Chemokine-like factor 1 promotes the migration of rat primary cortical neurons via the induction of

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  OBJECTIVE To investigate the potential roles of chemokine-like factor 1 (CKLF1), a newly cloned chemotactic cytokine, in the cortical neuronal migration.METHODS The organotypic brain slice cultures at 14 DIV were treated with CKLF1 alone or plus anti-CKLF1 antibody.By measuring the distance between the margin of brain slices and the leading population of migrating cells, we observed the effects of CKLF1 on the migration of cerebral cortical cell populations in rats.Wound-healing assays were further used to detect the chemotaxis of CKLF1 on the cortical neurons.Finally, actin polymerization in the cortical neurons was assessed by immunofluorescence staining.RESULTS On the brain slices, the extent of cell migration was markedly enhanced from 146.27 μm to 369.57 μm in response to CKLF1 treatment, which was neutralized by anti-CKLF1 antibody.As evidenced by wound-healing assays on the cortical neurons, the percentage of relative migration distance treated with CKLF1 at 200 nmol· L-1 and 2000 nmol· L-1 increased from 13.70% to 56.14% and 68.00%, respectively.By contrast,CKLF1 treatment at 20 nmol· L-1 failed significantly increased wound closure.F-actin immunostaining was present in neurons, which was identified by MAP2, a neuronspecific marker.Furthermore, the F-actin immunoreactivity was enhanced by 1.42 fold and F-actin was organized into thick bundles that formed filopodia in response to CKLF1 stimulation.CONCLUSION CKLF1 promotes the migration of rat primary cerebral cortical neurons in a concentration-dependent manner via the induction of actin polymerization.
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