The developmental stage and cell type dependent phosphorylation of eNOS in murine enteric mucosa and myenteric plexus

In order to clarify the developmental regulation of the eNOS activity in intestine by phosphorylation, we examined the immunohistochemical localizations of the eNOS phosphorylation sites at Ser(1177), Ser(116) and at Thr(495) in cells of the mouse enteric mucosa and myenteric plexus at E13.5, E14.5, E16.5, E18.5, E20.5 and P3. In addition, in cells of the E16.5 stage the protein levels of eNOS and the phosphorylation sites of eNOS at Ser(1177), Ser(116) and at Thr(495) were investigated by immunoblot. From E14.5 to P3, phosphorylation residues of eNOS at Ser(1177) and at Ser(116) were detected with different staining intensities in the enteric mucosa epithelium. In ganglion cells of the myenteric plexus Ser(116) was identified at E18.5 to P3. The absence of phosphorylated Thr(495) in cells of intestine during all developmental stages, was confirmed by immunoblot at E16.5. The immunoblot levels of eNOS and eNOS phosphorylated at Ser(1177) and at Ser(116) were comparable with the immunohistochemical results of E16.5 mouse intestine. It was concluded that development of epithelial cells of the enteric mucosa may be modulated by phosphorylation of eNOS at Ser(1177) and at Ser(116). The phosphorylation of eNOS in cells of the myenteric plexus is modulated at Ser(116). These data suggest that there is a developmental stage and cell type dependent phosphorylation of eNOS.