NEUROSCIENCE

Regulation of feeding by somatostatinneurons in the tuberal nucleusSarah Xinwei Luo1*, Ju Huang2*, Qin Li1,3*, Hasan Mohammad1, Chun-Yao Lee1,Kumar Krishna4, Alison Maun-Yeng Kok1, Yu Lin Tan1, Joy Yi Lim1, Hongyu Li1,Ling Yun Yeow1, Jingjing Sun2, Miao He5, Joanes Grandjean1, Sreedharan Sajikumar4,Weiping Han1, Yu Fu1,4†

The tuberal nucleus (TN) is a surprisingly understudied brain region.We found thatsomatostatin (SST) neurons in the TN, which is known to exhibit pathological or cytologicalchanges in human neurodegenerative diseases, play a crucial role in regulating feeding in mice.GABAergic tuberal SST (TNSST) neurons were activated by hunger and by the hunger hormone,ghrelin. Activation of TNSSTneurons promoted feeding, whereas inhibition reduced it viaprojections to the paraventricular nucleus and bed nucleus of the stria terminalis. Ablation ofTNSSTneurons reduced body weight gain and food intake.These findings reveal a previouslyunknown mechanism of feeding regulation that operates through orexigenic TNSSTneurons,providing a new perspective for understanding appetite changes.

The nucleus tuberalis lateralis (NTL; lateraltuberal nucleus) in the hypothalamus isrevealed by the “lateral eminences on theventral surface of the tuber cinereum” inhumans and primates (1), but the homol-

ogous structure in other animals is less well de-fined, and knowledge about theNTL is scarce (2).Pathological or cytological changes in the NTLhave been observed in several neurodegenerativeandneurological diseases (2–8).Deephypothalamiclesions that probablywent into theNTL inhumanpatients have led to hypophagia, and NTL pa-thology has been correlatedwith cachexia (4, 9–11),suggesting that the NTL plays a role in the reg-ulation of feeding, but this notion has neverbeen tested. The human NTL neighbors the tu-beromammillary nucleus (TMN) and is markedby dense staining with somatostatin (SST) anti-body (12). We therefore searched for SST-positiveneurons in the mouse tuberal nucleus (TN) andstudied their potential role in feeding regulation.We bred SST-Cremice with Ai14 reporter mice

(13), and SST neurons thus were labeled with thered fluorescent protein tdTomato. In the mousehypothalamus, SST neurons appeared in a scat-tered distribution in the medial-dorsal hypo-thalamus and in enriched clusters in the ventralhypothalamus. Specifically, SST neurons wereprevalent in the arcuate area (we refer to theseas ArcSST neurons) and the TN (TNSST neurons).

The arcuate cluster extended slightly anteriorly,and the TN cluster extended slightly posteriorly(Fig. 1A). The arcuate nucleus plays a critical rolein feeding regulation and contains well-studiedorexigenic agouti-related peptide (AgRP) neurons,tyrosine hydroxylase (TH)–positive neurons, andanorexic proopiomelanocortin (POMC) neurons(14–16). Immunohistochemistry showed that theArcSST neurons were distinct from POMC- andTH-positive neurons but exhibitedminor overlap(9.04 ± 2.16% of ArcSST neurons; n = 3mice) withAgRP neurons (Fig. 1B) (17). The TNSST neuronswere distinct and spatially segregated from lateralhypothalamic orexin- and melanin-concentratinghormone (MCH)–positive neurons (Fig. 1C), bothof which have been implicated in feeding andmetabolic regulation (18, 19). The TNSST did notshow obvious overlap with histaminergic neurons[revealed by staining for histidine decarboxylase(HDC)], the key neuronal subtype in the nearbyTMN region (20) (Fig. 1C and fig. S1). Therefore, asin humans, the TN inmouse ismarked by a densecluster of SST-positive neurons, which constitutea hypothalamic neuronal subtype that is distinctfrom the neurons currently known to supportfeeding and metabolic regulation. Either over-night fasting or intraperitoneal (i.p.) injectionof the hunger hormone, ghrelin—the levels ofwhich are increased by the hunger state (21)—induced robust expression of the immediate earlygene c-Fos in TNSST neurons (Fig. 1, D to G, andfig. S2), but not in ArcSSTneurons (fig. S3). Furtheranalysis showed no obvious difference in thepercentage of c-Fos–positive TNSST neurons atdifferent positions, indicating a homogeneousactivation of TNSST neurons (fig. S2). In acuteslices of SST-Cre::Ai14 mice, bath applicationof ghrelin excited 7 out of 17 TNSST neurons, asdetermined by whole-cell recording (Fig. 1, Hand I). Under the presence of synaptic blockers, 6out of 18 TNSST neurons were excited by ghrelinunder current-clamp conditions without the in-jection of depolarizing current, and 7 out of 16

