Supplementary Figure 6: NA2 and NA4-5 neurons display different pathways of development. | Nature Neuroscience

Supplementary Figure 6: NA2 and NA4-5 neurons display different pathways of development.

From: Visceral motor neuron diversity delineates a cellular basis for nipple- and pilo-erection muscle control

Supplementary Figure 6

(a) Triple immunohistochemistry for GFRα3, GFP (recapitulating Ret expression) and TRKA on P3, P5 and P11 SG sections from RetCFP mice. At P3, GFRα3 is expressed in a subset of TRKA+ neurons, while RetCFP and TRKA expression are mutually exclusive. Starting from P5, RetCFP expression is upregulated in a subset of GFRα3+TRKA+ neurons. (b) Triple immunohistochemistry for GFRα2, GFP (recapitulating Ret expression) and TRKA on P3, P5 and P11 SG sections from RetCFP mice. Neither GFRα2 nor RetCFP is expressed in noradrenergic TRKA+ neurons at P3 but GFRα2+RetCFP+ neurons are present at P5 and they increase in numbers at P11. GFRα2+TRKA- neurons are cholinergic neurons. (c) Triple immunohistochemistry for GFRα3, GFP (recapitulating Ret expression) and NPY on P11 SG sections from RetCFP mice. NA2 neurons express RetCFP+, NPY and GFRα3 (inset). (d) Triple immunohistochemistry for GFRα2, NPY and TH on P11 SG sections from RetCFP mice. NA4-5 neurons express GFRα2 and TH but are negative for NPY (inset). (e-f) NPY expression during development. (e) Quantification of NPY+ over TH+ neurons shows that the number of NPY+ neurons does not change between P3 and P11. t(10) = 0.4284, P = 0.6774; unpaired t test, n=6. (f) The percentage of RetCFP+NPY+ neurons increases from P3 to P11. n = 3 per stage, data are presented as mean ± s.e.m.. (g) Illustration summarizing these results. (a-d) are representative images from experiments carried out on at least two animals. Scale bars: 20 μm for all images.

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