. Na mgbakwunye, ọbụlagodi na NAcc, D1R na D2R mRNA na ọkwa protein anaghị enwe mmetụta. Suchdị nkọwa a yiri ihe ijuanya maka nnukwu mgbochi HDAC dị ka TSA nke metụtara klaasị I na II HDACs. Agbanyeghị, a kọọrọ TSA na-emetụta ngosipụta nke obere mkpụrụ ndụ ihe nketa na mkpụrụ ndụ mammalian30-32, gụnyere na ụmụ oke20.
Anyị gosipụtara ebe a na acetylation nke histone H3 na Lys14 na oxtr onye na - akwalite, mgbanwe na - esonyere na iji mkpụrụ ndụ ihe ana - eme ka ọ dịwanye gara nke gụnyere n’oge ọghe nke ụbụrụ33, 34, na-egosi ọkwa dị elu nke OTR mRNA na protein. Na nzaghachi na TSA, achorology histone na oxtr onye na - akwalite ndị mmadụ na - abawanye ma na - eme ka mmegharị ya dị na eriri mmadụ22, na-akwado ịchọta anyị oxtr enwere ike ịhazi epigenetically. Ebe ọ bụ na mgbochi nke OTR dị na NAcc zuru ezu iji gbochie mmetụta omume nke TSA, data anyị na-egosi na ngosipụta nke TSA na-ebute na OTR na NAcc n'oge njikọta ọnụ na-eme ka ọ dị mfe maka ịmepụta njikọ nke ibe. Ọzọkwa, awa 24 na-ebikọ ọnụ na mating, usoro a maara maka ịdabere na ntinye uche onye ibe na voirie ụmụ nwanyị, na-ebute mmụba yiri nke ahụ na Nkwupụta OTR na nwanyị nke NAcc. Nke a bụ na nkwekọrịta zuru oke na nnabata nke oxygentocin na onye na-anabata ya na neurobiology nke mmasị mmekọ ibe na vobe ụmụ nwanyị. Atinggwakọta akụ na ụba na-ebute mmụba na ọkwa extratolular oxygentocin na NAcc25, na mgbakwunye nke oxytocin n'ime NAcc na-eme ka mmasị mmekọ ibe bụrụ nke enweghị mating8. Ọzọkwa, ndị na-emegidekarị OTR na-egbochi ike mmadụ na ibe ya, ihe nchịkwa ma ọ bụ nnata adapụtara onwe ha8, 35. Ihe di nkpa, oria ojoo nke OTR na nwanyi nke NAC zuru oke iji kwado mmekorita nke onye choro ya.12, 13.
Na mgbakwunye na ịgbado ọrụ nke OTR, nsonaazụ anyị na-enyekwa ihe akaebe maka mmegharị nke ngosipụta mkpụrụ ndụ OTR site na usoro epigenetic n'oge nwoke na nwoke na-ebikọ ọnụ. N’ezie, n’agbanyeghi na ezughi oke ime ka ndi mmadu choro izo, ochoro ka n’ebughi mmadu mee obere oge, o gha eme ka ndi mmadu ghara ime ihe o choro.. Iji maa atụ, awa abụọ ekpughepụrụ nwa nwoke nye obere ihe ọ bụla ma ọ bụ nke anaghị adị oke na ntọhapụ oxygentocin na nwanyị nke nwanyị NAcc.25. Enwere ike ikwupụta na TSA ma ọ bụ NaB nwere ike ime ihe na - eme ka ọ bụrụ nwoke na nwoke na - ebikọ ọnụ, na - eme ka mmepe nke mmasị ibe. N'ụzọ na-akpali mmasị, ekwuputala ikike dị otú ahụ na akwara ebe klas I na II HDAC na-egbochi, gụnyere NaB na TSA, na-eme ka nchịkọta nke ihe mmụta na-esiteghị na ncheta ogologo oge na njikwa anụmanụ.36, 37. Na nkwado nke echiche a, ogologo oge ibikọ ọnụ (dịka., 48hours) nwere ike ime ka mmadụ ibe ya nwee mmasị ọbụlagodi na enweghị mating4. Ọ dịkwa mkpa ịmara na ibikọ ọnụ na mating sitere na ntanetị nwanyị nke NAcc usoro nhazi nke OTR na V1aR site na otu usoro epigenetic dị ka ndị ahụ hụrụ mgbe ebikọtara ya na ọgwụgwọ TSA, nke gosipụtara na TSA na mating metụtara otu ụzọ iji kwalite nhazi nke onye choro. Dị ka ihe dị mkpa, TSA anaghị eme ka OTR na V1aR dị na nwanyị bụ́ nwanyị na-ejikọtaghị nwoke. Na ngụkọta, nke a na-akwado echiche nke TSA na-eme ka enwee mmekọ nke onye ibe ya site na njikọta nke neuroadaptations endogenous neuroadaptations site na ibikọ ọnụ na nwoke, kama ime ka onwe ya rụọ ọrụ ndị a ma ọ bụ neuroadaptations dị iche iche.