TNSST neurons were excited by ghrelin with cur-rent injection for increasing baseline firing (fig.S4, A to F). We further characterized the TNSSTneurons by staining mRNA for ghrelin receptor(Ghsr), using fluorescence in situ hybridization(FISH); we found that 17.6% (120/682) of TNSSTneurons were Ghsr-positive, with the most caudalTN containing fewer Ghsr-positive SST neurons(fig. S4, G andH). Ghsr-postive and Ghsr-negativeTNSST neurons were similarly activated by over-night fasting (fig. S4, I and J), indicating that bothpopulations were involved in feeding regulation.We next injected Cre recombinase–inducible

adeno-associated viruses (AAVs) expressing che-mogenetic or optogenetic effectors into the TN ofSST-Cre mice (Fig. 2) and analyzed their eatingbehavior (fig. S5). After the mice habituated inbehavioral chambers for 4 days (fig. S6A), chemo-genetic activation of TNSST neurons by expres-sing the excitatory DREADD (designer receptorexclusively activated by designer drug) hM3D andinjection of CNO in the late morning (11 a.m.)dramatically promoted food consumptionwithinthe subsequent 3hours and significantly enhancedeating time and frequency (Fig. 2, A to E). Similarresults were found when CNO was injected at5 p.m., when mice were more spontaneously ac-tive (fig. S6, B and C). No effect of CNO on eatingbehavior was observed in control mice with greenfluorescent protein (GFP) expressed in TNSST neu-rons (Fig. 2, B to E, black traces). After express-ing Cre-dependent channelrhodopsin (ChR2)in TNSST neurons, we characterized the opto-genetic response of TNSST neurons in acute slices(fig. S7, A to F) and used a light intensity that canreliably excite TNSST neurons for in vivo experi-ments (22). In vivo unilateral optogenetic stim-ulation reliably and progressively promoted eatingas stimulation frequency was increased (fig. S7,G and H). Moreover, repeated optogenetic stim-ulation (20 Hz for 1 hour) reliably enhancedeating frequency, which required the presenceof chow (fig. S8A); in a control experiment, chemo-genetic activation of TNSST neurons did not elicitgnawing of a wood stick (fig. S8B). As reportedrecently (17), we also found that chemogeneticactivation of ArcSST promoted eating (fig. S6D).We further examined the necessity of TNSSTneurons in homeostatic eating by expressing theinhibitory DREADD k opioid receptor (KORD) inSST-Cre mice and injecting salvinorin B (SalB) toinhibit the activity of TNSSTneurons (Fig. 2F) (23).Injecting SalB at 10 p.m., at the start of the peakof homeostatic eating, significantly reduced cu-mulative eating time and eating frequency, butnot eating bout duration (Fig. 2, G to I), whichdid not correlate with eating frequency in homeo-static eating (fig. S5). SalB injection inGFP controlanimals did not produce a significant differencefrom vehicle-injected animals (Fig. 2, G to I, blacktraces). Inhibition of TNSST neurons by SalB alsosignificantly reduced food consumption in refeed-ing after overnight fasting (Fig. 2J). Becauseinjecting SalB at the peak of the animals’ darkcycle may cause stress and interfere with homeo-static eating, we expressed Cre-dependent ar-chaerhodopsin (ArchT) in the TN of SST-Cremice

