Immunohistochemical Detection of the Cytoskeletal Components in Gastric Parietal Cells

Abstract

Gastric parietal cells are characterized by intracellular canaliculi with many microvilli and tubulovesicles. These membranous structures are continuous and change in volume according to gastric acid secreting or resting state. Morphological changes are induced by cytoskeletal actin. Actin filaments are anchored to the cell membrane via ezrin. This study aimed to ascertain whether morphological changes were related to the nature of ezrin molecules, which were either phosphorylated or non-phosphorylated. Rat stomach tissues were rapid-frozen followed by freeze-substitution and embedding in paraffin for light microscopy. Also, the specimens were high pressure-frozen, freeze-substituted and embedded in Epon 812 or Lowicryl K4M for electron microscopy. Sections from paraffin or Lowicryl K4M were stained with anti-H⁺·K⁺-ATPase antibodies, anti-phosphorylated or non-phosphorylated ezrin antibodies, and anti-actin antibody. The H⁺·K⁺-ATPase was distributed on the intracellular canaliculi and tubulovesicles. The intracellular canaliculi were labeled with anti-phosphorylated ezrin antibodies in fed (gastric acid secreting state) rats. These were stained weakly in starved (gastric acid resting state) rats. Anti-nonphosphorylated ezrin antibodies hardly stained these organelles in either state. A possible explanation is that the phosphorylation of ezrin molecules is related to the reciprocal membranous transformation between intracellular canaliculi and tubulovesicles.