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. 2021 May 21;12:3020. doi: 10.1038/s41467-021-23294-7

Fig. 5. Neuronal cytoarchitectures are reproducibly patterned in close relationship to the gut tube.

Fig. 5

a Change in diameter of spinal cord (SC) compartment from days 13 to 22 (n = 47 EMLOs measured per line). b Change in TUJ1 density by normalized fluorescence of Z-stack volumes between days 13, 16, and 22 (n = 3 EMLOs per time point per line). c TUJ1 (red) and GATA4 (cyan) IF of F3.5.2 and H3.3.1 day 16 EMLOs. Peripheral ganglion formation (dotted circle) and neuronal bottleneck (red arrows) are appreciated. ImageJ inverted LUT (TUJ1) was used for black and white images. d Multi-dimensional visualization of gut tube-neuron interaction in H3.3.1 day 16 EMLO. Sagittal and transverse planes of gut tube subsumed by neural processes (left); end-on view of neuronal tunnel (TUJ1+, top) about gut tube (DAPI, bottom). e TUJ1/GATA4 of H3.3.1 day 22 EMLO. Maximally projected Z-stack (left, Z-total) is shown with two Z-slices (Z1, Z2). SC and ME labeled for orientation. Peripheral ganglia form in close proximity to gut tube anterior intestinal portal-like (AIP) region. Z2 inset depicts 3D undulating GATA4 exterior with interpenetrating neurons. High magnification TUJ1 reconstruction shown (right). For IF, GATA4 and TUJ1 staining was performed in each of the N = 11 H3.3.1 EMLO formation experiments with similar results c–e. Individual scale bars provided. Data reported as (mean ±s.e.m.).