Ostudylọ nyocha gosikwara ọrụ dị mkpa nke NAcc V1aR na usoro njikọ nke nwanyị, dị ka ụmụ anụmanụ si lebara anya nke TSA gosipụtara ọkwa V1aR dị elu, nke ihe mgbochi ya gbochiri mmasị mmekọ mmekọ TSA.. Ọzọkwa, mmetụta ndị a jikọtara ya na ọgwụ mgbochi sitos dị elu na avpr1a onye na - akwalite nkwalite, n’agbanyeghi oke ịrị elu nke V1aR mRNA nwere ike ịbụ n’ihi oge-adịghị mma. Ọ bụ ezie na anyị enweghị ike iwepụ usoro nke nkwụsi ike protein site na TSA site na acetylation nke protein na-abụghị histone38, nchọpụta a na - egosi na oxtr onye na-akwalite, TSA nwere ike ịkwalite avpr1a ntụgharị site na mpaghara histone acetylation. Agbanyeghị na ntinye aka nke AVP na njikọta njikọta ụmụ nwoke5, ọrụ ya na akparama ụmụ nwanyị ka na-arụ ụka. N'otu aka, ihe icv. Ntinye AVP na-eme ka enwee mmekorita nwoke na nwanyi na ntuli aka nke nwoke na nke nwanyi, nke gbochiri site na mgbochi nke ma obu V1aR ma obu OTR39. N’aka nke ozo, an icv. ogwu ogbugba nke V1aR na-egbochi udiri onye mmekorita nwoke na nwanyi, ma obughi nke nwanyi, prairie voles40. Agbanyeghị, ọmụmụ ihe ndị a niile ji icv. inje, na-egbochi nghọta ọzọ ọ bụla banyere ihe ndị metụtara ya. N'ebe a, anyị na-enye ihe akaebe mbụ na AVP neurotransmission n'ime NAcc nwere ike itinye aka na nhazi nke mmekọ na voles nwanyị, ebe ọtụtụ akwụkwọ na-akọwa ntinye ya na mpaghara dị iche iche dịka ventral pallidum, septum mpụta, akwa akwara nke stria teralis, na amygdala n’ime ụmụ nwoke5. Ya mere o gha buru ihe itughari iburu n’uche na mgbochi nke ma obu OTR ma obu V1aR n’ime nwanyi nke NAcc ezuru iji gbochie udiri onye obula n’abia usoro TSA, na-enye echiche na nhazi nke onye choro ya choro ka oru ya ma V1aR na OTR. Tihe ochoputa ya kwekọrọ, ma kwado, nlebara anya na preri nwoke na-eme ka uzo nke OTR na V1aR banye na LS di nkpa maka ochicho onye obula n’ebili.6. Ọzọkwa, nchọpụta nke mmụba dị na ọkwa abụọ nke OTR na V1aR na ụmụ anụmanụ a na-elekọta TSA na-akwado nkwado nke imegharị n'otu oge nke AVP na akwara oxygentocin maka njikọta ọnụ.
Na ntinye na oxygentocin na AVP, dopamine neurotransmission na NAcc modulates na mmeputa mmekorita nke nwanyi.9. Ọ bụ ezie na mating na-akpali ntọhapụ dopamine na NAcc9, mgbanwe dị iche iche na ọkwa nnabata a na-ahụ naanị mgbe ogologo oge dị ogologo karịa karịa karịa nke 24h- nke ibikọ ọnụ na mating, dị mkpa maka mmezi nke njikọkọ ọnụ.10. N'ikwekọ na nleba anya ndị a, praị nwanyị praịmarị mesoro TSA gosipụtara mmasị onye ibe na-enweghị nnukwu mgbanwe na ndị na-anabata dopamine D1R na D2R. Yabụ, na nnabata nke nnabata nnabata dopamine na-enye ihe akaebe ọzọ banyere njirimara nke TSA.
