I-wiring ye-hypocretin kunye ne-LC-NE neurons: impembelelo yokuvusa (2013)

Abstract

Ukulala kunye nokuvuka ziimeko ezimbini ezikhethekileyo ezijikelezayo kunye namaxesha e-ultradian kunye ne-circadian kubukumkani bezilwanyana. Ukuphaphama yimeko yokuqonda apho isilwanyana sinokubona kwaye sisebenzisana nendawo esingqongileyo. Emva kwexesha elide lokuphaphama, uxinzelelo lokulala luyanda kwaye lukhokelela ekuqaliseni kokulala okuphawulwa njengexesha lokungasebenzi kwesalamane kunye nokuma kwe-stereotyped kunye ne-sensory threshold ephezulu.

Kwizilwanyana ezanyisayo, ubuthongo budla ngokwahlulwa bubuthongo obunamaza acothayo (SWS, okanye ubuthongo be-NREM ebantwini), kunye nokulala kwamehlo okukhawulezayo (REM) (okukwabizwa ngokuba “yi-paradoxical sleep”). Ukuphaphama, i-SWS kunye nokulala kwe-REM ziimeko ezihlukeneyo zokuziphatha ezinokuthi zichazwe nge-electroencephalographic echanekileyo (EEG) kunye ne-electromyographic (EMG) iimpawu. Ngexesha lokuvuka, i-amplitude ephantsi, i-oscillations-frequency oscillations iphambili. I-SWS ibonakaliswe yi-high-amplitude i-oscillations ecothayo (0.5-4 Hz) i-predominance (njengoko ilinganiswe yi-EEG yobuninzi bamandla) ibonisa ubunzulu bokulala. Ukulala kwe-REM yimo enye yokuziphatha, ebonakaliswa ngokukhawuleza, i-oscillations yefrikhwensi edibeneyo, phakathi kwayo i-oscillations ye-theta (5-10 Hz) ilawula kwiimpuku, ihamba ne-atonia ye-muscle, kunye nokuguquguquka kwentliziyo kunye namazinga okuphefumla.

Nangona iimeko zokulala kunye nokuvuka zilula ngokomgangatho nangokobungakanani ukuzichaza, kuyamangalisa ukuba kube nzima ukuchaza ukuba kuthethwa ukuthini “ukuphaphama.” Igama elithi ukuvuswa lihlala libhekisa kwiqondo lokuphaphela kunye nokuphapha ngexesha lokuvuka, ukubonakalisa njengokunyuka kokusebenza kwemoto, ukuphendula kumagalelo oluvo, ukuphinda usebenze ngokweemvakalelo, kunye nokuphuculwa kwenkqubo yokuqonda.

Iindlela zobuchopho eziphantsi kombutho womjikelo wokuvuka kunye nenqanaba eliqhelekileyo lokuvuka lihlala lingacacanga kwaye izifundo ezininzi zeklasikhi zichonge inani labantu bee-neurons umsebenzi wabo ohambelana neemeko zokuziphatha ezahlukeneyo. Ekuqaleni kwakucingelwa ukuba ii-neuron ezisebenzayo ngaphambi kotshintsho lokuziphatha (okt, ii-neurons ezisebenzayo ngaphambi kokutshintsha kokulala ukuya-kuvuka) Khuthaza imeko ezayo, ngelixa ii-neurons ezisebenzayo ngexesha elithile (ukuvuka okanye ukulala) zibalulekile kugcina yona. Lo mbono wenziwa nzima ngakumbi ukuqonda ukuba i-neurons kwinethiwekhi ingabonisa umsebenzi onxulumene nomda welizwe ngenxa yoxhulumaniso kwezinye, ii-neurons ze-causal ezingaphezulu ngaphandle koxanduva ngokuthe ngqo kwiinguqu zelizwe. Nangona kunjalo, ngokuqhelekileyo kuye kwachazwa ukuba kukho i-neural populations edlala indima ebangela ukulala kunye / okanye iimeko zokuvusa. Abemi ekucingelwa ukuba bakhuthaza ukuvuswa babandakanya: i-hypocretin (i-hcrt-ekwabizwa ngokuba yi-"orexins") -i-neurons ebonisayo kwi-lateral hypothalamus, i-noradrenergic locus coeruleus (LC) -i-neurons ebonisa kwi-brainstem, i-serotoninergic dorsal raphe nuclei (DRN) kwi-brainstem, i-histaminergic tuberomammilary nucleus (TMN) kwi-hypothalamus yangasemva, i-cholinergic pedunculopontine (PPT) kunye ne-laterodorsal tegmental (LDT) nuclei kwi-midbrain, kunye ne-cholinergic neurons kwi-basal forebrain (Jones, 2003). Ngokwahlukileyo, ii-neurons ezithintelayo ezivela kwizakhiwo ze-hypothalamic zangaphambili ziyasebenza ngexesha le-SWS, ngelixa i-Melanin-Concentrating Hormone (MCH) neurons esuka kwi-lateral hypothalamus, kunye ne-glutamatergic kunye ne-GABAergic neurons evela kwi-brainstem iyasebenza ngexesha lokulala kwe-REM (Fort et al., 2009).

