A retrograde adeno-associated virus for collecting ribosome-bound mRNA from anatomically defined projection neurons

BLA (Basolateral amygdalar nucleus) MBA:295 projects to BST (Bed nuclei of the stria terminalis) MBA:351 · Mouse

After AAV9-Camk2a-TRAP injection into the anterodorsal/anterolateral/oval BNST, EGFP-L10a was expressed in Camk2a-positive basal amygdala neurons, indicating retrograde transduction of basal amygdala neurons innervating the BNST; TRAP of basal amygdala punches recovered mRNA with a higher Camk2a/GFAP ratio in the immunoprecipitate than in the supernatant, consistent with labelling of these Camk2a-positive projection neurons.

Figs. 5A–C, 6; Results, sections 4 and 5

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

BNST injection of the retrogradely transported AAV9-Camk2a-TRAP labelled Camk2a-positive neurons in the lateral hypothalamus, indicating a lateral hypothalamic projection to the BNST.

Figs. 5D–F; Results, section 4

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ENT (Entorhinal area) MBA:909 projects to HPF (Hippocampal formation) MBA:1089 · Mouse

After AAV9-Camk2a-TRAP was injected into the dorsal hippocampus, EGFP-L10a labelled cell bodies appeared in the entorhinal cortex, mainly in layer II with some in layer III and only occasional cells in layers V/VI, consistent with retrograde uptake by axon terminals of entorhinal projection neurons; these labelled cells were Camk2a-positive neurons and never GAD67-positive interneurons or GFAP-positive astrocytes.

Figs. 1G–K, 3A–C; Results, sections 1 and 3

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PVT (Paraventricular nucleus of the thalamus) MBA:149 projects to BST (Bed nuclei of the stria terminalis) MBA:351 · Mouse

BNST injection of AAV9-Camk2a-TRAP produced EGFP-L10a expression restricted to Camk2a-positive neurons in the paraventricular nucleus of the thalamus, consistent with retrograde transduction of thalamic neurons projecting to the BNST.

Figs. 5G–I; Results, section 4

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BLA (Basolateral amygdalar nucleus) MBA:295 projects to HPF (Hippocampal formation) MBA:1089 · Mouse

Injection of the retrogradely transported AAV9-Camk2a-TRAP into the hippocampus produced EGFP-L10a labelled somata in the basal amygdala; nearly all labelled cells were Camk2a-positive neurons, with none in GAD67-positive interneurons or GFAP-positive astrocytes.

Figs. 3D–F; Results, section 3

Made by an AI model reading the paper (claude-opus-5, extract@0.3.0); a second AI model (claude-opus-5) agrees.