RESEARCH

Luo et al., Science 361, 76–81 (2018) 6 July 2018 1 of 6

1Singapore Bioimaging Consortium, Agency for ScienceTechnology and Research (A*STAR), Singapore 138667.2Discipline of Neuroscience and Department of Anatomy,Histology and Embryology, School of Medicine, ShanghaiJiao Tong University, Shanghai, China 200025. 3Center forBrain Science, Key Laboratory of Magnetic Resonance inBiological Systems, Wuhan Institute of Physics andMathematics, Chinese Academy of Sciences, Wuhan, China430071. 4Department of Physiology, Yong Loo Lin School ofMedicine, National University of Singapore, Singapore 117597.5Institutes of Brain Science, State Key Laboratory of MedicalNeurobiology, Fudan University, Shanghai, China 200032.*These authors contributed equally to this work.†Corresponding author. Email: fu_yu@sbic.a-star.edu.sg

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and optogenetically inhibited these neurons byusing light from 10 p.m. to 12 a.m., without dis-turbing the mice’s activity (Fig. 2K). Again,optogenetic inhibition of TNSST neurons sig-nificantly reduced cumulative eating time andeating frequency, but not eating bout duration

(Fig. 2, L to N). The same optogenetic protocoldid not cause changes in eating behavior inGFP control mice (Fig. 2, L to N, black traces).To test whether TNSST neurons have long-termeffects on energy homeostasis, we injected AAV-flex-taCasp3-TEVp to express caspase-3 and ab-

late TNSST neurons in SST-Cremice (24) (fig. S9).Ablation of TNSST neurons reduced body weightgain by 56% over 10 weeks (Fig. 2O). We furtherstudied the impact of ablating TNSST neuronsby using another set of mice in metabolic cham-bers, 4 weeks after injecting viruses, when the

Luo et al., Science 361, 76–81 (2018) 6 July 2018 2 of 6

Fig. 1. The ventral hypothalamic SSTneuron clusters. (A) Serial coronalsections of SST-Cre::Ai14 mice showingdistinct SST neuron clusters in both thearcuate area and the tuberal nucleusof the hypothalamus. Scale bars, 500 mm.(B) Coronal sections of SST-Cre::Ai14mice stained for POMC, AgRP, and TH(the white arrow indicates a SST neuronthat is positive for AgRP). Scale bars,100 mm. IHC, immunohistochemistry.(C) Coronal sections of SST-Cre::Ai14 micestained for MCH, Orx (orexin), and HDC.Scale bars, 200 mm. (D) Coronal sectionsof SST-Cre::Ai14 mice that were (fasted)or were not (fed) subjected to overnightfasting and immunostained for c-Fos. Whitearrows indicate SST neurons that arec-Fos–positive. (E) Coronal sections ofSST-Cre::Ai14 mice after i.p. injection ofsaline or ghrelin, immunostained for c-Fos.White arrows indicate SST neurons thatare c-Fos–positive. Scale bars in (D) and (E),100 mm. (F) Percentage of c-Fos–positiveSST neurons under fed or fasted conditions(mean ± SEM, n = 3, unpaired t test, *P <0.02). (G) Percentage of c-Fos–positiveSST neurons after i.p. injection of saline orghrelin (mean ± SEM, n = 3, unpaired t test,*P < 0.02). (H) Whole-cell patch recordingtrace showing the change in firing frequencyafter bath application of ghrelin (100 nM).(I) Firing rate before and after ghrelinapplication (n = 17, paired t test, *P < 0.05).

