Phosphatidylcholine synthesis is elevated in neuronal models of Gaucher disease due to direct activation of CTP:phosphocholine cytidylyltransferase by glucosylceramide.
Glucosylceramide (GlcCer) accumulates in the inherited metabolic disorder, Gaucher disease, because of the defective activity of lysosomal glucocerebrosidase. We previously demonstrated that upon GlcCer accumulation, cultured hippocampal neurons exhibit modified growth patterns, altered endoplasmic reticulum density, and altered calcium release from intracellular stores. We here examined the relationship between GlcCer accumulation and phospholipid synthesis. After treatment of neurons with an active site-directed inhibitor of glucocerebrosidase, or in neurons obtained from a mouse model of Gaucher disease, [14C]methyl choline incorporation into [14C]phosphatidylcholine ([14C]PC) and [14C]sphingomyelin was elevated, as were [14C]CDP-choline levels, suggesting that CTP:phosphocholine cytidylyltransferase (CCT) is activated. Indeed, CCT activity was elevated in neurons that had accumulated GlcCer. GlcCer, but not galactosylceramide (GalCer), stimulated CCT activity in rat brain homogenates, and significantly higher levels of CCT were membrane associated in cortical homogenates from a mouse model of Gaucher disease compared with wild-type mice. Because CCT mRNA and protein levels were unaltered in either neurons or brain tissue that had accumulated GlcCer, it appeared likely that GlcCer activates CCT by a post-translational mechanism. This was verified by examination of the effect of GlcCer on CCT purified about 1200-fold from rat brain. GlcCer stimulated CCT activity, with stimulation observed at levels as low as 2.5 mol% and with maximal activation reached at 10 mol%. In contrast, GalCer had no effect. Together, these data demonstrate that GlcCer directly activates CCT, which results in elevated PC synthesis, which may account for some of the changes in growth rates observed upon neuronal GlcCer accumulation.
Bodennec
Jacques
J
Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel.
Pelled
Dori
D
Riebeling
Christian
C
Trajkovic
Selena
S
Futerman
Anthony H
AH
D000818:Animals; D001369:Axons / Q000378:metabolism; D001921:Brain / Q000187:drug effects / Q000201:enzymology; D002250:Carbon Radioisotopes; D002455:Cell Division; D002794:Choline / Q000031:analogs & derivatives* / Q000378:metabolism; D019916:Choline-Phosphate Cytidylyltransferase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005699:Galactosylceramides / Q000494:pharmacology; D005776:Gaucher Disease / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D005838:Genotype; D005962:Glucosylceramidase / Q000235:genetics / Q000378:metabolism; D005963:Glucosylceramides / Q000378:metabolism* / Q000494:pharmacology; D007294:Inositol / Q000031:analogs & derivatives* / Q000494:pharmacology; D050356:Lipid Metabolism; D051379:Mice; D009154:Mutation; D009474:Neurons / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D051381:Rats
D002250:Carbon Radioisotopes; D004791:Enzyme Inhibitors; D005699:Galactosylceramides; D005963:Glucosylceramides; D010713:Phosphatidylcholines; C017506:methylcholine; D007294:Inositol; D019916:Choline-Phosphate Cytidylyltransferase; D005962:Glucosylceramidase; D002794:Choline; C011423:conduritol epoxide