A method for in vitro studies on acid formation in human parietal cells. Stimulation by histamine, pentagastrin and carbachol

Abstract

Cells were isolated from human gastric mucosa on a large scale from gastric resection specimens and on a microscale from endoscopic biopsies by sequential incubations with pronase and collagenase. The accumulation of aminopyrine (AP) was used as an index of acid production in the parietal cells. Basal accumulation was about 0.2 pmol AP/10(4) parietal cells. Addition of histamine, db-cAMP, pentagastrin and carbachol increased the aminopyrine accumulation. Maximal accumulation was of the order of 1000-2800% of the control and was obtained after stimulation by 10(-4) M histamine and by 10(-3) M db-cAMP. Stimulation by pentagastrin and by carbachol reached 200 to 350% of the control. EC50 was 2 X 10(-6) M for histamine, 10(-8) M for pentagastrin, and 4 X 10(-6) M for carbachol. Human parietal cells were enriched from a mixture of gastric mucosal cells by isopycnic centrifugation on density gradients of Percoll. A parietal cell fraction with a purity of 83% was obtained. The density of human parietal cells was estimated to 1.06 g . ml-1.

The role of gastric histamine release in the acid secretory response to pentagastrin and methacholine in the dog

“…More recently, our group has been able to demonstrate in vivo in the intact dog that both pentagastrin and methacholine can stimulate the release of histamine from the stomach [15], and Chuang et al isolated an endocrine cell from the dog mucosa that contains histamine and responds to both gastrin and carbachol [16]. In drawing an analogy to human physiology, the dog model more closely approximates the human model in that human parietal cells are responsive to gastrin [27], and that histologically mast cells are the main storage sites for histamine, although the less abundant histamine containing ECL cells are probably the cells involved in acid secretion [28]. In accord with these observations, Man et al recently demonstrated that pentagastrin administration resulted in gastric histamine release in both dogs and people with duodenal ulcers [29].…”

Autoimmune Gastric Disease Chemical and Clinical Studies with Special Reference to H+, K+-ATPase

“…For instance vagal stimulation enhances gastrin release, partly by inhibition of somatostatin (157), a peptide that blocks gastrin release. Histamine plays an essential role in the stimulation of the parietal cell (130) and a 2 receptor, blocked by H2-receptor antagonists (132), has been identified on the parietal cell in all mammals investigated, including man (80,102). Histamine stimulation activates a membrane-associated enzyme, adenylate cyclase, which catalyzes the formation of cyclic adenosine monophosphate (CAMP) from ATP (Fig 1).…”

Host-specific differences in the physiology of acid secretion related to prostaglandins may play a role in gastric inflammation and injury

“…Mouse models have recently been accepted into the mainstream of research into gastric pathology, inasmuch as they rely on the availability of transgenic techniques and an extensive knowledge of the immune system. Aminopyrine accumulation is a reliable method for studying acid secretion in human (21) and animal parietal cells or gastric gland preparations (3), and we recently extensively characterized the pharmacology of acid secretion in the mouse (28) to meet the demand for knowledge in this area. The mouse model has been widely used in the study of Helicobacter-induced gastritis and revealed strain-dependent patterns (20).…”