Awake dynamics and brain-wide direct inputs of hypothalamic MCH and orexin networks

MRN (Midbrain reticular nucleus) MBA:128 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

The midbrain reticular nucleus was a major midbrain source of rabies-labelled inputs to MCH cells.

Fig. 5; Supplementary Fig. 5

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BST (Bed nuclei of the stria terminalis) MBA:351 functionally connects to LHA (Lateral hypothalamic area) MBA:194 · Mouse

Photostimulating ChR2-expressing BST VGAT axons in LH slices evoked fast postsynaptic currents in identified MCH cells that reversed near the chloride equilibrium potential, consistent with direct GABAergic inhibition; mean amplitude about 118 pA.

Fig. 6c,e

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NDB (Diagonal band nucleus) MBA:596 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

The diagonal band nucleus was among the cerebral nuclei contributing most input neurons to MCH cells.

Fig. 5; Supplementary Fig. 5

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ARH (Arcuate hypothalamic nucleus) MBA:223 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Rabies-labelled inputs to MCH cells included proopiomelanocortin-expressing neurons in the arcuate nucleus.

Supplementary Fig. 4c

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SO (Supraoptic nucleus) MBA:390 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

The supraoptic nucleus was a striking hotspot of rabies-labelled monosynaptic input neurons to MCH cells.

Figs 4b, 5; Supplementary Fig. 4

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sAMY (Striatum-like amygdalar nuclei) MBA:278 functionally connects to LHA (Lateral hypothalamic area) MBA:194 · Mouse

Optical activation of ChR2-expressing amygdala VGAT axon terminals produced short-latency currents in MCH neurons that reversed near chloride equilibrium, indicating direct GABAergic inhibition; mean amplitude about 115 pA.

Fig. 6d,e

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LS (Lateral septal nucleus) MBA:242 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Lateral septum was one of the main cerebral-nuclei sources of rabies-labelled inputs to MCH cells.

Figs 4b, 5; Supplementary Fig. 5

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CTX (Cerebral cortex) MBA:688 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Cortical areas contributed a minor share of rabies-labelled monosynaptic inputs to orexin cells.

Fig. 5; Supplementary Fig. 5

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BS (Brain stem) MBA:343 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Brainstem areas (pons, medulla) provided a smaller share of monosynaptic inputs to MCH cells.

Figs 4a, 5

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VTA (Ventral tegmental area) MBA:749 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Among midbrain areas, the VTA provided the most monosynaptic input neurons to MCH cells.

Fig. 5; Supplementary Fig. 5

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BS (Brain stem) MBA:343 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Brainstem areas contributed a smaller fraction of inputs to orexin cells.

Fig. 5; Supplementary Fig. 5

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RL (Rostral linear nucleus raphe) MBA:197 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

The rostral linear raphe nucleus was among the top midbrain sources of input to MCH cells.

Fig. 5; Supplementary Fig. 5

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TU (Tuberal nucleus) MBA:614 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

A high-magnification image shows rabies-labelled direct input neurons to MCH cells in the tuberal nucleus.

Fig. 3e

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MB (Midbrain) MBA:313 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Midbrain areas provided monosynaptic inputs to orexin cells.

Fig. 5; Supplementary Fig. 5

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HY (Hypothalamus) MBA:1097 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Cre-dependent monosynaptic rabies tracing from orexin cells in orexin-Cre mice labelled input neurons across hypothalamic regions.

Fig. 5; Supplementary Figs 4, 5

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BST (Bed nuclei of the stria terminalis) MBA:351 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Clusters of monosynaptic input cells to MCH neurons were found in the BST, the largest source among cerebral nuclei.

Figs 4b, 5; Supplementary Fig. 5

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CEA (Central amygdalar nucleus) MBA:536 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Rabies tracing revealed clusters of input neurons to MCH cells in the amygdala, with example labelled cells shown in the central amygdalar nucleus.

Figs 4b, 5; Supplementary Fig. 5

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CNU (Cerebral nuclei) MBA:623 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Cerebral nuclei were the largest extrahypothalamic source of monosynaptic inputs to orexin cells.

Fig. 5; Supplementary Fig. 5

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PA (Posterior amygdalar nucleus) MBA:780 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

The posterior amygdalar nucleus was one of the main cortical-subplate sources of direct input to MCH cells.

Fig. 5; Supplementary Fig. 5

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PAG (Periaqueductal gray) MBA:795 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

The periaqueductal gray contained many monosynaptic input neurons to MCH cells.

Figs 4b, 5; Supplementary Fig. 5

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MPO (Medial preoptic area) MBA:523 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

After PVH and LH, the medial preoptic area contributed the next largest share of hypothalamic monosynaptic inputs to MCH cells.

Fig. 5; Supplementary Fig. 5

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TH (Thalamus) MBA:549 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Thalamus supplied rabies-labelled inputs to orexin cells, ranked after cerebral nuclei.

Fig. 5; Supplementary Fig. 5

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TH (Thalamus) MBA:549 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Thalamus contributed input neurons to MCH cells, ranking after cerebral nuclei among extrahypothalamic large-scale areas.

Figs 4a, 5

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SUB (Subiculum) MBA:502 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Among cortical areas, the ventral subiculum held the most rabies-labelled input neurons to MCH cells.

Figs 4b, 5; Supplementary Fig. 5

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ACB (Nucleus accumbens) MBA:56 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Nucleus accumbens was among the cerebral nuclei providing the most monosynaptic input neurons to MCH cells.

Fig. 5; Supplementary Fig. 5

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PVH (Paraventricular hypothalamic nucleus) MBA:38 synapses onto LHA (Lateral hypothalamic area) MBA:194 · Mouse

Cre-dependent monosynaptic rabies tracing from MCH cells in Pmch-Cre mice labelled many input neurons in the PVH, one of the two most prominent hotspots, including oxytocin and vasopressin cells.

Figs 4b, 5; Supplementary Figs 4, 5

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