Perturbation of gastric mucosa in mice expressing the temperature-sensitive mutant of SV40 large T antigen. Potential for establishment of an immortalised parietal cell line

doi:10.1078/0171-9335-00249

European Journal of Cell Biology

Volume 81, Issue 5, May 2002, Pages 281-293

https://doi.org/10.1078/0171-9335-00249 Get rights and content

Summary

Gastric parietal cells have a unique secretory membrane system that undergoes a profound transformation when the parietal cell is stimulated to secrete acid. Understanding this process has been hindered by the lack of an immortalised parietal cell line. Here we have explored a strategy for the development of a parietal cell line by the generation of transgenic mice bearing the temperature-sensitive mutant of the SV40 large T antigen (SV40 tsA58) under the control of the regulatory sequences of the gastric H⁺/K⁺ ATPase β-subunit (H/Kβ-tsA58). Three H/Kβ-tsA58 transgenic mouse lines were established, namely 218, 224 and 228, all of which expressed the tsA58 T antigen in the gastric mucosa. Unexpectedly, the gastric mucosae of all lines were hypertrophic indicating that the temperature-sensitive large T antigen was partially active at 37 °C. Immunofluorescence together with light and electron microscopic studies revealed that mature parietal and zymogenic cells were absent in H/Kβ-tsA58 transgenic lines 218 and 224, and small undifferentiated cells were the dominant cell type in the gastric units. On the other hand, a few mature parietal cells were detected in line 228 together with an increased proportion of undifferentiated cells and, normally rare, pre-parietal cells. As line 228 represented a rich source of pre-parietal cells, gastric cells from line 228 were isolated and cultured at 33 °C, the permissive temperature for tsA58. Gastric epithelial cells, expressing the T antigen, were maintained in culture for over 6 weeks. Upon a temperature shift to 39 °C the cultured gastric cells developed characteristics of differentiated parietal cells, including the presence of a nascent canaliculus and dramatically increased production of the gastric H⁺/K⁺ ATPase β-subunit. Therefore, this system shows the potential to generate an immature parietal cell line that can be induced to differentiate in vitro.

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Perturbation of gastric mucosa in mice expressing the temperature-sensitive mutant of SV40 large T antigen. Potential for establishment of an immortalised parietal cell line

Summary

Trends Cell Biol.

J. Biol. Chem.

Brain Res.

Gastroenterology

J. Biol. Chem.

J. Biol. Chem.

Gastroenterology

J. Biol. Chem.

J. Biol. Chem.

Cytological transformations associated with parietal cell stimulation: critical steps in the activation cascade

J. Cell Sci.

A novel method for isolating mononuclear cells from the stomachs of mice with experimental autoimmune gastritis

Autoimmunity

Expression of a gastric autoantigen in pancreatic islets results in non-destructive insulitis after neonatal thymectomy

Eur. J. Immunol.

Isolation, identification and quantitative evaluation of specific cell types from the mammalian gastric mucosa

Cell Tissue Res.

Renal expression of the gene encoding the gastric H+-K+-ATPase beta-subunit

Am. J. Physiol.

Genetic ablation of parietal cells in transgenic mice:A new model for analyzing cell lineage relationships in the gastric mucosa

Proc. Natl. Acad. Sci. USA

Parietal cell culture – New models and directions

Annu. Rev. Physiol.

Primary culture of secretagogue-responsive parietal cells from rabbit gastric mucosa

Am. J. Physiol.

Differentiated mammalian cell lines immortalized by temperature-sensitive tumor viruses

Mol. Endocrinol.

Expression of rab11a N124I in gastric parietal cells inhibits stimulatory-recruitment of the H+-K+-ATPase

Am. J. Physiol.

Membrane and protein recycling associated with gastric HCl secretion

J. Intern. Med.

State of actin in gastric parietal cells

Am. J. Physiol.

Renal expression of the gene encoding the gastric H⁺-K⁺-ATPase beta-subunit

Expression of rab11a N124I in gastric parietal cells inhibits stimulatory-recruitment of the H^+-K⁺-ATPase