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Supplementary Information for Brain-wide functional architecture remodeling by alcohol dependence and abstinence Adam Kimbrough, Daniel J. Lurie, Andres Collazo, Max Kreifeldt, Harpreet Sidhu, Giovana Camila Macedo, Mark D’Esposito,Candice Contet, Olivier George Olivier George Email: [email protected] This PDF file includes: Figures S1 to S8 Tables S1 to S1 1 www.pnas.org/cgi/doi/10.1073/pnas.1909915117 Fig. S1. Withdrawal behaviors for separate control groups (alcohol-nondependent and naive mice) compared with alcohol-dependent mice. A. The number of irritable-like responses was significantly higher in alcohol-dependent mice (black bar) compared with both alcohol- nondependent mice (white bar) and naive mice (hashed bar). B. Alcohol-dependent mice (black bar) exhibited a significant increase in digging behavior compared with both alcohol- nondependent mice (white bar) and naive mice (hashed bar). *p < 0.05, vs. naive; #p < 0.05, vs. alcohol-nondependent. 2 Fig. S2. Saccharin intake in alcohol-dependent, nondependent, and naive (water) mice, expressed at mg/kg/24 h. Average saccharin intake over 7 days of testing was not different between alcohol-dependent (black bar), alcohol-nondependent (white bar), and naive (hashed bar) mice. 3 Count 0 40 80 120 −1 and Histogram and −0.5 Color K Color V alue 0 e 0.5 y 1 S Fig. Naive - Cortical amygdalar area posterior part 1 Entorhinal area medial part Entorhinal area lateral part Parasubiculum Lateral visual area - Agranular insular area ventral part 1 0.5 0 0.5 Color Key Color 3 Gustatory areas Piriform area . Postsubiculum Retrosplenial area ventral part Interbrain Interbrain region Cortical amygdalar area Piriform−amygdalar area Anterior cingulate area dorsal part Presubiculum Primary auditory area Prelimbic area Agranular insular area Subiculum Anterior cingulate area ventral part Perirhinal area Ectorhinal area Dorsal auditory area Primary somatosensory area mouth Agranular insular area posterior part Supplemental somatosensory area Ventral auditory area Orbital area medial part Infralimbic area posteromedial visual area Visceral area Temporal association areas Dorsal peduncular area Primary motor area Orbital area lateral part Pearson correlations for naive with for correlationsPearson naive mice Orbital area ventrolateral part Frontal pole cerebral cortex Primary somatosensory area lower limb Primary somatosensory area upper limb Taenia tecta Primary visual area Anterolateral visual area Primary somatosensory area trunk Secondary motor area Retrosplenial area dorsal part Field CA1 Field CA3 Field CA2 Dentate gyrus Posterior amygdalar nucleus Claustrum Lateral amygdalar nucleus Endopiriform nucleus Basolateral amygdalar nucleus Central amygdalar nucleus Intercalated amygdalar nucleus Caudoputamen Olfactory tubercle Nucleus accumbens Fundus of striatum Anterior amygdalar area Lateral septal complex Medial amygdalar nucleus Magnocellular nucleus Substantia innominata Diagonal band nucleus Medial septal nucleus Bed nuclei of the stria terminalis Globus pallidus external segment Paraventricular nucleus of the thalamus Medial habenula Medial geniculate complex Central medial nucleus of the thalamus Lateral habenula Anterodorsal nucleus Lateral posterior nucleus of the thalamus Lateral dorsal nucleus of thalamus Posterior complex of the thalamus Ventral anterior−lateral complex of the thalamus Mediodorsal nucleus of thalamus Ventral medial nucleus of the thalamus Parataenial nucleus Reticular nucleus of the thalamus Dorsal part of the lateral geniculate complex Anteroventral nucleus of thalamus Central lateral nucleus of the thalamus Tuberal nucleus Parasubthalamic nucleus Periventricular zone Suprachiasmatic nucleus Subparaventricular zone Median preoptic nucleus Anteroventral periventricular nucleus region labels.region Subthalamic nucleus Zona incerta Ventromedial hypothalamic nucleus Posterior hypothalamic nucleus Lateral hypothalamic area Dorsomedial nucleus of the hypothalamus Anterior hypothalamic nucleus Lateral preoptic area Paraventricular hypothalamic nucleus Periventricular hypothalamic nucleus posterior part Retrochiasmatic area Medial preoptic area Anteroventral preoptic nucleus Ventrolateral preoptic nucleus Medial preoptic nucleus Interpeduncular nucleus Ventral tegmental area Substantia nigra reticular part Midbrain reticular nucleus Substantia nigra compact part Pons motor related Inferior colliculus Pons Periaqueductal gray Superior colliculus motor related Pedunculopontine nucleus Culmen Olivary pretectal nucleus Superior colliculus sensory related Precommissural nucleus Medial pretectal area Medial pretectal area pretectal Medial Precommissur Super Oliv Culmen P Super P P Inf P Substantia nigr Midbr Substantia nigr V Inter n