scholarly journals Purification of Parietal and Chief Cells from the Gastric Mucosa

2002 ◽  
Vol 2 ◽  
pp. 1650-1653 ◽  
Author(s):  
John Graham

Acid-secreting parietal cells from the gastric mucosa are widely studied as a model in studies on ion transport. A discontinuous gradient of iodixanol has been found to be superior to earlier protocols using Nycodenz® and this method, which removes a significant amount of contaminating cells and mucus is a very useful prelude to further purification by elutriation.

2002 ◽  
Vol 2 ◽  
pp. 1643-1645 ◽  
Author(s):  
John Graham

Acid-secreting parietal cells from the gastric mucosa are widely studied as a model in studies on ion transport and the endocrine/paracrine ECL cells effectively control parietal cell function. Discontinuous gradients of iodixanol for the purification of ECL cells were subsequently simplified to the use of a density barrier. This technique is now commonly used following initial centrifugal elutriation.


1990 ◽  
Vol 1 (3) ◽  
pp. 259-268 ◽  
Author(s):  
P A Negulescu ◽  
A Harootunian ◽  
R Y Tsien ◽  
T E Machen

Regulation of cytosolic free Na (Nai) was measured in isolated rabbit gastric glands with the use of a recently developed fluorescent indicator for sodium, SBFI. Intracellular loading of the indicator was achieved by incubation with an acetoxymethyl ester of the dye. Digital imaging of fluorescence was used to monitor Nai in both acid-secreting parietal cells and enzyme-secreting chief cells within intact glands. In situ calibration of Nai with ionophores indicated that SBFI fluorescence (345/385 nm excitation ratio) could resolve 2 mM changes in Nai and was relatively insensitive to changes in K or pH. Measurements on intact glands showed that basal Nai was 8.5 +/- 2.2 mM in parietal cells and 9.2 +/- 3 mM in chief cells. Estimates of Na influx and efflux were made by measuring rates of Nai change after inactivation or reactivation of the Na/K ATPase in a rapid perfusion system. Na/K ATPase inhibition resulting from the removal of extracellular K (Ko) caused Nai to increase at 3.2 +/- 1.5 mM/min and 3.5 +/- 2.7 mM/min in parietal and chief cells, respectively. Na buffering was found to be negligible. Addition of 5 mM Ko and removal of extracellular Na (Nao) caused Nai to decrease rapidly toward 0 mM Na. By subtracting passive Na efflux under these conditions (the rate at which Nai decreased in Na-free solution containing ouabain), an activation curve (dNai/Nai) for the Na/K ATPase was calculated. The pump demonstrated the greatest sensitivity between 5 and 20 mM Nai. At 37 degrees C the pump rate was less than 3 mM/min at 5 mM Nai and 26 mM/min at 25 mM Nai, indicating that the pump has a great ability to respond to changes in Nai in this range. Carbachol, which stimulates secretion from both cell types, was found to stimulate Na influx in both cell types, but did not have detectable effects on Na efflux. dbcAMP+IBMX, potent stimulants of acid secretion, had no effect on Na metabolism.


1965 ◽  
Vol 208 (1) ◽  
pp. 18-23 ◽  
Author(s):  
Darwin Alonso ◽  
John B. Harris

In standard chloride solutions, methyl xanthines enhanced ion transport and oxygen consumption by frog gastric mucosa in vitro to a significantly greater extent than did histamine. Substituting glucuronate for chloride in the bathing solutions abolished ion transport, but left respiration unchanged. Caffeine and histamine had no effect on respiration in glucuronate solutions, but butyrate stimulated respiration to the same extent in chloride and glucuronate solutions. All of these effects appeared to be a function of the epithelial cells, since respiration by smooth muscle from the stomach was not affected by any of the compounds. It is postulated that the hydrogen ions of gastric secretion are derived from the dehydrogenation process during substrate oxidation. Following dehydrogenation, the methyl xanthines and histamine stimulate some step in the events that lead to the formation of hydrochloric acid. In the respiring, non-acid secreting mucosa, the hydrogen ions follow the usual intracellular pathways that lead to the formation of water.