Kwiminyaka yakutshanje, thina kunye nabanye siqalise ukusebenzisa itekhnoloji ye-optogenetic ngeemodeli ezahlukeneyo zemouse ukujongana nemibuzo efana nale Iinkqubo zokuvusa zikulawula njani ukuphaphama kunye nokuvuselela? Zisebenzisana njani ngokusebenzayo ukukhuthaza, ukugcina okanye ukwandisa ukuvuseleleka kwiimeko ezithile? Kuphononongo lwethu lwamva nje, besinomdla ngakumbi kwiineuron ezibonisa i-hcrt (de Lecea et al., 1998; Sakurai et al., 1998). I-hcrt zii-peptides ezimbini ze-neuro-excitatory (de Lecea et al., 1998; Sakurai et al., 1998) eveliswe kwi ~ 3200 neurons kwi-mouse lateral hypothalamus (~ 6700 kunye ne-50,000-80,000 kwi-rat kunye nengqondo yomntu, ngokulandelanayo) (de Lecea kunye ne-Sutcliffe, 2005; Modirrousta et al., 2005). Ezi neurons zifumana igalelo elisebenzayo kwiinkqubo ezininzi ezisasazwa kwi-cortex, i-limbic system, i-sub-cortical indawo ezibandakanya i-hypothalamus ngokwayo, i-thalamus, kunye nokunyuka kweengqikelelo ezivela kwi-brainstem cholinergic nuclei, i-reticular formation, i-midbrain raphe nuclei, kunye ne-periaqueductal grey. Ngapha koko, ezi projekthi ze-neurons kuyo yonke inkqubo ye-nervous central, kubandakanya ukuvusa kunye namaziko omvuzo wobuchopho, kwi-neurons ebonisa i-hcrt receptors (OX1R kunye ne-OX2R). I-afferent and efferent projection of hcrt neurons icebisa ukuba idlala indima kwimisebenzi emininzi ye-hypothalamic kuquka nokulawula umjikelo wokulala / wokuvuka kunye nokuziphatha okujoliswe kuko. Okubangela umdla kukuba, sifumanise ukuba uqikelelo olucacileyo olusuka kwi-hcrt neurons ukuya kwi-noradrenergic LC neurons lulamla inguquko yokulala ukuya kuvuka kunye nemiba engaphezulu yokuvuka.

Apha, sishwankathela iimvavanyo zamva nje ze-optogenetic ezivavanya i-hypothesis yokuba i-hcrt kunye ne-LC neurons zibangela utshintsho kunye nokugcinwa kwemeko yokuvuselela (Adamantidis et al., 2007; UCarter et al., 2009, 2010, 2012). Okokuqala, sigxininisa ngokufutshane kwaye sishwankathela iingxelo zangaphambili malunga nezi nkqubo zisebenzisa iindlela zendabuko zofuzo kunye ne-pharmacological. Okulandelayo, sidibanisa iziphumo zethu sisebenzisa i-optogenetic probes ukukhetha ukuvuselela okanye ukuthintela ezi nkqubo kwiimpuku ezihamba ngokukhululekileyo. Ekugqibeleni, sixoxa ngemibuzo engasonjululwanga kwaye siqikelele malunga nezahlulo ze-anatomical ezizayo kunye nokusebenza kweesekethe zokuvusa.

Hypocretins, ukuphaphama, kunye ne-narcolepsy

I-hcrt neurons ihlala ithule ngexesha lokuvuka okuthe cwaka, i-SWS, kunye nokulala kwe-REM kodwa ibonisa amazinga aphezulu okukhutshwa ngexesha lokuvuka okusebenzayo kunye noguquko lwe-REM yokulala ukuya-ukuvuka (Lee et al., 2005; Mileykovskiy et al., 2005; Takahashi et al., 2008; UHasani et al., 2009). Ukongezelela, babonisa amazinga aphezulu okukhutshwa ngexesha lokuvuswa okubangelwa yimpembelelo yendalo (umzekelo, i-auditory stimulus) (Takahashi et al., 2008) kunye nokuziphatha okujoliswe kuko (Mileykovskiy et al., 2005; Takahashi et al., 2008). Olu phononongo lucebisa ukuba i-hcrt neurons ithatha inxaxheba kutshintsho lokulala ukuya ekuvukeni, kunye nokwandiswa kokuphaphama okubonwa ngexesha lokuziphatha okujolise kwiinjongo ezahlukeneyo.