preoptic Medial V Antero area preoptic Medial area Retrochiasmatic P P Later Anter n Dorsomedial Later P V incer Zona Subthalamic n Antero n preoptic Median Subpar Supr P P T Centr Antero par Dorsal n Reticular P V n Mediodorsal V P Later Later n Anterodorsal Later Centr comple geniculate Medial haben Medial P Glob Bed n n septal Medial n band Diagonal Substantia innominata n Magnocellular am Medial Later Anter str of Fundus accumbens Nucleus Olf Caudoputamen am Intercalated Centr Basolater Endopir Later Claustr P gyr Dentate CA2 Field CA3 Field CA1 Field par dorsal area Retrosplenial Secondar Pr Anterolater Pr T Pr Pr cerebr pole Frontal v area Orbital later area Orbital Pr area peduncular Dorsal T Viscer posteromedial visual area Infr par medial area Orbital V Supplemental somatosensor Agr Pr auditor Dorsal area Ectorhinal P Anter Subiculum Agr Prelimbic area Pr Presubiculum Anter Pir Cor v area Retrosplenial P Pir Gustator Agr Later P later area Entorhinal medial area par Entorhinal Cor uber aenia tecta empor entr entrolater entromedial h entr entr entr edunculopontine n er ons ons motor related er ar oster er ar ar oster ar oster er ostsubiculum ar imar imar imar imar imar imar imar er actor if if alimbic area iaqueductal gr iv a iv asubthalamic n ataenial n a irhinal area asubiculum an an an tical am tical am tical ar or or ior colliculus peduncular n v v us pallidus e achiasmatic n entr entr al al preoptic area al h al dorsal n al poster al haben al septal comple al am al visual area al al tegmental area al medial n al anter al auditor ior h al later al medial n ior am al am ior cingulate area v ior cingulate area dorsal par ain ain reticular n al n entr entr y pretectal n pretectal y uclei of the str m−am area m ular insular ular area poster insular ular area ular insular area v ior colliculus sensor ior colliculus motor related al al area ior h ior comple ior am v v v y somatosensor y area visual y somatosensor y somatosensor y area motor y somatosensor y auditor y um a al al association areas entr entr entr if y tubercle y v ypothalamic area ypothalamic y areas y or icular h icular z ucleus entr al am icular h icular n y motor area motor y ypothalamic n ypothalamic ygdalar n ygdalar t of the later the t of ygdalar n ygdalar ypothalamic n ypothalamic ta ucleus of the thalam al al preoptic n ygdalar n ygdalar n m al al visual area ygdalar area ygdalar poster area ygdalar al al preoptic n al per al n ygdalar area ygdalar us al al n area ygdalar ygdalar n ygdalar ior−later ucleus iatum icular z ula entrolater ula ior n y area y ucleus of thalam ucleus al al n ucleus of the h area y ucleus ucleus ypothalamic n ypothalamic a compact par a reticular par n ygdalar ucleus of thalam ucleus y area y ucleus of thalam n ygdalar ucleus of the thalam ucleus one ypothalamic n ypothalamic ucleus of the thalam ucleus of the thalam ucleus ypothalamic n ypothalamic thalam the of x al par ucleus of the thalam ucleus iv xter a ucleus ucleus ucleus ucleus ucleus of the thalam ucleus ucleus y entr al al cor ucleus ucleus ucleus ucleus ia ter entr ucleus al par one al al comple nal nal segment x ucleus al al geniculate comple t icular n tr area y limb upper area y lo area y mouth area y al al par t ucleus ucleus 4 ucleus entr al al par ucleus entr te ucleus ucleus minalis x y related y t x t al al par al al par y area y t ucleus t t ior par ypothalam t t ucleus poster ucleus x of the thalam the of x us ucleus us ior par unk w us us us er er limb t t t t us us us us t us us x ior par us t Count 0 40 80 120 −1 and Histogram and −0.5 Color K Color V alue 0 e 0.5 y S Fig. - 1 1 Alcohol Nondependent Alcohol - Cortical amygdalar area posterior part 1 0.5 0 0.5 Color Key Color 4 Entorhinal area medial part Entorhinal area lateral part . Parasubiculum Interbrain Interbrain region Lateral visual area Agranular insular area ventral part Gustatory areas Piriform area Postsubiculum Retrosplenial area ventral part Cortical amygdalar area Piriform−amygdalar area Anterior cingulate area dorsal part Presubiculum Primary auditory area Prelimbic area Agranular insular area Subiculum Anterior cingulate area ventral part Perirhinal area Ectorhinal area Dorsal auditory area Primary somatosensory area mouth Agranular insular area posterior part Supplemental somatosensory area Ventral auditory area Orbital area medial part Infralimbic area Pearson correlations for alcohol nondependent mice with region labels.mice region with for correlationsPearson alcohol nondependent posteromedial visual area Visceral area Temporal association areas Dorsal peduncular area Primary motor area Orbital area lateral part Orbital area ventrolateral part Frontal pole cerebral cortex Primary somatosensory area lower limb Primary somatosensory area upper limb Taenia tecta Primary visual area Anterolateral