1956 ◽  
Vol 187 (3) ◽  
pp. 427-431 ◽  
Author(s):  
Joseph J. Vitale ◽  
Oscar M. Jankelson ◽  
Patricia Connors ◽  
D. Mark Hegsted ◽  
Norman Zamcheck

Effect of histamine on the activity of succinic oxidase and malic dehydrogenase was studied in guinea pig and human gastric mucosa. Human tissue was obtained through the surgical services of the Boston City Hospital. Control value for the succinic oxidase system of the proximal half of the guinea pig stomach was approximately 480 ( Qo2 (N) (µl O2/mg nitrogen/hr.)). After histamine, this value rose to 550 in 30 minutes with a simultaneous rise in titratable acidity of the stomach contents. Animals fasted for 72 hours had a Qo2 (N) of approximately 500 and after histamine a Qo2 (N) of 700 was observed. Spectrophotometric analysis of succinic dehydrogenase and cytochrome oxidase activities, two of the major components of the succinic oxidase system, revealed that both components are increased following histamine administration. Malic dehydrogenase, however, was not affected by histamine treatment. Succinic dehydrogenase was demonstrated by histochemical localization and was concentrated below the superficial mucous layer where parietal cells were abundant. Succinic oxidase activity of human gastric mucosa was demonstrable only in those specimens containing abundant parietal cells. This study confirms the view that HCl production by parietal cells is associated with aerobic metabolism and is perhaps under enzymatic control. The study suggests that the succinic oxidase system may be involved in the production or secretion of HCl.


1989 ◽  
Vol 256 (1) ◽  
pp. G254-G263 ◽  
Author(s):  
C. S. Chew ◽  
M. Ljungstrom ◽  
A. Smolka ◽  
M. R. Brown

A new procedure for isolation and primary culture of gastric parietal cells is described. Parietal cells from rabbit gastric mucosa are enriched to greater than 95% purity by combining a Nycodenz gradient separation with centrifugal elutriation. Cells are plated on the basement membrane matrix, Matrigel, and maintained in culture for at least 1 wk. Parietal cells cultured in this manner remain differentiated, cross-react with monoclonal H+-K+-ATPase antibodies, and respond to histamine, gastrin, and cholinergic stimulation with increased acid production as measured by accumulation of the weak base, [14C]aminopyrine. When stimulated, cultured cells undergo ultrastructural changes in which intracellular canaliculi expand and numerous microvilli are observed. These ultrastructural changes are similar to those previously found to occur in vivo and in acutely isolated parietal cells. Morphological transformations in living cells can also be observed with differential interference contrast optics in the light microscope. After histamine stimulation, intracellular canaliculi gradually expand to form large vacuolar spaces. When the H2 receptor antagonist, cimetidine, is added to histamine-stimulated cells, these vacuoles gradually disappear. The ability to maintain hormonally responsive parietal cells in primary culture should make it possible to study direct, long-term effects of a variety of agonists and antagonists on parietal cell secretory-related activity. These cultured cells should also prove to be useful for the study of calcium transients, ion fluxes, and intracellular pH as related to acid secretion in single cells, particularly since morphological transformations can be used to monitor "physiological" responses at the same time within the same cell.


1984 ◽  
Vol 16 (2) ◽  
pp. 55-61 ◽  
Author(s):  
I. Guldvog ◽  
A. Berstad
Keyword(s):  

2019 ◽  
Vol 16 (2) ◽  
pp. 0281
Author(s):  
AL –Nakeeb Et al.

          A histological study showed the wall of the stomach in Pica pica and Herpestes javanicus consists of four layers: mucosa, submucosa, muscularis externa and serosa. Also, the present study showed many  differences in the histological structures of the stomach for each in both types. The stomach of P. pica consists of two portions: the proventiculus and gizzard, while the stomach of H. javanicus consists of three portions: cardiac, fundic and pyloric regions. The mucosa layer formed short gastric folds, named plicae. In the proventiculus of P. pica, sulcus is found between each two plicae, but the folds called gastric pits in the gizzard, which are full with koilin. Lamina properia in both types contained gastric glands (straight simple tubular glands) named superficial glands, as well as another gastric gland found in the submucosa layer of the proventiculus in P. pica only named deep gastric glands. The gastric gland in the stomach of H. javanicus contained: mucous neck cells and parietal cells positive to AB/PAS stains in cardiac portion, as well as chief cells in fundic portion, but pyloric portion had just mucous neck cells. Muscularis externa in both types formed two muscle layers: inner and outer layer.


1967 ◽  
Vol 124 (4) ◽  
pp. 1070-1073 ◽  
Author(s):  
A. Gerald ◽  
J. DeGraff ◽  
R. Lev ◽  
G. B. Jerzy Glass

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