Ukuvalwa okanye ukucinezelwa komqondiso we-hcrt kubonisa imfuneko ye-hcrt yemfezeko yeendawo zokuziphatha kwiimpuku, iimpuku, izinja, abantu, kunye ne-zebrafishes (Sakurai, 2007; Yokogawa et al., 2007). Ngokwenene, obona bungqina bunyanzelekileyo bokulahleka komsebenzi buvela kwikhonkco phakathi kokusilela kwe-hcrt kunye neempawu ze-narcolepsy (Peyron et al., 2000; Saper et al., 2010). Izigulana ze-narcoleptic ezine-cataplexy zinokungabikho ngokupheleleyo hcrt imibhalo yofuzo kwi-hypothalamus kunye namanqanaba angabonwayo okanye angabonakaliyo e-hcrt kwi-cerebrospinal fluid (Thannickal et al., 2000; Sakurai, 2007; Yokogawa et al., 2007). Izinja ze-Doberman narcoleptic zinotshintsho OX2R, kunye nazo zonke iimpuku ezenziwe ngokwemfuza ngokucinywa kwe hcrt, OX2R, okanye iiseli ze-hcrt zibonisa ukubanjwa kokuziphatha okufana ne-cataplexy, uphawu lwe-narcolepsy (Jones, 2003; Sakurai, 2007; Sehgal kunye neMignot, 2011). Okubalulekileyo, ukuhlangulwa kofuzo lwe hcrt Ukubonakaliswa kofuzo kunciphisa iimpawu ze-narcolepsy kwiimpuku (Liu et al., 2011; UBlanco-Centurion et al., 2013).

I-Intracerebroventricular (icv) infusion ye-hcrt peptides okanye i-hcrt agonists ibangela ukwanda kwexesha elichithwe liphapheme kunye nokuncipha kwe-SWS kunye nokulala kwe-REM [ukuhlaziywa kwe-Sakurai (2007)]. Inaliti ye-stereotactic ye-peptide kwi-LC, LDT, i-basal forebrain, okanye i-hypothalamus esecaleni yonyusa ukuphaphama kunye nomsebenzi we-locomotor uhlala unxulunyaniswa nokuncipha okuphawulweyo kwi-SWS kunye nokulala kwe-REM (Hagan et al., 1999). Kutshanje, i-genetic dis-inhibition ye-hcrt neurons isebenzisa i-GABA-B ekhethiweyo yokususwa kofuzo lwe-receptor kuphela kwi-hcrt neurons ibangele ukuqhekeka okukhulu kokulala / ukuvuka ngexesha lokukhanya kunye nobumnyama ngaphandle kokubonisa ukungaqhelekanga kubude obupheleleyo bokulala / ukuvuka okanye iimpawu. yecataplexy (Matsuki et al., 2009). Ngokudibeneyo, ezi datha zibonisa ukuba i-hcrt peptides ibalulekile ukuchaza imida phakathi kokulala kunye nokuvuka, njengoko kuboniswa kukuqhekeka kokulala kunye nokuvuka kwimodeli yezilwanyana ze-narcolepsy.

Nangona kubhalwe ngokubanzi ukuba umsebenzi webhayoloji we-hcrt peptides uyimfuneko ukugcina ukuvuswa okufanelekileyo kunye nokulala, kuhlala kungacaci ukuba yeyiphi i-hcrt receptors ezimbini, i-OX1R, okanye i-OX2R, inoxanduva lwebhayoloji yeziphumo ze-hcrt ekuvukeni, kunye nokuzinza kokulala. kunye nokulawula ithoni yemisipha. OX1R I-mRNA ibonakaliswa kwiindawo ezininzi zobuchopho, ngakumbi i-LC, i-raphe nuclei, i-LDT ngelixa OX2R I-mRNA ibonisa ipateni ehambelanayo yokubonakalisa kwi-cerebral cortex, i-raphe nuclei, kunye ne-dorsomedial kunye ne-posterior (kwi-tuberomammillary nucleus) i-hypothalamus (Trivedi et al., 1998; UMarcus et al., 2001; UMieda et al., 2011). Ke, kucetyiwe ukuba ulawulo lokuphaphama kunye nokulala-ukuvuka kwe-NREM kuxhomekeke kakhulu kwi-OX2R (Mochizuki et al., 2011) ngelixa i-dysregulation yokulala kwe-REM (eyodwa kwi-narcolepsy-cataplexy) iphumela ekulahlekeni komqondiso nge-OX1R kunye ne-OX2R (i-Mieda et al., 2011). Nangona kunjalo, iimpembelelo zabo kulawulo lwe-narcolepsy, ngakumbi i-cataplexy kunye nokuhlaselwa kokulala, zihlala zingacacanga. Izinja ezine-narcolepsy ezifuywayo zithwala utshintsho olungenanto kwi OX2R gene (Lin et al., 1999) kunye nomfuziselo wemouse ohambelanayo, i OX2R Iimpuku ze-KO, zibonisa iimpawu ezincinci kunezinja (Willie et al., 2003). Nangona i-OX1R ithatha inxaxheba kulawulo lokuvusa (uMieda et al., 2011), igalelo layo kwiimpawu ze-narcolepsy lihlala libonakaliswa ngakumbi.

