The chemical estimation of adrenaline-like substances in blood

Weil‐Malherbe, H.; Bone, A. D.

doi:10.1042/bj0510311

Cited by 450 publications

(60 citation statements)

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“…Perchloric acid was added into the su pernatant to give a final concentration of 0.4 N, and cal phostin C in the sample was then removed by chloroform extraction, because this compound interfered with the sub sequent determination of catecholamines. Catechol amines were measured by the ethylenediamine condensa tion method (11). Free Ca 21 concentration in the medium was measured with a Cat+-sensitive electrode constructed with the neutral ligand ETH 1001 (12).…”

Section: Nicotinicmentioning

confidence: 99%

Calphostin C, a Potent and Specific Inhibitor of Protein Kinase C, Reduces Phorbol Ester-Induced but Not Primary Ca2+-Induced Catecholamine Secretion from Digitonin-Permeabilized Bovine Adrenal Medullary Cells

Isosaki

Minami

Nakashima

1994

Japanese Journal of Pharmacology

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ABSTRACT-Calphostin C, a protein kinase C inhibitor, reduced phorbol ester-induced enhancement of catecholamine secretion but not primary Ca"-induced secretion from digitonin-permeabilized bovine adrenal medullary cells, indicating that this compound selectively inhibited protein kinase C-dependent secretion.Keywords: Adrenal medulla, Protein kinase C, Secretion Nicotinic agonist-induced catecholamine secretion from bovine adrenal medullary cells is triggered by a rise in the cytosolic Ca2+ concentration as a consequence of an increased permeability of the plasma membrane to extracellular Ca 2+. The sequence of the events between a rise in the cytosolic Ca 21 concentration and exocytotic catecholamine secretion remains poorly understood. Sev eral lines of evidence indicate that Ca 2+-activated, phos pholipid-dependent protein kinase (protein kinase C) is involved in the process of exocytosis (1-3). Ca 21 influx into the cells causes both the translocation of protein kinase C molecules to the membrane and diacylglycerol accumulation from the breakdown of phosphoinositides, which results in the activation of protein kinase C (2). Active phorbol esters that can replace diacylglycerol in stimulating protein kinase C also potentiate Ca 2+-in duced catecholamine secretion from electropermeabilized and digitonin-permeabilized adrenal medullary cells (1, 3). Moreover, a variety of protein kinase C inhibitors such as neomycin, spermine, calmidazolium and polymyx in B have been reported to depress not only phorbol ester induced enhancement of catecholamine secretion but also Ca2+-induced secretion (4), suggesting that protein ki nase C plays an essential role in secretion. In contrast, pro tein kinase inhibitors such as staurosporine and K-252a, a pseudosubstrate peptide of protein kinase C, and mild treatment with trypsin were found to reduce only catechol amine secretion enhanced by phorbol ester without effect on Ca" -induced secretion (5 8). Calphostin C has been reported to be a novel and potent inhibitor of protein kinase C and about 1000 times more specific to protein kinase C than other protein kinases such as cAMP-depend ent protein kinase and tyrosine-specific protein kinase (9). Thus, the effect of calphostin C on Ca 2+-dependent and phorbol ester-dependent catecholamine secretion from digitonin-permeabilized bovine adrenal medullary cells was examined in the present study.Bovine adrenal medullary cells were isolated by sequen tial digestion of adrenal medullary slices with collagenase as reported previously (10), purified by a Percoll (Phar macia, Uppsala, Sweden) gradient and maintained as monolayer cultures in Eagle's minimal essential medium containing 10°70 heat-inactivated calf serum, 100 units/ml penicillin, 100 pg/ml streptmycin, 0.3 pg/ml amphoteri cin B and 10 pM cytosine arabinoside. The cells were plat ed in 24-well cluster plates at a density of 8 x 105 cells/well. The cultured cells were washed with 0.5 ml of sodium glutamate medium consisting of 150 mM monoso dium glutamate and 10 mM Na PIPES (pH...

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Section: Nicotinicmentioning

confidence: 99%

Calphostin C, a Potent and Specific Inhibitor of Protein Kinase C, Reduces Phorbol Ester-Induced but Not Primary Ca2+-Induced Catecholamine Secretion from Digitonin-Permeabilized Bovine Adrenal Medullary Cells

Isosaki

Minami

Nakashima

1994

Japanese Journal of Pharmacology

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“…Noradrenaline was eluted with 20 ml of 3 N sodium acetate buffer (pH 6.0) containing 7% ethanol and then dopamine was eluted with 15 ml of 3 N sodium acetate (pH 6.0) containing 25% ethanol. Noradrenaline in the eluate was determined by trihydroxyindole method (34) and dopamine by ethylendiamine condensing method (35).…”

mentioning

confidence: 99%

Function and Metabolism of Catecholamines in the Brain

Matsuoka

1964

Japanese Journal of Pharmacology

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“…Catecholamines were determined in the incubation medium and in the superfusate by ethylenediamine condensation method (13). When slices were incubated with 32P and acetylcholine (0.1 mM) for 20 min (eserine was not added), 32P incorporation into PA, PC and PI were stimulated by acetylcholine (Fig.…”

Section: Methodsmentioning

confidence: 99%