Quantitative aspects of the acetylcholine-induced release of dopamine-β-hydroxylase and catecholamines from the bovine adrenal medulla

Serck‐Hanssen, Guldborg; Helle, Karen B.; Sommersten, B.; Pihl, K.E.

doi:10.1016/0306-3623(80)90070-1

Cited by 5 publications

(2 citation statements)

References 20 publications

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“…Inactivation of released DBH activity has been suggested to occur in Tyrode solution at 37 'C (half-inactivation time, 15 min) (Serck-Hanssen, Helle, Sommersten & Pihl, 1980). This is not the case in the present experiments, because the perifusion solution of the cells was changed with a half time of about 1-5 min, and the fractions were collected on ice.…”

Section: Discussionmentioning

confidence: 67%

Time course of release of catecholamine and other granular contents from perifused adrenal chromaffin cells of guinea‐pig.

Ito¹

1983

The Journal of Physiology

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SUMMARY1. Experiments were carried out to investigate the time course of the release of catecholamine, dopamine-,8-hydroxylase (DBH) and adenine nucleotides from isolated chromaffin cells of guinea-pig adrenal gland.2. When the isolated chromaffin cells were incubated with medium containing acetylcholine (ACh) (0-1 mM), veratridine (0-1 mM) or scorpion (Leiurus quinquestriatus) venom, (10 ,ug/ml.), catecholamine was released into the medium. Catecholamine secretion induced by veratridine or scorpion venom was inhibited by tetrodotoxin (1 gM) but not by atropine (0-1 mM) plus hexamethonium (041 mM). On the other hand, the secretary response to ACh was abolished by the cholinergic blocking drugs but not by tetrodotoxin.3. DBH was released together with catecholamine into the medium in which cells were suspended with these drugs. The ratio of catecholamine (n-mole) to DBH activity (n-mole/hr) appearing in the supernatant was 7-08 + 055, 6-60 + 0X27 and 8X91 + 0 47 for ACh, veratridine and scorpion venom, respectively. These values were close to that found in the lysate of chromaffin granules obtained from guinea-pig adrenal glands (7-37 + 0-39).4. The application of ACh or veratridine to perifused chromaffin cells was found to cause a parallel increase in catecholamine and DBH secretion in the perifusion medium without corresponding amounts of phenylethanolamine-N-methyltransferase leakage. However, DBH secretion tended to last for a longer period than catecholamine secretion.5. Adenine nucleotides were released from perifused chromaffin cells together with catecholamine, by ACh and veratridine. ATP added to the perifusion medium was metabolized to ADP and AMP, of which the ratio (ATP, 21-6 %; ADP, 34 %; AMP, 17-9 %) was close to those of adenine nucleotides released from the cells.6. The secretion ofadenine nucleotides induced by both secretagogues ceased much faster than the catecholamine secretion, so that the molar ratio of catecholamine to adenine nucleotides was gradually increased during and after stimulation. 7. The results indicate that catecholamine secretion is accompanied with a simultaneous release of DBH and ATP from adrenal chromaffin cells. Therefore, it is suggested that the delayed output of DBH, unlike catecholamine secretion, in perfused adrenal glands results from the presence of a diffusion barrier for this protein. The releasable secretary granules of isolated chromaffin cells are suggested to be heterogeneous with respect to the ratio of catecholamine to ATP.

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Section: Discussionmentioning

confidence: 67%

Time course of release of catecholamine and other granular contents from perifused adrenal chromaffin cells of guinea‐pig.

Ito¹

1983

The Journal of Physiology

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“…Dopamine-/I-hydroxylase (DBH) has been reported to be released with CA in the same proportion as is found in the lysate of isolated chromaffin granules from perfused adrenal glands of calf (Viveros, Arqueros & Kirshner, 1968), cat (Sorimachi & Yoshida, 1979) and guinea-pig (Ito, Nakazato & Ohga, 1980), and in isolated chromaffin cells of guineapig (Ito, 1983). However, in bovine adrenal glands, DBH activity released from chromaffin cells is inactivated in the plasma, and thus the ratio of DBH to CA in the evoked secretory response is markedly lower than that contained in chromaffin granules (Aunis, Serck-Hanssen & Helle, 1978;Serck-Hanssen, Helle, Sommersten & Pihl, 1980). Furthermore, no proportional release of CA and DBH is obtained in response to splanchnic nerve stimulation or ACh in perfused cat adrenal glands (Dixon, Garcia & Kirpekar, 1975) or in adrenal glands of conscious calves (Edwards, Furness & Helle, 1980).…”

Section: Introductionmentioning

confidence: 88%

CATECHOLAMINES AND DOPAMINE‐β‐HYDROXYLASE SECRETION FROM PERFUSED GOAT ADRENAL GLANDS

Yamada

Nakazato

et al. 1988

Exp Physiol

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SUMMARYSplanchnic nerve stimulation caused a frequency-dependent increase in catecholamine (CA) secretion which attained a maximum at 10 Hz in perfused goat adrenal glands. Acetylcholine (ACh) and nicotine also caused a dose-dependent increase in CA secretion. The percentage of three catecholamines released (adrenaline, 51-56 %; noradrenaline, 42-47 %; dopamine, 1 3-2 %) were not significantly different among three different stimuli. These values were almost the same as those found in the extract of adrenal gland and in purified chromaffin granules. The response to ACh was completely abolished by hexamethonium alone. Pilocarpine was ineffective in increasing CA secretion. Dopamine-,?-hydroxylase (DBH) activity increased with the increase in CA secretion in response to splanchnic nerve stimulation and ACh, though the declining phase of DBH was much slower than that of CA. The ratios of CA to DBH activity released for 25 min during and after splanchnic nerve stimulation and ACh infusion were 0 40 + 0-07 and 0 50 + 0-07, respectively. The ratio was 0 41 + 0-06 in chromaffin granules.The results indicate that only nicotinic receptors are involved in CA secretion and that DBH may be useful as an index of exocytosis in goat perfused adrenal glands.

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Occurrence and Mechanism of Exocytosis in Adrenal Medulla and Sympathetic Nerve

Winkler¹

1988

Handbook of Experimental Pharmacology

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Quantitative aspects of the acetylcholine-induced release of dopamine-β-hydroxylase and catecholamines from the bovine adrenal medulla

Cited by 5 publications

References 20 publications

Time course of release of catecholamine and other granular contents from perifused adrenal chromaffin cells of guinea‐pig.

Time course of release of catecholamine and other granular contents from perifused adrenal chromaffin cells of guinea‐pig.

CATECHOLAMINES AND DOPAMINE‐β‐HYDROXYLASE SECRETION FROM PERFUSED GOAT ADRENAL GLANDS

Occurrence and Mechanism of Exocytosis in Adrenal Medulla and Sympathetic Nerve

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