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TNSST-ablated mice did not show significantbody weight differences from the control micebut gained body weight significantly moreslowly (fig. S10). TNSST ablation resulted in mildbut significantly reduced daily and dark-cyclefood intake, reduced prandial drinking, mild-ly reduced respiratory exchange ratio, similartotal energy expenditure, and mild changes inactivity (fig. S10). Because both positive and nega-tive valence mechanisms appear to play a role

in controlling feeding (25–28), we explored themotivational valence of TNSST in mice. In the ab-sence of food, chemogenetic activation of theTNSSTneurons, pairedwithCNO injection, eliciteda significant preference for the chamber (Fig. 2P).To explore the circuit mechanisms of feeding

regulation by TNSST neurons, we examined theprojection pattern of TNSST neurons by injectingAAV-FLEX-GFP into the TN of SST-Cre mice(Fig. 3A). Similarly to AgRP neurons (29), TNSST

neurons projected to various brain regions, in-cluding the paraventricular nucleus (PVN), thebed nucleus of the stria terminalis (BNST), thecentral amygdala (CeA), and the periaqueductalgray (PAG), with sparse projection to the para-ventricular nucleus of the thalamus (PVT) andthe parabrachial nuclei (PBN) (Fig. 3A). By in-jecting the retrograde tracers CTB488 and CTB647into the BNST and PVN, respectively, in SST-Cre:Ai14 mice, we found that the majority (87.7%)

Luo et al., Science 361, 76–81 (2018) 6 July 2018 3 of 6

Fig. 2. The TNSSTneu-rons regulate eating.(A) AAV-DIO-hM3D-mCherry was bilaterallyinjected into SST-Cremice (DIO,double-floxedinverse orientation).(B) Food consump-tion, (C) cumulativeeating time, (D) meaneating bout duration,and (E) eating frequencyduring the 3 hoursafter CNO injection at11 a.m. (n = 7, **P <0.01, paired two-tailedt test). SST-Cre micewere injected withAAV-FLEX-GFP as acontrol (n = 6, P > 0.5,paired two-tailedt test). n.s., notsignificant. (F) AAV-DIO-KORD-mCitrinewas bilaterally injectedinto SST-Cre mice.(G) Cumulative eatingtime, (H) eatingfrequency, and(I) eating bout durationduring the 3 hoursafter SalB injection(n = 6, *P < 0.05,paired two-tailedt test). SST-Cre micewere injected withAAV-FLEX-GFP as acontrol (n = 8, P > 0.5,paired two-tailedt test). (J) SST-Cremice were injected withAAV-DIO-KORD-mCitrine, and foodconsumption afterovernight fasting wasmeasured [n = 7, *P <0.05, one-way analysisof variance (ANOVA)]. (K) AAV-FLEX-ArchT-GFP was bilaterally injected intoSST-Cre mice. (L) Cumulative eating time, (M) eating frequency, and(N) eating bout duration during 2 hours of yellow light illumination (n = 6, *P <0.05, **P < 0.01, paired two-tailed t test). SST-Cre mice were injected withAAV-FLEX-GFP as a control (n = 7, P > 0.5, paired two-tailed t test). (O) Bodyweight gain on a normal chow diet for SST-Cre mice bilaterally injected in theTN with either AAV-FLEX-GFP as a control (n = 4) or AAV-flex-taCasp3-TEVp(n= 5) for ablating TNSSTneurons (ablation versus control: repeatedmeasures

two-way ANOVA, F = 24.21, P <0.001). (P) Experimental design of conditionedplace preference with chemogenetic activation of TNSSTneurons. A heat mapshows the percent occupancy time before and after conditioning sessions. Redbar, chemogenetic activation side.The bar graph shows the change in occu-pancy time after conditioning sessions for SST-Cre mice injected bilaterallywith AAV-DIO-hM3D-mCherry (n = 4) or AAV-FLEX-GFP (n = 3) (**P < 0.01,one-way ANOVA). Bold traces, means ± SEM; faint traces, individual mice. 3v,third ventricle. LED, light-emittingdiode. Scale bars in (A), (F), and (K), 500 mm.

