Abstract
The lateral hypothalamus is a major integrative hub with a complex architecture characterized by intricate and overlapping cellular populations expressing a large variety of neuro-mediators. In rats, the subfornical lateral hypothalamus (LHsf) was identified as a discrete area with very specific outputs, receiving a strong input from the nucleus incertus, and involved in defensive and foraging behaviors. We identified in the mouse lateral hypothalamus a discrete subfornical region where a conspicuous cluster of neurons express the mu opioid receptor. We thus examined the inputs and outputs of this LHsf region in mice using retrograde tracing with the cholera toxin B subunit and anterograde tracing with biotin dextran amine, respectively. We identified a connectivity profile largely similar, although not identical, to what has been described in rats. Indeed, the mouse LHsf has strong reciprocal connections with the lateral septum, the ventromedial hypothalamic nucleus and the dorsal pre-mammillary nucleus, in addition to a dense output to the lateral habenula. However, the light input from the nucleus incertus and the moderate bidirectional connectivity with nucleus accumbens are specific to the mouse LHsf. A preliminary neurochemical study showed that LHsf neurons expressing mu opioid receptors also co-express calcitonin gene-related peptide or somatostatin and that the reciprocal connection between the LHsf and the lateral septum may be functionally modulated by enkephalins acting on mu opioid receptors. These results suggest that the mouse LHsf may be hodologically and functionally comparable to its rat counterpart, but more atypical connections also suggest a role in consummatory behaviors.
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The datasets generated and analyzed during the current study are available from the corresponding author on reasonable request.
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Abbreviations
- 3N:
-
Oculomotor N
- 3V:
-
3Rd ventricle
- A24a:
-
Anterior cingulate area 24a
- A24b:
-
Anterior cingulate area 24b
- A25:
-
Anterior cingulate area 25
- A32:
-
Anterior cingulate area 32
- AAA:
-
Anterior amygdaloid area
- an:
-
Anterior commissure
- aca:
-
Anterior commissure, anterior limb
- Acb:
-
N accumbens
- AcbC:
-
N accumbens, core
- AcbSh:
-
N accumbens, shell
- ACo:
-
Anterior cortical amygdaloid N
- AD:
-
Anterodorsal thalamic N
- AH:
-
Anterior hypothalamic area
- AHiA:
-
Amygdalohippocampal area, anterior part
- AI:
-
Agranular insular cortex
- AOM:
-
Anterior olfactory N, medial part
- Aq:
-
Cerebral aqueduct
- Arc:
-
Arcuate hypothalamic N
- ATg:
-
Anterior tegmental N
- Bar:
-
Barrington’s N
- BDA:
-
Biotin dextran amine
- BLA:
-
Basolateral amygdaloid N, anterior
- BLP:
-
Basolateral amygdaloid N, posterior
- BMA:
-
Basomedial amygdaloid N, anterior
- BMP:
-
Basomedial amygdaloid N, posterior
- BST:
-
Bed N of the stria terminalis
- BSTIA:
-
BST, intra-amygdaloid
- BSTLD:
-
Lateral BST, dorsal
- BSTLP:
-
Lateral BST, posterior
- BSTLV:
-
Lateral BST, ventral
- BSTMA:
-
Medial BST, anterior part
- BSTMP:
-
Medial BST, posterior
- BSTMPI:
-
Medial BST, postero-intermediate
- BSTMPL:
-
Medial BST, posterolateral
- BSTMPM:
-
Medial BST, posteromedial part
