Receptor subtype involved and mechanism of norepinephrine-induced stimulation of glutamate uptake into primary cultures of rat brain astrocytes.
Glutamate uptake into rat brain astrocytes is potently stimulated by addition of norepinephrine (NE). This effect is mediated by alpha 1-adrenergic receptors expressed by these cells (Hansson and Rönnbäck: Life Sci 44:27, 1989; Brain Res 548:215, 1991). The present study was undertaken in order to identify the adrenergic receptor subtype involved, and to determine the sequence of events following receptor activation. NE increased glutamate uptake rates in a dose- and time-dependent manner (EC50 = 6 microM). Both, the selective alpha 1-receptor antagonist prazosin (IC50 = 2.5 microM) and the alpha 1b-adrenergic receptor subtype specific alkylating agent chloroethyl-clonidine (CEC, 100 microM) prevented NE (100 microM) evoked stimulation of glutamate uptake. Furthermore, omission of Ca2+ from the extracellular medium had no significant influence on NE-induced increase in glutamate uptake, indicating that the stimulatory effect is mediated by alpha 1b-adrenergic receptors. Treatment of cells with pertussis toxin (PTX) for 24 h or with 12-O-tetradecanoylphorbol-13-acetate (TPA) for 30-45 min prior to NE addition abolished the NE-mediated effect on glutamate uptake. Addition of TPA alone resulted in a rapid increase of glutamate uptake, which declined to control levels when TPA was applied 30 min prior to uptake initiation by glutamate. The increase in glutamate uptake elicited by TPA and NE added at the same time showed no additivity of the stimulatory effect resulting from treatment with each agent alone.(ABSTRACT TRUNCATED AT 250 WORDS)
Fahrig
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Department of Experimental Neurology, Troponwerke GmbH&Co.KG, Köln, Federal Republic of Germany.
D027322:Amino Acid Transport System X-AG; D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D001464:Barium / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects; D001921:Brain / Q000166:cytology; D002478:Cells, Cultured; D003000:Clonidine / Q000031:analogs & derivatives / Q000494:pharmacology; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D006023:Glycoproteins / Q000378:metabolism*; D009638:Norepinephrine / Q000494:pharmacology*; D037342:Pertussis Toxin; D051381:Rats; D011942:Receptors, Adrenergic, alpha / Q000145:classification / Q000187:drug effects* / Q000378:metabolism; D015290:Second Messenger Systems / Q000187:drug effects; D013268:Stimulation, Chemical; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D027322:Amino Acid Transport System X-AG; D005971:Glutamates; D006023:Glycoproteins; D011942:Receptors, Adrenergic, alpha; D010566:Virulence Factors, Bordetella; D001464:Barium; D018698:Glutamic Acid; C051731:chlorethylclonidine; D037342:Pertussis Toxin; D003000:Clonidine; D013755:Tetradecanoylphorbol Acetate; D009638:Norepinephrine