Okubalulekileyo, umsebenzi kwezinye iinkqubo zokuvusa uphazamiseka kakhulu ngexesha le-cataplexy. I-LC neurons iyayeka ukukhupha (Gulyani et al., 1999) kunye ne-serotoninergic neurons zinciphisa kakhulu umsebenzi wazo (Wu, 2004), ngelixa iiseli ezikwi-amygdala (Gulyani et al., 2002) kunye ne-TMN ibonise inqanaba elongeziweyo lokudubula (uJohn et al., 2004). Lo mbutho ucebisa ukuba zombini i-OX1R (LC, raphe), kunye ne-OX2R (TMN, i-raphe) ibandakanyeka ekugcineni ithoni ye-muscle efanelekileyo. Uphononongo lwakutsha nje luye lwaqaqambisa indima yeenkqubo zecholinergic ezitshintshiweyo ekuqaliseni i-cataplexy kwiimpuku ze-narcoleptic (Kalogiannis et al., 2011, 2010). Ke ngoko, injongo ebalulekileyo, engasonjululwanga kukuchonga i-wiring esebenzayo ye-hcrt neurons, kunye ne-dynamics yokukhululwa kwe-synaptic kwii-terminals ze-hcrt ukucacisa ngokuchanekileyo uqikelelo olusezantsi (de Lecea et al., 2012) ezilawula ukuvuseleleka, iimeko zokulala, ithoni yezihlunu, kunye nokuziphatha okujoliswe kuko.

I-locus coeruleus, i-norepinephrine, kunye ne-arousal

I-LC ikufuphi ne-4^th i-ventricle kwi-brainstem kwaye iqulethe i-neurons eyenza i-monoamine norepinephrine (NE). Nangona ezinye iiseli ezine zabemi nazo zivelisa i-NE (i-A1, A2, A5, kunye namaqela eeseli ze-A7), i-LC ivelisa ~ 50% ye-NE yengqondo iyonke kwaye kuphela komthombo kwi-cortex. Zininzi ii-receptors ze-NE ezisebenzayo ezifumaneka kuyo yonke ingqondo, kunye ne-α1 kunye ne-β receptors zihlala zibangela amandla e-postsynaptic e-excitatory kunye ne-α2 receptors edla ngokubangela amandla okuthintela i-postsynaptic. Ii-receptors ze-α2 zifumaneka kakhulu kwi-LC neurons (iBerridge kunye ne-Waterhouse, 2003) ngokwabo kwaye basebenze njenge-autoreceptors ezithintelayo ukucinezela umsebenzi ongaphakathi.

Ukurekhodwa kwizilwanyana eziziphetheyo eziphaphileyo kubonisa ukuba i-LC neurons ivutha umlilo kwi-1-3 Hz ngexesha lokuvuka, umlilo omncinci ngexesha lokulala kwe-SWS, kwaye zithe cwaka ngexesha lokulala kwe-REM (Aston-Jones kunye neBloom, 1981; Jones, 2003; Saper et al., 2010). I-LC iphinda iqhume ngokukhawuleza kwi-8-10 Hz ngexesha lokubonisa izinto ezivuselelayo ezinokunyusa ixesha lokuvuka. Njenge-hcrt neurons, utshintsho kwisantya sokukhupha lulandela utshintsho kutshintsho lokulala ukuya kuvuka (Aston-Jones kunye neBloom, 1981), ecebisa ukuba ezi seli zibalulekile kutshintsho lokuvuka okanye ingqalelo.

Okubangela umdla kukuba, izilonda zomzimba ze-LC azifuni utshintsho olungaguqukiyo kwi-cortical EEG okanye izalathisi zokuziphatha zokuvuka (Lidbrink, 1974; UBlanco-Centurion et al., 2007). Ukukhutshwa kwemfuza ye-dopamine beta-hydroxylase, i-enzyme efunekayo kwi-NE synthesis, nayo ayiphazamisi iindawo zokulala (uHunsley et al., 2006). Oku kuphakamisa ubukho be-neural circuitry engafunekiyo, ngaphandle kwesakhiwo se-LC, ukuxhasa umsebenzi we-cortical kunye neendlela zokuphuhlisa imbuyekezo, ngokulandelanayo. Nangona kunjalo, iinaliti eziphambili ze-pharmacological antagonists ye-α1 kunye ne-β noradrenergic receptors (iBerridge kunye ne-España, 2000) okanye i-agonists ye-inhibitory α2 autoreceptors (De Sarro et al., 1987) zineziphumo ezininzi zokuthomalalisa. Ulawulo oluphakathi lwe-NE ngqo kwi-ventricles okanye i-forebrain ikhuthaza ukuphaphama (uSegal noMandell, 1970; I-Flicker kunye neGeyer, 1982). Ukuvuselela i-neurons kwi-LC usebenzisa i-microinjections yendawo ye-cholinergic agonist (bethanechol) ivelisa ukusebenza ngokukhawuleza kwe-forebrain EEG kwiigundane ze-halothane-anesthetized (i-Berridge kunye ne-Foote, 1991). Kutshanje, inkqubo ye-LC-NE ibonakaliswe ibalulekile ekugcineni ukonyuka kwenwebu ye-cortical neurons ekuvukeni xa kuthelekiswa neendawo zokulala (iConstantinople kunye neBruno, 2011). Zithathiwe kunye, ezi zifundo zithetha ukuba inkqubo ye-LC-NE yenza i-cortical isebenze kwaye inyuse i-membrane ye-cortical enokuthi inyuse ukuvuswa.