- BSTMV:
-
Medial BST, ventral
- cb:
-
Cerebellum
- cc:
-
Corpus callosum
- CeC:
-
Central amygdaloid N, capsular
- CeL:
-
Central amygdaloid N, lateral
- CeM:
-
Central amygdaloid N, medial
- CGA:
-
Central gray, alpha part
- CGB:
-
Central gray, beta part
- CGRP:
-
Calcitonin gene-related peptide
- CM:
-
Central medial thalamic N
- CnF:
-
Cuneiform N
- cp:
-
Cerebral peduncle
- CPu:
-
Caudate putamen
- CTB:
-
Cholera toxin B subunit
- DEn:
-
Dorsal endopiriform N
- DG:
-
Dentate gyrus
- Dk:
-
N of Darkschewitsch
- DLL:
-
Dorsal N of the lateral lemniscus
- dlPAG:
-
Dorsolateral periaqueductal gray
- DM:
-
Dorsomedial hypothalamic N
- dmPAG:
-
Dorsomedial periaqueductal gray
- DR:
-
Dorsal raphe N
- DRC:
-
Dorsal raphe N, caudal
- DRD:
-
Dorsal raphe N, dorsal
- DRL:
-
Dorsal raphe N, lateral
- DRV:
-
Dorsal raphe N, ventral
- DTg:
-
Dorsal tegmental N
- DTT:
-
Dorsal tenia tecta
- EA:
-
Extended amygdala
- f:
-
Fornix
- fmi:
-
Forceps minor of the corpus callosum
- fr:
-
Fasciculus retroflexus
- hbc:
-
Habenular commissure
- HDB:
-
N of the horizontal limb of the diagonal band
- I:
-
Intercalated nuclei of the amygdala
- Im:
-
Intercalated nucleus of the amygdala, main part
- ic:
-
Internal capsule
- IC:
-
Inferior colliculus
- ICjM:
-
Island of Calleja, major island
- IF:
-
Interfascicular N
- IPN:
-
Interpeduncular N
- IsRt:
-
Isthmic reticular formation
- JPLH:
-
Juxtaparaventricular lateral hypothalamus
- La:
-
Lateral amygdaloid N
- LaDL:
-
Lateral amygdaloid N, dorsolateral part
- LaV:
-
Lateral amygdaloid N, ventral part
- LA:
-
Lateroanterior hypothalamic N
- LC:
-
Locus coeruleus
- LDTg:
-
Laterodorsal tegmental N
- LH:
-
Lateral hypothalamic area
- LHAa:
-
Lateral hypothalamic area, anterior region (Rat)
- LHAd:
-
Lateral hypothalamic area, dorsal region (Rat)
- LHAjv:
-
Lateral hypothalamic area, juxtaventromedial region (Rat)
- LHAsfa:
-
Lateral hypothalamic area, subfornical region, anterior zone (Rat)
- LHAsfp:
-
Lateral hypothalamic area, subfornical region, posterior zone (Rat)
- LHb:
-
Lateral habenular N
- LHbL:
-
Lateral habenular N, lateral part
- LHbM:
-
Lateral habenular N, medial part
- LHsf:
-
Lateral hypothalamus, subfornical region (mouse)
- ll:
-
Lateral lemniscus
- lPAG:
-
Lateral periaqueductal gray
- LPBC:
-
Lateral parabrachial N, central
- LPBD:
-
Lateral parabrachial N, dorsal
- LPBE:
-
Lateral parabrachial N, external
- LPBI:
-
Lateral parabrachial N, internal
- LPBS:
-
Lateral parabrachial N, superior
- LPBV:
-
Lateral parabrachial N, ventral
- LPO:
-
Lateral preoptic area
- LS:
-
Lateral septum
- LSD:
-
Lateral septal N, dorsal
- LSI:
-
Lateral septal N, intermediate
- LSV:
-
Lateral septal N, ventral
- LV:
-
Lateral ventricle
- MA3:
-
Medial accessory oculomotor N
- MD:
-
Mediodorsal thalamic N
- MeA:
-
Medial amygdaloid N
- Me5:
-
Mesencephalic trigeminal N and tract
- MeAD:
-
Medial amygdaloid N, anterodorsal
- MeAV:
-
Medial amygdaloid N, anteroventral
- MePD:
-
Medial amygdaloid N, posterodorsal
- MePV:
-
Medial amygdaloid N, posteroventral
- MHb:
-
Medial habenular N
- MiTg:
-
Microcellular tegmental N
- ml:
-
Medial lemniscus
- mlf:
-
Medial longitudinal fasciculus
- ML:
-
Medial mammillary N, lateral
- MM:
-
Medial mammillary N, medial
- MnPO:
-
