Stereologic description of the changing expression of constitutive nitric oxide synthase and heme oxygenase in the enteric plexuses of the pig small intestine during development

Abstract

The similarities between heme oxygenase-2 (HO-2) and nitric oxide synthase (nNOS) and the transient expression of nNOS during development led us to investigate whether both systems are similarly affected by changes that occur during development and by regional differences along the small intestine. By combining NADPH diaphorase histochemistry and HO-2 immunohistochemistry on whole-mount preparations and by using stereologic methods, a qualitative and quantitative description of HO-2 and nNOS expression was obtained. Examinations were carried out on the small intestine of fetal, 1-2-day and 5-6-week-old pigs. In all age groups, three enteric plexuses were distinguished. The presence of HO-2-immunoreactive (HO-2-IR) and NADPH diaphorase-positive neurons corresponded to earlier morphological and physiological reports. Nevertheless, the total number of nitrergic neurons remained constant or decreased in the enteric plexuses, whereas the total number of HO-2-IR neurons displayed an overall increase. Changing concentrations of glucocorticoids, target-derived signals, presynaptic input, and an effect of HO-2 activity on nNOS synthesis are likely to play roles in the observed developmental changes. The numerical density of HO-2-IR neurons remained relatively constant along the intestinal tract; in contrast, the nitrergic neurons were most numerous in the inner submucous and myenteric plexus in the duodenum and ileum, respectively. It is believed that the duodenal nitrergic neurons in the inner submucous plexus could be involved in the regulation of duodenal secretion processes, whereas the region-dependent density in the myenteric plexus possibly forms the morphological basis for a regionally different participation of NO in the relaxation of the small intestine.