Regulation and glucocorticoid-independent induction of lipocortin I in cultured astrocytes.
Stimulation of prostaglandin (PG) release in rat astroglial cultures by various substances, including phorbol esters, melittin, or extracellular ATP, has been reported recently. It is shown here that glucocorticoids (GCs) reduced both basal and stimulated PGD2 release. Hydrocortisone, however, did not inhibit ATP-, calcium ionophore A23187-, or tetradecanoyl phorbol acetate (TPA)-stimulated arachidonic acid release, and only TPA stimulations were affected by dexamethasone. GC-mediated inhibition of PGD2 release thus appeared to exclude regulation at the phospholipase A2 (PLA2) level. Therefore, the effects of GCs on the synthesis of lipocortin I (LC I), a potent, physiological inhibitor of PLA2, were studied in more detail. Dexamethasone was not able to enhance de novo synthesis of LC I in freshly seeded cultures and failed to increase LC I synthesis in 2-3-week-old cultures. It is surprising that LC I was the major LC synthesized in those cultures, and marked amounts accumulated with culture time, reaching plateau levels at approximately day 10. In contrast, LC I was barely detectable in vivo. This tonic inhibition of PLA2 is the most likely explanation for unsuccessful attempts to evoke PG release in astrocyte cultures by various physiological stimuli. GC receptor antagonists (progesterone and RU 38486) given throughout culture time reduced LC I accumulation and simultaneously increased PGD2 release. Nonetheless, a substantial production of LC I persisted in the presence of antagonists. Therefore, LC I induction did not seem to involve GC receptor activation. This was confirmed in serum- and GC-free brain cell aggregate cultures. Here also a marked accumulation of LC I was observed.(ABSTRACT TRUNCATED AT 250 WORDS)
Gebicke-Haerter
P J
PJ
Institute of Pharmacology, University of Freiburg, F.R.G.
Schobert
A
A
Dieter
P
P
Honegger
P
P
Hertting
G
G
D000818:Animals; D017302:Annexins; D001253:Astrocytes / Q000378:metabolism*; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002449:Cell Aggregation; D002478:Cells, Cultured; D005938:Glucocorticoids / Q000494:pharmacology*; D010740:Phospholipases / Q000037:antagonists & inhibitors; D015230:Prostaglandin D2 / Q000037:antagonists & inhibitors; D011965:Receptors, Glucocorticoid / Q000037:antagonists & inhibitors; D013997:Time Factors; D015854:Up-Regulation
D017302:Annexins; D002135:Calcium-Binding Proteins; D005938:Glucocorticoids; D011965:Receptors, Glucocorticoid; D010740:Phospholipases; D015230:Prostaglandin D2