Ischemia-induced changes in catecholamine release and their mechanisms: a study using cultured bovine adrenal chromaffin cells.
Ischemia-induced changes in neurotransmitter release and their mechanisms were examined using cultured bovine adrenal chromaffin cells. When the cells were incubated in glucose-free media equilibrated with 0% O2/100% N2 (ischemia), ATP content decreased and reached the minimum level within 40 min. Control incubation was done in media equilibrated with 21% O2 in N2. After 10-min incubation under ischemic conditions, basal catecholamine (CA) release was elevated and the elevation persisted up to 90 min. High K(+)-evoked CA release was transiently enhanced at 10 min, but after that, it decreased to reach the minimum level at 60 min. At 10 min, cytosolic free Ca2+ concentration ([Ca2+]i) and 45Ca2+ uptake of the resting cells (basal values) and high K(+)-evoked increases in these two parameters were unchanged, but CA release from permeabilized cells in response to Ca2+ in media was augmented. After 60-min incubation under ischemic conditions, basal [Ca2+]i was elevated: the elevation was observed even in the absence of extracellular Ca2+. In contrast, high K(+)-evoked increases in [Ca2+]i and in 45Ca2+ uptake were suppressed, but basal 45Ca2+ uptake into intact cells and CA release from permeabilized cells were unchanged. These results suggest that in an early phase (10 min) of ischemia, both basal and stimulation-evoked CA release are augmented because of increased sensitivity of exocytotic machinery to Ca2+. In the late phase (60 min), basal CA release is augmented because of an increase in basal [Ca2+]i, which is due to accumulation of Ca2+ derived from intracellular Ca2+ pools: stimulation-evoked CA release is suppressed because of inhibition of stimulation-evoked increase in [Ca2+]i, which is due to functional disturbance of voltage-dependent Ca2+ channels.
Department of Neurosurgery, Kyoto University Faculty of Medicine, Japan.
A
A
Itoh
S
S
Miwa
K
K
Koshimura
Y
Y
Akiyama
Y
Y
Takagi
S
S
Yamagata
H
H
Kikuchi
T
T
Masaki
D000255:Adenosine Triphosphate / Q000378:metabolism; D000313:Adrenal Medulla / Q000187:drug effects / Q000378:metabolism*; D000704:Analysis of Variance; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002395:Catecholamines / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D007511:Ischemia / Q000378:metabolism*; D007700:Kinetics; D008954:Models, Biological; D011188:Potassium / Q000494:pharmacology; D011336:Probability
D002395:Catecholamines; D000255:Adenosine Triphosphate; D011188:Potassium; D002118:Calcium