Ukusetyenziswa kwe-Perinatal Western Diet Ukusetyenziswa Kukhokelela kwiPlastiki enobuchule kunye ne-GABAergic Intshentshi kwiinguqu ze-Hypothalamus kunye neNzuzo yoNyaka ukusuka ekuzalweni koBuchule boSondo kwi-Rat (2017)

. 2017; 8: 216.

Ipapashwe kwi-intanethi 2017 Aug 29. doi:  I-10.3389 / fendo.2017.00216

PMCID: PMC5581815

Abstract

Ukusetyenziswa koomama ngaphambi kokubeleka kokutya okuxineneyo kwamandla kwandisa umngcipheko wokutyeba ebantwaneni. Oku kuhambelana nokugqithiswa kokutya okunencasa okusetyenzisiweyo kwipropati yayo ye-hedonic. Indlela esisiseko enxulumanisa ukutya kukamama okhulelweyo kunye nokukhetha inzala ngamafutha ayikaqondwa kakuhle. Kolu phononongo, sijolise ekufundeni ifuthe lokutya okunamafutha aphezulu / okuneswekile ephezulu [ukutya kwasentshona (WD)] ngexesha lokumitha kunye nokuncancisa kwindlela yomvuzo olawula ukondla kwinzala yempuku ukusuka ekuzalweni ukuya ekukhuleni ngokwesondo. Senze ulandelelwano olude lwe-WD kunye noLawulo lwenzala ngamaxesha amathathu abalulekileyo (ubuntwaneni, ukufikisa, kunye nokuba mdala) kwaye sigxininise ekuphandeni ifuthe lokuvezwa kwe-perinatal ekutyeni okunencasa kwi (i) ukhetho lwamafutha, (ii) iprofayili yokubonisa imfuza. , kunye (iii) utshintsho lwe-neuroanatomical/architectural ye-mesolimbic dopaminergic networks. Sibonise ukuba ukondla kwe-WD okukhawulelwe kwixesha lokubeleka kunempembelelo ecacileyo ehlala ixesha elide kumbutho we-homeostatic kunye neesekethe zobuchopho be-hedonic kodwa hayi kukhetho lwamafutha. Sibonise ixesha elithile lokuzivelela kokukhetha amanqatha esinxibelelana nawo kunye neesayino ezithile zeemolekyuli zengqondo. Kwinzala ephuma kumadama esondliwe e-WD, sabona ngexesha lobuntwana ubukho bokuthanda amanqatha okuhambelana nokubonakaliswa okuphezulu kofuzo oluphambili olubandakanyekayo kwiinkqubo ze-dopamine (DA); ebusheni, ukukhetha okunamafutha aphezulu kumaqela omabini, kuncitshiswe ngokuthe ngcembe ngexesha lovavanyo lweentsuku ze-3 kwiqela le-WD kwaye linxulunyaniswa nokuncipha kokubonakaliswa kofuzo oluphambili olubandakanyekayo kwiinkqubo ze-DA zeqela le-WD elinokuthi licebise indlela yokubuyisela ukubakhusela. ukusuka ekuvezweni ngakumbi kwamafutha aphezulu; kwaye ekugqibeleni ebudaleni, ukhetho lwamanqatha awayefana nokulawula iimpuku kodwa ahambelana nokuguqulwa okunzulu kwiijenethi eziphambili ezibandakanyekayo kwinethiwekhi ye-γ-aminobutyric acid, i-serotonin receptors, kunye ne-polysialic acid-NCAM-exhomekeke kuhlengahlengiso lwe-hypothalamus. Lilonke, ezi datha zityhila ukuba i-WD kamama, ithintelwe kwixesha lokubeleka, ayinayo impembelelo ezinzileyo kwi-homeostasis yamandla kunye nokukhethwa kwamafutha kamva ebomini nangona uhlengahlengiso olomeleleyo lwe-hypothalamic homeostatic kunye nendlela yomvuzo ebandakanyeka ekuziphatheni kokutya kwenzeka. Eminye imifuniselo esebenzayo iya kufuneka ukuqonda ukubaluleka kwezi sekethe zihlengahlengiswa.

Internet: umvuzo, i-DOHaD, ukhetho lokutya, isondlo, i-γ-aminobutyric acid, uluhlu lwe-TaqMan oluphantsi koxinaniso

intshayelelo

Imekobume yobomi bokuqala kunye neziganeko ngoku ziqondwa kakuhle ukuba negalelo kwimpilo nakwisifo esihlala sihleli kamva ebomini (-). Umbono wokuprintwa kwe-metabolic ucetyisiwe ukuchaza ukuba utshintsho kwimeko yesondlo kunye nehomoni ngexesha lokubeleka lunokuthi lubeke inzala ekutyebeni kakhulu kunye ne-pathologies ehambelana nayo kamva. Umba obalulekileyo wendlela yethu yokuphila occidental kukutya ngokugqithisileyo njengesiphumo sokusetyenziswa kokutya okuxineneyo kwamandla. Ewe, abantu abavezwa ngokutya koomama kolu hlobo lokutya basengozini enkulu yokutyeba kakhulu kunye nesifo se-metabolic syndrome (, ). Izifundo ezininzi zibonise ukuba ukutya okunamafutha aphezulu kumama (HFD) ngokumitha kunye nokuncancisa kunempembelelo yexesha elide kwimetabolism yabantwana (-). Ukongeza kwiindlela ezibandakanyekayo kulawulo lwe-metabolic, iinkqubo zomvuzo wobuchopho nazo zidlala indima ebalulekileyo ekuziphatheni kokutyisa (, ). I-Mesolimbic dopamine (DA) neurotransmission, efundwe ngokunzulu kumxholo womvuzo kunye nokulutha, iguqulwa kukutyeba okubangelwa kukutya kubo bobabini abantu (-) kunye nezilwanyana (-). Uqikelelo lweDA luyaphuhla, inxalenye enkulu, emva kokubeleka (), kwaye ngoko ke uphuhliso lwabo lunokuchaphazeleka kukutya kwangaphambili. Kule minyaka imbalwa idlulileyo, iimvavanyo zeempuku zibonise ukuba ukuthatha i-HFD kamama kuphucula ukondla kwe-hedonic kwinzala (, ). Nangona olu qwalaselo lubandakanya utshintsho oluthile kumsebenzi wenkqubo yeDA (-), idatha encinci iyafumaneka malunga ne-ontogeny kunye nokulungiswa kweendlela zomvuzo ngexesha lobomi bokuqala (). Ngaphezu koko nokuba indlela engeyiyo i-DA yokubonakalisa inxalenye yenkqubo yomvuzo efana ne-GABA (γ-aminobutyric acid) inkqubo inokuchaphazeleka ngoxinzelelo lwezondlo lwe-perinatal alubhalwanga. Enyanisweni, i-GABA neurons ibonakala idlala indima ephambili kumvuzo kunye nokungafuni. Indawo ye-ventral tegmental (VTA) I-GABA neurons ifumana ipateni efanayo yegalelo kwiindawo ezahlukeneyo zobuchopho.), kunye nezifundo zokuziphatha ezisekelwe kwi-optogenetic zamva nje zibalaselisa indima enkulu ye-VTA GABA kwindawo yokuchaswa kwemeko () kunye nokuziphatha kokugqiba umvuzo (). I-Nucleus accumbens (NAc) yenziwa ikakhulu luqikelelo lwe-GABAergic medium spiny neurons kwaye isebenza njenge-limbic-motor interface edibanisa imiqondiso evela kwi-limbic system kwaye iyijike isebenze. ngokusebenzisa imveliso kwi-ventral pallidum (VP) kunye nezinye ii-motor effectors (). Kwaye ekugqibeleni, i-hypothalamus eyenziwa ludibaniso oluninzi lwe-GABA kwi-LH () kwaye i-arcuate nucleus, idibanisa imiqondiso yendlala kunye nokuhlutha ().

Olu phononongo lujolise ekuchongeni impembelelo yokutya kwasentshona koomama (WD) kwinzala yempuku ukusuka ekuzalweni ukuya ekukhuleni ngokwesondo (i) ekuthandeni amanqatha (ii) kwiprofayile yegene yenkqubo ye-DA, inkqubo ye-GABAergic kunye neplastiki ye-hypothalamus. , kunye (iii) notshintsho lwe-neuroanatomical/architectural ye-mesolimbic dopaminergic networks for the same time. Ngoko ke sivavanye, kwisifundo sexesha elide (ukusuka ekulumleni, i-P25, ukuya ekukhuleni ngokwesondo, i-P45 kunye nobuntu obudala, i-P95), umphumo we-WD yomama ekukhuleni kwesisindo somzimba kunye nokuphuhliswa kwezicubu ze-adipose zenzala egcinwe phantsi kwe-chow rhoqo emva kokulunyulwa. Ngokuhambelanayo, senze uvavanyo lokukhetha amanqatha olulandelwa luhlalutyo oluzinikeleyo lwe-transcriptomic kunye nohlalutyo lwecandelo eliphambili (PCA) lokukhetha abamakishi bokutya, ukhetho kunye neenkqubo zokulawula ezikhuthazayo. Iziphumo zethu ziye zatyebisa kakhulu iziphumo zakutsha nje ezijolise kwinkqubo yesondlo senkqubo yeDA.

Impahla nenkqubo

Statement Ethics

Zonke iimvavanyo zenziwe ngokuhambelana nezikhokelo zekomiti yentlalontle yezilwanyana zendawo, i-EU (umyalelo 2010/63/EU), i-Institut National de la Recherche Agronomique (eParis, eFransi), kunye neSebe lezilwanyana zaseFransi (A44276). Iprothokholi yokulinga yamkelwa yikomiti yeenqobo ezisesikweni yeziko yaze yabhaliswa phantsi kwereferensi ye-APAFIS 8666. Onke amanyathelo okhuseleko athatyathwa ukunciphisa uxinzelelo kunye nenani lezilwanyana ezisetyenziswa kuthotho lwemifuniselo.

