Rescue of injury-induced neurofilament loss by BDNF gene transfection in primary septo-hippocampal cell cultures.
We employed primary septo-hippocampal cell cultures to determine the ability of liposome-mediated BDNF gene transfection to facilitate recovery of neurofilament loss caused by depolarization injury. After BDNF gene transfection in uninjured cultures, RT-PCR and immunohistochemical staining confirmed increases in BDNF mRNA and protein in transfected cells. Three days after depolarization injury, Western blot and immunohistochemical analyses detected significant loss of neurofilament proteins in non-transfected cultures, while BDNF transfection produced marked increases in neurofilament proteins following either pre-injury transfection or transfection 24 h following injury. Immunohistochemical studies also detected enhanced immunolabeling of BDNF and total neurofilament protein (phosphorylated and non-phosphorylated) in injured neurons following BDNF transfection or administration of exogenous BDNF protein, compared to untransfected, injured controls.
Hayes
R L
RL
Department of Neurosurgery, University of Texas Houston Health Science Center 77030, USA.
Yang
K
K
Whitson
J S
JS
Xue
J J
JJ
Kampfl
A
A
Mu
X S
XS
Zhao
X
X
Faustinella
F
F
Clifton
G L
GL
D000818:Animals; D015153:Blotting, Western; D019208:Brain-Derived Neurotrophic Factor; D002478:Cells, Cultured; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism*; D007150:Immunohistochemistry; D009414:Nerve Growth Factors / Q000096:biosynthesis / Q000235:genetics*; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics*; D009454:Neurofibrils / Q000502:physiology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D012194:RNA-Directed DNA Polymerase; D051381:Rats; D014162:Transfection
D019208:Brain-Derived Neurotrophic Factor; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D012194:RNA-Directed DNA Polymerase