Dissection of insular cortex layer 5 reveals two sublayers with opposing modulatory roles in appetitive drinking behavior

Isocortex MBA:315 projects to CEAm (Central amygdalar nucleus, medial part) MBA:559 · Mouse

CTB injections mostly covering the ipsilateral CeM labeled DgI cells chiefly in L5b and L2/3, with few labeled cells in L5a.

Fig. S3A, S3C

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Isocortex MBA:315 projects to CEAm (Central amygdalar nucleus, medial part) MBA:559 · Mouse

GFP-labeled axons from the L5b population formed dense arbors in the ipsilateral CeM.

Fig. 3I-J; Fig. S4B

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Isocortex MBA:315 projects to VP (Ventral posterior complex of the thalamus) MBA:709 · Mouse

GFP-labeled insular axon arbors were observed in the parvicellular ventral posterior thalamus.

Fig. 1F

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Isocortex MBA:315 projects to BMA (Basomedial amygdalar nucleus) MBA:319 · Mouse

GFP-labeled axons from the L5b population formed dense arbors in the ipsilateral basomedial amygdala.

Fig. 3I

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Isocortex MBA:315 projects to CEA (Central amygdalar nucleus) MBA:536 · Mouse

Intersectional Cre-dependent GFP labeling of L5a cells, using retrograde Cre from the contralateral CeA, revealed axon arbors confined to the RGS14-positive CeL/CeC on both sides.

Fig. 3A-E; Fig. S4A

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Isocortex MBA:315 projects to BMA (Basomedial amygdalar nucleus) MBA:319 · Mouse

GFP-labeled insular axon arbors were present in the basomedial amygdala after anterograde AAV labeling.

Fig. 1B

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Isocortex MBA:315 projects to PSTN (Parasubthalamic nucleus) MBA:364 · Mouse

Cre-dependent GFP labeling of L5b cells, using retrograde Cre from the PSTh, showed dense axon arbors in the ipsilateral PSTh.

Fig. 3F-H

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Isocortex MBA:315 projects to CEA (Central amygdalar nucleus) MBA:536 · Mouse

After AAV-GFP injection into the middle-posterior insula, dense GFP axon arbors appeared in the central amygdala of both hemispheres. Contralateral arbors were confined to the PKC-delta-positive CeL/CeC.

Fig. 1B-D

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Isocortex MBA:315 projects to MD (Mediodorsal nucleus of thalamus) MBA:362 · Mouse

Anterogradely labeled insular axon arbors were found in the mediodorsal thalamic nucleus.

Fig. 1B

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Isocortex MBA:315 projects to CEA (Central amygdalar nucleus) MBA:536 · Mouse

CTB injected into the contralateral CeA labeled DgI cells mainly in L5a, most of which were NECAB1-positive. CTB in the ipsilateral CeL/CeC labeled cells in L5a and L2/3, some of them double-labeled from the contralateral CeA.

Fig. 2B-G; Fig. S3A-B

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Isocortex MBA:315 projects to BST (Bed nuclei of the stria terminalis) MBA:351 · Mouse

GFP-labeled axons from the L5a population reached the BNST on both sides, limited to the oval (STLD) and juxtacapsular (STLJ) subdivisions.

Fig. S4C

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Isocortex MBA:315 projects to CEAm (Central amygdalar nucleus, medial part) MBA:559 · Mouse

Ipsilateral GFP-labeled arbors from the L5a population stayed within the CeL/CeC and did not extend into the CeM.

Fig. 3E

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Isocortex MBA:315 projects to RE (Nucleus of reuniens) MBA:181 · Mouse

Anterogradely labeled insular axon arbors were found in the nucleus reuniens.

Fig. 1B

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Isocortex MBA:315 projects to ACB (Nucleus accumbens) MBA:56 · Mouse

Labeled insular axons were also detected in the nucleus accumbens.

Fig. S1B-E

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Isocortex MBA:315 projects to NTS (Nucleus of the solitary tract) MBA:651 · Mouse

Labeled insular axons were also detected in the nucleus of the solitary tract.

Fig. S1B-E

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Isocortex MBA:315 projects to PVT (Paraventricular nucleus of the thalamus) MBA:149 · Mouse

Anterogradely labeled insular axon arbors were found in the paraventricular thalamic nucleus.

Fig. 1B

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Isocortex MBA:315 projects to CEA (Central amygdalar nucleus) MBA:536 · Mouse

Only sparse L5b-derived GFP terminals appeared in the CeL/CeC, in contrast to the dense CeM innervation.

Fig. 3I-J

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Isocortex MBA:315 projects to PAG (Periaqueductal gray) MBA:795 · Mouse

Labeled insular axons were also detected in the periaqueductal gray.

Fig. S1B-E

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Isocortex MBA:315 projects to CEAm (Central amygdalar nucleus, medial part) MBA:559 · Mouse

The ipsilateral insular projection to the central amygdala covered the CeM next to the CeL/CeC. The contralateral projection did not reach the CeM.

Fig. 1C

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Isocortex MBA:315 projects to PB (Parabrachial nucleus) MBA:867 · Mouse

Labeled insular axons were also detected in the parabrachial nucleus.

Fig. S1B-E

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Isocortex MBA:315 projects to PSTN (Parasubthalamic nucleus) MBA:364 · Mouse

Dense insular GFP axon arbors filled the ipsilateral parasubthalamic nucleus. Few labeled fibers reached the contralateral side.

Fig. 1F

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Isocortex MBA:315 projects to BST (Bed nuclei of the stria terminalis) MBA:351 · Mouse

Insular GFP axons innervated the lateral BNST. The ipsilateral side received axons in STLD, STLP and STLV, while the contralateral side received them only in STLD.

Fig. 1E

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Isocortex MBA:315 projects to PSTN (Parasubthalamic nucleus) MBA:364 · Mouse

CTB injected into the ipsilateral posterior lateral hypothalamus including the PSTh labeled DgI cells mostly in L5b, nearly all CTIP2-positive. None of these cells were double-labeled with CTB from the contralateral CeA.

Fig. 2B-H

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Isocortex MBA:315 projects to BST (Bed nuclei of the stria terminalis) MBA:351 · Mouse

L5b-derived GFP axons in the ipsilateral BNST were found in STLP and STLV but not in STLD/STLJ. No such axons were seen contralaterally.

Fig. S4D

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