Izilwanyana kunye nokutya

Izilwanyana zagcinwa kumjikelezo we-12 h / 12 h ukukhanya / ubumnyama kwi-22 ± 2 ° C kunye nokutya kunye namanzi. ad adum. Amashumi amathathu anesibini eempuku zaseSprague-Dawley (ubunzima bomzimba: 240–290 g) ngosuku lokumitha 1 (G1) zathengwa ngqo kuJanvier (Le Genest Saint Isle, France). Bahlaliswa ngabanye kwaye bondliwa nokuba kukutya okulawulwayo (CD) (i-5% yamafutha enkomo kunye ne-0% ye-sucrose) kwi-16 yabo okanye i-WD (i-21% yamanqatha enyama yenkomo kunye ne-30% ye-sucrose) kwi-16 kubo ngexesha lokumitha kunye ne-lactation. (bona iTheyibhile Itheyibhile1: 1: ukubunjwa kokutya kwipesenti kcal ukusuka ku-ABdiet Woerden, eNetherlands). Ekuzalweni, ubungakanani benkunkuma bulungelelaniswe ukuya kumantshontsho asibhozo kwilitha nganye kunye ne-1: 1 yendoda ukuya kumfazi. Sagcina amadama ali-12 kwali-16 kunye nenkunkuma eyenziwe ngamadoda ama-4 namabhinqa ama-4 kwiqela ngalinye. Ekulunyulweni (P21), inzala ezalelwe kwiCD kunye namadama e-WD yagcinwa kwi-chow eqhelekileyo kude kube sekupheleni kovavanyo (Amanani (Amanani1A, B).1A,B). Ubunzima bomzimba wenjana barekhodwa ekuzalweni kwaye emva koko yonke imihla ngentsimbi ye-10:00 kusasa ukuya kwi-P21 (ukulunyulwa). Emva kokulunyulwa kwaye kude kube sekupheleni kovavanyo, iimpuku zanikwa ubunzima rhoqo ngeentsuku ezi-3. Sibonisa idatha kwinzala yamadoda kuphela. Iimpuku ezingamamazi zazisetyenziselwa olunye uphononongo (Umfanekiso (Umzobo11).

1 Table 

Ukubunjwa kokutya kwipesenti ye-kcal ukusuka kwicandelo ngalinye lokutya koomama okulawulwa ngexesha lokukhulelwa kunye ne-lactation kunye nokutya okusemgangathweni kwinzala.
Umzobo 1 

Uyilo lo vavanyo. (A) Umzobo wesicwangciso soyilo lokufunda. Iigundane ezingamashumi amathathu anesibini ze-SPD zabasetyhini ngexesha lokukhulelwa kwe-1 (G1) zondliwa ngokutya okulawulwayo kwi-16 yazo okanye ukutya kwasentshona kwabanye ngexesha lokukhulelwa kunye nexesha lokuncancisa. Ekulunyulweni, inzala ...

Ukuziphatha (Uvavanyo lokuKhetha iibhotile ezimbini)

Amaxesha amathathu ophuhliso abalulekileyo ahlolisisiwe (i-P21 ukuya kwi-P25: intsha, i-P41 ukuya kwi-P45: ukufikisa kunye ne-P91 ukuya kwi-P95: umntu omdala osemtsha). Amantshontsho angama-24 angamadoda (n = I-12 ngeqela ngalinye) zikhethwe ngokungenamkhethe kwaye zifakwe kwikheji nganye ukwenza uvavanyo lweebhotile ezimbini zasimahla (Amanani (Amanani1A, B)1A,B) (-). Olu vavanyo lusetyenziselwe ukufunda ngokuthe ngqo umtsalane kwincasa yamafutha ngokuyisusa kwincasa eswiti kwaye kangangoko kunokwenzeka kwisiphumo sokutya kwekhalori. Enyanisweni, i-1% yokusetyenziswa kwesisombululo seoli yengqolowa idibaniswa nokutya kwe-0.09 kcal / ml kuphela. Emva kosuku olunye lokuhlala kubukho beebhotile ezimbini, uvavanyo lwenziwa ngaphezulu kweentsuku ezi-2 kwi-P25 kwaye ngaphezulu kweentsuku ezi-4 kwi-P41 kunye ne-P91 (Umfanekiso. (Umfanekiso1A) 1A). Ngokweenkcukacha, xa elunyulwa (P21), amantshontsho angama-24 ahlaliswa eyedwa kangangeentsuku ezi-2 (Umfanekiso (Umfanekiso1A):1A): usuku lwe-1, isigaba sokuhlala, usuku lwe-2, iigundane zanikwa i-bottle-bottle free choice phakathi kwe-emulsion ye-1% yeoli yengqolowa kwi-0.3% xanthan gum (Sigma Aldrich, St. Quentin Fallavier, France) kunye ne-xanthan gum solution ( 0.3%). Kwi-P41 kunye ne-P91, amantshontsho angama-24 asetyenzisiweyo kwaye ukhetho lwasimahla lweebhotile ezimbini lwacetywa iintsuku ezintathu ezilandelelanayo. Ukusetyenziswa kwesisombululo se-xanthan gum kunye nesisombululo se-taste (ioli ye-corn 1%) yabhalwa imihla ngemihla ngo-11: 00 ekuseni ngeentsuku ze-3 (P45 kunye ne-P95). Ukuma kweebhotile ezimbini kwaguqulwa imihla ngemihla ukunqanda ukukhetha okukhethiweyo. Amanqaku okukhetha amanqatha abalwe njengomlinganiselo "wesisombululo se-fat" umthamo osetyenzisiweyo kumthamo opheleleyo osetyenziswe kwi-24 h. Zonke iigundane zagcinwa phantsi kokutya okuqhelekileyo kwe-chow kulo lonke uvavanyo lokuziphatha.

Ukuqokelelwa kwezicubu kunye neSampulu yeGazi

Ngomhla emva komhla wokugqibela wovavanyo lweebhotile ezimbini zasimahla, isiqingatha seempuku (n = 6 ngeqela ngalinye) baxhaswe ngokukhawuleza phakathi kwe-09:00 kunye ne-12:00 am ngu-CO.2 ukuphefumla. Igazi laqokelelwa kwiityhubhu kunye ne-EDTA (Laboratoires Léo SA, St Quentin en Yvelines, France) kunye ne-centrifuged kwi-2,500 g imizuzu eyi-15 kwi-4°C. Iplasma yayikhenkceke kwi-−20°C. Amalungu kunye nendawo yokugcina i-fat retroperitoneal nganye yachithwa kwaye yabekwa ubunzima. Ingqondo yasuswa ngokukhawuleza kwaye yafakwa kwi-matrix yengqondo (WPI, Sarasota, FL, USA rat 300-600 g). Okokuqala i-hypothalamus yachithwa [ngokwePaxinos's atlas coordinates: -1.0 ukuya -4.5 mm ukusuka eBregma ()] ngoko, kwi-rat nganye, iziqwenga ezimbini ze-coronal ze-2 mm ubukhulu kwinqanaba le-NAc kunye nenye enye kwinqanaba le-VTA zifunyenwe. Iisampulu ze-NAc ekunene kunye nesobunxele kunye ne-VTA ekunene kunye nesobunxele (iisampulu ezine zizonke kwisilwanyana ngasinye) zifunyenwe ngokukhawuleza kusetyenziswa iipuntshi ezimbini ezahlukeneyo ze-biopsy (i-Stiefel Laboratories, i-Nanterre, eFransi) (ububanzi be-4 mm ye-NAc kunye ne-3 mm kwi-ventral midbrain). Iisampulu zaxhwilwa emkhenkceni kwinitrogen engamanzi zaze zagcinwa ku-−80°C ukuze kugqitywe ukuchazwa kofuzo yi-TaqMan low-density array (TLDA).

Ezinye iimpuku (n = 6 ngeqela ngalinye) i-anesthetized enzulu kunye ne-pentobarbital (150 mg / kg ip) kwaye ixutywe nge-transcardial physiological saline perfusion elandelwa yi-ice-cold 4% paraformaldehyde kwi-phosphate buffer (PB), pH 7.4. Ubuchopho bususwe ngokukhawuleza, bufakwe kwisilungiso esifanayo kwi-1 h kwi-4 ° C, kwaye ekugqibeleni igcinwe kwi-25% ye-PB sucrose ye-24-48 h. Ubuchopho ke babukhenkceke kwi-isopentane ku-−60°C, kwaye ekugqibeleni bagcinwa ku-−80°C de busetyenziswe. I-NAc, i-hypothalamus kunye ne-VTA ziye zasikwa zaba ngama-20 µm amacandelo e-coronal ene-cryostat (i-Microm, i-Microtech, i-Francheville, i-France). Iintlobo ezimbini okanye ezintathu zeesilayidi zeglasi ze-10 eziqukethe amacandelo e-4-6 zenziwa kwindawo nganye yobuchopho. Kwisilayidi seglasi nganye amacandelo othotho ahlulwe ngama-200 µm (Umfanekiso (Umzobo66).

Umzobo 6 

Ubungakanani be-TH / NeuN positive neurons kwindawo ye-ventral tegmental (VTA) kunye ne-TH density fibers kwi-nucleus accumbens (NAc) ukusuka elunyulwe ukuya ebudaleni kwinzala evela kwi-western diet (WD) okanye i-control diet (CD) yokutya amadama. (A) Isicwangciso esivela ePaxinos kunye neWatson's ...

