Muscarinic receptor regulation of Ca2+ mobilization in a human salivary cell line. 1989

X J He, and X Z Wu, and R B Wellner, and B J Baum
Clinical Investigations and Patient Care Branch, National Institute of Dental Research, Bethesda, MD 20892.

We have studied receptor-mediated Ca2+ mobilization in an established exocrine epithelial cell line (HSG-PA) derived from a human submandibular gland. These cells possess a single class of high-affinity muscarinic cholinergic receptors identified using [3H]-quinuclidinyl-benzilate (Kd = 0.17 +/- 0.07 nmol/l; Bmax = 37 +/- 2 fmol/mg protein; n = 3). The muscarinic agonist carbachol elicits a concentration dependent increase of [3H]-inositol trisphosphate in HSG-PA cells (100 mumol/l; greater than 2 fold by 30 s). Carbachol also results in a rapid, approximately 5-fold increase in cytosolic [Ca2+]. This response is made up of two components, one arising from the release of intracellular Ca2+ (La3+ insensitive; independent of extracellular [Ca2+]), the other from the entry of extracellular Ca2+ (La3+ sensitive; dependent on extracellular [Ca2+]). These Ca2+ mobilizing mechanisms are completely blocked by the muscarinic antagonist atropine (10 mumol/l) but unaffected by several voltage-dependent Ca2+ channel antagonists (verapamil, nifedipine, diltiazem) and by membrane depolarization (incubation in 55 mmol/l KCl). These results demonstrate that HSG-PA cells respond to muscarinic stimulation by mobilizing Ca2+ from an intracellular store and via a receptor-operated Ca2+ entry pathway.

UI MeSH Term Description Entries
D007295 Inositol Phosphates Phosphoric acid esters of inositol. They include mono- and polyphosphoric acid esters, with the exception of inositol hexaphosphate which is PHYTIC ACID. Inositol Phosphate,Phosphate, Inositol,Phosphates, Inositol
D011869 Radioligand Assay Quantitative determination of receptor (binding) proteins in body fluids or tissue using radioactively labeled binding reagents (e.g., antibodies, intracellular receptors, plasma binders). Protein-Binding Radioassay,Radioreceptor Assay,Assay, Radioligand,Assay, Radioreceptor,Assays, Radioligand,Assays, Radioreceptor,Protein Binding Radioassay,Protein-Binding Radioassays,Radioassay, Protein-Binding,Radioassays, Protein-Binding,Radioligand Assays,Radioreceptor Assays
D011976 Receptors, Muscarinic One of the two major classes of cholinergic receptors. Muscarinic receptors were originally defined by their preference for MUSCARINE over NICOTINE. There are several subtypes (usually M1, M2, M3....) that are characterized by their cellular actions, pharmacology, and molecular biology. Muscarinic Acetylcholine Receptors,Muscarinic Receptors,Muscarinic Acetylcholine Receptor,Muscarinic Receptor,Acetylcholine Receptor, Muscarinic,Acetylcholine Receptors, Muscarinic,Receptor, Muscarinic,Receptor, Muscarinic Acetylcholine,Receptors, Muscarinic Acetylcholine
D002118 Calcium A basic element found in nearly all tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes. Coagulation Factor IV,Factor IV,Blood Coagulation Factor IV,Calcium-40,Calcium 40,Factor IV, Coagulation
D002217 Carbachol A slowly hydrolyzed CHOLINERGIC AGONIST that acts at both MUSCARINIC RECEPTORS and NICOTINIC RECEPTORS. Carbamylcholine,Carbacholine,Carbamann,Carbamoylcholine,Carbastat,Carbocholine,Carboptic,Doryl,Isopto Carbachol,Jestryl,Miostat,Carbachol, Isopto
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D013363 Submandibular Gland One of two salivary glands in the neck, located in the space bound by the two bellies of the digastric muscle and the angle of the mandible. It discharges through the submandibular duct. The secretory units are predominantly serous although a few mucous alveoli, some with serous demilunes, occur. (Stedman, 25th ed) Submaxillary Gland,Gland, Submandibular,Gland, Submaxillary,Glands, Submandibular,Glands, Submaxillary,Submandibular Glands,Submaxillary Glands
D015544 Inositol 1,4,5-Trisphosphate Intracellular messenger formed by the action of phospholipase C on phosphatidylinositol 4,5-bisphosphate, which is one of the phospholipids that make up the cell membrane. Inositol 1,4,5-trisphosphate is released into the cytoplasm where it releases calcium ions from internal stores within the cell's endoplasmic reticulum. These calcium ions stimulate the activity of B kinase or calmodulin. 1,4,5-InsP3,Inositol 1,4,5-Triphosphate,Myo-Inositol 1,4,5-Trisphosphate,1,4,5-IP3,Myoinositol 1,4,5-Triphosphate

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