Median preoptic N
- MnR:
-
Median raphe N
- MO:
-
Medial orbital cortex
- MPA:
-
Medial preoptic area
- MPB:
-
Medial parabrachial N
- MPL:
-
Medial paralemniscal N
- MPO:
-
Medial preoptic N
- mRt:
-
Mesencephalic reticular formation
- MS:
-
Medial septal N
- mt:
-
Mammillothalamic tract
- MT:
-
Medial terminal N
- MTu:
-
Medial tuberal hypothalamic N
- N:
-
Nucleus
- NI:
-
N Incertus
- ot:
-
Optic tract
- Pa:
-
Paraventricular hypothalamic N
- PAG:
-
Periaqueductal gray
- PBP:
-
Parabrachial pigmented N of the VTA
- pc:
-
Posterior commissure
- PDR:
-
Posterodorsal raphe N
- PH:
-
Posterior hypothalamic N
- PIF:
-
Parainterfascicular N of the VTA
- PIL:
-
Posterior intralaminar thalamic N
- Pir:
-
Piriform cortex
- PLCo:
-
Posterolateral cortical amygdaloid area
- pm:
-
Principal mammillary tract
- PMCo:
-
Posteromedial cortical amygdaloid area PMD: premammillary N, dorsal part
- PMnR:
-
Paramedian raphe N
- PMV:
-
Premammillary N, ventral part
- PN:
-
Paranigral N of the VTA
- PoT:
-
Posterior thalamic nuclear group, triangular
- PR:
-
Prerubral field
- PrC:
-
Precommissural N
- PT:
-
Paratenial thalamic N
- PTg:
-
Pedunculotegmental N
- PV:
-
Paraventricular thalamic N
- PVA:
-
Paraventricular thalamic N, anterior
- RCh:
-
Retrochiasmatic area
- RChL:
-
Retrochiasmatic area, lateral
- Re:
-
Reuniens thalamic N
- RLi:
-
Rostral linear N
- RM:
-
Retromammillary N
- RMC:
-
Red N, magnocellular part
- RMM:
-
Retromammillary N, medial
- rPAG:
-
Rostral periaqueductal gray
- RPC:
-
Red N, parvicellular part
- Rt:
-
Reticular thalamic N
- SCh:
-
Suprachiasmatic N
- scp:
-
Superior cerebellar peduncle
- Shi:
-
Septohippocampal N
- Shy:
-
Septohypothalamic N
- sm:
-
Stria medullaris
- SNC:
-
Substantia nigra, compact part
- SNR:
-
Substantia nigra, reticular part
- SO:
-
Supraoptic N
- sox:
-
Supraoptic decussation
- SST:
-
Somatostatin
- st:
-
Stria terminalis
- StHy:
-
Striohypothalamic N
- Su3:
-
Supraoculomotor periaqueductal gray
- Su3C:
-
Supraoculomotor cap
- Sub:
-
Submedius thalamic N
- VDB:
-
N of the vertical limb of the diagonal band
- vlPAG:
-
Ventrolateral periaqueductal gray
- VMH:
-
Ventromedial hypothalamic N
- VMHC:
-
Ventromedial hypothalamic N, central part
- VMHDM:
-
Ventromedial hypothalamic N, dorsomedial part
- VMHVL:
-
Ventromedial hypothalamic N, ventrolateral part
- VMPO:
-
Ventromedial preoptic N
- VO:
-
Ventral orbital cortex
- VP:
-
Ventral pallidum
- vsc:
-
Ventral spinocerebellar tract
- VTA:
-
Ventral tegmental area
- VTAR:
-
Ventral tegmental area, rostral part
- VTg:
-
Ventral tegmental N
- xscp:
-
Decussation of the superior cerebellar peduncle
- ZI:
-
Zona incerta
References
Ardianto C, Yonemochi N, Yamamoto S, Yang L, Takenoya F, Shioda S, Nagase H, Ikeda H, Kamei J (2016) Opioid systems in the lateral hypothalamus regulate feeding behavior through orexin and GABA neurons. Neuroscience 320:183–193. https://doi.org/10.1016/j.neuroscience.2016.02.002
Baldo BA, Gual-Bonilla L, Sijapati K, Daniel RA, Landry CF, Kelley AE (2004) Activation of a subpopulation of orexin/hypocretin-containing hypothalamic neurons by GABAA receptor-mediated inhibition of the nucleus accumbens shell, but not by exposure to a novel environment. Eur J Neurosci 19(2):376–386
