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Fig. 2. Immunochemical investigation of cGMP formation and the expression of NOS
in the embryonic enteric nervous system. (A,B) Confocal
fluorescence images of migrating enteric neurons on the midgut at 65% E in a
so-called `tissue blot' preparation. Neurons were marked immunocytochemically
for anti-cGMP (red) and anti-acetylated 
-tubulin (green). (C)
Merged image of A and B. m, gut musculature. Scale bar: 50 µm. (D)
Immunoblot analysis with an antibody against universal NOS in homogenates of
different parts of embryonic gut and in the CNS (65-70% E). The antibody
recognizes a protein of 
135-140 kDa and a second, somewhat lighter
protein band in the fore- and midgut cytosolic fraction. Bottom lanes provide
an acetylated -tubulin band as loading control. For quantification, the
ratio of the NOS signal to the -tubulin signal was calculated and
averaged for all probed blots (mean NOS ratios, n=4, s.e.m. of the
ratios are: ±0.07 MG, ±0.04 MG-M, ±0.01 FG, ±0.08
CNS). The NOS ratio directly below the lanes corresponds to the actual signals
of the shown western blot. MG, midgut; MG-M, membranous part of midgut (both
midgut homogenates included adjacent hindgut); FG, foregut; CNS, central
nervous system, including ventral nerve cord.
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