Sonic hedgehog is associated with H+-K+-ATPase- containing membranes in gastric parietal cells and secreted with histamine stimulation

Yana Zavros, Melissa A. Orr, Chang Xiao, Danuta H. Malinowska

Research output: Contribution to journalArticlepeer-review

35 Scopus citations

Abstract

Sonic hedgehog (Shh) is found within gastric parietal cells and processed from a 45-kDa to a 19-kDa bioactive protein by an acid- and protease-dependent mechanism. To investigate whether Shh is associated with the parietal cell membrane compartment that becomes exposed to both acid and proteolytic enzymes during acid secretion, the cellular location of Shh within resting and stimulated gastric parietal cells was examined. Immunofluorescence microscopy of rabbit stomach sections showed that Shh colocalized predominantly with parietal and pit, not chief/zymogen or neck, cell markers. In resting and histaminestimulated rabbit gastric glands Shh was expressed only in parietal cells close to H+-K+-ATPase-containing tubulovesicular and secretory membranes with some colocalizing with γ-actin at the basolateral membrane. Gastric gland microsomal membranes were prepared by differential and sucrose gradient centrifugation and immunoisolation with an anti-H+-K+-ATPase-α subunit antibody. The 45- and 19-kDa Shh proteins were detected by immunoblot in immunopurified H +-K+-ATPase-containing membranes from resting and stimulated gastric glands, respectively. Incubating glands with a high KCl concentration removed Shh from the membranes. Histamine stimulated 19-kDa Shh secretion from gastric glands into the medium. In human gastric cancer 23132/87 cells cultured on permeable membranes, histamine increased 19-kDa Shh secretion into both apical and basolateral media. These findings show that Shh is a peripheral protein associated with resting and stimulated H+-K +-ATPase-expressing membranes. In addition, Shh appears to be expressed at or close to the basolateral membrane of parietal cells.

Original languageEnglish (US)
Pages (from-to)G99-G111
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume295
Issue number1
DOIs
StatePublished - Jul 2008
Externally publishedYes

Keywords

  • 23132/87 cells
  • Pepsinogen A
  • Processing
  • Rabbit gastric glands
  • Transepithelial electrical resistance
  • Tubulovesicles

ASJC Scopus subject areas

  • Physiology
  • Hepatology
  • Gastroenterology
  • Physiology (medical)

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