Uhlalutyo lwePlasma yeBiochemical

I-plasma ye-EDTA eqokelelwe kwi-P25, i-P45, kunye ne-P95 iigundane zisetyenziselwa ukulinganisa i-glucose ye-plasma, i-NEFA (i-non-esterified fatty acids), i-insulin kunye ne-leptin. I-Glucose kunye ne-NEFA zilinganiswe ngokusebenzisa i-colorimetric reaction reactions nge-kits ezithile (i-glucose kunye ne-NEFA PAP 150 kits, i-BioMérieux, i-Marcy-l'Etoile, eFransi). IiHormones zavavanywa ngeekhithi ezithile ze-ELISA ngokulandela imiyalelo yomenzi we-insulin kunye ne-leptin (i-rat/mouse insulin ELISA kit, i-rat leptin ELISA kit, i-Linco Research, iSt. Charles, MO, USA).

Immunohistochemistry

Iislayidi zeglasi eziqulethe i-serial VTA kunye namacandelo e-NAc aqale avaliwe kwi-3-4 h kwaye emva koko afakwe ubusuku bonke kwi-4 ° C kunye nomxube we-antibodies elandelayo: i-mouse anti-NeuN (1: 500; IgM; iMillipore Bioscience Research Reagents, Merk, USA) kunye nomvundla ochasene ne-TH (1: 1,000; iMillipore Bioscience Research Reagents, Merk, USA). Emva kokufakwa kunye ne-antibodies yokuqala kunye nokuhlamba okulandelayo kunye ne-PB, amacandelo afakwe kumxube we-antibodies yesibini: i-Alexa 488 conjugated donkey anti-mouse IgM kunye ne-Alexa 568-conjugated donkey anti-rabbit IgG (1: 500; Invitrogen, ThermoFisher Shaltific, Walter Shamtific , MA, USA) kwi-2 h. Amacandelo axhonywe kwi-superfrost kunye nezilayidi zegolide (ThermoFisher Scientific, Waltham, MA, USA), zomiswe ngomoya, kwaye zagqunywa ngeProLong™ Gold antifade reagent (Invitrogen, ThermoFisher Scientific, Waltham, MA, USA).

I-TH Neurons Bala kwi-VTA

Kwimpuku nganye, iiseli ezine-TH zibalwa njengoko kuchaziwe ngaphambili () kumanqanaba amathathu ahlukeneyo e-rostrocaudal ye-VTA: kwinqanaba lokuphuma kwi-nerve yesithathu (umgama onxulumene neBregma: -5.3 mm), i-200 µm rostral kunye ne-200 µm caudal kule nqanaba (Amanani (Amanani6A) 6A). Kwicala lasekhohlo nelasekunene, umfanekiso ofakwe ekhompyutheni oquka yonke i-VTA esuka kwisixhobo sokufikelela kwitheminali ukuya kumda osecaleni we-mesencephalon ufunyenwe kusetyenziswa × 40 ukukhulisa i-NanoZoomer-XR Digital slide scanner. C12000 (uHamamatsu, eJapan). Umgca uzotywe ujikeleze umjikelezo we-VTA kwicandelo ngalinye. Imida yakhethwa ngokuhlolisisa imilo yeeseli kunye nokubhekisela kwi-atlas ye-Paxinos kunye ne-Watson. I-neuron ye-dopaminergic yachazwa njenge-NeuN (+) / TH (+) i-immunoreactive cell body ene-nucleus ebonakalayo ngokucacileyo. Ukusebenzisa i-software ye-NIH Image J (i-plugin counter counter), iiseli ze-NeuN (+) / TH (+) zibalwe ngabantu ababini abahlukeneyo abangenalo ulwazi lwamaqela ezilwanyana. Iimpazamo zokubalwa kweeseli eziqhekezayo zalungiswa kusetyenziswa ifomula ye-Abercrombie (), apho N = n[t/(t + d)] ((N = inani elipheleleyo leeseli; n = inani leeseli ezibaliweyo; t = ubukhulu becandelo; kwaye d = i-cell diameter), kwaye le nto yokulungisa yayiyi-0.65. Idatha ichazwa njengentsingiselo [NeuN(+)/TH(+) ekhohlo nasekunene VTA] ± SEM.

I-TH Fiber Density kwi-NAc

Umxholo weprotheyini ye-TH kwii-terminals ze-dopaminergic nerve ze-NAc zaqikelelwa luhlalutyo lwe-anatomical densitometric yamacandelo e-TH angenawo ama-immunolabeled. Uxinaniso lwe-TH lwefibers lubalulwe kumanqanaba amathathu angenasizathu ecaleni kwe-rostrocaudal axis ye-NAc (Bregma 2.20, 1.70, kunye ne-1.20 mm) (Umfanekiso (Umzobo6B) .6B). Ngokufutshane, umfanekiso ofakwe edijithali obandakanya yonke i-striatum kunye ne-NAc efunyenwe kusetyenziswa × 40 ukukhulisa iskena sesilayidi seNanoZoomer-XR Digital C12000 (Hamamatsu, eJapan) zafunyanwa. Kwi-NAc enikeziweyo, umgca wawuzotywe ujikeleze i-nucleus yonke ukuchaza indawo yomlinganiselo we-optical density (OD) (Figure). (Umzobo6B) .6B). Ixabiso elifunyenweyo liqhelekile ngexabiso le-OD elilinganiswe ukusuka kummandla ojikelezayo ozotywe kwi-corpus callosum (ummandla ongangcoliswanga kwi-TH immunochemistry) yecandelo elifanayo usebenzisa i-software ye-NIH Image J. Idatha ichazwa njengentsingiselo ye-OD ratio (ixabiso le-OD kwixabiso le-NAc / OD kwi-corpus callosum yamacandelo amathathu) ± SEM.

Ukuchazwa kweGene nguTLDA kunye neTaqMan

I-RNA yayibekwe yodwa kwi-NAc ekhenkcezayo, iisampulu ze-VTA ezityetyisiweyo, kunye ne-hypothalamus, kusetyenziswa i-NucleoSpin RNA/protein kit (Macherey-Nagel, Hoerdt, France). Iyonke i-RNA yangeniswa kwi-DNase yokugaya ngokulandela imiyalelo yomenzi, ubungakanani buqikelelwe yi-260/280 nm i-UV yokufunxa, kwaye umgangatho wavavanywa kusetyenziswa i-Agilent 2100 Bioanalyzer System, inombolo yengqibelelo ye-RNA (RIN) emva koko yabalwa. Iisampulu ezine-RIN engaphantsi kwe-8 zalahlwa. Imicrogram enye ye-RNA iyonke yakhutshelwa umva kwi-cDNA kusetyenziswa Ikhithi ye-RT enomthamo oPhezulu (Ii-Applied Biosystems, i-Foster City, i-CA, e-USA) kumthamo opheleleyo we-10 µl.

Njengoko kuchaziwe ngaphambili (), i-TLDA yikhadi le-384-well micro-fluidic apho i-384 ye-PCRs yexesha langempela inokwenziwa (i-Applied Biosystems, i-Foster City, i-CA, i-USA). Sisebenzise i-TLDA eyilwe ngokukodwa eyenzelwe ukugubungela iintsapho ezahlukeneyo zemfuza ezifanelekileyo kwiplastiki kunye nokulawulwa kokutya. Ikhadi ngalinye lesiko lalungiswa njenge-2 × i-4-isampulu yokulayisha imigca equkethe i-2 × i-48 yamagumbi okuphendula (i-reference: 96a). Iseti yemfuza engama-92 (Itheyibhile S1 kwi-Supplementary Material) kunye nejene zogcino lwezindlu ezine (18S, Gapdh, Polr2a, kunye nePpia) zaphononongwa. I-PCR yexesha langempela yaqhutywa kusetyenziswa i-Life Technologies TaqMan reagents kwaye iqhutywe kwi-ABI Prism 7900HT inkqubo yokufumanisa ulandelelwano (i-Applied Biosystems, i-Foster City, i-CA, i-USA). Idatha ye-fluorescence ekrwada yaqokelelwa ngePCR kusetyenziswa isoftwe ye-SDS 2.3 (Applied Biosystems, Foster City, CA, USA), ethe yavelisa ngakumbi imijikelo ye-threshold Ct ngomiselo oluzenzekelayo lwesiseko kunye nomda. Emva kokucoca kusetyenziswa i-ThermoFisher cloud App (i-ThermoFisher, e-USA) ukucalula ii-PCR ezibalekayo, iimvavanyo ngesampulu nganye bezi. n = 6 (n = 5 kwiqela le-WD kwi-P25). Emva koko idatha yahlaziywa nge-ThermoFisher Cloud App (i-ThermoFisher, e-USA) malunga nobungakanani obuhambelanayo. Ubungakanani obunxulumeneyo bokubonakaliswa kofuzo (RQ) lwalusekwe kwindlela yokuthelekisa i-Ct kusetyenziswa i-equation RQ = 2−ΔΔCt, apho i-ΔΔCt yofuzo enye ekujoliswe kuyo yayiyeyayo ukwahluka kwe-Ct ethatyathwe kwisampulu yesilinganisi kwaye yenziwe yesiqhelo ngolawulo olungapheliyo. Ngokuchanekileyo, sinqume eyona gene yogcino lwendlu oluzinzile usebenzisa i-geNorm algorithm (i-ThermoFisher Cloud App RQ, iThermoFisher, eU.SA). Phakathi kwemizila emine yokugcina indlu, i-Gapdh yachazwa njengolawulo olungapheliyo lwe-NAc kunye ne-hypothalamus, kunye ne-Ppia ye-VTA kwaye oku kwakuyinyaniso kuzo zonke iisampuli ukusuka kumaxesha amathathu ahlalutyiweyo. Umboniso womzobo wokubonakaliswa kofuzo wawuyilwe ngesandla ukunika umbala omnye kwi-10% yokunyuswa kwembonakalo yemfuza xa kuthelekiswa neqela leCD. Ukwahluka okubalulekileyo, usebenzisa i-non-parametric Wilcoxon yovavanyo lwenqanaba esayiniweyo, yaqatshelwa ngeenkwenkwezi.