Berthoud HR, Munzberg H (2011) The lateral hypothalamus as integrator of metabolic and environmental needs: from electrical self-stimulation to opto-genetics. Physiol Behav 104(1):29–39. https://doi.org/10.1016/j.physbeh.2011.04.051
Bester H, Besson JM, Bernard JF (1997) Organization of efferent projections from the parabrachial area to the hypothalamus: a Phaseolus vulgaris-leucoagglutinin study in the rat. J Comp Neurol 383(3):245–281
Bodnar RJ (2019) Endogenous opioid modulation of food intake and body weight: implications for opioid influences upon motivation and addiction. Peptides 116:42–62. https://doi.org/10.1016/j.peptides.2019.04.008
Bonnavion P, Mickelsen LE, Fujita A, de Lecea L, Jackson AC (2016) Hubs and spokes of the lateral hypothalamus: cell types, circuits and behaviour. J Physiol 594(22):6443–6462. https://doi.org/10.1113/JP271946
Canteras NS (2002) The medial hypothalamic defensive system: hodological organization and functional implications. Pharmacol Biochem Behav 71(3):481–491. https://doi.org/10.1016/s0091-3057(01)00685-2
Canteras NS, Simerly RB, Swanson LW (1995) Organization of projections from the medial nucleus of the amygdala: a PHAL study in the rat. J Comp Neurol 360(2):213–245. https://doi.org/10.1002/cne.903600203
Comoli E, Ribeiro-Barbosa ER, Canteras NS (2000) Afferent connections of the dorsal premammillary nucleus. J Comp Neurol 423(1):83–98. https://doi.org/10.1002/1096-9861(20000717)423:1%3c83::aid-cne7%3e3.0.co;2-3
Dobolyi A, Irwin S, Makara G, Usdin TB, Palkovits M (2005) Calcitonin gene-related peptide-containing pathways in the rat forebrain. J Comp Neurol 489(1):92–119. https://doi.org/10.1002/cne.20618
Dobolyi A, Palkovits M, Usdin TB (2010) The TIP39-PTH2 receptor system: unique peptidergic cell groups in the brainstem and their interactions with central regulatory mechanisms. Prog Neurobiol 90(1):29–59. https://doi.org/10.1016/j.pneurobio.2009.10.017
Dong HW, Swanson LW (2004) Projections from bed nuclei of the stria terminalis, posterior division: implications for cerebral hemisphere regulation of defensive and reproductive behaviors. J Comp Neurol 471(4):396–433. https://doi.org/10.1002/cne.20002
Erbs E, Faget L, Scherrer G, Matifas A, Filliol D, Vonesch JL, Koch M, Kessler P, Hentsch D, Birling MC, Koutsourakis M, Vasseur L, Veinante P, Kieffer BL, Massotte D (2015) A mu-delta opioid receptor brain atlas reveals neuronal co-occurrence in subcortical networks. Brain Struct Funct 220(2):677–702. https://doi.org/10.1007/s00429-014-0717-9
Fillinger C, Yalcin I, Barrot M, Veinante P (2018) Efferents of anterior cingulate areas 24a and 24b and midcingulate areas 24a’ and 24b’ in the mouse. Brain Struct Funct 223(4):1747–1778. https://doi.org/10.1007/s00429-017-1585-x
Georgescu D, Zachariou V, Barrot M, Mieda M, Willie JT, Eisch AJ, Yanagisawa M, Nestler EJ, DiLeone RJ (2003) Involvement of the lateral hypothalamic peptide orexin in morphine dependence and withdrawal. J Neurosci 23(8):3106–3111
Goto M, Swanson LW, Canteras NS (2001) Connections of the nucleus incertus. J Comp Neurol 438(1):86–122
Goto M, Canteras NS, Burns G, Swanson LW (2005) Projections from the subfornical region of the lateral hypothalamic area. J Comp Neurol 493(3):412–438. https://doi.org/10.1002/cne.20764