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of PVN-projecting TNSST neurons projected tothe BNST, and the majority (67.8%) of BNST-projecting TNSST neurons projected to the PVN(Fig. 3B). Using a similar method, we found that36.1% of CeA-projecting TNSST neurons projectedto the BNST, and 45.7% of BNST-projecting TNSSTneurons projected to the CeA; 7.4% of BNST-projecting TNSST neurons projected to the PAG,

and 30.5% of PAG-projecting TNSST neuronsprojected to the BNST; and, lastly, 13.8% of CeA-projecting TNSST neurons projected to the PAG,and 29.6% of PAG-projecting TNSST neuronsprojected to the CeA (Fig. 3, C to E, and fig. S11).Therefore, in contrast to AgRP neurons, differentsubpopulations of which project to differentbrain regions in a one-to-one pattern (29), the

TNSST neurons showed a one-to-many projectionpattern.We then optogenetically stimulated TNSST

axon terminals in different brain regions andexamined their roles in feeding regulation. Stim-ulating the TNSST terminals in the PVN andBNST strongly promoted feeding, but stimulat-ing the terminals in the CeA and PAG had no

Luo et al., Science 361, 76–81 (2018) 6 July 2018 4 of 6

Fig. 3. The TNSSTneurons regulatefeeding through projections to the PVNand BNST. (A) Top left, diagram of asagittal brain section summarizing theaxonal projections of TNSST neurons ofSST-Cre mice injected with AAV-FLEX-GFP.Other panels show GFP-expressing axonterminals in different brain regions. Scalebars, 200 mm. (B to E) Left, diagramsillustrating the injection of retrogradetracers in different brain regions ofSST-Cre::Ai14 mice. Center, red TNSSTneurons took up CTB647 or CTB488injected in different projection sites.Yellow arrows indicate the TNSST neuronspositive for both CTB647 and CTB488.Green arrows indicate the TNSST neuronspositive only for CTB488. Blue arrowsindicate the TNSST neurons positive onlyfor CTB647. Scale bar, 50 mm. Right, Venndiagrams illustrate the quantificationsfrom three mice. (F to I) Top, diagramsshowing optogenetic stimulation of differentaxon terminals of TNSST neurons injectedwith AAV-DIO-ChR2-mCherry. Bottom,bar graphs showing the change in eatingfrequency after optogenetic stimulation(bars, means ± SEM; black lines, individualmice; *P < 0.05, **P < 0.01, pairedtwo-tailed t test).

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effect on feeding (Fig. 3, F to I, and fig. S12). Tobetter understand the neural transmission be-tween TNSST neurons and their downstreamneurons, we performed FISH for SST and Gad1or Gad2, the two major enzymes for producingg-aminobutyric acid (GABA) (30). We found that91.4% of TNSST neurons expressed Gad1, and87.8% expressed Gad2 (Fig. 4, A and B). We fur-ther confirmed by immunohistochemistry thatthe majority of TNSST neurons produced GABA(Fig. 4C). Moreover, no TNSST neurons werepositive for VGlut2 (Fig. 4D), the major gluta-matergic neuron marker in the hypothalamus(25). To directly examine the contribution ofGABA signaling on the effect of activating TNSSTneurons, we locally injected CNO with or with-out the GABAA receptor antagonist bicucullineinto the PVN of SST-Cre mice that had beeninjected with AAV-DIO-hM3D in the TN. Localinjection of CNO into the PVN stimulated feedingsimilarly to i.p. injection of CNO (1.43 ± 0.14 gversus 1.20 ± 0.11 g chow; P = 0.27), but co-injecting bicuculline into the PVN dramaticallyreduced this effect (1.20 ± 0.11 g versus 0.43 ±0.05 g; P < 0.01) (Fig. 4E). Therefore, TNSSTneurons stimulated feeding through the inhibi-tion of downstream neurons, a mechanism sim-ilar to that of AgRP neurons (31, 32). Despite thissimilar effect in promoting feeding, ablatingAgRP neurons leads to severe starvation (33), butwe found that ablating TNSST neurons resultedin reduced body weight gain—an effect that iscomparable to that of ablating lateral hypo-thalamic or zona incerta GABAergic neurons

(28, 34)—suggesting a functional differentia-tion between AgRP and other hypothalamicorexigenic neurons.Our results show that, as in humans (12), the

TN in mice is marked by SST-positive neurons.This fact provided us with genetic access to studythe function of this previously enigmatic brainregion. We found that TNSST neurons are re-quired formaintaining normal bodyweight gain,and TNSST neurons project to various brain re-gions and promote feeding by inhibiting down-streamneurons. Taken together, our results revealan important role of the TN in energy homeosta-sis and point to a previously unknown circuitmechanism of feeding regulation that operatesthrough orexigenic TNSSTneurons (Fig. 4F). Thesefindings promise a better understanding of theappetite and body weight changes.

REFERENCES AND NOTES

1. W. E. Le Gros Clark, J. Beattie, G. Riddoch, N. M. Dott,The Hypothalamus: Morphological, Functional, Clinical andSurgical Aspects (Oliver and Boyd, 1938).

2. D. F. Swaab, Handb. Clin. Neurol. 79, xi–xii (2003).3. H. P. Kremer, R. A. Roos, G. Dingjan, E. Marani, G. T. A. M. Bots,

J. Neuropathol. Exp. Neurol. 49, 371–382 (1990).4. H. P. Kremer, Prog. Brain Res. 93, 249–261 (1992).5. J. A. van de Nes, W. Kamphorst, R. Ravid, D. F. Swaab,

Acta Neuropathol. 96, 129–138 (1998).6. H. Braak, E. Braak, Brain Res. 802, 119–124 (1998).7. H. Braak, E. Braak, Neuropathol. Appl. Neurobiol. 15, 13–26

(1989).8. K. Kovacs, H. L. Sheehan, Fertil. Steril. 37, 83–89 (1982).9. L. E. White, R. F. Hain, Arch. Pathol. 68, 275–281 (1959).10. K. Lewin, D. Mattingly, R. R. Millis, BMJ 2, 629–630

(1972).

11. N. Kamalian, R. E. Keesey, G. M. ZuRhein, Neurology 25, 25–30(1975).

12. H. J. Timmers, D. F. Swaab, J. A. van de Nes, H. P. Kremer,Brain Res. 728, 141–148 (1996).

13. L. Madisen et al., Nat. Neurosci. 13, 133–140 (2010).14. G. J. Morton, T. H. Meek, M. W. Schwartz, Nat. Rev. Neurosci.

15, 367–378 (2014).15. E. Gropp et al., Nat. Neurosci. 8, 1289–1291 (2005).16. X. Zhang, A. N. van den Pol, Nat. Neurosci. 19, 1341–1347

(2016).17. J. N. Campbell et al., Nat. Neurosci. 20, 484–496 (2017).18. T. Sakurai et al., Cell 92, 573–585 (1998).19. B. B. Whiddon, R. D. Palmiter, J. Neurosci. 33, 2009–2016

(2013).20. H. Haas, P. Panula, Nat. Rev. Neurosci. 4, 121–130 (2003).21. M. Tschöp, D. L. Smiley, M. L. Heiman, Nature 407, 908–913

(2000).22. A. M. Aravanis et al., J. Neural Eng. 4, S143–S156 (2007).23. E. Vardy et al., Neuron 86, 936–946 (2015).24. C. F. Yang et al., Cell 153, 896–909 (2013).25. J. H. Jennings, G. Rizzi, A. M. Stamatakis, R. L. Ung,

G. D. Stuber, Science 341, 1517–1521 (2013).26. J. N. Betley et al., Nature 521, 180–185 (2015).27. Y. Chen, Y. C. Lin, C. A. Zimmerman, R. A. Essner, Z. A. Knight,

eLife 5, e18640 (2016).28. X. Zhang, A. N. van den Pol, Science 356, 853–859

(2017).29. J. N. Betley, Z. F. Cao, K. D. Ritola, S. M. Sternson, Cell 155,

1337–1350 (2013).30. M. G. Erlander, N. J. Tillakaratne, S. Feldblum, N. Patel,

A. J. Tobin, Neuron 7, 91–100 (1991).31. D. Atasoy, J. N. Betley, H. H. Su, S. M. Sternson, Nature 488,

172–177 (2012).32. A. S. Garfield et al., Nat. Neurosci. 18, 863–871 (2015).33. S. Luquet, F. A. Perez, T. S. Hnasko, R. D. Palmiter, Science

310, 683–685 (2005).34. J. H. Jennings et al., Cell 160, 516–527 (2015).

