title: Netrin-1 rescues neuron loss by attenuating secondary apoptosis in ipsilateral thalamic nucleus following focal cerebral infarction in hypertensive rats.
abstract: Neurological deficit following cerebral infarction correlates with not only primary injury, but also secondary neuronal apoptosis in remote loci connected to the infarction. Netrin-1 is crucial for axonal guidance by interacting with its receptors, deleted in colorectal cancer (DCC) and uncoordinated gene 5H (UNC5H). DCC and UNC5H are also dependence receptors inducing cell apoptosis when unbound by netrin-1. The present study is to investigate the role of netrin-1 and its receptors in ipsilateral ventroposterior thalamic nucleus (VPN) injury secondary to stroke in hypertensive rats. Renovascular hypertensive Sprague-Dawley rats underwent middle cerebral artery occlusion (MCAO). Continuous intracerebroventricular infusion of netrin-1 (600 ng/d for 7 days) or vehicle (IgG/Fc) was given 24h after MCAO. Neurological function was evaluated by postural reflex 8 and 14 days after MCAO. Then, immunoreactivity was determined in the ipsilateral VPN for NeuN, glial fibrillary acidic protein, netrin-1 and its receptors (DCC and UNC5H2), apoptosis was detected with Terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP-biotin nick-end labeling (TUNEL) assay, and the expressions of caspase-3, netrin-1, DCC, and UNC5H2 were quantified by western blot analysis. MCAO resulted in the impaired postural reflex after 8 and 14 days, with decreased NeuN marked neurons and increased TUNEL-positive cells, as well as an up-regulation in the levels of cleaved caspase-3 and UNC5H2 protein in the ipsilateral VPN, without significant change in DCC or netrin-1 expression. By exogenous netrin-1 infusion, the number of neurons was increased in the ipsilateral VPN, and both TUNEL-positive cell number and caspase-3 protein level were reduced, while UNC5H2 expression remained unaffected, simultaneously, the impairment of postural reflex was improved. Taken together, the present study indicates that exogenous netrin-1 could rescue neuron loss by attenuating secondary apoptosis in the ipsilateral VPN after focal cerebral infarction, possibly via its receptor UNC5H2, suggesting that relative insufficiency of endogenous netrin-1 be an underlying mechanism of secondary injury in the VPN post stroke.
authors.affiliation: Department of Neurology, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China.
authors.forename: S-J
authors.initials: SJ
authors.lastname: Liao
authors.forename: Q
authors.initials: Q
authors.lastname: Gong
authors.forename: X-R
authors.initials: XR
authors.lastname: Chen
authors.forename: L-X
authors.initials: LX
authors.lastname: Ye
authors.forename: Q
authors.initials: Q
authors.lastname: Ding
authors.forename: J-S
authors.initials: JS
authors.lastname: Zeng
authors.forename: J
authors.initials: J
authors.lastname: Yu
mesh_terms: D000818:Animals; D034961:Antigens, Nuclear / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D020520:Brain Infarction / Q000150:complications / Q000378:metabolism*; D000075389:DCC Receptor; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006978:Hypertension, Renovascular / Q000150:complications / Q000378:metabolism*; D008297:Male; D009414:Nerve Growth Factors / Q000378:metabolism / Q000494:pharmacology*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D000075388:Netrin-1; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011956:Receptors, Cell Surface / Q000378:metabolism; D013787:Thalamic Nuclei / Q000187:drug effects* / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000378:metabolism / Q000494:pharmacology*; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology
chemical_list: D034961:Antigens, Nuclear; D000075389:DCC Receptor; C501912:Dcc protein, rat; D005904:Glial Fibrillary Acidic Protein; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; C000617354:Ntn1 protein, rat; C546339:Rbfox3 protein, rat; D011956:Receptors, Cell Surface; D025521:Tumor Suppressor Proteins; C524887:Unc5b protein, rat; D000075388:Netrin-1