Cytochrome b558 (p22phox) in the guinea-pig adrenal medulla

Paraganglionic cells are sensitive to hypoxia, and the involvement of a plasmalemmal cytochrome b558-like protein in oxygen sensing by these cells has been suggested, but neither the identity of the immunoreactive protein detected by immunohistochemistry nor its anticipated subcellular (i.e., plasmalemmal) localization were directly proven. Thus, we extended these studies to the largest paraganglion, i.e., the adrenal medulla, in the guinea-pig, which, due to its size and accessibility, allowed us to address both of these issues utilizing antisera raised against synthetic peptides of the small (22 kD) subunit of cytochrome b558, p22phox. Cytochrome b558 was originally identified in granulocytes and macrophages, and antisera against this phagocyte p22phox were utilized. Immunoreactivity to p22phox was observed in all adrenal medullary endocrine cells, and the identity of the immunoreactive protein to the small cytochrome b558-subunit was confirmed by Western blotting. Immuno-electron microscopy of ultrathin cryosections and of resin-embedded tissue demonstrated its subcellular localization in the dense core vesicles of endocrine A-cells but not in the plasma membrane. In conclusion, the present study documents the presence of the small subunit of cytochrome b558 in guinea-pig adrenal medullary cells, but its subcellular vesicular localization does not support the initial interpretation of cytochrome b558 serving as a plasmalemmal oxygen sensor.