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Effect of Lentiviral shRNA of Nogo Receptor on Rat Cortex Neuron Axon Outgrowth

Published online by Cambridge University Press:  02 December 2014

Shengming Xu
Affiliation:
Department of Orthopaedics, Shanghai Changzheng Hospital
Mingyuan Liu
Affiliation:
Department of Neurology, Shanghai Changzheng Hospital
Tao Zhang
Affiliation:
Department of Orthopaedics, Shanghai sixth People's Hospital, Shanghai
Bitao Lv
Affiliation:
Department of Orthopaedics, Shanghai Changzheng Hospital
Baifeng Liu
Affiliation:
Department of Orthopaedics, Daqing People's Hospital, Saertu Development Zone, Daqing City, China
Wen Yuan*
Affiliation:
Department of Orthopaedics, Shanghai Changzheng Hospital
*
Department of Orthopaedics, Shanghai Changzheng Hospital, 415 Fengyang Road, Shanghai, 200003, China
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Abstract:

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Background and Aims:

Axon growth is crucial for injured neural tissue to recover; however it is difficult to achieve in general. Axon outgrowth is inhibited by the activation of the Nogo receptor (NgR) by one of three different ligands. The present study aimed to suppress the inhibitory effect of the three inhibitory proteins to facilitate axon outgrowth.

Methods:

A lentiviral vector, siNgR199 (that has the capacity to interfere with the gene of NgR expression), was constructed for suppressing the gene transcription of NgR. Rat cortex neurons and oligodendrocytes were prepared to observe the effect of siNgR199 on facilitating axon outgrowth.

Results:

After transfection, the lentiviral siRNA of NgR remained in target neurons for almost two weeks whereas the conventional siRNA of NgR remained in neurons less than five days. Lentivirus-mediated delivery of exogenous small interfering RNA (siNgR199) targeting NgR significantly reduced the expression of this receptor and promoted axon outgrowth. In contrast, provision of naked siRNA targeting NgR (NgRsiRNA) showed less inhibitory effect on NgR protein expression and did not affect axon outgrowth.

Conclusions:

Lentiviral siRNA of NgR effectively suppresses the expression of NgR in cultured neurons that facilitates the axon outgrowth. The data implicate that lentiviral siRNA of NgR has therapeutic potential in facilitating the recovery of injured neural tissue.

Résumé:

Résumé:Contexte et objectif:

La croissance axonale est cruciale pour la guérison du tissu nerveux lésé. Cependant, elle est difficile à réaliser. La régénération axonale est inhibée par l'activation du récepteur Nogo (NgR) par l'un de trois ligands différents. Le but de cette étude était de supprimer l'effet inhibiteur des trois protéines inhibitrices pour faciliter la régénération axonale.

Méthodes:

Un vecteur lentiviral, siNgR199 (qui peut interférer avec l'expression du gène NgR), a été construit pour supprimer la transcription du gène NgR. Des neurones corticaux et des oligodendrocytes de rat ont été préparés pour observer l'effet de siNgR199 sur la régénération axonale.

Résultats:

Après transfection, l'ARNsi lentiviral de NgR est demeuré dans les neurones cibles pendant près de deux semaines alors que l'ARNsi conventionnel de NgR est demeuré dans les neurones moins de cinq jours. La livraison médiée par le lentivirus de petits ARNsi (siNgR199) exogènes interférents ciblant NgR a diminué significativement l'expression de ce récepteur et favorisé la régénération axonale. Par contre, l'ARNsi nu ciblant NgR a eu moins d'effet inhibiteur sur l'expression de la protéine NgR et n'a pas influencé la régénération axonale.

Conclusions:

L'ARNsi lentiviral de NgR supprime efficacement l'expression de NgR dans des neurones en culture, facilitant la régénération axonale. Ces données sont compatibles avec un effet thérapeutique potentiel de l'ARNsi lentiviral de NgR pour faciliter la récupération de tissus nerveux lésés.

Type
Original Article
Copyright
Copyright © Canadian Neurological Sciences Federation 2011

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