Uhlalutyo lweSatisati

Iziphumo zicaciswa njengee-SEM eziphantsi kwiitheyibhile nakumanani. Uvavanyo lweMann-Whitney non-parametric lusetyenziselwe uhlalutyo lobunzima boMzimba ngamaxesha ahlukeneyo, ukhetho lwamafutha, kunye nomlinganiselo we-OD ofunyenwe kwi-immunohistochemistry.

Ukuvavanya ukubaluleka kweentsuku ze-3 ezikhethiweyo zamanqatha, senze uhlalutyo lwamanani ekholomu ngosuku ngalunye. Kwiqela ngalinye, ukusetyenziswa kwesisombululo samafutha kunye nesisombululo sokulawula kwavavanywa kusetyenziswa uvavanyo lwe-Wilcoxon olungena-parametric. Siqhathanise ixabiso elikhethiweyo elikhethiweyo kunye nexabiso le-hypothetical ye-50% (umgca obomvu onamachaphaza). Ukwahluka okuphawulekayo kwaphawulwa ngeenkwenkwezi ezibomvu. Sisebenzise uvavanyo olufanayo lohlalutyo lwexabiso le-qPCR RQ; sithelekisa ixabiso eliphakathi kwe-RQ kunye nexabiso le-hypothetical ye-1. Ukwahluka okuphawulekayo kwaphawulwa nge-asterisk (Umfanekiso (Umzobo44).

Umzobo 4 

Ukubonakaliswa kofuzo oluhambelanayo kwi-nucleus accumbens (NAc), indawo ye-ventral tegmental (VTA), kunye ne-hypothalami evela kwi-perinatal-western diet feeded rats kunye ne-perinatal-control diet feeded rats ngamaxesha amathathu. Ubungakanani obungaxeshanye bokubonakaliswa kofuzo kwi ...

Uhlalutyo lwesampulu yeplasma, senze uvavanyo olungeyoparametric kaMann kunye noWhitney. Inani leeseli ze-TH-positive zahlalutywa ngeendlela ezimbini ze-ANOVA kunye ne p ixabiso libaliwe. Ngenxa yobuninzi beemvavanyo eziphunyeziweyo, iBonferroni iposi ulungiso lusetyenzisiwe lulandele olu vavanyo kuphela. Uhlalutyo lwamanani lwenziwa kusetyenziswa isoftware yePrism 6.0 (GraphPad Software Inc., La Jolla, CA, USA).

I-PCA engalawulwayo yenziwa kuqala kwiiparamitha ze-130 (i-TLDA, ukuziphatha, kunye nedatha ye-plasma) ngexesha elihlukeneyo kwi-punches ye-brain biopsy (VTA, NAc, kunye ne-hypothalamus) ukujonga isakhiwo ngokubanzi sesethi yedatha (oko kukuthi, i-PCA yesithathu yehlabathi ngexesha elinye). I-PCA inokuchazwa njengentelekelelo ye-orthogonal yedatha kwisithuba somda esisezantsi somda, ukuze umahluko wedatha eqikelelweyo yandiswe kwisithuba esingaphantsi. Siqale sahluza imizila yemfuza engachazwanga okanye engachazwanga kancinane (Figure (Figure5) .5). Amaxabiso enzala asuka kumadama e-CD awondliwayo kunye namadama awondliwa yi-WD avele ngemibala eyahlukeneyo kwizicwangciso ze-PCA zomntu ngamnye ukuze ubone ukuba la maqela mabini okulinga ahlulwe kakuhle ngamacandelo e-PCA angagadwanga. Olu hlalutyo lwahlula amaqela emfuza abonakaliswa ngokwahlukileyo phakathi kwamaqela amabini abantwana. Emva koko, ii-PCA ezigxininisiweyo zenziwa kwiqela elihlukeneyo labamakishi be-mRNA: iplastiki (i-cell adhesion, cytoskeleton, neurotrophic factor, synaptogenesis, kunye ne-transcription regulatory), indlela ye-DA, indlela ye-GABAergic, i-epigenetic modulators (i-histone deacetylase kunye ne-histone acetyl transferase). Ezi PCAs zigxininisekile zivumela umntu ukuba abone ngeso lengqondo ngaxeshanye ulungelelwaniso phakathi kokutya komama kunye nabanye abamakishi kunye nolungelelwaniso phakathi kohlobo oluthile losapho. Umlinganiselo osemgangathweni wawusetyenziselwa uhlalutyo lwe-PCA kunye ne-PCA egxininisekileyo: +++: ukuhlukana okuhle kakhulu; ++: Ukwahlula okulungileyo kunye negundane elinye kwicala elingalunganga lokwahlula kwePCA; +: ukwahlukana okuhle kakhulu kunye neempuku ezimbini (enye kwiqela ngalinye) kwicala elingalunganga, −: akukho kwahlulwa kucacileyo.

Umzobo 5 

Uhlalutyo lwecandelo eliphambili (PCA). Inqaku lokuchithachitha iploti yePCA (A, B). (A) I-PCA ye-Global ukusuka kwi-nucleus accumbens (NAc) iisampulu ze-P95 zamadoda. Oonxantathu abaMnyama bahambelana nenzala evela kulawulo lokutya (CD) amadama abondliwe kunye nonxantathu obomvu bahambelana nenzala ...

iziphumo

Ubunzima bomzimba kunye nokukhula

Ukuthathwa kwe-WD yoomama ngexesha lokumitha (ukusuka kwi-G1 ukuya kwi-G21) akuzange kuchaphazele ubunzima bomzimba wabantwana ekuzalweni (Umfanekiso (Figure2) 2) (CD: 6.55 ± 0.07 g vs WD: 6.54 ± 0.05 g p = 0.9232) (Amanani (Amanani2A, B).2A,B). Ubunzima bomzimba ukusuka ekuzalweni ukuya elunyulwani lube ngama-21% aphezulu kubantwana abazalwa kumadama e-WD kunabantwana abaphuma kumadama e-CD anobunzima bomzimba obuphezulu kakhulu ekulumleni kwinzala ezelwe kumadama e-WD (36.19 ± 0.90 g vs 47.32 ± 1.48 g p <0.001) (Umzobo (Umfanekiso2C) 2C). Ukususela ekulumleni ukuya ekupheleni kovavanyo (P95), iigundane zondliwa ngokutya okusemgangathweni kwe-chow kunye nobunzima bomzimba buhlala buphezulu kwinzala evela kumadama e-WD kunenzala yamadama e-CD. Kwiinkcukacha: ngexesha lokufikisa (P39) (Amanani (Amanani2A, D),2A,D), CD: 176.8 ± 3.3 g vs WD: 192.2 ± 3.3 g p = 0.0016 kunye ne-P93 (umdala omdala) (Amanani (Amanani2A,E)2A,E) CD: 478 ± 9.9 g vs WD: 508.6 ± 10.3 g p = 0.0452.

Umzobo 2 

Ukuvela kobunzima bomzimba wenzala ukusuka ekuzalweni ukuya ebudaleni. (A) Ubunzima bomzimba usuku 0 ukuya kusuku 100. Ixesha lokuncancisa kumaxesha abomvu nasemva kokulunyulwa (c) ebuntwaneni, (d) ukufikisa, kunye (e) nolutsha olungwevu. Kwigophe lokukhula, inzala eyindoda evela kulawulo lokutya ...

IiHormones kunye namanqaku eMetabolic ngexesha elahlukileyo leXesha

I-Plasma leptin, i-insulin, i-glucose, kunye nokugxilwa kwe-NEFA kulinganiswa kwi-P25, i-P45, kunye ne-P95. Kuyo yonke iminyaka, i-plasma glucose, i-NEFA kunye namanqanaba e-leptin abantwana be-WD ayengahlukanga ngokwezibalo kwi-CD yenzalo (Itheyibhile (Itheyibhile2,2, n = 6 kwiqela ngalinye). Siye sabona ukwanda okubonakalayo kokubekelwa amanqatha (i-retroperitoneal fat mass ratio) kwinzala evela kumadama e-WD awondliwe kwi-P25 kuphela (p = 0.0327, uvavanyo lukaMann noWhitney).

2 Table 

I-retroperitoneal fat mass ratio kunye ne-plasma dosage: i-glucose; insulin, NEFA, kunye leptin.

Impembelelo ye-Perinatal WD kwi-Fat Preference ukusuka elunyulweni ukuya ebudaleni

Ukuphonononga impembelelo ye-WD ekukhethweni kwamafutha, sasebenzisa i-paradigm yokukhetha iibhotile ezimbini kwiindawo ezintathu ezahlukeneyo ngexesha lokukhula. Olu vavanyo lwalusetyenziselwa ukufunda ngokukhethekileyo ukhetho lwencasa yamanqatha ngokuphepha kangangoko kunokwenzeka umphumo we-metabolic wokungena kwawo. phakathi kwamaqela (Amanani S1A–C kwiMathiriyeli eyoNgezelelweyo). Ngaphezu koko, umehluko wokusetyenziswa kwe-1% yesisombululo se-oyile yengqolowa kubangela ukunyuka kweekhalori nge-1% kwiigundane ze-WD kwi-P25 (WD: 4.9% vs CD: 3.9% yeekhalori ezidliwayo) kunye ne-0.5% kwiigundane ze-CD kwi-P45 (WD: I-2% vs CD: 2.5% yeekhalori ezityiweyo) (Amanani S1D–F kwiMathiriyeli eyoNgezelelweyo). Kwi-P25, amantshontsho avela kumadama e-CD awakhethi mafutha (44.87 ± 9.8%), p = 0.339); kwelinye icala iimpuku ze-WD zibonakalisa ukhetho lwamafutha (75.12 ± 8.04%, p = 0.039 ilandela uvavanyo lwe-Wilcoxon olusayiniweyo, iinkwenkwezi ezibomvu). Ngaphezu koko kukho umahluko wezibalo phakathi kwamaqela amabini ane p = 0.0347 (uvavanyo lweMann noWhitney, i-hash tag emnyama) (Umfanekiso (Umzobo33A).

