Regulation of hydrogen ion secretion in rat gastric mucosal parietal cells isolated by centrifugal elutriation

Conference · Sat Mar 01 04:00:00 EST 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5498233

Thompson, W J; Black, E W; Strada, S J

To study the second messengers, cyclic AMP (cA) and calcium as regulators of gastric acid secretion, the authors have developed a method to isolate and enrich parietal cells (PC) from rat gastric mucosa by centrifugal elutriation. After mucosal dispersion with Pronase/EDTA in KRB buffer containing glucose and albumin, cells are elutriated with a four step washout procedure. The PC fraction is 60-80% pure with 95% viability and yields of 40-60%. PC cells are used to identify calcium/calmodulin and cA-mediated protein phosphorylations. CA level is measured with a solid phase radioimmunoassay using sample acetylation and hydrogen ion production is determined indirectly by /sup 14/C-aminopyrine (AP) accumulation. PC-rich (PCR) and PC-poor (PCP) fractions show similar basal cA content (380 fmol/million cells) but have markedly different responses to secretagogues in both rate and magnitude. AP and cA correlate with stimulation induced by forskolin (F) and histamine (H) but not carbachol. F increases cA 14 fold linearly for 60 min in PCP and 180 fold in PCR. H gives a weak 1 fold cA increase in PCP with little change in AP, but shows a 10 fold increase in cA and an 8 fold increase in AP in PCR. No changes in cA are observed with carbachol induced AP accumulation. These data support the role of cA in regulation of acid secretion and demonstrate the utility of centrifugal elutriation as a method to obtain parietal cells from rat gastric mucosa.

• Research Organization: Univ. of South Alabama College of Medicine, Mobile
• OSTI ID: 5498233
• Report Number(s): CONF-8604222-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:3
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALINE EARTH METALS
AMINES
AMINO ACIDS
AMP
ANIMAL CELLS
ANIMALS
AZOLES
BIOLOGICAL MATERIALS
BODY FLUIDS
CALCIUM
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
CHELATING AGENTS
CHEMICAL REACTIONS
EDTA
ELEMENTS
GASTRIC ACID
HETEROCYCLIC COMPOUNDS
HISTAMINE
IMIDAZOLES
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MATERIALS
MEMBRANES
METALS
MUCOUS MEMBRANES
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORYLATION
PROTEINS
RADIOASSAY
RADIOIMMUNOASSAY
RADIOIMMUNOLOGY
RATS
RODENTS
SECRETION
TRACER TECHNIQUES
VERTEBRATES