Scanning EM of resting gastric parietal cells reveals a network of cytoplasmic tubules and cisternae connected to the intracellular canaliculus.

Abstract

Using a recently developed fixation technique for parietal cells (Sugai et al., Acta Anat Nippon 1995:70:S79, 1999:74:S101), we have reinvestigated the organization of the cytoplasmic membrane system in the resting stomach by ultra-high-resolution scanning electron microscopy (SEM). Rat gastric mucosae were microwave-fixed in cacodylate buffer [334 milliosmoles/kg H(2)O (mOsm)], to which 1.0% glutaraldehyde and 0.5% formaldehyde were added. Specimens examined by transmission electron microscopy (TEM) of thin sections revealed cytoplasm packed with tubular membranes similar to images detected by rapid-freeze/freeze-substitution fixation which is generally considered to cause minimal structural alterations. To render the cytoplasmic membranes visible by SEM, fixed mucosae were frozen, fractured, and the exposed cytoplasm of parietal cells was macerated by the aldehyde-osmium-DMSO-osmium procedure. With much of the cell matrix and filaments removed, SEM revealed numerous 30-60 nm tubules which formed a meshwork and also small cisternae. The cytoplasmic surface of the tubules was smooth while some cisternal areas had attached polyribosomes. Vesicles or isolated tubules were not found in appropriately macerated parietal cells. The cytoplasmic surface of the intracellular canaliculus was smooth except for round openings representing the bases of macerated microvilli. In favorable sites connections of the tubular membranes to the canaliculi were clearly visible. Stereo pair views were particularly useful to demonstrate these continuities. Connections between these two membrane compartments suggest the probability of rapid membrane transposition.