Umzobo 3 

Inguquko ekhulayo yokukhetha amafutha ukusuka elunyulweni ukuya ebudaleni. (A) Usuku lokuqala olukhethwa ngamafutha kwi-P25, i-P45, kunye ne-P95. Iiseti ezahlukeneyo zezilwanyana zazisetyenziswa ngexesha ngalinye (n = 6/iqela/ixesha lexesha). (B) Iintsuku ezintathu ezilandelelanayo zamafutha ...

Kwi-P45 kunye ne-P95, amaqela amabini anokhetho olubalulekileyo lwamafutha, oko kukuthi, ahluke kakhulu kwixabiso lethiyori ye-50% (kwi-P45, CD: 80.68 ± 2.2% p = 0.0005 kunye ne-WD: 78.07 ± 3.25% p = 0.0005; kwi-P95, CD: 74.84 ± 8.4% p = 0.0425 kunye ne-WD: 69.42 ± 8.9% p = 0.109 ilandela iWilcoxon yovavanyo lwenqanaba esayiniweyo, inkanyezi ebomvu) (Umfanekiso (Umfanekiso3A) 3A). Amaxabiso ala maqela mabini ayengabonakali emva kosuku olunye lwenkcazo-ntetho (kwi-P45 p = 0.7857 kunye ne-P95 p = 0.9171 Mann-Whitney uvavanyo) (Figure (Umzobo33A).

Ukwazi ukuba iigundane zilawula njani ukusetyenziswa kwamafutha ngokuhamba kwexesha, siphindaphinda inkcazo yamanqatha kwiintsuku ezintathu ezilandelelanayo kwi-P45 kunye ne-P95 (Amanani (Amanani3B, C).3B,C). Okubangela umdla kwi-P45, ngamadoda kuphela aphuma kumadama e-WD athe aphulukana nokukhethwa kwesisombululo samafutha (Umfanekiso (Umfanekiso3B)3B) (usuku lwesithathu: 53.12 ± 8.36% p = 0.851 ilandela iWilcoxon yovavanyo lwenqanaba esayiniweyo). Nangona kunjalo, kwi-P95 (ubudala obudala) zonke izilwanyana zikhetha amanqatha ngaphandle kwe-evolution ngexesha lovavanyo lweentsuku ze-3 (Figure (Umzobo33C).

Isishwankathelo, kule modeli, siye sabona, kwasekuqaleni (ebuntwaneni), ukukhetha amanqatha kwimpuku ezondliwa ngamadama e-WD ngokungenamdla oqhubekayo ngexesha lokufikisa. Asibonanga mahluko phakathi kwala maqela mabini eempuku ekukhuleni kwethu.

ISiginitsha yeMolekyuli yePlastiki yeBrain kunye neeSekethe ze-GABA zokuHlaziywa ngokutsha kwi-Hypothalamus kunye neeNdlela zoMvuzo

Ukufumanisa ukuba ingaba i-WD yomama ngexesha lokubeleka kunye nokuncancisa kunempembelelo kwi-hypothalamus kunye neendlela zomvuzo wenzala, silinganise ukubonakaliswa kwezinto ezininzi eziphambili zeplastiki yobuchopho, imodeli yobuchopho, kunye nabamakishi beesekethe ze-neuronal ezibandakanyeka kukutya kunye ne-epigenetic. abalawuli. Sisebenzise i-TLDA ukuhlalutya ubuninzi babo kwiindawo ezahlukeneyo zengqondo (okt, hypothalamus, VTA, kunye ne-NAc) (Itheyibhile S1 kwi-Supplementary Material) ngamaxesha amathathu. Uvavanyo lwenziwa emva kovavanyo lweebhotile ezimbini kwi-P25, P45, kunye ne-P95 (Umfanekiso. (Figure1) 1) kumadoda amathandathu azalwe kumadama e-WD kunye namadoda amathandathu azalwe kumadama e-CD.

Kwi-P25 kwi-hypothalamus, imizila yemfuza emihlanu evela kwiindidi ezilishumi elinesithathu ezahlukeneyo zibonise inqanaba elisezantsi kakhulu le-mRNA yokubonisa ikakhulu kwiziphawuli zeplastiki kunye neempawu ze-GABA ezisukela phakathi -20% (Gfap) kunye -40% (Gabra5) kumantshontsho asuka kumadama ondliwe eWD xa kuthelekiswa neempuku ezisuka. Amadama ondla iCD. Kwindlela yomvuzo we-biopsies (i-VTA kunye ne-NAc), iijini ezimbini zibonise amanqanaba aphezulu enkcazo ye-mRNA (D2R kunye ne-Gabra1), oko kukuthi, ukusayinwa kwe-DA kunye ne-GABA receptors kunye nohlobo olunye lwentetho ephantsi (Hcrtr2) (okt, i-orexin 2 receptor) kwi-NAc. , ngelixa iijini ezine zibonise inqanaba lokubonisa i-mRNA ephezulu kakhulu (Imephu2, i-Gabara1, i-Hcrtr1, kunye ne-Hcrtr2) (oko kukuthi, iimpawu zeplastiki, i-GABA receptors kunye ne-serotoninergic receptors) kwi-VTA (Figure (Umzobo44).

Kwi-P45 kwi-hypothalamus, imizila yemfuza emihlanu evela kwiindidi ezilishumi elinesithathu ezahlukeneyo zibonise inqanaba lokuthetha elisezantsi le-mRNA eliphakathi -20% (Fos) kunye -50% (FosB) kumantshontsho asuka kumadama atyiswe i-WD xa kuthelekiswa neempuku ezisuka kumadama e-CD. Kwi-P45 kumvuzo we-biopsies yendlela yomvuzo, iijini ezine zibonise inqanaba lenkcazo ye-mRNA ephezulu (Gfap, Dat, Cck2r, kunye ne-Kat5) kunye neentlobo ezimbini zentetho ephantsi (i-Fos kunye ne-FosB) kwi-NAc ngelixa iintlobo ezintathu zibonisa inqanaba lokubonisa elisezantsi le-mRNA (Arc, I-FosB, kunye ne-Th) kunye ne-gene enye inqanaba eliphezulu (Gabrg2) kwi-VTA.

Kwi-P95 kwi-hypothalamus, iijini ezingama-20 ezivela kwiindidi ezilishumi elinesithathu ezahlukeneyo zibonise inqanaba eliphezulu lokubonisa i-mRNA ukusuka +20 kunye +40% (Syt4 ukuya kwiGjd2) kunye ne-3 yemfuza ebonisa intetho ephantsi ye-mRNA (FosB, D1r, kunye neGabarb1) kumantshontsho asuka kwi-WD ondliwe amadama xa kuthelekiswa neempuku ezivela kumadama abondliwe eCD. Kwi-P95 kwi-biopsies yendlela yomvuzo, i-12 yofuzo ibonise inqanaba eliphezulu lokubonisa i-mRNA phakathi kwe-20 kunye ne-40% (i-Syn1 ukuya kwi-Hcrt1) kunye ne-1 gene ibinzana eliphantsi (Th) kwi-NAc, i-6 yemfuza ibonise inqanaba lokubonisa i-mRNA ephezulu (Ncam1 , i-Gja1, i-Gjd2, i-Gabra5, i-Htr1a, kunye ne-Htr1b), kunye ne-6 yemfuza ebonisa inqanaba lokubonisa elisezantsi le-mRNA (Cntf, Igf1, Fos, Socs3, Gabrb2, kunye ne-Hdac3) kwi-VTA.

Emva koko senza i-PCA emithathu engajongwanga ehambelana ne-biopsies emithathu yobuchopho sisebenzisa zonke iiparameters ezibaliweyo (okt, idosi yeplasma, idatha yokuziphatha kunye nokwahluka kwenkcazo ye-mRNA). Ukwahlula okucacileyo kwamaqela amabini kwafunyanwa kuphela kwi-P95 ye-NAc kunye ne-VTA (Itheyibhile (Itheyibhile33).

3 Table 

Uhlalutyo lwecandelo eliphambili (PCA) synthesis: uhlalutyo lwekhwalithi ye-PCA yokwahlula kweqela le-PCA yehlabathi kunye ne-PCA egxininisekileyo.

Ngokwesangqa se-PCA yokulungelelanisa kunye nedatha ye-TLDA (emele ubuninzi bezinto eziguquguqukayo ezibandakanyiweyo kule PCA), sichaze iintsapho ze-gene ezinokuthi zibe noxanduva lokwahlula kwaye zenze i-PCA egxininisiweyo (Amanani). (Amanani5A,B,5A,B, umzekelo). I-PCA egxininisiweyo ibonise ukuba kwiimpawu ze-P25 ze-DA kwi-NAc kunye namanqaku eplastiki kwi-hypothalamus inokwahlula amaqela amabini enzalo (Itheyibhile (Itheyibhile33 isishwankathelo). Alukho lucalucalulo lunjalo olwathi lwafunyanwa kwi-P45. Nangona kunjalo, uhlalutyo olufanayo kwi-P95 lubonakalise ukuba abamakishi abahlukeneyo benkqubo ye-GABA kwi-NAc kunye ne-hypothalamus, kunye neempawu zeplastiki (kwi-hypothalamus, i-NAc kunye ne-VTA) kunye ne-epigenetic regulators (kuphela kwi-NAc) igalelo lokuhlula amaqela amabini ezilwanyana (kwi-hypothalamus, i-NAc kunye ne-VTA) Umzobo (Umfanekiso 5; 5; Itafile Table33).

