Biochemical analysis of myelin lipids and proteins in a model of methyl donor pathway deficit: effect of S-adenosylmethionine.
S-Adenosylmethionine (SAMe) is the methyl donor to numerous acceptor molecules. We used cycloleucine (CL), which prevents the conversion of methionine to SAMe by inhibiting ATP-l-methionine-adenosyltransferase (MAT), to characterize the lipid and protein changes induced in peripheral nerve and brain myelin in rats during development. We also investigated the effect of exogenous SAMe by administering SAMe-1,4-butane disulfonate (SAMe-SD4). CL was given on days 7, 8, 12, and 13 and SAMe-SD4 was given daily from day 7; the animals were killed on day 18. CL accumulates in the brain reaching a concentration within 24 h compatible with its ID(50) in vitro and interacting with methionine metabolism; brain MAT activity and SAMe levels were lower and methionine levels higher than in controls. CL significantly reduced brain and nerve weight gains, brain myelin content, proteins, phospholipids, and galactolipids. Among phospholipids in nerve and brain, only sphingomyelin was significantly increased, by 35-50%. Sciatic nerve protein analyses showed some significant changes: protein zero in sciatic nerve remained unchanged but the 14.0- and 18.5-kDa isoforms of myelin basic protein showed a dramatic increase. Among the main proteins, in purified brain myelin, the proteolipid protein and dimer-20 isoform decreased after CL. SAMe-SD4 highlights some sensitive parameters by counteracting, at least partially, some alterations of PL--particularly galactolipids and sphingomyelins--and proteins induced by CL. The partial beneficial effects might also be explained by the age-related limited bioavailability of exogenous SAMe, a finding, to our knowledge, not yet reported elsewhere. This study demonstrates that availability of methyl donors is closely related to the formation of myelin components.
Bianchi
R
R
Department of Molecular Biochemistry and Pharmacology, Istituto di Ricerche Farmacologiche Mario Negri, Milan, 20157, Italy.
Calzi
F
F
Savaresi
S
S
Sciarretta-Birolo
R
R
Bellasio
R
R
Tsankova
V
V
Tacconi
M T
MT
D000818:Animals; D000833:Animals, Suckling; D015153:Blotting, Western; D001923:Brain Chemistry / Q000187:drug effects; D002073:Butanes / Q000494:pharmacology; D003515:Cycloleucine / Q000493:pharmacokinetics; D003720:Densitometry; D038983:Galactolipids; D006017:Glycolipids / Q000032:analysis; D007274:Injections, Intraperitoneal; D008745:Methylation; D008970:Molecular Weight; D004676:Myelin Basic Protein / Q000032:analysis* / Q000378:metabolism*; D009186:Myelin Sheath / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D012436:S-Adenosylmethionine / Q000494:pharmacology*; D012584:Sciatic Nerve / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D013109:Sphingomyelins / Q000032:analysis*; D013451:Sulfonic Acids / Q000494:pharmacology
D002073:Butanes; D038983:Galactolipids; D006017:Glycolipids; D004676:Myelin Basic Protein; D013109:Sphingomyelins; D013451:Sulfonic Acids; D003515:Cycloleucine; C046888:butane; D012436:S-Adenosylmethionine