I-Optogenetic dissection ye-hcrt kunye ne-LC-NE yokulawula ukuvuswa

Umsebenzi we-hcrt kunye ne-LC-NE neurons ihambelana nokutshintsha kokulala ukuya kuvuka, nangona kunjalo, kuye kwaba nzima ukukhetha ukuvuselela okanye ukuthintela i-hcrt ethile kunye ne-LC-NE yabemi kunye nesisombululo sexeshana esihambelana nokulala okanye ukuvuka, kunye nokufezekisa. ukukhethwa kwendawo ukukhangela ezo seli ngaphandle kokuchaphazela iiseli ezijikelezileyo okanye iifibers-of-passage. Kwiinzame zokuqonda ngcono amandla exeshana ee-neural circuits of wakefulness, kutshanje sisebenzisa i-optogenetics ukubuyisela umva nangokukhetha umsebenzi we-hcrt kunye ne-LC neurons kwizilwanyana ezihamba ngokukhululekileyo (Adamantidis et al., 2007; UCarter et al., 2009, 2010, 2012). I-Optogenetics isebenzisa iimolekyuli ze-actuator opsin (umzekelo, i-channelrhodopsin-2 (ChR2) okanye i-halorhodopsin-NpHR) ngokukhetha ukusebenzisa okanye ukuthulisa iiseli ezijoliswe kumfuzo, ngokulandelelanayo, ngokudanyaza kokukhanya kwi-wavelength ethile (uBoyden et al., 2005). Olunye ulwazi malunga netekhnoloji ye-optogenetic inokufumaneka kwezinye iiphononongo ezininzi ezigqwesileyo (Zhang et al., 2006; Miesenbock, 2009; Scanziani kunye noHäusser, 2009; Yizhar et al., 2011; Deisseroti, 2012).

Ukuhambisa ezi actuators kwi-hcrt okanye kwi-LC neurons, sasebenzisa i-lentiviral kunye ne-cre-dependent adeno-associated viral (AAV) izixhobo zokuhambisa i-gene, ngokulandelanayo, phantsi kolawulo lwabakhuthazi abathile beeseli (Adamantidis et al., 2007). Ukuhambisa ukukhanya kwi-hcrt okanye kwi-LC field, siye sayila i-optical-neural interfaces apho iifayili ze-optical zafakwa ngokungapheliyo kwi-skull ye-mouse, njengoko kuchazwe kwenye indawo (Adamantidis et al., 2005, 2007; Aravanis et al., 2007; UZhang et al., 2010). Ukusebenzisa esi sicwangciso, sakwazi ukulawula umsebenzi we-hcrt neural zombini in vitro kwaye kwi vivo ngovuselelo oluchanekileyo lwemillisecond (Adamantidis et al., 2007). Ukuchaneka okuphezulu kwexesha kunye nendawo yokuvuselela kusivumele ukuba silinganise uluhlu lwe-physiological of hypocretin neuron discharge rates (1-30 Hz) (Hassani et al., 2009). Ewe, sisebenzise oololiwe bokukhanya be-pulse kuvuselelo lwethu lwe-optogenetic olusekwe kwiparamitha kuhlalutyo lwenyani lwe-hcrt neurons. kwi vivo (oku kuyinyaniso kulawulo lwe-optogenetic lwe-LC-NE neurons echazwe ngezantsi). Sifumene ukuba ukuvuselela unilateral optical optical stimulation of hcrt neurons kwandisa amathuba okutshintshela ekuvukeni ukusuka nokuba yi-SWS okanye ukulala kwe-REM (Umfanekiso (Figure1A) .1A). Okubangela umdla kukuba, uvuselelo oluphezulu lwe-optical frequency (5-30 Hz light pulse trains) lunciphise ukubambezeleka ukuba luvuke ngelixa oololiwe be-1 Hz bengazange, becebisa ukukhutshwa kwe-synaptic exhomekeke rhoqo kwi-neurotransmitter (glutamate) kunye ne-neuromodulators, kubandakanya i-hcrt okanye i-dynorphin ukusuka kwii-terminals. Saye sabonisa ukuba iziphumo zokuvuselela i-hcrt neurons zinokuvaleka ngokutofa kwe-OX1R okanye ngokucinywa kwemfuza ye-hcrt gene, ebonisa ukuba i-hcrt peptides ilamla, ubuncinci, i-optogenetically-induced sleep-to-wake transitions. Ezi ziphumo zibonisa ukuba ukukhululwa kwe-hcrt kwi-hcrt-expressing neurons kuyimfuneko kwiipropati zokukhuthaza ukuvuka kwezi neurons. Okubalulekileyo, ezi ziphumo zibonisa ikhonkco le-causal phakathi kwe-hcrt neuron activation kunye nokuguquka kokulala ukuya kuvuka, ngokuhambelana nezifundo zangaphambili ezihambelanayo. Oku kwaxhaswa ngakumbi kukuba ukuthuliswa kwe-optical ye-hcrt neurons kukhuthaza i-SWS (Tsunematsu et al., 2011).