Olu hlalutyo lubonisa ifuthe elihlala lihleli lokutya okuhlala ixesha elide kwiimpawu ze-GABAergic kunye neplastiki kunye neempawu ze-epigenetic kuzo zombini iindlela ze-homeostatic kunye nomvuzo obandakanyekayo kwindlela yokuziphatha yokondla.

I-Immunohistochemistry ye-TH Cells Confirmed Transcript Analysis

Ngenxa yokuba siye sabona umahluko kwi-TH mRNA kwi-NAc kunye ne-VTA ngamaxesha ahlukeneyo ophuhliso, sijolise ekunxibelelaniseni ezi ziphumo kunye ne-TH immunostaining. Inani leeseli ze-TH / NeuN ezintle zahlalutywa kwi-VTA apho imizimba yeeseli ze-dopaminergic ikhoyo kwaye i-OD ye-TH immunolabeling yalinganiswa kwisiphelo se-nerve esikwi-NAc. Iiseli ze-TH (+) zazincinci kwi-VTA ye-WD xa kuthelekiswa neegundane ze-CD kwi-P45 kuphela (Amanani (Amanani6A,C,E;6A,C,E; Umzobo S2A kwiMathiriyeli eyoNgezelelweyo). Kwakungekho ukusebenzisana okubalulekileyo phakathi kwenqanaba lecandelo kunye ne-TH / NeuN yokulinganisa ngamaxesha amathathu (P25 p = 0.9991, P45 p = 0.9026, kunye ne-P95 p = 0.9170). Kwi-P45 kuphela, umahluko wezibalo wafunyanwa phakathi kwamaqela amabini enzalo (p = 0.0002) (Umzobo (Umfanekiso6E).6E). Ukongeza, asibonanga mahluko kwi-OD ye-TH immunostaining kwi-NAc kwi-P25 kunye ne-P45 phakathi kwamaqela amabini (amaxabiso omlinganiselo we-OD kwi-P25: 1.314 ± 0.022 kwi-CD vs 1.351 ± 0.026 kwi-WD, p = 0.2681; Amaxabiso omlinganiselo we-OD kwi-P45: 1.589 ± 0.033 kwi-CD vs 1.651 ± 0.027 kwi-WD, p = 0.1542). Nangona kunjalo, ukwehla okukhulu kwe-OD ye-TH nerve endings kwafunyanwa kwi-NAc kwiqela le-WD kwi-P95 (amaxabiso omlinganiselo we-OD kwi-p95: 1.752 ± 0.041 kwi-CD vs 1.550 ± 0.046 kwi-WD, p = 0.0037) (Amanani (Amanani6B,D,F;6B,D,F; Umzobo S2B kwiMathiriyeli eyoNgezelelweyo).

ingxoxo

Kolu phononongo, sicinge ukuba ukondleka kokubeleka koomama kuya kuba nefuthe kwinkqubo yophuhliso lweendlela zomvuzo ezibandakanyeka kwi-homeostasis yamandla, ukhetho lokutya, kunye nokutya kokutya kwenzala. Sihlolisise ngokubanzi impembelelo yokuthatha i-WD yomama ukususela ekuzalweni ukuya elunyulwe kwi-GABA, i-serotonin, kunye neendlela ze-DA zeendawo ezithile zengqondo (VTA, NAc, kunye ne-hypothalamus) kwinzala, ukususela ebuntwaneni ukuya ebudaleni. Iziphumo zethu zibonisa ukuba ukusetyenziswa kokutya, okutyebileyo ngamanqatha kunye neswiti, kuthintelwe ngokungqongqo kwixesha lokuzala kunempembelelo ekukhetheni amanqatha kwangoko (ubuntwana) kwinzala ehambelana notshintsho kwiprofayili yokubonakaliswa kofuzo kunye notshintsho lwe-neuroanatomical / lwezakhiwo ze-mesolimbic. iinethiwekhi ze-dopaminergic. Nangona kunjalo, xa inzala yayigcinwe phantsi kokutya kwe-chow, sabona kwi-WD yokufikisa yondla iigundane ilahleko eqhubekayo yokutsalela amanqatha adityaniswa nokunciphisa ukubonakaliswa kofuzo lwenkqubo ye-DA kunye nokwehliswa okuncinci kwe-TH-positive neurons kwi-VTA. . Kamva ebomini ukhetho lwamafutha lwalungahlukanga phakathi kwamaqela nangona iplastiki ebalulekileyo yothungelwano lwe-GABAergic kunye nenethiwekhi yamandla ye-homeostasis ye-hypothalamus ichongiwe kwimpuku evela kumadama esondliwe e-WD (Umfanekiso (Umzobo77).

Umzobo 7 

I-Graphical abstract. I-NAc, i-nucleus accumbens; I-VTA, indawo ye-ventral tegmental.

Impembelelo yokuqala ye-perinatal-WD intake esiyibonile kolu phononongo bubunzima bomzimba obunyukileyo benzala ekulumleni kodwa akukho mahluko ekuzalweni. Enyanisweni, izilwanyana zeqela le-WD zifumana i-21% ubunzima ngaphezu kweCD ekupheleni kwexesha lokuncancisa. Uphononongo lwangaphambili lubonelele ngeziphumo ezingqubanayo malunga nokutshintsha kobunzima bokuzalwa kwinzala evela kumadama esondliwe yi-WD: ubunzima bomzimba obuphezulu (, ), ubunzima bomzimba obuphantsi (, , ) okanye akukho mahluko (, ). Idatha yethu ihambelana nohlalutyo lwamva nje lwe-meta-regression () kuqhutywe kupapasho lovavanyo lwe-171 olugqibe ukuba ukubonakaliswa kwe-HFD yomama akuzange kuchaphazele ubunzima bokuzalwa kwenzala kodwa kubangele ukwanda komzimba ekupheleni kwexesha lokuncancisa. Ubunzima bomzimba obuphezulu benzala ye-WD mhlawumbi bubonisa utshintsho ekubunjweni kobisi kunye / okanye ukuveliswa kobisi olubonakalisiweyo kupapasho lwangaphambili (, ). Ngokuhambelana nobunzima bomzimba wabo obuphezulu, i-retroperitoneal fat ratio ye-WD inzala yayiphezulu kakhulu kunaleyo ye-CD inzala ekupheleni kwexesha lokuncancisa (P25, Itheyibhile Uludwe 2), 2), ekwahambelana nezifundo zangaphambili (, ). Nangona kunjalo, i-adiposity ephezulu ayizange iqhubeke kwi-P45 kunye ne-P95, kunye nezinye iiparitha ze-metabolic njenge-insulin, i-NEFA, kunye ne-glucose plasma azizange zihluke phakathi kwamaqela. Iziphumo zethu zibonise ukuba ngaphandle kokutyeba okucacileyo komama ngexesha lokumitha kunye nokuncancisa, ukutya ngokwako akwanelanga ukwenza iziphumo ezihlala zihleli kwi-metabolic kwinzala., , ).

Kuye kwaxelwa ukuba ukuthatha i-HFD ye-perinatal ihambelana ngokufanelekileyo kunye nenzala ekhetha ukutya okunencasa (). Kwisifundo sethu, senze uphando olude olujolise ekuvavanyeni ukhetho lwamafutha kwinzala elunyulwe kwi-chow eqhelekileyo.

Impembelelo yePerinatal WD ebuntwaneni (emva kokulunyulwa)

Amantshontsho eempuku atya ukutya okuqinileyo kwiintsuku eziyi-19-20 emva kokuzalwa () xa iindlela zabo zomvuzo wobuchopho zingekavuthwa (). Ke ngoko bekunomdla kakhulu ukufunda ukuthanda kwabo kwangethuba ngamanqatha kwaye banxibelelanise oku kukhetha kwangoko kunye nohlalutyo lwemibhalo yobuchopho. Nje emva kokulunyulwa, siye sabona ukhetho lwamafutha kwinzala ye-WD engazange ingqinwe kwiimpuku zeCD. Oku kuhambelana nezinye iingxelo ezibonisa ikhonkco phakathi kokungondleki kwe-perinatal kunye nokukhethwa kokutya okunencasa kunye nokukhethwa okuphantsi kwamafutha ebusheni bokulawula iigundane ().

I-PCA yehlabathi ayizange ivumele ukucalulwa kweqela leenjana ngokubhekiselele kukutya komama kuloo minyaka. Nangona kunjalo, xa i-PCA ekujoliswe kuyo, kuphela kubamakishi be-DA, yenziwa, safumana ulwahlulo oluhle lwamaqela. Enyanisweni, kukho ukwanda okuphawulekayo kwintetho ye-D2 receptor mRNAs kwi-NAc kwi-WD pups. Le postsynaptic D2 overexpression kwi-NAc inokubandakanyeka ngokuyinxalenye kwinkuthazo ephezulu yamafutha (). Imbalwa eminye imibhalo eguqulweyo kwi-WD pups xa kuthelekiswa ne-CD pups, njengokunyuka kwe-alpha 1 GABAA subunit kwi-NAc kunye ne-VTA kunye nokuncipha kwe-alpha 5 GABAA subunit kwi-hypothalamus ebonisa ukuhlengahlengiswa kwe-GABAA receptors kule nuclei.

