An inhibitory circuit from central amygdala to zona incerta drives pain-related behaviors in mice

CEA (Central amygdalar nucleus) MBA:536 functionally connects to ZI (Zona incerta) MBA:797 · Mouse

Optical stimulation of ChR2-expressing central amygdala terminals also elicited inhibitory postsynaptic currents in 8 of 15 VGAT-negative zona incerta neurons, with paired-pulse ratios indistinguishable from those of VGAT-positive cells.

Figure 2D

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to LHA (Lateral hypothalamic area) MBA:194 · Mouse

Sparse axonal terminals from PKCδ central amygdala neurons were seen in the lateral hypothalamus of all five brains.

Results, Table 1, Figure 1C-D

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to PAG (Periaqueductal gray) MBA:795 · Mouse

Sparse axonal terminals from PKCδ central amygdala neurons were observed in the periaqueductal grey in three of the five brains.

Results, Table 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to BST (Bed nuclei of the stria terminalis) MBA:351 · Mouse

Cre-dependent mCherry or ChrimsonR-tdTomato expressed in PKCδ-positive central amygdala neurons produced labelled axon terminals in the bed nucleus of the stria terminalis, with moderate-to-dense labelling consistently present in all five brains examined.

Figure 1C-D, Table 1, Results (Identification of CeA-PKCδ neuronal efferent targets)

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to PPN (Pedunculopontine nucleus) MBA:1052 · Mouse

Sparse axonal terminal labelling from PKCδ central amygdala neurons was present in the pedunculopontine tegmental nucleus in three of five brains.

Results, Table 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to Isocortex MBA:315 · Mouse

No labelled axon terminals from PKCδ central amygdala neurons were detected in any cortical region in any of the five brains surveyed.

Results (Identification of CeA-PKCδ neuronal efferent targets), Table 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y functionally connects to ZI (Zona incerta) MBA:797 · Mouse

In mice with cre-dependent ChR2 in PKCδ central amygdala neurons, whole-cell recordings from 30 zona incerta neurons across 8 mice yielded no light-evoked postsynaptic currents despite robust terminal labelling, leaving the monosynaptic PKCδ-to-ZI connection unresolved.

Author response (attempted cre-dependent ChR2 recordings)

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to LDT (Laterodorsal tegmental nucleus) MBA:162 · Mouse

Sparse terminal labelling from PKCδ central amygdala neurons was seen in the laterodorsal tegmental nucleus in three of five brains.

Results, Table 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to LC (Locus ceruleus) MBA:147 · Mouse

Sparse terminal labelling from PKCδ central amygdala neurons was found in the locus coeruleus in four of five brains.

Results, Table 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to LPO (Lateral preoptic area) MBA:226 · Mouse

Sparse terminal labelling from PKCδ central amygdala neurons was observed in the lateral preoptic area of all five brains.

Results, Table 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to MRN (Midbrain reticular nucleus) MBA:128 · Mouse

Sparse-to-moderate terminal labelling from PKCδ central amygdala neurons was present in the midbrain reticular formation in four of five brains.

Results, Table 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to GPe (Globus pallidus, external segment) MBA:1022 · Mouse

Sparse axonal terminal labelling from PKCδ central amygdala neurons was found in the globus pallidus of all five brains examined.

Results, Table 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to PB (Parabrachial nucleus) MBA:867 · Mouse

Dense terminal labelling from PKCδ central amygdala neurons was present in the lateral parabrachial nucleus in three of five brains, with sparse labelling in one and no labelling in another.

Results, Table 1, Figure 1C-D

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CEA (Central amygdalar nucleus) MBA:536 functionally connects to ZI (Zona incerta) MBA:797 · Mouse

After ChR2-EYFP expression in central amygdala neurons, blue-light stimulation of their terminals in the zona incerta evoked monosynaptic (TTX/4-AP resistant) inhibitory postsynaptic currents in 12 of 20 VGAT-positive zona incerta neurons; the currents were reduced by the GABA-A antagonist bicuculline and absent or small near the chloride reversal potential.

Figure 2C-F

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to CEA (Central amygdalar nucleus) MBA:536 · Mouse

Labelled terminals from PKCδ neurons were seen locally within the central amygdala itself at moderate-to-dense levels in all five injected brains.

Results, Table 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to SI (Substantia innominata) MBA:342 · Mouse

Anterograde labelling of PKCδ central amygdala neurons revealed sparse-to-moderate terminal labelling in the substantia innominata in all five brains analysed.

Results, Table 1, Figure 1C-D

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to STN (Subthalamic nucleus) MBA:470 · Mouse

Sparse-to-moderate labelling of PKCδ central amygdala terminals was observed in the subthalamic nucleus in three of five injected brains.

Results, Table 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to PSTN (Parasubthalamic nucleus) MBA:364 · Mouse

Sparse-to-moderate terminal labelling from PKCδ central amygdala neurons was found in the parasubthalamic nucleus in four of the five brains analysed.

Results, Table 1, Figure 1C-D

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to VTA (Ventral tegmental area) MBA:749 · Mouse

Sparse-to-moderate terminal labelling from PKCδ central amygdala neurons was seen in the ventral tegmental area in three of the five brains.

Results, Table 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to ZI (Zona incerta) MBA:797 · Mouse

Cholera toxin B conjugated to Alexa 647 injected into the zona incerta of PKCδ-cre::Ai9 mice retrogradely labelled central amygdala neurons, about 19% of which were also PKCδ-tdTomato positive; these double-labelled cells lay in the capsular and lateral subdivisions.

Figure 2A-B, Figure 2—figure supplement 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to ZI (Zona incerta) MBA:797 · Mouse

Viral cre-dependent fluorophore/opsin expression in PKCδ central amygdala neurons produced sparse-to-moderate axon terminal labelling in the zona incerta in four of five brains, concentrated in the middle and ventral sectors between 1.46 and 2.70 mm posterior to bregma.

Figure 1C-D, Figure 1—figure supplement 2, Table 1

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CEAl PKC-delta-expressing neurons nt-g513b58b0y projects to SNr (Substantia nigra, reticular part) MBA:381 · Mouse

Sparse-to-moderate axonal terminal labelling from PKCδ central amygdala neurons was present in the substantia nigra in four of five brains.

Results, Table 1, Figure 1C-D

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CEA (Central amygdalar nucleus) MBA:536 projects to ZI (Zona incerta) MBA:797 · Mouse

Retrograde CTB-647 injected in the zona incerta labelled neurons across all central amygdala subnuclei (capsular, lateral, medial) along the rostro-caudal extent, with the highest densities between 1.06 and 1.70 mm posterior to bregma.

Figure 2—figure supplement 1

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