Groenewegen HJ, Ahlenius S, Haber SN, Kowall NW, Nauta WJ (1986) Cytoarchitecture, fiber connections, and some histochemical aspects of the interpeduncular nucleus in the rat. J Comp Neurol 249(1):65–102. https://doi.org/10.1002/cne.902490107
Hahn JD, Swanson LW (2010) Distinct patterns of neuronal inputs and outputs of the juxtaparaventricular and suprafornical regions of the lateral hypothalamic area in the male rat. Brain Res Rev 64(1):14–103. https://doi.org/10.1016/j.brainresrev.2010.02.002
Hahn JD, Swanson LW (2015) Connections of the juxtaventromedial region of the lateral hypothalamic area in the male rat. Front Syst Neurosci 9:66. https://doi.org/10.3389/fnsys.2015.00066
Jennings JH, Ung RL, Resendez SL, Stamatakis AM, Taylor JG, Huang J, Veleta K, Kantak PA, Aita M, Shilling-Scrivo K, Ramakrishnan C, Deisseroth K, Otte S, Stuber GD (2015) Visualizing hypothalamic network dynamics for appetitive and consummatory behaviors. Cell 160(3):516–527. https://doi.org/10.1016/j.cell.2014.12.026
Kincheski GC, Mota-Ortiz SR, Pavesi E, Canteras NS, Carobrez AP (2012) The dorsolateral periaqueductal gray and its role in mediating fear learning to life threatening events. PLoS ONE 7(11):e50361. https://doi.org/10.1371/journal.pone.0050361
Lantos TA, Gorcs TJ, Palkovits M (1995) Immunohistochemical mapping of neuropeptides in the premamillary region of the hypothalamus in rats. Brain Res Brain Res Rev 20(2):209–249. https://doi.org/10.1016/0165-0173(94)00013-f
Martinez RC, Carvalho-Netto EF, Amaral VC, Nunes-de-Souza RL, Canteras NS (2008) Investigation of the hypothalamic defensive system in the mouse. Behav Brain Res 192(2):185–190. https://doi.org/10.1016/j.bbr.2008.03.042
Mendez IA, Ostlund SB, Maidment NT, Murphy NP (2015) Involvement of endogenous enkephalins and beta-endorphin in feeding and diet-induced obesity. Neuropsychopharmacology 40(9):2103–2112. https://doi.org/10.1038/npp.2015.67
Millhouse OE (1973) The organization of the ventromedial hypothalamic nucleus. Brain Res 55(1):71–87
Paxinos G, Franklin KBJ (2012) Paxinos and Franklin’s the mouse brain in stereotaxic coordinates, 4th edn. Academic Press
Pecina S, Berridge KC (2005) Hedonic hot spot in nucleus accumbens shell: where do mu-opioids cause increased hedonic impact of sweetness? J Neurosci 25(50):11777–11786. https://doi.org/10.1523/JNEUROSCI.2329-05.2005
Petrovich GD, Risold PY, Swanson LW (1996) Organization of projections from the basomedial nucleus of the amygdala: a PHAL study in the rat. J Comp Neurol 374(3):387–420. https://doi.org/10.1002/(SICI)1096-9861(19961021)374:3%3c387::AID-CNE6%3e3.0.CO;2-Y
Reppucci CJ, Petrovich GD (2016) Organization of connections between the amygdala, medial prefrontal cortex, and lateral hypothalamus: a single and double retrograde tracing study in rats. Brain Struct Funct 221(6):2937–2962. https://doi.org/10.1007/s00429-015-1081-0
Reppucci CJ, Petrovich GD (2018) Neural substrates of fear-induced hypophagia in male and female rats. Brain Struct Funct 223(6):2925–2947. https://doi.org/10.1007/s00429-018-1668-3
Risold PY, Swanson LW (1997) Connections of the rat lateral septal complex. Brain Res Brain Res Rev 24(2–3):115–195