Ihe omuma anyi diri oge mbu ihe mejuputara anyi na akwara ozi nke ejikota onu, ma tugharia na TSA mee ka onodu “ikike ichoro” n’azu nwanyi, na eleghari ihe ndi mmadu anya na nnabata, ma na-akwalite mmezi nke mmekorita ndi mmadu. eduga na ibe ibe. Ya mere, a na-anwale ịkọwa na mmasị nke onye mmekọ TSA nwere ike nwekwuo ike ma duga na nkekọ na-adịgide. Agbanyeghi HDACs a kapịrị ọnụ ka ga-amata, ọ ga-abụ ihe na-atọ ụtọ inyochakwu nsonaazụ nke TSA na omume ndị ọzọ metụtara atụmatụ ndụ otu nwanyị na preirie voles, dịka ịkpa oke na nlekọta nne na nna. Ideringtụle mkpa nke akpọrọ praị dị na i modelomi usoro akwara ozi na ejikọta mmadụ na ibe ya5, na HDAC na-ekwe nkwa na-egbochirịrị ule ndị dị n'ụlọ ọgwụ24, 41, 42, data anyị na-emeghe ụzọ maka ohere ọgwụ ọhụụ iji metụta omume mmekọrịta mmadụ.
ụzọ
Ndị isi
Mmekọahụ naïve nwanyi prairie voles (Microtus ochrogaster) site na ogige ozuzu ụlọ nyocha ka a kwụpụrụ site na ụbọchị 21 ma werekwa nwanne na nwoke na ibe ya na mkpuchi plastik (12 × 28 × 16 cm). ad libitum. Echebere mkpuchi niile n'okpuru 14: 10 h light-dark wareeor, na ọnọdụ okpomọkụ dị ihe dị ka 20 ° C. E kenyere anụmanụ niile aka na otu ụdị nyocha mgbe ha ruru ụbọchị 70 – 90. Onu ogugu anumanu eji eme ihe sitere na omumu ihe omumu gara aga nke ndi otu anyi na ndi ozo, tinyere ikike nyocha. Kọmitii Na-ahụ Maka Anụmanụ na Nlekọta Anụmanụ kwadoro Mahadum Florida.
Drugs
Sodium butyrate (NaB), nke etisasịwo na saline, na Trichostatin A (TSA), nke etisasịwo na mmiri ozuzu mmiri (CSF, BioFluids, Rockville, MD) bụ ndị a zụtara na Sigma-Aldrich (St Louis, MO). NaB gbawara intraperitoneally (ip) na onodu ogwu nke 600 mg / kg, nke amaara ka eme ka achota histone n’ime otutu uzo n’ime akwara.43, 44. N'otu aka ahụ, ụdị-iji maka TSA sitere na ọrụ ndị gara aga na-ekpebi ịdịmma ya na iwebata ihe omume Acetylation obodo na ọdịiche na nkwụpụta mkpụrụ ndụ na akwara19, 20. Nhọrọ V1aR na-anabata antagonist V1aRA, d(CH2)5[Tyr (Me)] AVP, na OTA antagonist OTA (B), [d(CH2)5, Tyr (mụ)2, Nkem4, Tyr-NH29] -OVT), ka enwetara site na Bachem (Torrance, CA). Ihe nke abuo karie, Otr antagonist nke ozo, OTA (T), dGly-NH2-d(CH2)5 [Tyr (mụ)2, Nkem4] OVT45, nke Dr. Maurice Manning (Mahadum nke Toledo, OH) nyere. A họọrọ ndị na-emegide ndị a na usoro ọgwụgwọ ha dabere n'ọmụmụ ihe ọmụmụ gara aga na-egosi nhọta ha maka V1aR ma ọ bụ OTR, n'otu n'otu35, 39, 46-49.
