Cationic lipid-mediated NGF gene transfection increases neurofilament phosphorylation.
We examined the effect of cationic lipid-mediated gene transfection of nerve growth factor (NGF) in primary septo-hippocampal cell cultures. Rat NGF cDNA was subcloned into a pUC19-based plasmid containing a CMV promoter. Two days after NGF gene transfection in primary cell cultures, ELISA confirmed increases in NGF protein secretion from transfected cells. To study the biological effect of cationic lipid-mediated NGF gene transfection, we analyzed the amount of neurofilament protein from NGF-transfected cell cultures. Western blot and immunohistochemical analyses detected significant increases in the phosphorylated form of neurofilament proteins in the cultures after cationic lipid-mediated NGF cDNA transfection. Cationic lipid-mediated NGF cDNA transfection did rot cause significant changes in the total amount of neurofilament protein. Our studies suggest that cationic lipid-mediated NGF gene transfection can increase neurofilament phosphorylation but not total neurofilament protein.
Yang
K
K
Department of Neurosurgery, University of Texas Health Sciences Center at Houston 77030, USA.
Xue
J J
JJ
Qiu
Y H
YH
Perez-Polo
J R
JR
Whitson
J
J
Faustinella
F
F
Kampfl
A
A
Zhao
X
X
Iwamoto
Y
Y
Mu
X S
XS
Clifton
G
G
Hayes
R L
RL
D000818:Animals; D002478:Cells, Cultured; D007150:Immunohistochemistry; D008055:Lipids / Q000494:pharmacology*; D009414:Nerve Growth Factors / Q000235:genetics* / Q000378:metabolism; D016900:Neurofilament Proteins / Q000187:drug effects*; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection / Q000187:drug effects*
D008055:Lipids; D009414:Nerve Growth Factors; D016900:Neurofilament Proteins