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Fig. 4. Pds5B^-/- mice display multiple defects in the peripheral nervous system. (A) Summary of carotid nerve and superior cervical ganglion (SCG) abnormalities in Pds5B-deficient mice. SCG AML, SCG delayed migration or abnormal localization; TCN, thin carotid nerve; CNM, carotid nerve missing. ^*P<0.05 and ^**P<0.001 determined by a Z-test for significant differences in incidence between WT and Pds5B homozygotes. (B-G) TOH whole-mount staining of sympathetic neurons of neonate mice. (B,C) Abnormal caudal location of SCG in Pds5B^-/- mice compared to wild-type mice (SCG, arrows). Dashed circle indicates the expected normal location of SCG (C). Note the severe reduction or absence of carotid nerves in Pds5B^-/- (E,G) compared to WT mice (D,F). Carotid nerves are indicated by arrows in D and E, and by two dashed lines in F and G. ey, eye; co, cochlea. Scale bars: 400 µm (B-E); 200 µm (F,G). (H,I) E12.5 enteric neurons in the distal bowel of whole gut demonstrated by TuJ1 immunohistochemistry (H, wild type; I, Pds5B^-/-). ^*, ileocecal junction. Note that the enteric neuron migration wave front did not pass the ileocecal junction in Pds5B^-/- mice (I). Scale bars: 10 µm (H,I). (J,K) TuJ1 staining of P0 paraffin sections of distal colon from wild-type (J) and Pds5B^-/- mice (K), showing the lack of clustered enteric neuron cell bodies in Pds5B^-/- mice. Scale bars: 20 µm. (Insets) High magnification of TuJ1 staining of P0 distal colons. Scale bars: 10 µm. Arrowheads point to the clustered enteric neuron cell bodies. (L,M) Tuj1 whole-mount staining of E18.5 guts from wild-type mice (L) and Pds5B^-/- (M). Wild-type mice show normal formation of enteric plexus with clustered neuronal cell bodies (arrowheads) in the mid-colon, whereas Pds5B^-/- mice lack enteric neuron cell bodies and show only a few thin nerve fibers (arrows). Scale bar: 100 µm. (N) Schematic representation of ENS defects in Pds5B^-/- mice [E12.5: Pds5B^-/- (n=7) and WT (n=9); P0: Pds5B^-/- (n=12) and WT (n=13)]. The scale at the top corresponds to the percentage of the respective intestinal segment (small or large intestine) successfully colonized by neurons.

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