Characterization of Hevin (SPARCL1) Immunoreactivity in Postmortem Human Brain Homogenates. (15th July 2021)

• Record Type: Journal Article
• Authors: Nuñez-delMoral, Amaia; Brocos-Mosquera, Iria; Vialou, Vincent; Callado, Luis F.; Erdozain, Amaia M.
• Is Part Of: Neuroscience. Volume 467(2021)
• Page Start: 91
• Page End: 109
• Publication Date: 2021-07-15
• Subjects: PFC prefrontal cortex -- HIP hippocampus -- CAU caudate nucleus -- CB cerebellum -- ECM extracellular matrix -- CSF cerebrospinal fluid -- ADAMTS4 a disintegrin and metalloproteinase with thrombospondin motifs 4 -- MMP-3 matrixmetalloproteinase 3 -- EndoF3 endoglycosidase F3 -- EndoH endoglycosidase H -- PNGaseF N-glycosidase F -- CNS central nervous system -- WT wild-type -- P2 membrane-enriched fraction -- PMD postmortem delay -- SLF SPARC-like fragment, IKβα, NFKB inhibitor alpha -- VGLUT1 vesicular glutamate transporter 1 -- NR2A N-methyl D-aspartate receptor subtype 2A -- PSD95 postsynaptic density protein 95; hevin -- SPARCL1 -- human brain -- postmortem -- Western blot; Neurochemistry -- Periodicals; Neurophysiology -- Periodicals; Neurology -- Periodicals; Neurochimie -- Périodiques; Neurophysiologie -- Périodiques; Neurochemistry; Neurophysiology; Electronic journals; Periodicals; Electronic journals; 612.8
• DOI: 10.1016/j.neuroscience.2021.05.017
• Languages: English
• ISSNs: 0306-4522

Abstract

Highlights: Two forms of protein hevin, differentially glycosylated, are detected in human brain: 130 kDa and 100 kDa. Hevin expression is higher in prefrontal cortex compared to hippocampus, caudate and cerebellum. Hevin is strongly expressed in membrane-enriched fraction compared to cytosolic fraction. Hevin protein is detected in cerebrospinal fluid and serum, but absent from any blood cell type. Abstract: Hevin is a matricellular glycoprotein that plays important roles in neural developmental processes such as neuronal migration, synaptogenesis and synaptic plasticity. In contrast to other matricellular proteins whose expression decreases when development is complete, hevin remains highly expressed, suggesting its involvement in adult brain function. In vitro studies have shown that hevin can have different post-translational modifications. However, the glycosylation pattern of hevin in the human brain remains unknown, as well as its relative distribution and localization. The present study provides the first thorough characterization of hevin protein expression by Western blot in postmortem adult human brain. Our results demonstrated two major specific immunoreactive bands for hevin: an intense band migrating around 130 kDa, and a band migrating around 100 kDa. Biochemical assays revealed that both hevin bands have a different glycosylation pattern. Subcellular fractionation showed greater expression in membrane-enriched fraction than in cytosolic preparation, and a higher expression in prefrontal cortex (PFC) compared to hippocampus (HIP), caudate nucleus (CAU) and cerebellum (CB). We confirmed that a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS4) and matrixmetalloproteinase 3 (MMP-3) proteases digestion led to an intense double band with similar molecular weight to that described as SPARC-like fragment (SLF). Finally, hevin immunoreactivity was also detected in human astrocytoma, meningioma, cerebrospinal fluid and serum samples, but was absent from any blood cell type.