ACKNOWLEDGMENTS

We thank M. Stryker for critical reading of the manuscript.Funding: This work was supported by an A*STAR Investigatorshipprovided by the Biomedical Research Counsel (BMRC) of A*STAR

Luo et al., Science 361, 76–81 (2018) 6 July 2018 5 of 6

Fig. 4. The TNSST neurons are GABAergic.FISH for (A) SST and Gad1 or (B) SSTand Gad2 in C57BL/6J mice. Scale bars,200 mm. (C) Immunohistochemical stainingfor GABA in SST-Cre::Ai14 mice. Scalebars, 200 mm. (D) FISH for SST and VGlut2in C57BL/6J mice. Scale bar, 200 mm.(E) SST-Cre mice were injected bilaterallywith AAV-DIO-hM3D-mCherry in the TN,and CNO was injected either i.p. or byinfusion cannula implanted over the PVNwith or without bicuculline (Bic). Thesame animal was subjected to all threedifferent injections on different days,and the amount of food consumption3 hours after drug injection was analyzed(n = 6, **P < 0.01, paired t test). Graybars, means ± SEM; black lines, individualmice. (F) TNSST neurons in the mouseTN can be activated by hunger signals,including ghrelin. Activation of TNSSTneurons stimulates feeding throughinhibiting downstream neurons in thePVN and BNST, a mechanism similar tothe activation of AgRP neurons in thearcuate area.

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(1530700142 to Y.F.), intramural funding from A*STAR BMRC(to W.H.), the National Natural and Science Foundation of China(81771215 to J.H.), and National Medical Research Council CollaborativeResearch Grants (NMRC/CBRG/0099/2015 and NMRC-OFIRG-0037-2017 to K.K. and S.S.). Author contributions: Y.F. conceived of theproject and wrote the manuscript. S.X.L., J.H., Q.L., and Y.F. designedand implemented the study. H.M. performed retrograde tracing andlocal CNO injection. C.-Y.L., K.K., and S.S. contributed electrophysiological

recordings. A.M.-Y.K., Y.L.T., and M.H. performed immunohistochemicalstaining. J.Y.L., and H.L. performed the metabolic chamber study.J.S. helped with in vivo optogenetic stimulation. L.Y.Y. and J.G.performed magnetic resonance imaging. All authors discussed theresults and commented on the manuscript. Competing interests:The authors declare no competing interests. Data and materialsavailability: All data deeded to evaluate the conclusions of the paperare present in the paper and the supplementary materials.

SUPPLEMENTARY MATERIALS

www.sciencemag.org/content/361/6397/76/suppl/DC1Materials and MethodsFigs. S1 to S12References (35–37)

18 November 2017; accepted 8 May 201810.1126/science.aar4983

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Regulation of feeding by somatostatin neurons in the tuberal nucleus

Yu FuJoy Yi Lim, Hongyu Li, Ling Yun Yeow, Jingjing Sun, Miao He, Joanes Grandjean, Sreedharan Sajikumar, Weiping Han and Sarah Xinwei Luo, Ju Huang, Qin Li, Hasan Mohammad, Chun-Yao Lee, Kumar Krishna, Alison Maun-Yeng Kok, Yu Lin Tan,

DOI: 10.1126/science.aar4983 (6397), 76-81.361Science 

, this issue p. 76; see also p. 29Sciencecircuits via projections to other hypothalamic nuclei.systemic metabolic balance. This newly described regulatory center is extensively connected with other feeding control

controlhunger hormone. Loss- and gain-of-function experiments indicated that these cells are necessary and sufficient to -aminobutyric acid) (see the Perspective by Diano). These neurons were activated by food deprivation orγ(GABA,

GABAergic somatostatin neurons in the tuberal nucleus are functionally involved in the regulation of feeding in mice found thatet al.The tuberal nucleus, an area of the hypothalamus, has not been studied in great detail. Luo

Neurons that regulate feeding

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