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    Elsevier

    Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology

    Volume 128, Issue 2 , February 2001 , Pages 349-357
    Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology

    Characterization of oxyntic glands isolated from the rat gastric mucosa

    Author links open overlay panel LeilaAzerkan PerBengtsson KarinTømmerås Zhao-QiLi SvenMårdh

    https://doi.org/10.1016/S1095-6433(00)00309-3 Get rights and content

    Abstract

    A simple and reproducible method for isolating oxyntic glands from the rat gastric mucosa was developed. The mucosa was incubated with pronase and EGTA, and then treated mechanically to release glands that were separated from single cells by sedimentation. Parietal cells were identified by immunostaining using a monoclonal antibody against H,K-ATPase. The glandular cells appeared morphologically intact. By careful control of the conditions of gland isolation, long glandular structures comprising hundreds of cells surrounding the lumen were obtained. Intraperitoneal injection of Br-deoxyuridine in the rat 1.5 h before the isolation procedure resulted in glands with a labeling of cells in their neck region. The glands were viable, as demonstrated by their ability to respond to various hormones. Histamine dose-dependently stimulated the acid formation which was measured as the accumulation of [14C]aminopyrine. At 100 μM histamine the accumulation was increased 5–10-fold. At 100 nM, pentagastrin potentiated the histamine stimulated accumulation by approximately 40% but pentagastrin alone did not stimulate. The oxyntic glands obtained by the present procedure appear useful for studies on cell physiology, including regulation of acid secretion, cellular interactions, and possibly also differentiation and proliferation mechanisms since long glandular fragments that contained the proliferative zone could be isolated.

    Keywords

    Acid secretion
    Acridine orange
    Aminopyrine accumulation
    Br-deoxyuridine
    Gastric mucosa
    Histamine stimulation
    H,K-ATPase
    Pentagastrin stimulation
    Pronase
    Rat oxyntic glands
    Copyright © 2001 Elsevier Science Inc. All rights reserved.