 

Umzobo 1

I-Optogenetic dissection yeesekethe ezivuselelayo zobuchopho. (A) Ukukhuthazwa kwe-hcrt neurons kunye ne-ChR2 kubangela ukuncipha kwe-latency yokulala ukuya ku-wake kwi-10 Hz kodwa kungekhona i-1 Hz (idatha esuka kwi-Adamantidis et al., 2007). (B) Ukukhuthazwa kwe-LC neurons kunye neChR2 kubangela ngokukhawuleza ...

Ezi ziphumo zisanda kuqinisekiswa nguSasaki kunye nabasebenzisana nabo (Sasaki et al., 2011), osebenzise indlela ye-pharmacogenetic ebizwa ngokuba yi-Designer Receptors Exclusively Activated yi-Designer Drugs (DREADDs) ukuba isebenze kwaye icinezele umsebenzi we-hcrt neural. Itekhnoloji ye-DREADD ivumela ukumodareyithwa kwe-bimodal yomsebenzi we-neural kunye nesisombululo sexeshana seeyure ezininzi (Dong et al., 2010). Baye bafumanisa ukuba ukusebenza kwe-hcrt neural activity kwandisa ukuphaphama ngelixa ukucinezelwa komsebenzi we-hcrt kukhuthaza i-SWS.

Kwisifundo sesibini (uCarter et al., 2009), sibonise ukuba ulawulo lwe-hcrt yokuguqulwa kokulala-ukuvuka kuphantsi kokuxhomekeka kweenkqubo zokulala ze-homeostasis ekubeni i-hcrt-mediated sleep-to-wake iinguqu zivaliwe ngokunyuka koxinzelelo lokulala (okubangelwa kukungabikho kokulala). Nangona kunjalo, impembelelo ye-optogenetic stimulations ye-hcrt iqhubekile kwi-histamine decarboxylase knockout iimpuku (iimpuku ezingakwaziyo ukwenza i-histamine) icebisa ukuba enye into ekujoliswe kuyo ukuba inkqubo ye-histaminergic inoxanduva lwesiphumo se-hcrt. Okokugqibela, sibonise ukuba amaziko asezantsi avuselelayo anje nge-LC neurons omabini awonyusa umsebenzi wawo (njengoko kulinganiswe yintetho ye-c-Fos) ekuphenduleni i-hcrt optogenetic stimulation. Ngenxa yokuba umsebenzi wangaphambili ubonise impembelelo evuyisayo ye-hcrt kwi-LC NE neurons (uBourgin et al., 2000), siye saphanda uxhulumaniso lwe-hcrt-LC kwaye sijolise kuphando lwethu lokulinga kwi-noradrenergic LC njengento ekujoliswe kuyo entsha ye-optogenetic manipulation.

