A nose-to-brain circuit underlies anxiety regulation by nasal afferent frequency in mice

PERI (Perirhinal area) MBA:922 projects to BLAp (Basolateral amygdalar nucleus, posterior part) MBA:311 · Mouse

Flp-dependent EGFP expressed in perirhinal parvalbumin interneurons of PV-FlpO mice revealed long-range axons with dense terminals in the posterior basolateral amygdala.

Fig. 5A

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MOBopl (Main olfactory bulb, outer plexiform layer) MBA:244 projects to PERI (Perirhinal area) MBA:922 · Mouse

After retrograde tracer injection in the perirhinal cortex, labelled bulbar somata showed essentially no overlap with cholecystokinin immunostaining that marks tufted cells, arguing that tufted cells do not provide this projection.

Fig. 4C

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MOB (Main olfactory bulb) MBA:507 projects to ECT (Ectorhinal area) MBA:895 · Mouse

The ectorhinal cortex was among the top five limbic-cortical regions containing labelled axons after anterograde tracer injection into the olfactory bulb.

SI Appendix, Fig. S6B

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PERI (Perirhinal area) MBA:922 projects to MS (Medial septal nucleus) MBA:564 · Mouse

Anterograde tracing from perirhinal parvalbumin interneurons showed axonal labelling in the medial septum.

SI Appendix, Fig. S14A

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MOB (Main olfactory bulb) MBA:507 projects to PERI (Perirhinal area) MBA:922 · Mouse

Injection of an anterograde AAV expressing EGFP into the olfactory bulb produced labelled fibres in the anterior perirhinal cortex, which ranked among the densest limbic-cortical targets by cell-density quantification. Fibre terminals in PRC colocalized more densely with parvalbumin-positive than with somatostatin-positive interneurons.

SI Appendix, Fig. S6 A–C; SI Appendix, Fig. S9 C and D

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MOB (Main olfactory bulb) MBA:507 projects to AIp (Agranular insular area, posterior part) MBA:111 · Mouse

Mapping of fluorescent fibres after olfactory bulb anterograde tracer injection showed labelling confined to the perirhinal cortex, with no extension into the neighbouring posterior agranular insular area.

SI Appendix, Fig. S6D

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MOBmi (Main olfactory bulb, mitral layer) MBA:236 projects to PERI (Perirhinal area) MBA:922 · Mouse

Retrograde AAV11-hSyn-mCherry injected into the perirhinal cortex labelled numerous somata located in the mitral cell layer of the olfactory bulb, identifying mitral cells as the source of the bulbar input to PRC.

Fig. 4 A–C

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MOB (Main olfactory bulb) MBA:507 projects to ENTl (Entorhinal area, lateral part) MBA:918 · Mouse

Anterograde viral tracing from the olfactory bulb revealed labelled projections in the lateral entorhinal cortex, one of the five main limbic-cortical targets identified.

SI Appendix, Fig. S6 A and B

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MOB (Main olfactory bulb) MBA:507 projects to AIp (Agranular insular area, posterior part) MBA:111 · Mouse

Injection of the retrograde tracer AAV11 into the posterior agranular insular area produced no retrogradely labelled cells in the olfactory bulb, indicating this insular area receives no direct bulbar input.

SI Appendix, Fig. S6 E and F

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PERI (Perirhinal area) MBA:922 projects to NDB (Diagonal band nucleus) MBA:596 · Mouse

Anterograde tracing from perirhinal parvalbumin interneurons showed axonal labelling in the vertical limb of the diagonal band of Broca.

SI Appendix, Fig. S14A

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MOB (Main olfactory bulb) MBA:507 projects to PIR (Piriform area) MBA:961 · Mouse

Anterograde EGFP tracing from the olfactory bulb labelled dense projections in both the anterior and posterior piriform cortex, which together with PRC received the highest input density among limbic cortical areas.

SI Appendix, Fig. S6 A–C

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PERI (Perirhinal area) MBA:922 synapses onto BLAp (Basolateral amygdalar nucleus, posterior part) MBA:311 · Mouse

Anterograde transsynaptic WGA-iCre expressed in perirhinal parvalbumin neurons drove Cre-dependent EGFP in postsynaptic pBLA cells; almost all of these labelled pBLA neurons were glutamatergic (Vglut1- or Vglut2-positive).

SI Appendix, Fig. S13 A and B

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CL:0000207 functionally connects to PERI (Perirhinal area) MBA:922 · Mouse

Optogenetic activation of channelrhodopsin-expressing olfactory sensory neurons in the nasal cavity changed perirhinal local field potential high-gamma power in a frequency-dependent way (increase at 2 Hz, decrease at 7 Hz) and, by fibre photometry, produced corresponding increases or decreases in calcium activity of perirhinal parvalbumin interneurons.

Fig. 2 A–C; Fig. 3 C–F

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PERI (Perirhinal area) MBA:922 projects to BLAp (Basolateral amygdalar nucleus, posterior part) MBA:311 · Mouse

Retrograde AAV11-hSyn-fDIO-mCherry (and separately retroAAV-hSyn-Cre) injected in the posterior basolateral amygdala of PV-FlpO mice labelled parvalbumin-positive neurons in the perirhinal cortex, confirming a PRC PV projection to pBLA.

Fig. 5B and 5I

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PERI (Perirhinal area) MBA:922 projects to DG (Dentate gyrus) MBA:726 · Mouse

Anatomical mapping of perirhinal parvalbumin neuron axons revealed robust projections to the dentate gyrus.

SI Appendix, Fig. S14A

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MOB (Main olfactory bulb) MBA:507 synapses onto PERI (Perirhinal area) MBA:922 · Mouse

Anterograde transsynaptic ScAAV2/1-hSyn-Cre delivered to the olfactory bulb of PV-FlpO mice drove Cre/Flp-dependent reporter and actuator expression in parvalbumin-positive perirhinal neurons, and bulbar anterograde label colocalized with retrogradely identified PV cells in PRC, showing that bulbar axons contact PRC PV interneurons.

Fig. 4E; Fig. 5B

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PERI (Perirhinal area) MBA:922 projects to LS (Lateral septal nucleus) MBA:242 · Mouse

Flp-dependent anterograde labelling of perirhinal parvalbumin neurons showed robust axonal projections to the lateral septum.

SI Appendix, Fig. S14A

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