Extended Data Fig. 3: Casp3-mediated ablation of RAm Nts neurons.
From: A brainstem circuit for phonation and volume control in mice
a, b, smFISH labeling of Nts in brainstem of NtsCre mice injected in RAm with AAV-FLEX-GFP (a, mock ablation) or AAV-FLEX-Casp3 (b, ablation condition) for RAm Nts neuron ablation experiments (see Fig. 2). Insets, magnified view of Sp5 and RAm regions. Note ablation of RAm Nts+ neurons in Casp3 mice with sparing of Sp5. Scale bars, 200 µm. Inset scale bars, 50 µm. c, Percent of RAm Nts+ neurons remaining following AAV-FLEX-Casp3 injection relative to GFP mock ablation controls. n = 868 total RAm Nts neurons scored in 4 mice per condition. d, e, smFISH of Nts+ subpopulation of Amb motor neurons in GFP (d) and Casp3 (e) conditions above. Red arrowheads, Nts+ Amb motor neurons, marked by expression of Vacht (vesicular acetylcholine transporter). White arrowheads, Nts- Amb motor neurons. Note sparing of Amb Nts+ neurons in Casp3 condition, indicating AAV8 approach was selective for RAm interneurons and avoided nearby motor neurons. Scale bars, 25 µm. All data shown as mean ± S.D.
