Downregulation of Nogo-A expression of oligodendrocytes in vitro with antisense oligodeoxynucleotides.
OBJECTIVE
To investigate the effect of antisense oligonucleotides (ODN) to Nogo-A mRNA expression in oligodendrocytes and to establish the base for further research of repair of optic nerve injury.


METHODS
(1) Oligodendrocytes were obtained by inoculating the optic nerve of newborn (2 days) rats and were identified with galactocerebroside(GC) antibody immunocytochemical stain. (2) In order to observe the effects of antisense ODN on cultured cells, we set up five groups, including the groups of three concentration of antisense Nogo-A ODN (2 microM, 5 microM, 10 microM), a group with the random sequence added to the medium and the control group. Reverse transcription-polymerase chain reaction (RT-PCR) was adopted to study the effects of ODN on the expression of Nogo-A in oligodendrocytes.


RESULTS
(1) Three days after inoculation, a few of round or fusiform shape cells migrated from optic nerve tissue; About 11 days later, the coverlips were completely covered by the cells; The cells identified with GC antibody immunocytochemical stain were positive cells. (2) The result of RT-PCR study showed that antisense Nogo-A ODN could significantly and specifically inhibit the expression of Nogo-A after 24 hours (P < 0.01). Random sequence has no effect on Nogo-A expression.


CONCLUSIONS
Antisense Nogo-A ODN can effectively and specifically inhibit the expression of Nogo-A.
Department of Ophthalmology, Daping Hospital, Research Institute of Surgery, Third Military Medical University, Chongqing 400042, China.
Xijia
X
Peng
Shaozhang
S
Liu
Jian
J
Ye
Rongdi
R
Yuan
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D015536:Down-Regulation; D009185:Myelin Proteins / Q000096:biosynthesis* / Q000235:genetics; D000070798:Nogo Proteins; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism*; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D009900:Optic Nerve / Q000166:cytology / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D009185:Myelin Proteins; D000070798:Nogo Proteins; D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; C000603197:Rtn4 protein, rat