Proteomic analysis of plasma biomarkers for murine neonatal HI brain injury. Proteomic analysis of plasma biomarkers for murine neonatal HI brain injury.

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Proteomic analysis of plasma biomarkers for murine neonatal HI brain injury. Proteomic analysis of plasma biomarkers for murine neonatal HI brain injury. A, Plasma proteins collected from unchallenged (UN), low-dose LPS (0.3 mg/kg, IP)-exposed, and HI- or LPS/HI-injured P11 mice were separated by SDS-PAGE and analyzed as depicted (n = 3 for each). Also shown are representative tetrazolium chloride–stained brains in each group, showing cerebral infarction (pale color) in only HI- or LPS/HI-injured animals. B, Venn diagram showing the number of unique proteins in the depicted conditions compared with the unchallenged animals. C, Heatmap and hierarchical clustering of the 12 examined animals based on the signal intensity of HI- and LPS/HI-associated proteins. Shown is the log2 scale of fold change. D, E, Immunoblot showing specific OPN induction in plasma and the ipsilateral hemisphere (right, R*) of mice after HI or LPS/HI insult. In contrast, although cystatin C showed increased plasma levels after HI and LPS/HI injury, it was not elevated in the brain after these injuries. *, Nonspecific anti–cystatin C band in the brain. rOPN, recombinant mouse OPN used as positive control. F, Immunoblotting showing that cerebral stroke (thrombosis) also induced high levels of plasma OPN, whereas ICV injection of phosphate saline (ICV-PBS), unilateral carotid artery ligation (RCCAO), i.p. injection of 3 mg/kg LPS (IP-LPS[10×]), and intratracheal application of 0/3 mg/kg LPS (IT-LPS) lacked this effect. The numbers of mouse pups examined in each condition are indicated. **, p < 0.01; ***, p < 0.001 by t-test compared with unchallenged animals. Yikun Li et al. eNeuro 2017;4:ENEURO.0253-16.2016 ©2017 by Society for Neuroscience