Genetic Ablation of the ClC-2 Cl-Channel disrupts mouse gastric parietal cell acid secretion

Meghali Nighot, Prashant Nighot, Thomas Ma, Danuta H. Malinowska, Gary E. Shull, John Cuppoletti, Anthony T. Blikslager

Research output: Contribution to journalArticle

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Abstract

The present studies were designed to examine the effects of ClC-2 ablation on cellular morphology, parietal cell abundance, H/K ATPase expression, parietal cell ultrastructure and acid secretion using WT and ClC-2-/-mouse stomachs. Cellular histology, morphology and proteins were examined using imaging techniques, electron microscopy and western blot. The effect of histamine on the pH of gastric contents was measured. Acid secretion was also measured using methods and secretagogues previously established to give maximal acid secretion and morphological change. Compared to WT, ClC-2-/-gastric mucosal histological organization appeared disrupted, including dilation of gastric glands, shortening of the gastric gland region and disorganization of all cell layers. Parietal cell numbers and H/K ATPase expression were significantly reduced by 34% (P<0.05) and 53% (P<0.001) respectively and cytoplasmic tubulovesicles appeared markedly reduced on electron microscopic evaluation without evidence of canalicular expansion. In WT parietal cells, ClC-2 was apparent in a similar cellular location as the H/K ATPase by immunofluorescence and appeared associated with tubulovesicles by immunogold electron microscopy. Histaminestimulated [H+] of the gastric contents was significantly (P<0.025) lower by 9.4 fold (89%) in the ClC-2-/-mouse compared to WT. Histamine/carbachol stimulated gastric acid secretion was significantly reduced (range 84-95%, P<0.005) in ClC-2-/-compared to WT, while pepsinogen secretion was unaffected. Genetic ablation of ClC-2 resulted in reduced gastric gland region, reduced parietal cell number, reduced H/K ATPase, reduced tubulovesicles and reduced stimulated acid secretion.

Original languageEnglish (US)
Article numberA1344
JournalPloS one
Volume10
Issue number9
DOIs
StatePublished - Sep 17 2015

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Gastric Parietal Cells
Proton-Translocating ATPases
Ablation
stomach
Gastric Mucosa
secretion
Adenosine Triphosphatases
Gastrointestinal Contents
Acids
gastric mucosa
acids
mice
Histamine
Stomach
Electron Microscopy
Cell Count
Electron microscopy
cells
histamine
Pepsinogen A

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Nighot, Meghali ; Nighot, Prashant ; Ma, Thomas ; Malinowska, Danuta H. ; Shull, Gary E. ; Cuppoletti, John ; Blikslager, Anthony T. / Genetic Ablation of the ClC-2 Cl-Channel disrupts mouse gastric parietal cell acid secretion. In: PloS one. 2015 ; Vol. 10, No. 9.
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abstract = "The present studies were designed to examine the effects of ClC-2 ablation on cellular morphology, parietal cell abundance, H/K ATPase expression, parietal cell ultrastructure and acid secretion using WT and ClC-2-/-mouse stomachs. Cellular histology, morphology and proteins were examined using imaging techniques, electron microscopy and western blot. The effect of histamine on the pH of gastric contents was measured. Acid secretion was also measured using methods and secretagogues previously established to give maximal acid secretion and morphological change. Compared to WT, ClC-2-/-gastric mucosal histological organization appeared disrupted, including dilation of gastric glands, shortening of the gastric gland region and disorganization of all cell layers. Parietal cell numbers and H/K ATPase expression were significantly reduced by 34{\%} (P<0.05) and 53{\%} (P<0.001) respectively and cytoplasmic tubulovesicles appeared markedly reduced on electron microscopic evaluation without evidence of canalicular expansion. In WT parietal cells, ClC-2 was apparent in a similar cellular location as the H/K ATPase by immunofluorescence and appeared associated with tubulovesicles by immunogold electron microscopy. Histaminestimulated [H+] of the gastric contents was significantly (P<0.025) lower by 9.4 fold (89{\%}) in the ClC-2-/-mouse compared to WT. Histamine/carbachol stimulated gastric acid secretion was significantly reduced (range 84-95{\%}, P<0.005) in ClC-2-/-compared to WT, while pepsinogen secretion was unaffected. Genetic ablation of ClC-2 resulted in reduced gastric gland region, reduced parietal cell number, reduced H/K ATPase, reduced tubulovesicles and reduced stimulated acid secretion.",
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Genetic Ablation of the ClC-2 Cl-Channel disrupts mouse gastric parietal cell acid secretion. / Nighot, Meghali; Nighot, Prashant; Ma, Thomas; Malinowska, Danuta H.; Shull, Gary E.; Cuppoletti, John; Blikslager, Anthony T.

In: PloS one, Vol. 10, No. 9, A1344, 17.09.2015.

Research output: Contribution to journalArticle

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AU - Nighot, Prashant

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