Adrenoceptors in brain: cellular gene expression and effects on astrocytic metabolism and [Ca(2+)]i.
Recent in vivo studies have established astrocytes as a major target for locus coeruleus activation (Bekar et al., 2008), renewing interest in cell culture studies on noradrenergic effects on astrocytes in primary cultures and calling for additional information about the expression of adrenoceptor subtypes on different types of brain cells. In the present communication, mRNA expression of alpha(1)-, alpha(2)- and beta-adrenergic receptors and their subtypes was determined in freshly isolated, cell marker-defined populations of astrocytes, NG2-positive cells, microglia, endothelial cells, and Thy1-positive neurons (mainly glutamatergic projection neurons) in murine cerebral cortex. Immediately after dissection of frontal, parietal and occipital cortex of 10-12-week-old transgenic mice, which combined each cell-type marker with a specific fluorescent signal, the tissue was digested, triturated and centrifuged, yielding a solution of dissociated cells of all types, which were separated by fluorescence-activated cell sorting (FACS). mRNA expression in each cell fraction was determined by microarray analysis. alpha(1A)-Receptors were unequivocally expressed in astrocytes and NG2-positive cells, but absent in other cell types, and alpha(1B)-receptors were not expressed in any cell population. Among alpha(2)-receptors only alpha(2A)-receptors were expressed, unequivocally in astrocytes and NG-positive cells, tentatively in microglia and questionably in Thy1-positive neurons and endothelial cells. beta(1)-Receptors were unequivocally expressed in astrocytes, tentatively in microglia, and questionably in neurons and endothelial cells, whereas beta(2)-adrenergic receptors showed tentative expression in neurons and astrocytes and unequivocal expression in other cell types. This distribution was supported by immunochemical data and its relevance established by previous studies in well-differentiated primary cultures of mouse astrocytes, showing that stimulation of alpha(2)-adrenoceptors increases glycogen formation and oxidative metabolism, the latter by a mechanism depending on intramitochondrial Ca(2+), whereas alpha(1)-adrenoceptor stimulation enhances glutamate uptake, and beta-adrenoceptor activation causes glycogenolysis and increased Na(+), K(+)-ATPase activity. The Ca(2+)- and cAMP-mediated association between energy-consuming and energy-yielding processes is emphasized.
Hertz
Leif
L
Department of Clinical Pharmacology, College of Basic Medical Sciences, China Medical University, Shenyang, PR China. leifhertz@xplornet.ca
Lovatt
Ditte
D
Goldman
Steven A
SA
Nedergaard
Maiken
M
D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D001923:Brain Chemistry / Q000235:genetics* / Q000502:physiology*; D020013:Calcium Signaling / Q000502:physiology*; D002469:Cell Separation; D002478:Cells, Cultured; D005434:Flow Cytometry; D015870:Gene Expression / Q000502:physiology; D005947:Glucose / Q000378:metabolism; D006003:Glycogen / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic / Q000502:physiology; D046228:Microarray Analysis; D008928:Mitochondria / Q000378:metabolism; D010084:Oxidation-Reduction; D019289:Pyruvic Acid / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D011941:Receptors, Adrenergic / Q000096:biosynthesis* / Q000235:genetics*; D018340:Receptors, Adrenergic, alpha-1 / Q000096:biosynthesis / Q000235:genetics; D018341:Receptors, Adrenergic, alpha-2 / Q000096:biosynthesis / Q000235:genetics; D011943:Receptors, Adrenergic, beta / Q000096:biosynthesis / Q000235:genetics
D011941:Receptors, Adrenergic; D018340:Receptors, Adrenergic, alpha-1; D018341:Receptors, Adrenergic, alpha-2; D011943:Receptors, Adrenergic, beta; D012313:RNA; D019289:Pyruvic Acid; D006003:Glycogen; D005947:Glucose