Simultaneous detection of histamine release and lactate production in rat mast cells induced by compound 4880 using 1H NMR

Allergen‐induced histamine secretion associated with lactate production in mast cells detected by 1H NMR

“…However, in this study we did not detect any significant increase in lactate without exocytosis, as reported previously (2). Thus, basal glycolysis seems inactive in mast cells at room temperature, even though the mast cells might be under hypoxic conditions in the NMR tube.…”

“…2d), indicating negative nuclear Overhauser effects on the histamine resonances, as reported previously (1,2). This suggested that the histamine molecule was in a certain "bound" state in the granule matrix.…”

“…We also detected an increase in a sharp resonance at 1.3 ppm, and this resonance was assumed to correspond to the methyl protons of lactate (2). In order to confirm the assignment, the cell-free supernatant was obtained by centrifugation at 1000 rpm.…”

“…We also detected histamine in cells suspended in buffer prepared with H,O and found that histamine secretion in mast cells induced by compound 48/80 (condensation product of N-methyl-p-methoxy-phenethylamine with formaldehyde) was associated with the appearance of a large aliphatic resonance in mast cell suspension. We tentatively assigned it to the methyl protons of lactate (2).…”

Expression of kininogens in the connective tissue‐type mast cells of the rat

“…For preparation of the mast cell extracts, the peritoneal cavity of rats was first washed with Hank's balanced salt solution (HBSS) containing 0·1% bovine serum albumin (BSA), and the washing medium was centrifuged at 450 g for 15 min to recover the peritoneal cells. These cells were separated by centrifugation on density gradients of Percoll, and the mast cells obtained were washed in the 0·1% BSA/HBSS, pelleted and stored at −80° 9 . Purity of the mast cells thus prepared was > 98%, and > 96% were viable.…”