Regulation of hydrogen ion secretion in rat gastric mucosal parietal cells isolated by centrifugal elutriation
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-; TRN: 86-026538
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
GASTRIC ACID
SECRETION
PROTEINS
PHOSPHORYLATION
AMP
ANIMAL CELLS
CALCIUM
CARBON 14 COMPOUNDS
EDTA
HISTAMINE
MUCOUS MEMBRANES
RADIOIMMUNOASSAY
RATS
TRACER TECHNIQUES
ALKALINE EARTH METALS
AMINES
AMINO ACIDS
ANIMALS
AZOLES
BIOLOGICAL MATERIALS
BODY FLUIDS
CARBOXYLIC ACIDS
CHELATING AGENTS
CHEMICAL REACTIONS
ELEMENTS
HETEROCYCLIC COMPOUNDS
IMIDAZOLES
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MATERIALS
MEMBRANES
METALS
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
RADIOASSAY
RADIOIMMUNOLOGY
RODENTS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques