title: Human mast cell line-1 (HMC-1) cells exhibit a membrane capacitance increase when dialysed with high free-Ca(2+) and GTPγS containing intracellular solution.
abstract: An increase in cytosolic free calcium concentration [Ca(2+)]i initiates the exocytotic activity in various types of secretory cells. The guanosine 5'-O-[3-thio]triphosphate (GTPγS), a non-hydrolysable analogue of GTP (guanosine 5'-triphosphate), is an effective secretagogue for different cell types of different species, like mast cells, neutrophils or eosinophils. Consequently, the internal administration of GTPγS causes degranulation of mouse and rat mast cells. Regarding rat mast cells, it is proved that Ca(2+) can cooperate with GTP or GTPγS in accelerating and increasing amplitude of the secretory response. All the previous studies with respect to capacitance recordings and mast cells were performed using mouse or rat mast cells, usually derived from peritoneum or the rat basophilic leukaemia cell line RBL. In this study, we applied the capacitance measurement technique to the human mast cell line-1 (HMC-1) cells, an immature cell line established from a patient with mast cell leukaemia. Patch-clamp dialysis experiments revealed that high intracellular free Ca(2+) and GTPγS concentrations are both required for considerable capacitance increases in HMC-1 cells. During degranulation of HMC-1 cells, the total membrane capacitance (Cm) increase appeared continuously and, in some cases, as a discrete capacitance change, developing in a stepwise manner. Then, we tested the effect of latrunculin B upon HMC-1 cell capacitance increase as well as of some classic mast cell stimulators like PMA, A23187 and IL-1β in hexosaminidase release. Finally, we could conclude that the HMC-1 cell line represents a suitable model for the study of human mast cell degranulation.
authors.affiliation: Department of Toxicology, Hannover Medical School, Carl Neuberg Str. 1, 30625 Hannover, Germany; Department of Neurology, Hannover Medical School, Carl Neuberg Str. 1, 30625 Hannover, Germany. Electronic address: Balletta.Andrea@mh-hannover.de.
authors.forename: Andrea
authors.initials: A
authors.lastname: Balletta
authors.affiliation: Department of Cellular Imaging and Electron Microscopy, Leibniz Institute of Molecular Pharmacology, Robert Rössle Str. 10, 13125 Berlin, Germany. Electronic address: lorenz@fmp-berlin.de.
authors.forename: Dorothea
authors.initials: D
authors.lastname: Lorenz
authors.affiliation: Department of Toxicology, Hannover Medical School, Carl Neuberg Str. 1, 30625 Hannover, Germany. Electronic address: Rummel.Andreas@mh-hannover.de.
authors.forename: Andreas
authors.initials: A
authors.lastname: Rummel
authors.affiliation: Department of Toxicology, Hannover Medical School, Carl Neuberg Str. 1, 30625 Hannover, Germany. Electronic address: Gerhard.Ralf@mh-hannover.de.
authors.forename: Ralf
authors.initials: R
authors.lastname: Gerhard
authors.affiliation: Department of Toxicology, Hannover Medical School, Carl Neuberg Str. 1, 30625 Hannover, Germany. Electronic address: Bigalke.Hans@mh-hannover.de.
authors.forename: Hans
authors.initials: H
authors.lastname: Bigalke
authors.affiliation: Department of Neurology, Hannover Medical School, Carl Neuberg Str. 1, 30625 Hannover, Germany. Electronic address: Wegner.Florian@mh-hannover.de.
authors.forename: Florian
authors.initials: F
authors.lastname: Wegner
mesh_terms: D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D015550:Cell Degranulation / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology*; D025802:Electric Capacitance; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology; D016209:Interleukin-8 / Q000378:metabolism; D008407:Mast Cells / Q000187:drug effects / Q000502:physiology*; D006435:Renal Dialysis; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D053778:Thiazolidines / Q000494:pharmacology
chemical_list: D019086:Bridged Bicyclo Compounds, Heterocyclic; D053583:Interleukin-1beta; D016209:Interleukin-8; D053778:Thiazolidines; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D000001:Calcimycin; C037068:latrunculin B; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium
keywords: Conductance; Exocytosis; Free Ca(2+); GTPγS; Human mast cell; Membrane capacitance