Stereotaxic cannulation na microinjection
Ejiri ọgwụ sodium nwanyi sodium pentobarbital (1mg / 10g), na 26 igwe igwe anaghị agba nchara nduzi cannulae (Plastics Otu, Roanoke, VA), etinyere na stereotaxically late (ebumnuche; mgbochi imi na −2.5 mm, 0.6 rostral, mpụta 1.0 mm, na 2.6 mm ventral to bregma) ma ọ bụ saịtị-dị iche iche na NAcc (bilaterally; imi imi na −2.5 mm, 1.7 mm rostral, ± 1.0 mm bilateral, na 4.5 mm ventral to bregma). Mgbe ụbọchị 3 gbakere, ndị ọ natara natara microinjections nke CSF ma ọ bụ CSF nwere ọtụtụ TSA. Mgbe ejiri ndị antagonists maka OTR ma ọ bụ V1aR mee ihe, a na-etinye ha oge nkeji 30 tupu TSA. Ejiri agịga 33 nke gbatịrị 1 mm n'okpuru ntuziaka cannula banye ebe a chọrọ, na ntụtụ ọkụ ọkụ nke 500 nL n'ime ventricle mpụta (icv.) ma ọ bụ 200 nL kwa akụkụ n'ime NAcc. Ejiri agịga ahụ na Hamilton Syringe (Hamilton, Reno, NV) site na polyethylene-20 tubing na ịda mbà n'obi na-eme nwayọ, na-achọ minit 1 kwa oge. Na ngwụsị nke nnwale ahụ, ejiri nkwatu ngwa ngwa wee chụọ ihe niile n'ọkụ na ụbụrụ amịpụtara iji nyochaa ebe ndị mmadụ na-elele anya site na nyocha. Ewepuru ihe ndi nwere uzo nke edoputaghi ya na nyocha.
Inwe mmekorita nwoke na ibe ya
Ngwa ngwa ip., icv., ma obu intra-NAcc inje ogwu nke ogwu, umu nwanyi noo na nwoke dika nwoke nke 6 awa n’enweghi nmekorita. Ekwuputaghi na enweghị mating ahụ bụ site na inyocha omume e sere na vidio ahụ. Maka nyocha nke neuroadaptations kpatara site na ibikọ ọnụ na mating, ụmụ nwanyị estrogen-primed (2µg kwa ụbọchị, ip., maka ụbọchị 3) nwoke na nwoke biri n'otu oge elekere 6 ma ọ bụ 24, wee gosipụta na ọnụnọ nke mating a posteriori na vidiyo vidiyo (sitere na 6 ruo 11 ụfụ n'oge awa 6 izizi nke ibikọ ọnụ).
Emere nnabata mmasị onye mmekọ ahụ ozugbo ịgbatịkọta ọnụ nke 6-hour, dị ka akọwara na mbụ11. Na nkenke, akụrụngwa nke ụlọ ime ụlọ atọ nwere ọnụ ụlọ na-anọpụ iche nke ejikọtara na ngịga abụọ yiri ibe ha, nke ọ bụla nwere anụmanụ na-akpali akpali - nwoke "ọbịa" amabeghị ma ọ bụ nwoke "ibe" maara nke ọma ejiri oge ahụ. Subjectsmụ nwanyị nwere onwe ha ịgagharị na ngwa ahụ n'oge nyocha nke 3, na ụmụ nwoke na-akpali akpali ka ejikọtara n'ime ngịga ha, na-ekwe ka enweghị mmekọrịta ọ bụla. A na-enyocha vidio ahụ dum na oge nke isiokwu ahụ n'akụkụ n'akụkụ na ma ọ bụ onye òtù ọlụlụ ma ọ bụ onye ọbịbịa ka onye nyocha a zụrụ azụ na-amaghị maka ìgwè ndị dị ndụ. A kọwapụtara mmasị onye ọ bụla dị ka ndị na-etinye oge dị ukwuu n'inwe ahụ ma soro onye ọbịbịa, dị ka mkpebi abụọ, nke abụọ si kpebie. t-wa kacha. Na ntinye, ngwa ulo ato nke nwere fotobeam-sensọ, na-enye ohere mkpebi nke ọrụ nwetagoro egosi site na ọnụọgụ nke ntinye nwanyị gosipụtara n'ime ụlọ mkpali. Ntube akara a na - enyere anyị aka ijikwa mmetụta ọgwụ ọjọọ na omume ụmụ nwanyị, dịka ọrụ izugbe, nchekasị, ma ọ bụ mgbanwe nke gburugburu ebe a na - eme akwụkwọ akụkọ, dịka otu anyị na ndị ọzọ si eme.12.