Kwisifundo sesithathu (uCarter et al., 2010), sijolise ngokwemfuza i-LC-NE neurons ngenaliti ye-stereotaxic ye-Cre recombinase-exhomekeke kwi-adeno-associated virus (rAAV) kwiigundane ezinkqonkqoza ngokukhethayo ezichaza iCre kwi-tyrosine hydroxylase (TH) neurons (Atasoy et al., 2008; Tsai et al., 2009). Sifumene ukuba zombini i-NpHR kunye ne-ChR2 yayisebenza kwaye inokuthintela kwaye isebenze, ngokulandelelana, i-LC-NE neurons zombini. in vitro kwaye kwi vivo (Umzobo (Figure1B) .1B). Sifumene ukuba i-optogenetic low frequency (1-10 Hz) yokuvuselela i-LC-NE neurons ibangele ngokukhawuleza (ngaphantsi kwe-5 s) ukuguquka kokulala ukuya kuvuka ukusuka kwi-SWS kunye nokulala kwe-REM. Ukuvuselela i-LC neurons ngexesha lokuvuka kwandisa umsebenzi we-locomotor kunye nexesha elipheleleyo elichithwe liphapheme, liqinisekisa umphumo onamandla wokuvusa. Ngokwahlukileyo, ukuthuliswa kwe-NpHR-mediated ye-LC-NE neurons yehlisile ubude beziqephu zokuvuka kodwa ayizange ithintele utshintsho lokulala ukuya kuvuka xa izilwanyana zilele. Kuthatyathwe kunye, olu phononongo lubonise ukuba ukusebenza kwe-LC-NE neurons kuyimfuneko yokugcina ubude obuqhelekileyo bokuvuka (uvavanyo lwe-NpHR), kwaye ngokwaneleyo ukukhuthaza utshintsho olukhawulezileyo lokulala, ukuphaphama okuqhubekayo, kunye nokunyuka kwe-locomotor arousal. Ke, sicebise ukuba i-LC-NE neurons isebenze njengenkqubo yokulungisa ngokukhawuleza ukukhuthaza utshintsho lokulala kunye nokuvusa ngokubanzi. Okubangela umdla kukuba, sifumene ukuba ukusebenza okubonakalayo kwe-LC-NE neurons kubangela ukubanjwa kwe-locomotor (uCarter et al., 2010). Ukubanjwa kokuziphatha okunjalo kwabelana ngeempawu eziqhelekileyo kunye ne-cataplexy, catatonia okanye ukuziphatha okukhenkcezayo zombini kwiimodeli zezilwanyana kunye nezigulana ezingabantu (i-Scammell et al., 2009). Iindlela ezinokwenzeka zinokubandakanya ukuncitshiswa kwe-NE kwiitheminali ze-synapse ze-LC-NE okanye i-LC-NE overexcitation ye-brainstem motor nuclei enokukhokelela ekukhubazekeni. Uphononongo olongezelelweyo luyafuneka ukutyhila iindlela ezisisiseko.

Kuphononongo lwethu lwamva nje (uCarter et al., 2012), siye savavanya i-hypothesis yokuba i-LC yomsebenzi wamasango i-hcrt yempembelelo ye-neuron ekutshintsheni kokulala ukuya ekuvukeni. Ngenxa yokuba i-hcrt kunye ne-LC neural populations zibekwe kwimimandla yobuchopho eyahlukeneyo, kunokwenzeka ngokwasemzimbeni ukufikelela kuzo zombini izakhiwo ngaxeshanye kwisilwanyana esinye. Ke ngoko sithathe indlela emibini ye-optogenetic yokuvuselela i-hcrt neurons ngelixa sithintela okanye sivuselela i-noradrenergic LC neurons ngexesha lokulala kwe-SWS. Sifumene ukuba ukuthulisa i-LC neurons ngexesha lovuselelo lwe-hcrt luvalwe i-hcrt-mediated sleep-to-wake utshintsho (Figure (Umzobo1C) .1C). Ngokwahlukileyo koko, safumanisa ukuba ukonyusa i-excitability ye-LC neurons ngokusebenzisa i-step-function opsin (SFO) activation-oko kunyuswa kweeseli ekujoliswe kuzo (Berndt et al., 2009)—ngexesha lovuselelo lwe-hcrt (usebenzisa i-LC stimulation protocol eyodwa enganyusi iinguqu zokulala ukuya kuvuka) iphuculwe i-hcrt-mediated sleep-to-wake transitions (Figure (Umfanekiso1D) 1D). Sithathiwe kunye, iziphumo zethu zibonisa ukuba i-LC isebenza njengento efunekayo kunye neyoneleyo ye-downstream effect kwi-hcrt-mediated SWS-to-wake transitions ngexesha elingasebenziyo.

hcrt kunye ne-LC-NE inkqubo ye-dynamics

Kuzo zonke izifundo zethu zovavanyo, siye sabona ukuba i-optogenetic manipulation ye-hcrt kunye ne-LC-NE neurons ichaphazela ukulala-ukuvusa iinguqu ezinobunzima bexeshana ezahlukeneyo (Adamantidis et al., 2007; UCarter et al., 2009, 2010, 2012). Ukusetyenziswa kwe-acute optical activation ye-hcrt neurons kubangela ukulala-ukuvuka utshintsho kwixesha le-10-30 s, ngelixa ukuvuselela i-LC neurons kubangela ukuguquka kokulala-ukuvuka ngaphantsi kwe-5 s. Enye ingcaciso kukuba i-hcrt neurons inokusebenza njenge-upstream integrator of arousal ngexesha lemisebenzi enxulumene ne-hypothalamic ngelixa inkqubo ye-LC-NE isebenza njengeyona nto iphambili yokuvusa, uxinzelelo kunye nokuqwalaselwa. Nangona kunjalo, iinkqubo ze-neuronal effector zinokwenzeka ukuba zingafuneki kwaye zenziwe zisebenze ngeeseti ezahlukeneyo zamagalelo. Ke ngoko, asinakukhupha ukuba ukuthintela ezinye iinkqubo zokuvusa, ezinjenge-histaminergic kunye neenkqubo ze-cholinergic, nazo ziyakuchaphazela kakhulu iinguqu zokuziphatha ze-hcrt kwezinye iimeko zovavanyo.

