title: Lentiviral-Mediated Netrin-1 Overexpression Improves Motor and Sensory Functions in SCT Rats Associated with SYP and GAP-43 Expressions.
abstract: Spinal cord injury (SCI), as a major cause of disability, usually causes serious loss of motor and sensory functions. As a bifunctional axonal guidance cue, netrin-1 can attract axons via the deleted in colorectal cancer (DCC) receptors and repelling others via Unc5 receptors, but its exact role in the recovery of motor and sensory function has not well been studied, and the mechanisms remains elusive. The aim of this experiment is to determine whether lentiviral (LV)-mediated overexpression of netrin-1 or RNA interference (RNAi) can regulate the functional recovery in rats subjected to spinal cord transection (SCT). Firstly, two lentiviral vectors including Lv-exNtn-1 (netrin-1 open reading frame (ORF)) and Lv-shNtn-1 (netrin-1 sh) were constructed and injected into spinal cords rostral and caudal to the transected lesion site. Overexpressing netrin-1 enhanced significantly locomotor function, and reduced thermal and mechanical stimuli in vivo, compared with the control, while silencing netrin-1 did not significantly change the situation. Western blot and immunostaining analysis confirmed that netrin-1 ORF treatment not only effectively increased the expression level of netrin-1, also up-regulated the level of synaptophysin (SYP) in spinal cord rostral to the lesion, but also enhanced growth-associated protein-43 (GAP-43) expression in spinal cord caudal to the lesion site. Comparatively, knockdown of netrin-1 did not give rise to positive findings in our experimental condition. These findings therefore pointed that Lv-mediated netrin-1 overexpression could promote motor and sensory functional recoveries following SCT, and the underlying mechanisms were associated with SYP and GAP-43 expressions. The present study therefore provided a novel strategy for the treatment of SCI and explained the possible mechanisms for the functional improvement.
authors.affiliation: Institute of Neuroscience and Department of Anatomy, Kunming Medical University, Kunming, 650000, China.
authors.forename: Xue Fei
authors.initials: XF
authors.lastname: Han
authors.affiliation: Institute of Neuroscience and Department of Anatomy, Kunming Medical University, Kunming, 650000, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Anesthesia, Institute of Neurological Disease, Translational Neuroscience Center, West China Hospital, Sichuan University, Chengdu, 610041, People's Republic of China.
authors.forename: Liu Lin
authors.initials: LL
authors.lastname: Xiong
authors.affiliation: Department of Anesthesia, Institute of Neurological Disease, Translational Neuroscience Center, West China Hospital, Sichuan University, Chengdu, 610041, People's Republic of China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Institute of Neuroscience and Department of Anatomy, Kunming Medical University, Kunming, 650000, China.
authors.forename: Piao
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Department of Anesthesia, Institute of Neurological Disease, Translational Neuroscience Center, West China Hospital, Sichuan University, Chengdu, 610041, People's Republic of China. xiaqj2005@126.com.
authors.forename: Qing Jie
authors.initials: QJ
authors.lastname: Xia
authors.affiliation: Department of Anesthesia, Institute of Neurological Disease, Translational Neuroscience Center, West China Hospital, Sichuan University, Chengdu, 610041, People's Republic of China. tinghua_neuron@263.net.
authors.forename: Ting Hua
authors.initials: TH
authors.lastname: Wang
authors.affiliation: Institute of Neuroscience and Department of Anatomy, Kunming Medical University, Kunming, 650000, China. byc71206@163.com.
authors.forename: Ying Chun
authors.initials: YC
authors.lastname: Ba
mesh_terms: D000818:Animals; D019054:Evoked Potentials, Motor / Q000187:drug effects / Q000502:physiology*; D005260:Female; D019922:GAP-43 Protein / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D016086:Lentivirus; D000075388:Netrin-1 / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D016716:PC12 Cells; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D020127:Recovery of Function / Q000187:drug effects / Q000502:physiology; D012677:Sensation / Q000187:drug effects / Q000502:physiology*; D013119:Spinal Cord Injuries / Q000235:genetics / Q000378:metabolism* / Q000628:therapy; D016708:Synaptophysin / Q000096:biosynthesis* / Q000235:genetics; D013904:Thoracic Vertebrae
chemical_list: D019922:GAP-43 Protein; D016708:Synaptophysin; C569016:Syp protein, rat; D000075388:Netrin-1
keywords: GAP-43; Netrin-1; Neurobehavior; RNA interference and overexpression; Rats; SYP; Spinal cord transection