Sakanaka M, Magari S (1989) Reassessment of enkephalin (ENK)-containing afferents to the rat lateral septum with reference to the fine structures of septal ENK fibers. Brain Res 479(2):205–216
Sakanaka M, Magari S, Shibasaki T, Inoue N (1989) Co-localization of corticotropin-releasing factor- and enkephalin-like immunoreactivities in nerve cells of the rat hypothalamus and adjacent areas. Brain Res 487(2):357–362
Sukhov RR, Walker LC, Rance NE, Price DL, Young WS 3rd (1995) Opioid precursor gene expression in the human hypothalamus. J Comp Neurol 353(4):604–622. https://doi.org/10.1002/cne.903530410
Swanson LW (2004) Brain maps: structure of the rat brain. A laboratory guide with printed and electronic templates for data, models and schematics, 3rd edn. Elsevier, Amsterdam
Swanson LW, Cowan WM (1979) The connections of the septal region in the rat. J Comp Neurol 186(4):621–655. https://doi.org/10.1002/cne.901860408
Swanson LW, Sanchez-Watts G, Watts AG (2005) Comparison of melanin-concentrating hormone and hypocretin/orexin mRNA expression patterns in a new parceling scheme of the lateral hypothalamic zone. Neurosci Lett 387(2):80–84. https://doi.org/10.1016/j.neulet.2005.06.066
Thompson RH, Swanson LW (2010) Hypothesis-driven structural connectivity analysis supports network over hierarchical model of brain architecture. Proc Natl Acad Sci U S A 107(34):15235–15239. https://doi.org/10.1073/pnas.1009112107
Urstadt KR, Stanley BG (2015) Direct hypothalamic and indirect trans-pallidal, trans-thalamic, or trans-septal control of accumbens signaling and their roles in food intake. Front Syst Neurosci 9:8. https://doi.org/10.3389/fnsys.2015.00008
Vertes RP (2004) Differential projections of the infralimbic and prelimbic cortex in the rat. Synapse 51(1):32–58. https://doi.org/10.1002/syn.10279
Vertes RP, Fortin WJ, Crane AM (1999) Projections of the median raphe nucleus in the rat. J Comp Neurol 407(4):555–582
Vertes RP, Linley SB, Hoover WB (2015) Limbic circuitry of the midline thalamus. Neurosci Biobehav Rev 54:89–107. https://doi.org/10.1016/j.neubiorev.2015.01.014
Williams RG, Dockray GJ (1983) Distribution of enkephalin-related peptides in rat brain: immunohistochemical studies using antisera to met-enkephalin and met-enkephalin Arg6Phe7. Neuroscience 9(3):563–586. https://doi.org/10.1016/0306-4522(83)90175-6
Zahm DS, Parsley KP, Schwartz ZM, Cheng AY (2013) On lateral septum-like characteristics of outputs from the accumbal hedonic “hotspot” of Pecina and Berridge with commentary on the transitional nature of basal forebrain “boundaries.” J Comp Neurol 521(1):50–68. https://doi.org/10.1002/cne.23157
Acknowledgements
We thank the Chronobiotron (UMS3415) for animal housing and animal care, and the imaging platform of INCI (UPS3156) for their assistance.
Funding
This work was supported by the Centre National de la Recherche Scientifique (contract UPR3212), the University of Strasbourg and the NeuroTime Erasmus Mundus Joint Doctorate Program.
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Ugur, M., Doridot, S., la Fleur, S.E. et al. Connections of the mouse subfornical region of the lateral hypothalamus (LHsf). Brain Struct Funct 226, 2431–2458 (2021). https://doi.org/10.1007/s00429-021-02349-x
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DOI: https://doi.org/10.1007/s00429-021-02349-x