Astrocyte beta1-adrenergic receptor immunoreactivity and agonist induced increases in [Ca2+]i: differential results indicative of a modified membrane receptor.
Antibodies against the C-terminus of the beta1-adrenergic receptor were used for staining cultured astrocytes from the rat cerebral cortex. Immunoreactivity was found to be localized exclusively to an intracellular organelle structure similar to the Golgi complex, with no staining of the plasma membrane. The astrocytes stained positive with BODIPY CGP 12177, a FITC-conjugated beta-adrenergic receptor agonist, and this staining was blocked by the beta1-adrenergic antagonist atenolol, indicating that these receptors are expressed on the surface of the astrocytes. The presence of functional plasma membrane beta1-adrenergic receptors was further verified using microspectrofluorometry for measurements of intracellular calcium changes upon beta-adrenergic agonist stimulation. Intracellular immunoreactivity confined to the organelles was also found in astrocytes from mixed astroglial-neuronal cultures. In contrast, the neurons in these cultures showed a strong labeling of the cell bodies by the beta1-adrenergic receptor antibodies. Thus, the beta1-adrenergic receptor antibody, which stains the cell bodies of the neurons, recognizes the astroglial receptors only intracellularly, although functional beta1-adrenergic receptors are present on the astroglial surface. Taken together, these data suggest that the beta1-adrenergic receptors observed intracellularly might be processed on their passage to the surface to a modified form of the final plasma membrane receptor, which is not recognized by the antibodies.
Thorlin
T
T
Institute of Clinical Neuroscience, Göteborg University, Sweden. thorleif.thorlin@anatcell.gu.se
Anders
P
P
Persson
I
I
Eriksson
P S
PS
Rönnbäck
L
L
Hansson
E
E
D000316:Adrenergic alpha-Agonists / Q000494:pharmacology*; D058665:Adrenergic beta-1 Receptor Agonists; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D018920:Coculture Techniques; D005456:Fluorescent Dyes; D007150:Immunohistochemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D018342:Receptors, Adrenergic, beta-1 / Q000378:metabolism*
D000316:Adrenergic alpha-Agonists; D058665:Adrenergic beta-1 Receptor Agonists; D005456:Fluorescent Dyes; D018342:Receptors, Adrenergic, beta-1; D002118:Calcium