Differences in the stimulation of the phosphoinositide cycle by amine neurotransmitters in cultured rat forebrain neurones and astrocytes.
In this study, we compared the stimulation by carbachol (CCh), noradrenaline (NA), and histamine (HA) of phosphoinositide hydrolysis in rat forebrain neuronal and glial cultures. When Ca2+ was omitted from the stimulation buffer (low microM extracellular Ca2+), amine-induced [3H]inositol phosphate accumulation was reduced to a higher extent in astrocytes (70-80% for CCh and NA and 100% for HA) than in neurones (around 50-60% for all the amines). Furthermore, guanosine 5'-[gamma-thio]trisphosphate (GTP[S]) stimulation of phosphoinositidase C (PIC) in membranes was 5-fold higher in neurones than in astrocytes. These results indicate differences in the mechanism of PIC stimulation in the two cell types. After 30 min stimulation in the presence of 10 mM Li+, a higher accumulation of [3H]inositol 4-monophosphate and [3H]inositol 1,4-bisphosphate than of [3H]inositol 1/3-monophosphate occurred for all agonists in neurones, whereas the opposite was observed in astrocytes. Moreover, in these cells stimulation for 5 min in the absence of Li+ produced a 2-3-fold accumulation of all metabolites of the 3-kinase pathway of inositol-1,4,5-trisphosphate metabolism but not of those of the 5-phosphatase pathway. Thus, regardless of the amine receptor stimulated, the 3-kinase route appeared to prevail in astrocytes and the 5-phosphatase pathway in neurones. The histamine response in neurones differed from that of the other agonists in that it rapidly declined. Taken together these results indicate that the heterogeneity in amine stimulation of the phosphoinositide cycle previously observed in brain slices could arise to a great extent from the cellular diversity of this preparation and be related to the differential contribution of the amine receptors located in neurones and astrocytes.
Oliva
A M
AM
Institut de Biologia Fonamental Vicent Villar Palasi and Departament de Bioquímica i Biologia Molecular, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Bas
N
N
García
A
A
D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology*; D002478:Cells, Cultured; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006632:Histamine / Q000494:pharmacology*; D009474:Neurons / Q000378:metabolism*; D009638:Norepinephrine / Q000494:pharmacology*; D010716:Phosphatidylinositols / Q000378:metabolism*; D016548:Prosencephalon / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
D010716:Phosphatidylinositols; D006632:Histamine; D006160:Guanosine Triphosphate; D002217:Carbachol; D002118:Calcium; D009638:Norepinephrine