RNA na mmịpụta protein
A na-eji ụmụ nwanyị achụrụ ụmụ nwanyị àjà ngwa ngwa, a na-ewepụta ụbụrụ ozugbo ma na-ajụkwa oyi n’elu mmiri akọrọ. Akpọnyere akụkụ coronal (200 µm) na cryostat na ntu oyi nke etinyebere na slides microscope. Ewere ọnya anụ ahụ na dayameta nke 1 mm ka site na NAcc dum na caudate putamen, nke ikpeazụ bụ mpaghara nchịkwa, ma chekwaa na −80 ° C ruo mgbe edoziri. A na-ewepụta ngụkọta RNA na protein site na iji usoro TRI-Reagent dịka ntuziaka onye nrụpụta (Molecular Research Center, Cincinnati, OH) si kwuo.
Nyochapụta protin site na Western-blot
Na-eso nkewa na 10% polyacrylamide gel (15% maka akụkọ), a na-ebufe protein na mkpocellulose membranes ma tinye ya na ọgwụ mgbochi ndị a: mgbochi-OTR (sc-8102, 1: 1000), -V1aR (sc-18096, 1: 500), -D1R (sc-33660, 1: 1000), -D2R (sc-9113, 1: 1000, Santa Cruz Biotechnology, Santa Cruz, CA), -actin (A2066, 1: 1000, Sigma Aldrich. St Louis, MO), ma ọ bụ anti-acetyl histone H3 (Lys14, # 06 – 911, 1: 1000) na mkpokọta H3 (# 05 – 928, 1: 1000, Millipore, Temecula, CA). A kwadoro ọgwụ mgbochi niile maka iji ha na ụmụ mmadụ, oke, na ụmụ oke, nke prairie voles na-ekerịta pasent dị elu nke usoro ọmụmụ (sitere na 81 ruo 96%). Egosiputa membranes na ECL (ECL SuperSignal West Dura), Pierce Biotechnologies, Rockford, IL) ma kpughee ihe nkiri Fuji XAR (Fuji Film, Tokyo, Japan). Emere ihe nchoputa site na iji ngwanrọ AIS 6.0 Image (Nyocha Onyonyo, St. Catharines, Ontario, Canada), na akara niile bụ ihe edoziri n'ime otu akpụkpọ ahụ iji mee ihe, belụsọ maka mgbaama acetyl-H3 nke ahaziri nke ọma na ngụkọta ihe ngosi H3. Ekwesịrị igosipụta data nke ọma ka pasent nke anụmanụ anụmanụ ejiri CSF.
Ihe mmeghari polymerase na-eme ka ihe dị n'otu (RT-PCR)
A haziri 0.5 µg nke RNA mkpokọta maka njikọta ijikọ DNA, wee nyochazie dịka esi kọwaa ya na mbụ.50 na iji usoro nicotinamide adenine dinucleotide dehydrogenase (NADH) nicotinamide. Emere mmeghachi omume niile na njem ndị njem na kwenyesiri ike na ha bụ eziokwu site na ịgbaze usoro nyocha na nkewa na 2% Agarose gel. Usoro nke primers ji mee ihe dị ka ndị a: 5'-TCCAAGGCCAAAATCCGCACGG-3 '(Fwd) na 5'-GGCAGAAGCTTCCTTGGGGGG-3' (Rev) maka OTR, 5'-GAGGTGAACAATGGCACTAAAACCGGGXGGXGGXXGGXXGGXCGGXCXGX (Rev) maka AVP3aR, 5'- TTAACAACAATGGGGGGGGGGGG XXX '(N'ihi) na 3'-GGCATGAGGGATCAGGTAAA-1' (Rev) maka D5R, 3'-GTGAAGGCGCTGTAGGGGGGGGGGGGTGGGGGGGCGGXCGGXCG ) maka D5R, na 3'-CTATTAATCCCCGCCTGACC-1 '(N'ihi na) na 5'-GGAGCTCGATTTGTTTTGCT-3' (Rev) maka NADH. Ekwesịrị igosipụta data nke ahaziri dịka pasent nke anụmanụ anụmanụ ejiri CSF.