Impembelelo ye-Perinatal WD kwi-Adolescence

Kwi-P45, siye sabona ukhetho olufanayo lwamafutha aphezulu kuwo omabini amaqela ngosuku lokuqala lokubonisa kodwa, umdla, iigundane ze-WD ngokuthe ngcembe zaphulukana nomdla wazo wamanqatha emva kokubonisa ngokuphindaphindiweyo. Ubutsha lixesha elibalulekileyo lohlengahlengiso lwe-neurobehavioral oluyimfuneko ekusebenzeni kwengqondo ubomi obude (), kunye nezifundo ezahlukeneyo zibonise ukuba semngciphekweni okubonakalayo kwimpembelelo eyingozi yokuqonda ukutya okunamafutha (-). Esi siphumo sichasene ngokucacileyo nomsebenzi wangaphambili weqela likaMuhlhausler (, ) apho iigundane ezincinci (iiveki ze-6) zibonise ukukhetha okucacileyo kokutya okungenamsoco. Nangona kunjalo, kupapasho lwabo iparadigm yovavanyo yayahlukile kuba iimpuku zazinokufikelela simahla kuzo zombini i-chow eqhelekileyo kunye nokutya okungenamsoco ukusuka elunyulweni ukuya kwidini (iiveki ezi-6).

Ngokuhambelanayo, silinganise ukwanda kwe-Dat mRNA kwi-NAc kunye nokuncipha kwe-Th mRNA kwi-VTA eyaqinisekiswa yi-immunohistochemistry ebonisa ukunciphisa i-TH (+) inombolo yeeseli kwi-VTA yeegundane ze-WD. Emva komsebenzi okhutshelweyo ophakamileyo wenkqubo ye-DA ekulumleni, umsebenzi oncitshisiweyo kwi-P45 unokuchaza umdla ophantsi wokutya okunencasa okubonwe kwiimpuku zethu ze-WD. Kufuneka kwakhona kuqatshelwe ukuba ukuhla okucwangcisiweyo kwe-Fos kunye ne-FosB mRNA yokubonakalisa kwii-nuclei ezahlukeneyo esizihlalutyileyo kunokuba luphawu lokuncipha komsebenzi wobuchopho emva kokuvezwa kwe-WD kamama.

Iigundane ze-WD ezifikisayo zibonise ukungakhathali ngokukhawuleza kwamafutha okuchasene nokuziphatha kwabo kwangaphambili. Ukusetyenziswa kokutya “okuqhelekileyo” kwasebuntwaneni kubonakala “kubakhusela” ekuthandeni amanqatha agqithisileyo xa befikisa. Ngokuchasene noko xa iimpuku zinokufikelela simahla ekutyeni okungenamsoco emva kokulunyulwa, njengakwiNgcaciso. (, ), babonisa ebusheni bathanda kakhulu amafutha. Esi siphumo sicebisa ukuba iiveki ezi-3 ze-chow yokutya emva kokulunyulwa zinokuthi zilungelelanise iisekethe kwaye zenze inzala efikisayo ingakhathali kumngeni wamafutha.

Impembelelo ye-Perinatal WD kubantu abadala

Iigundane zabantu abadala azisabonisi umahluko wokukhetha amanqatha, nasemva kokuphindaphindwa kwamafutha njengoko sele kuchaziwe (, ). Ngokuhambelanayo, siye sabona ukuncipha kwe-Th mRNA kunye neprotheni kwi-NAc, kunye notyekelo lokunciphisa ukubonakaliswa kwe-Dat mRNA kwi-VTA. UNef kunye nomntu osebenza naye () sele ichaze umsebenzi ophantsi wenkqubo ye-DA kwiigundane zabantu abadala ezondliwa ngexesha lokubeleka kunye ne-HFD, kunye nempendulo ephosakeleyo ye-DA kwi-amphetamine elinganiswe nge-microdialysis kunye nokwenyuka kwenkuthazo yomvuzo wamafutha (jonga itafile eshwankathela idatha ye-qPCR yakutshanje kule modeli, Itheyibhile S2 kwiMathiriyeli eyoNgezelelweyo). Umda omnye we-TH quantification (mRNA kunye ne-immunohistochemistry) kwi-NAc ivela kwinto yokuba iiseli ze-NAc zinokubonisa i-Th mRNA kunye neprotheyini kwaye emva koko zinokuthatha icala kwi-DA yefibers quantification., ). Nangona kunjalo, ukusetyenziswa kwe-TH immunostaining kwi-NAc ikakhulu kutyhile iitheminali ze-axon ezixineneyo ezivela kwi-midbrain DA neurons (VTA kunye ne-SNc). Ngokuqhelekileyo, i-TH ebonisa i-neurons kwi-striatum kunye ne-NAc inokubonwa kuphela kwizilwanyana ezinobungozi ze-DA () kwaye ke ngoko ayinakubonwa kumacandelo ethu amajoni omzimba. Kolu phononongo, siye sabona ukonyuka okunamandla kwi-opioid receptor kwi-NAc xa amanye amaqela, aneemodeli ezahlukeneyo, ebonisa ukuncipha kokubonakaliswa kwi-ventral striatum yegundane evezwe kwangoko kwi-HFD (ngexesha lokuncancisa kunye nokuzala) (, ) okanye akukho tshintsho (). Olu hlengahlengiso, olulinganiswe kwinqanaba le-mRNA kuphela, lunokubonisa umsebenzi omncinci we-hypo yeesekethe ze-DA ezinxulumene novakalelo oluphezulu lwe-opioid () mhlawumbi akwanelanga ukuba nefuthe kuvavanyo lokuziphatha esilwenzileyo. Ezi ngqikelelo kufuneka ziqinisekiswe kusetyenziswa iindlela ezisebenzayo. Kwiphepha lamva nje, elinomzekelo ofanayo, uRomani-Perez et al., Abakwazanga ukubona ukwanda okukhulu kwenkuthazo kwiibhokisi ezisebenzayo zokulungisa inzala ye-HFD kodwa baqaphela ukubambezeleka okufutshane ukufikelela kwibhokisi yenjongo kwi-paradigm yovavanyo lwe-runway.). Ngaphandle kokungabikho kokukhethwa kwamafutha okuhlala ixesha elide kwiimeko zethu zovavanyo, sifumanise ukuba ukuthathwa kwe-WD yoomama be-perinatal kunefuthe elihlala ixesha elide kwezinye iisekethe ze-cerebral ikakhulu zilamlwa yi-GABA yokulungiswa kwakhona kwi-NAc kunye ne-Hypothalamus. I-NAc ithathwa njenge "sensory sentinel" yokuziphatha okugqibeleleyo (). Uphononongo lwakutsha nje lubonise ukuba ukutya kwacinezelwa kukuthintelwa kwe-GABA-ikhupha i-LH neurons.). U-O'Connor et al. ibonise ukuba i-NAc D1R neurons (i-GABAergic projecting neurons) inqanda i-LH VGAT neurons ukuba iyeke ukutya.). Ezi zilingo zityhila isiphaluka se-GABA (NAc / Hypothalamus) enokuthi ibe noxanduva lokulawula impendulo yokuziphatha. Le nkqubo ye-ventral striatum-hypothalamic incedisa enye isekethe ebandakanya i-nucleus stria terminalis ye-GABA-ekhupha i-VGAT ekhupha i-neuron kwi-glutamate ekhupha i-Vglut LH neurons kunye nokuvinjelwa ngokuthe ngqo kwe-LH vglut2 ikhuthaza ukutya (). Elinye icandelo elibalulekileyo lesekethe yokulawula ukutya ebandakanya iqokobhe le-NAc yi-GABA-ikhulula inhibitory projection kwi-VP (). Ezi datha zibonisa indima ebalulekileyo ye-GABA yokubonisa kwi-interplay phakathi kwe-hypothalamus kunye ne-NAc ukukhuthaza ukutya. Kuphononongo lwethu, asikwazanga ukucalula inani lee-neurons ezibandakanyekayo kwi-GABA yokulungiswa kwakhona kunye nendlela ezi zilungiso ezinokutshintsha ngayo amanethiwekhi. Nangona kunjalo, indima ephambili yeesekethe ze-GABA ifanele umdla ngakumbi. Ngokukodwa, kuya kuba nomdla kakhulu ukwenza ezinye iimvavanyo ezisebenzayo zezi zijikelezo ze-GABA usebenzisa iindlela ze-electrophysiological (). Siphinde sabona ukulawulwa kwehlabathi jikelele kwe-mRNA transcript ye-5HT1a kunye ne-5HT1b receptors kwii-nuclei ezintathu ezifundiweyo. Uninzi lweprojekithi zemicu ye-serotonin zivela kwi-dorsal raphe nucleus (DRN) kunye ne-median raphe nucleus (MRN). Idatha yakutsha nje evela kwi vivo ukurekhodwa kunye nezifundo zokucinga zibonise indima entle ye-5HT kumvuzo (). Iifiber ze-5HT ezivela kwi-DRN zibandakanyeka kulawulo lokunyanzeliswa (). Ukwandisa i-5HT1a kwi-VTA kunye ne-NAc inokuba yindlela yokubuyisela inokulawula ukungahambi kakuhle. Kwi-hypothalamus, izifundo ze-pharmacological zibonisa ukuba i-5HT1a i-receptor subtypes inokucinezela ukuziphatha kokutya okubangelwa yi-serotonin stimulation (, ). Ukunyuka kwe-5HT1a kunye ne-b receptors kwi-hypothalamus kunokubangela intshukumo yokutya-ecinezelayo ye-serotonin kwaye ke ngoko inokwenza indlela yokuhlawula. Ezi ngcinga kufuneka ziqinisekiswe ngokwenza imifuniselo efanelekileyo yokusebenza.