Ngaphandle kwezi ziphumo zexesha elifutshane, kukwanika umdla ukuba imifuniselo yefotostimulation egciniweyo (okt, engaguqukiyo) ye ~ 1-4 h ye-hcrt neurons yonyusa iinguqu zokulala ukuya kuvuka ngaphandle kokutshintsha ubude bexesha lokuvuka, ngelixa ixesha elide. ifotostimulation ye-LC-NE neurons yonyusa kakhulu ixesha lokuvuka. Ezi ziphumo zibonisa ukuba inkqubo ye-hcrt inokulawula imida yokulala ngelixa i-neurons ye-LC-NE inokulawula ixesha lokuvuka ngokunyusa amandla e-cortical membrane kunye nokunciphisa i-cortical EEG.

I-hypothalamic localization ye-hcrt neurons ithetha ukuba ezi seli zinendima ebalulekileyo yokuvuselela ngexesha leenkqubo ze-homeostatic, kubandakanya ukuziphatha ngokwesondo, ukutya, ukusabela koxinzelelo kunye nokukhuthaza. Ngaphandle kokulawula ukuphaphama, iinkqubo ezivuselelayo zikwathatha inxaxheba ekuziphatheni kokufuna umvuzo, izenzo zesondo, iimpendulo zokubhabha okanye ukulwa, njl njl. Oku kuphelelwa amandla kunokuba kudibanise umsebenzi wokuvusa kuyo yonke indaleko kunye neendlela ezahlukeneyo zobuchopho ezixhasa ukuphaphama kunye nokuziphatha okunxulumene nokuvuselela okuyimfuneko ukusinda. Ngokomzekelo, ukusebenza kwenkqubo ye-LC-NE kwandisa ukuvusa kwaye kubangela ukuziphatha okuxhalabisayo (u-Itoi noSugimoto, 2010). Ngokwahlukileyo, inkqubo ye-neuropeptide S (NPS), i-peptide eveliswa yi-neuronal ethintelweyo ye-ventral kwi-LC, nayo yonyusa ukuvuswa kodwa. kuncipha ixhala (uPape et al., 2010). Ke, ukuxhasa imisebenzi eyahlukeneyo yokuziphatha, iisekethe ezivuselelayo kufuneka zibe zifikelele kwinqanaba eliphezulu lokuchazwa, ngokunokwenzeka ngokudibanisa okukhethiweyo koqhagamshelo lwabo oluhambelanayo kunye nolusebenzayo, ukukhupha amandla okuhambisa / iimodyuli kunye nomsebenzi ohambelanayo kunye nezinye iisekethe ezivuselelayo.

Iimpembelelo

Kwiminyaka emihlanu edlulileyo, ukudibanisa kwe-optogenetics, imodeli yemouse eyenziwe ngofuzo, kunye nohlalutyo lwe-EEG / EMG yokulala lunikeze isethi ekhethekileyo kunye nenamandla yezixhobo zokuqonda ngakumbi igalelo leenkqubo ze-hcrt kunye ne-LC ekuvukeni, kunye ezinye iiseli zemithambo-luvo ezilawula amaqondo obuthongo kunye nokuvuka. Ukujolisa kwi-optogenetic probes kwezinye ii-neurons ezisengqondweni kuya kugqiba indima yabo kunye neyokudibanisa kwimida yokulala / yokuvuka. Ngaphaya koko, ezi zixhobo ziya kusivumela ukuba sijonge indlela yobuchopho ephantsi kwe-wake states ngokusekwe kuqikelelo lwe-anatomical, i-synaptic neurotransmission, kunye ne-dynamics yokukhululwa kwe-transmitter. Ukukwazi ukujolisa kunye nokulawula ezi sekethe ngokuchaneka okuphezulu kwexesha (<1 s) ngakumbi kuvumela ithuba lokuphanda indima yabo kuluhlu olubanzi lweendlela zokuziphatha ezinjengokutya, umlutha, uxinzelelo, ingqalelo, kunye nokuvuswa ngokwesondo. Ekugqibeleni, ezi zifundo zinokutyhila iindlela ze-pathophysiological zokuphazamiseka kwengqondo okunje ngoxinzelelo olungapheliyo, ukuba likhoboka, ukunqongophala kwengqwalasela, kunye noxinzelelo.

Imibulelo

Ucaphulo