Extended Data Figure 3. Amb^Cardiac markers are expressed in other brainstem parasympathetic nuclei.

In situ hybridization data (from Allen Brain Atlas¹⁵) showing expression (dark purple) of the indicated Amb^Cardiac-specific genes (Celf6, Kcna5, Tbx3, Dgkb, from Fig. 1) in parasympathetic neurons across the brainstem. Top row, Amb; middle row, dorsal motor nucleus of the vagus (10N); bottom row, lacrimal and salivatory nuclei (Lac/Sal). Note the genes are expressed in a subset of Amb neurons (top row panels), as predicted by scRNAseq (Fig. 1), but also in parasympathetic neurons of the dorsal motor nucleus of vagus (10N) that innervate thoracic and abdominal viscera (second row panels). Note absence of expression in the adjacent somatic motor neurons of the hypoglossal nucleus (12N). All four genes are also expressed in parasympathetic neurons of the facial nerve controlling the lacrimal and salivary glands (Lac/Sal) (third row panels) but had absent or lower expression in the adjacent facial motor nucleus (7N). Allen Brain Atlas images obtained from available postnatal day 56 sections (Celf6, Kcna5, Dgkb) and embryonic day 18.5 sections (Tbx3). Amb sections are sagittal except for Celf6 (coronal). 10N and Lac/Sal sections are sagittal. Bar of 200 μm applies to all sections.