Olu tshintsho lwenethiwekhi lunxulunyaniswa nohlengahlengiso lweempawu zeplastiki njenge-Ncam mRNA. Kwi-hypothalamus yamagundane abantu abadala, siye sabona ukunyuka kwe-Ncam1 kunye ne-St8sia4 imibhalo ebonisa kunye nokunyuka kwe-polysialic acid (PSA) yokubonisa. I-PSA yi-cell-surface glycan emodareyitha ukusebenzisana kweeseli ukuya kwiseli. I-Polysialylation ye-cell adhesion proteins ibandakanyeka kwiinkqubo ezahlukeneyo ezixhomekeke kwi-synaptic plasticity kwinkqubo ye-nervous central kwaye kuye kwaxelwa ukuba iyafuneka kwi-adaptive synaptic plasticity yeesekethe zokutya ngexesha lokulinganisa amandla abukhali (, ). Ukongeza abanye abalawuli bentsebenziswano yeeseli kunye ne-synaptogenesis banokubandakanyeka kule plastikhi ye-hypothalamic.

Ukuqukumbela (Umfanekiso (Figure7), i-7), ukufakwa kwe-WD yomama kunempembelelo ehlala ixesha elide kwintlangano yeesekethe ze-homeostatic kunye ne-hedonic ezilawula ukuziphatha kokutya kwinzala. Ngohlalutyo lwamaxesha amathathu abalulekileyo, sakwazi ukubonisa inguquko ecacileyo yokukhetha amanqatha anxibelelene neesayino ezithile zeemolekyuli zengqondo. Ngexesha lobuntwana, ukhetho lwamafutha lunokunxulunyaniswa nomsebenzi ophezulu wenkqubo yeDA. Ukufikisa, okuphawulwa ngokuguquguquka kokuthandwa kwamafutha, kwanxulunyaniswa nokubonakaliswa okusezantsi kweempawu zenkqubo ye-DA ezicebisa indlela yombuyekezo. Inqaku elinomdla kakhulu lokwazisa lelokuba, kule modeli, ukutya okunesondlo emva kokulunyulwa kunokukhusela impuku efikisayo kwimikhwa eyingozi yokutya ngokunciphisa umnqweno wabo wamafutha. Nangona ekukhuleni kwamaqela amabini anokhetho oluphezulu olufanayo lwamafutha, iigundane ezisuka kumadama e-WD ondliwe zibonise ukulungiswa okunzulu kweesekethe ze-GABA. Ziziphi iziphumo zolu hlobo lweplastiki luhlala luhleli? Ngaba ukutya okune-obesogenic ebaxiweyo ngexesha lokufikisa kuya kuphinda kusebenze le nkqubo yomvuzo ibuthuntu? Imibuzo enjalo inokuba luncedo ekulandeleni isondlo sabasanda kuzalwa kunye nabantwana abakhulele kumazwe asentshona.

Statement Ethics

Zonke iimvavanyo zenziwe ngokuhambelana nezikhokelo zekomiti yentlalontle yezilwanyana zendawo, i-EU (umyalelo 2010/63/EU), i-Institut National de la Recherche Agronomique (eParis, eFransi), kunye neSebe lezilwanyana zaseFransi (A44276). Iprothokholi yokulinga yamkelwa yikomiti yeenqobo ezisesikweni yeziko yaze yabhaliswa phantsi kwereferensi ye-APAFIS 8666. Onke amanyathelo okhuseleko athatyathwa ukunciphisa uxinzelelo kunye nenani lezilwanyana ezisetyenziswa kuthotho lwemifuniselo.

Umbhali Wemivuzo

I-JP kunye ne-PB yenze uvavanyo kwaye ithatha inxaxheba kwingxoxo nokubhala. U-TM wenze i-PCA kwaye wathatha inxaxheba kwingxoxo nokubhala. I-SN ibe negalelo kuyilo lovavanyo kwaye ithathe inxaxheba kwingxoxo. I-PP ibe negalelo kuyilo lovavanyo, ithathe inxaxheba kwiingxoxo, kwaye ibhale umbhalo-ngqangi. I-VP yenzelwe kwaye yenza iimvavanyo, yahlalutya idatha, kwaye yabhala umbhalo wesandla.

Ukuxabana kweNkcazo yeNzala

Ababhali bavakalisa ukuba uphando lwenziwe ngokungabikho naluphi na ulwalamano lwezorhwebo okanye lwezezimali olubhekiswa njengengxabano yenzuzo.

Imibulelo

Ababhali bangathanda ukuvuma uGuillaume Poupeau kunye noBlandine Castellano ngokunyamekela izilwanyana kulo lonke uphando, u-Anthony Pagniez ngoncedo lwakhe kwi-mRNA extraction kunye ne-TLDA, u-Isabelle Grit ngoncedo lwakhe kuhlalutyo lweesampuli zeplasma, kunye no-Alexandre Benani noMarie-Chantal Canivenc. kwingxoxo yabo eluncedo kunye noyilo lweTLDA.

Imihlathi

 

Inkxaso-mali. Olu phando luxhaswe ngummandla we-des pays de la Loire isibonelelo se-PARIMAD (VP), isibonelelo sesiseko se-LCL (VP kunye nePP), isiseko seSanteDige (VP) kunye ne-INRA Metaprogram DIDIT (SN, VP, PP).

 

 

Izinto ezongezelelweyo

Isixhobo esongezelelweyo sale nqaku sinokufumaneka kwi-Intanethi apha http://journal.frontiersin.org/article/10.3389/fendo.2017.00216/full#supplementary-material.

Umfanekiso weS1

Ukuthathwa kwamandla apheleleyo kwioli yombona enebhotile. (A) Iikhalori ezithathwa kwibhotile ye-oyile yombona iiyure ezingama-24 nge-P25 kumantshontsho asuka kwi-western diet (WD) etyisa amadama namantshontsho asuka kwi-control diet (CD) kumadama atyiswayo. (B) Iikhalori ezithathwa kwibhotile yeoli yengqolowa ye-24 h kwi-P45 (usuku lwesithathu lovavanyo lwebhotile). (C) Iikhalori ezithathwa kwibhotile yeoli yengqolowa ye-24 h kwi-P95 (usuku lwesithathu lovavanyo lwebhotile). Ngeephaneli (A-C), idatha ichazwa njengentsingiselo ± SEM, akukho mahluko wamanani (p > 0.05) yabonwa, kulandela uvavanyo lukaMann noWhitney olungeyo-parametric, kuyo yonke iminyaka. (D) Ipesenti yeekhalori ezithatha kwibhotile yeoli yengqolowa thelekisa kunye neekhalori ezipheleleyo zokuthatha (ibhotile yeoli yengqolowa + i-chow standard diet) ye-24 h kwi-P25 kwi-WD pups kunye ne-CD pups. (E) Ipesenti yeekhalori ezithathwa kwibhotile yeoli yengqolowa thelekisa kunye ne-calories ethathwayo (ibhotile yeoli yengqolowa + ukutya okuqhelekileyo kwe-chow) kwi-24 h kwi-P45 (usuku lwesithathu lovavanyo lwebhotile) kwi-WD pups kunye ne-CD pups. (F) Ipesenti yeekhalori ezithatha kwibhotile yeoli yengqolowa ithelekisa ne-calorie eyonke ithathwayo (ibhotile yeoli yengqolowa + ukutya okuqhelekileyo kwe-chow) kwi-24 h kwi-P95 (usuku lwesithathu lovavanyo lwebhotile) kwi-WD pups kunye ne-CD pups. Ngeephaneli (D,E), idatha ibonakaliswe kwipesenti yokuthatha ikhalori iyonke akukho mahluko wezibalo (p > 0.05) yabonwa, ilandela i-chi-square kunye nokulungiswa kweYates, kuyo yonke iminyaka.

Umfanekiso weS2

Iifotomicrographs ezimeleyo ze-TH immunostaining kwi-nucleus accumbens (NAc) kunye ne-ventral tegmental area (VTA) ngamaxesha amathathu ahlukeneyo. (A) I-Photomicrograph ye-TH / NeuN immunostaining kwinqanaba le-VTA, -5.30 mm ukusuka eBregma. Ukuleyibhile okubomvu yeye-NeuN, kwaye enye eluhlaza yeye-TH. Utolo olumhlophe lubonisa ukuphuma komthambo wesithathu. (B) Ifotomicrograph ye-TH immunostaining kwinqanaba le-NAc, + 1.70 mm ukusuka eBregma. Ileyibheli eluhlaza yeye-TH. Utolo olumhlophe lubonisa i-commissure yangaphambili.

Ithebula leS1

Uluhlu lwejene yoxinaniso oluphantsi lwe-TaqMan kunye neekhowudi zobomi ezihambelana nobugcisa bobomi.

Ithebula leS2

Isishwankathelo sedatha epapashiweyo malunga ne-dopamine pathway transcripts expression. Iimpawu ezibomvu zihambelana nexesha lobuntwaneni, eziluhlaza okwesibhakabhaka ukuya ebusheni, kunye nabamnyama ukuya kumntu omdala. =: ihambelana nentetho ekhutshiweyo efanayo phakathi kwamaqela, +: ihambelana nenkcazo ekhutshelweyo ephezulu kumantshontsho asuka kwi-caloric diet [ukutya okungenamsoco, ukutya okusentshona (WD), okanye ukutya okunamafutha aphezulu (HFD)] amadama awondliwe, kunye -: ihambelana nokuchazwa okubhaliweyo okuphantsi kwi-pups ukusuka kwi-caloric yokutya ephezulu (ukutya okungenamsoco, i-WD okanye i-HFD) amadama ondliwe.

Ucaphulo

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