Nkọwapụta nke Chromatin
E nyochara Aconelation XoneNUMX (Lys3) na NAcc na caudate putamen tissue punches site na iji Magna ChIP protein G Tissue Kit (Millipore, Temecula, CA) na-agbaso ntuziaka onye nrụpụta. Na nkenke, mgbe agbasachara ya na 14% formaldehyde, chụrụ chronatin site na iji Misonix XL-1 na ibepu nke 2000 – 200 bp. Emechara nweta nghọta nke acetylated histone H600 (Lys3) na 14µg nke mgbochi acetyl-H10 (Lys3) antibody (Millipore) chi abọọ na 14 ° C. Hes ghachaa nsị, nzere site na beads na nloghachi nke njikọta njikọ, a na - ejikwa DNA mee ka ihe dị ọcha ma nyochaa nyocha nke RT-PCR n'elu ikpo okwu iCycler (lee nke dị n'elu) ya na usoro dị omimi n'ime nke emere site n'omume INPUT. Emere ndị isi ala ahụ iji mee ka mpaghara 4 bp dị ogologo dị n'etiti 236 bp dị elu nke akara exon mbụ maka praịm cha cha OTRoxtr, Ntinye GenbankAF079980), ma ọ bụ 192 bp-ogologo mpaghara dị 141 bp elu elu nke mbụ exon nzuzo maka prairie vole V1aR (avpr1a, Ntinye GenbankAF069304). Usoro ndị ahụ bụ ndị a: 5'-CTCCGGAGCCGGGGAAAGT-3 '(Fwd) na 5'-ACCGCTTCCCCGAGAGTAGGG-3' (Rev) maka oxtr, na 5'-GGTGGACCAGCCAGACCCCA-3 '(Fwd) na 5'-TGCAGAGCCAGGCGCTTTCC-3' (Rev) maka avpr1a. Ejiri usoro INPUT dị iche iche wee kwado ọkwa ọ bụla, yana egosikwa data dịka ọnụọgụ nke anụmanụ CSF mesoro.
Nyocha nyocha na nhazi data
Maka nyocha nke mmasị onye mmekọ, a na-agụpụ anụmanụ ndị gosipụtara omume ịlụ oke n'oge ebikọ ma ọ bụ na cannulae na-ezighi ezi. Maka nyocha ndị ọzọ nke molekul, enwere ike karịa otu isi ihe data n'otu kọmpụta na - achọpụta ihe mgbe enwere ya. Ọtụtụ n'ime nyocha ndị ahụ bụ ndepụtaghachi, belụsọ mgbe nsonaazụ doro anya. Ejiri oge ejiri na-agagharị site na anụ na -akpata ma anụmanụ nwere ike ime n'oge nnabata nke onye mmekọ ahụ ejirila njikọta abụọ jikọrọ ọnụ. t-wa kacha. Ejiri akara okpukpo abụọ tụọ ntụpọ locomotion t-ogodi (maka otu abuo) ma obu uzo ANOVA (nke kariri otu abuo), na mgbe okwesiri, Fischer's PLSD post-hoc Emere nnwale a na ntinye dị mkpa nke P <0.05. Mgbe nkwenye nke iwu zuru oke, ejiri data abụọ tụlee data ndị ọzọ niile t-nche iche nha ma ọ bụ nha anya dịgasị iche nwalere tupu oge a. Emere nyocha nyocha niile site na iji StatView sọftụwia (SAS Institute). Mgbe atulere data na njikwa ha (% nke CSF, Saline ma ọ bụ Mating otu), a duziri nyocha nke ndekọ ọnụ ọgụgụ ahụ.
Nkweta
Nkwado sitere na National Institute of Mental Health (NIMH) nyere MHR21-083128 nye MK na ZW, MHR01-058616 nye ZW Anyị na-ekelekwa Dr. Maurice Manning maka onyinye mmesapụ aka nke Oagon antagonist OTA (T).
Ihe odide ala
Esemokwu mmasị: Ndị na-ede akwụkwọ ekwupụtaghị mmasị ọ bụla.
Onye nyere onyinye
HW, FD na YL mere nnwale ahụ. HW na FD nyochara data ahụ. HW, FD, ZW na MK mere nchọpụta ahụ. FD, ZW na MK dere akwụkwọ ahụ. Ndị edemede niile kwurịtara nsonaazụ ya ma kwuo banyere ihe odide ahụ.
References
;