Reduced phospholipase C-beta activity and isoform expression in the cerebellum of TS65Dn mouse: a model of Down syndrome.
Agonist- and guanine-nucleotide-stimulated phospholipase C-beta (PLC) activity was characterized in crude plasma membrane preparations from cerebral cortex, hippocampus and cerebellum of Ts65Dn mice, a model for Down syndrome, and their control littermates. The levels of expression of PLC-beta((1-4)) isoforms and G-protein alpha(q/11) subunits were also quantified by Western blot analysis to establish their contribution to the patterns of PLC functioning. PLC activity regulated by G-proteins and muscarinic and 5-HT(2) receptors presented a regional distribution in both control and Ts65Dn mice. In both groups of mice, the intensity of PLC responses to maximal activation by calcium followed the sequence cerebellum > cortex > hippocampus. Both basal and maximal PLC activities, however, were significantly lower in cerebellar membranes of Ts65Dn than in control mice. This difference was mostly revealed in crude plasma membranes prepared from cerebellum at the level of G-protein-dependent-PLC activity because the concentration-response curve to GTPgammaS showed a reduction of the maximal effect in Ts65Dn mice, with no change in sensitivity (EC(50)). Western blot analysis showed a heterogeneous distribution of PLC-beta((1-4)) isoforms in both groups of mice. The levels of PLC-beta4 isoform, however, were significantly lower in the cerebellum of Ts65Dn than in control mice. We conclude that the cerebellum of Ts65Dn mice has severe deficiencies in PLC activity stimulated by guanine nucleotides, which are specifically related to a lower level of expression of the PLC-beta4 isoform, a fact that may account for the neurological phenotype observed in this murine model of Down syndrome.
Ruiz de Azúa
I
I
Department of Pharmacology, Faculty of Pharmacy, University of the Basque Country, Alava, Spain.
Lumbreras
M A
MA
Zalduegui
A
A
Baamonde
C
C
Dierssen
M
M
Flórez
J
J
Sallés
J
J
D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D002531:Cerebellum / Q000187:drug effects / Q000201:enzymology* / Q000503:physiopathology; D002540:Cerebral Cortex / Q000201:enzymology / Q000503:physiopathology; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004314:Down Syndrome / Q000201:enzymology* / Q000235:genetics / Q000503:physiopathology; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics*; D019204:GTP-Binding Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000235:genetics*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000493:pharmacokinetics; D007527:Isoenzymes / Q000172:deficiency* / Q000235:genetics; D008297:Male; D051379:Mice; D008818:Mice, Neurologic Mutants; D018721:Muscarinic Agonists / Q000494:pharmacology; D018727:Muscarinic Antagonists / Q000494:pharmacology; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000235:genetics / Q000378:metabolism / Q000493:pharmacokinetics; D054799:Phospholipase C beta; D020033:Protein Isoforms / Q000172:deficiency / Q000235:genetics; D044402:Receptor, Serotonin, 5-HT2A; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000378:metabolism; D011985:Receptors, Serotonin / Q000187:drug effects / Q000378:metabolism; D012702:Serotonin Antagonists / Q000494:pharmacology; D017366:Serotonin Receptor Agonists / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D013347:Subcellular Fractions / Q000187:drug effects / Q000201:enzymology; D014316:Tritium / Q000493:pharmacokinetics; D010738:Type C Phospholipases / Q000172:deficiency* / Q000235:genetics
D007527:Isoenzymes; D018721:Muscarinic Agonists; D018727:Muscarinic Antagonists; D019269:Phosphatidylinositol 4,5-Diphosphate; D020033:Protein Isoforms; D044402:Receptor, Serotonin, 5-HT2A; D011976:Receptors, Muscarinic; D011985:Receptors, Serotonin; D012702:Serotonin Antagonists; D017366:Serotonin Receptor Agonists; D014316:Tritium; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D010738:Type C Phospholipases; D054799:Phospholipase C beta; C518043:Plcb4 protein, mouse; D019204:GTP-Binding Proteins; D002118:Calcium