Stable transfection of GM1 synthase gene into GM1-deficient NG108-15 cells, CR-72 cells, rescues the responsiveness of Trk-neurotrophin receptor to its ligand, NGF.
Previous studies from this laboratory and others have suggested the evidences that acidic glycosphingolipid, ganglioside GM1 (GM1), is an endogenous regulator of high affinity nerve growth factor receptor, Trk, which is an essential factor for the normal development and differentiation of neuronal cells by forming a complex with Trk. The present study was aimed to examine whether Trk expressed in cells that are deficient in endogenous GM1 due to the mutation of GM1 synthase gene (NG-CR72 cells) is responsive to its ligand nerve growth factor and how genetic restoration of GM1 synthase gene by a stable transfection of the gene affects the function of the Trk protein. The data clearly showed that (1) confocal lazor microscopic studies disclosed NG-CR72 cells are really deficient in GM1, (2) stable transfection of GM1 synthase cDNA into these cells (NG-CR72G cells) restores the expression of GM1 in the cells, and (3) Trk protein is expressed in NG-CR72 cells but its location seemed not to be on the plasma membrane, whereas we clearly observed that the Trk protein is expressed on the plasma membrane in NG-CR72G cells. (4) NGF did not elicit the autophosphorylation of the Trk protein in GM1 deficient NG-CR72 cells but did elicit the activation of the Trk protein in NG-CR72G cells with an activation of mitogen activated protein kinase. These studies strongly suggested that GM1 is necessary for the normal expression of the Trk protein function and for normal targeting of the Trk protein to the plasma membrane.
Mutoh
Tatsuro
T
2nd Department of Internal Medicine, Faculty of Medicine, Fukui Medical University, Japan. mutoh@fmsra.fukui-med.ac.jp
Hamano
Tadanori
T
Yano
Shigeaki
S
Koga
Hiroshi
H
Yamamoto
Hiroko
H
Furukawa
Koichi
K
Ledeen
Robert W
RW
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D006602:Hexosyltransferases / Q000235:genetics*; D018613:Microscopy, Confocal; D020932:Nerve Growth Factor / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding; D051381:Rats; D020917:Receptor, trkA / Q000378:metabolism*; D014162:Transfection
D020932:Nerve Growth Factor; C014866:G(M1)ganglioside synthetase; D006602:Hexosyltransferases; D020917:Receptor, trkA