Neonatal Hypoxia-Ischemia alters Brain-Derived Contactin-2-Positive Extracellular Vesicles in the Mouse Plasma

Shirelle X Liu; Daniela G Villacis Calderon; Zia L Maxim; Montana M Beeson; Raghavendra Rao; Phu V Tran

doi:10.1016/j.neuroscience.2024.03.014

Neonatal Hypoxia-Ischemia alters Brain-Derived Contactin-2-Positive Extracellular Vesicles in the Mouse Plasma

Neuroscience. 2024 Mar 19:545:141-147. doi: 10.1016/j.neuroscience.2024.03.014. Online ahead of print.

Authors

Shirelle X Liu¹, Daniela G Villacis Calderon¹, Zia L Maxim¹, Montana M Beeson¹, Raghavendra Rao¹, Phu V Tran²

Affiliations

¹ Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.
² Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA. Electronic address: tranx271@umn.edu.

PMID: 38513760
DOI: 10.1016/j.neuroscience.2024.03.014

Abstract

Neonatal encephalopathy (NE) impairs white matter development and results in long-term neurodevelopmental deficits. Leveraging prior findings of altered neuronal proteins carried by brain-derived extracellular vesicles (EVs) that are marked by a neural-specific cell surface glycoprotein Contactin-2 (CNTN2) in NE infants, the present study aimed to determine the correlation between brain and circulating CNTN2⁺-EVs and whether NE alters circulating CNTN2⁺-EV levels in mice. Brain tissue and plasma were collected from postnatal day (P)7, 10, 11, 15 mice to determine the baseline CNTN2 correlation between these two compartments (n = 4-7/time point/sex). NE was induced in P10 pups. Brain and plasma samples were collected at 1, 3, 6, 24, and 120 h (n = 4-8/time point/sex). CNTN2 from brain tissue and plasma EVs were quantified using ELISA. ANOVA and linear regression analyses were used to evaluate changes and correlations between brain and plasma CNTN2⁺-EVs. In baseline experiments, CNTN2 in brain tissue and plasma EVs peaked at P10 with no sex-difference. Brain and plasma CNTN2⁺-EV showed a positive correlation across early postnatal ages. NE pups showed an elevated CNTN2 in brain tissue and EVs at 1 h and only in brain tissue at 24 h. NE also abolished the positive plasma-brain correlation. The findings establish a link for central CNTN2 and its release into circulation during early postnatal life. The immediate elevation and release of CNTN2 following NE highlight a potential molecular response shortly after a brain injurious event. Our findings further support the utility of circulating brain-derived EVs as a possible bioindicator of NE.

Keywords: biomarker; contactin-2 (CNTN2); extracellular vesicle; hypoxia-ischemia (HI); neonatal encephalopathy (NE).