Linking emotional valence and anxiety in a mouse insula-amygdala circuit

AI (Agranular insular area) MBA:95 projects to ACB (Nucleus accumbens) MBA:56 · Mouse

Dense eYFP-labelled axons from aIC glutamatergic neurons were seen in the nucleus accumbens core. aIC-BLA collaterals in ipsilateral NAc were comparable in density to the BLA, with dense synaptophysin-labelled contacts.

Fig. 3d-e, 3i-j

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AI (Agranular insular area) MBA:95 functionally connects to BLA (Basolateral amygdalar nucleus) MBA:295 · Mouse

Photostimulating ChETA-expressing insular terminals evoked EPSCs in BLA neurons that partly survived TTX+4AP, showing a monosynaptic input, and were abolished by AP5+NBQX, showing it is glutamatergic. IPSCs were polysynaptic, and the synapses showed short-term depression.

Fig. 4a-g

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AIp (Agranular insular area, posterior part) MBA:111 projects to ACB (Nucleus accumbens) MBA:56 · Mouse

Dense axonal fibers from pIC glutamatergic neurons were seen in the nucleus accumbens core.

Fig. 3e; Supplementary Fig. 3f-g

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AI (Agranular insular area) MBA:95 projects to CEAm (Central amygdalar nucleus, medial part) MBA:559 · Mouse

aIC axons were found in the medial central amygdala, though CeM had the lowest density of aIC axons and of aIC-BLA collaterals among the amygdala nuclei examined.

Fig. 3d, 3i-j; Results, synaptic properties section

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AI (Agranular insular area) MBA:95 functionally connects to CEAm (Central amygdalar nucleus, medial part) MBA:559 · Mouse

Optogenetic activation of insular axons evoked EPSCs in CeM neurons that persisted under TTX+4AP and were blocked by AP5+NBQX, showing direct glutamatergic input. IPSCs disappeared under TTX+4AP, so they were polysynaptic.

Fig. 4a-g

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AIp (Agranular insular area, posterior part) MBA:111 projects to CEAm (Central amygdalar nucleus, medial part) MBA:559 · Mouse

CTB injected into the CeM labelled posterior insula neurons, which were recorded as pIC-CeM neurons.

Fig. 4h-n

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AI (Agranular insular area) MBA:95 projects to CEAm (Central amygdalar nucleus, medial part) MBA:559 · Mouse

CTB injected into the CeM labelled anterior insula neurons, which were recorded as aIC-CeM neurons.

Fig. 4h-n

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AI (Agranular insular area) MBA:95 projects to CEAl (Central amygdalar nucleus, lateral part) MBA:551 · Mouse

Anterograde eYFP labelling from aIC glutamatergic neurons showed a substantial amount of axons in the lateral central amygdala. Collaterals of aIC-BLA neurons in ipsilateral CeL had density similar to the BLA, and synaptophysin-mCherry puncta were dense there.

Fig. 3d-e, 3h-j

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AIp (Agranular insular area, posterior part) MBA:111 projects to BLA (Basolateral amygdalar nucleus) MBA:295 · Mouse

Neurons retrogradely labelled by BLA CTB injections were found in the posterior insula and recorded as pIC-BLA neurons.

Fig. 4h-k

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AIp (Agranular insular area, posterior part) MBA:111 projects to BLA (Basolateral amygdalar nucleus) MBA:295 · Mouse

Anterograde eYFP labelling of posterior insula glutamatergic neurons revealed axonal fibers in the BLA. The authors say this contradicts strictly segregated aIC-BLA and pIC-CeA pathways.

Fig. 3e; Supplementary Fig. 3f

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AI (Agranular insular area) MBA:95 projects to BLA (Basolateral amygdalar nucleus) MBA:295 · Mouse

CTB injected into the BLA labelled aIC neurons, which were patched for intrinsic properties. CAV2-Cre injected into the BLA drove Cre-dependent expression in aIC neurons projecting to the BLA.

Fig. 4h-k; Fig. 5g-h

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AI (Agranular insular area) MBA:95 projects to BLA (Basolateral amygdalar nucleus) MBA:295 · Mouse

With eYFP expressed in aIC glutamatergic neurons, the BLA held the densest axonal labelling of all twelve targets and served as the normalization reference. Collateral mapping of aIC-BLA neurons with synaptophysin-mCherry showed the densest boutons in the ipsilateral BLA, plus substantial axons and boutons in the contralateral BLA.

Fig. 3c-e, 3f-k

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