Seladin-1/DHCR24 Is Neuroprotective by Associating EAAT2 Glutamate Transporter to Lipid Rafts in Experimental Stroke.

Abstract

BACKGROUND AND PURPOSE:

3β-Hydroxysteroid-Δ24 reductase (DHCR24) or selective alzheimer disease indicator 1 (seladin-1), an enzyme of cholesterol biosynthetic pathway, has been implicated in neuroprotection, oxidative stress, and inflammation. However, its role in ischemic stroke remains unexplored. The aim of this study was to characterize the effect of seladin-1/DHCR24 using an experimental stroke model in mice.

METHODS:

Dhcr24(+/-) and wild-type (WT) mice were subjected to permanent middle cerebral artery occlusion. In another set of experiments, WT mice were treated intraperitoneally either with vehicle or U18666A (seladin-1/DHCR24 inhibitor, 10 mg/kg) 30 minutes after middle cerebral artery occlusion. Brains were removed 48 h after middle cerebral artery occlusion for infarct volume determination. For protein expression determination, peri-infarct region was obtained 24 h after ischemia, and Western blot or cytometric bead array was performed.

RESULTS:

Dhcr24(+/-) mice displayed larger infarct volumes after middle cerebral artery occlusion than their WT littermates. Treatment of WT mice with the seladin-1/DHCR24 inhibitor U18666A also increased ischemic lesion. Inflammation-related mediators were increased after ischemia in Dhcr24(+/-) mice compared with WT counterparts. Consistent with a role of cholesterol in proper function of glutamate transporter EAAT2 in membrane lipid rafts, we found a decreased association of EAAT2 with lipid rafts after ischemia when DHCR24 is genetically deleted or pharmacologically inhibited. Accordingly, treatment with U18666A decreases [(3)H]-glutamate uptake in cultured astrocytes.

CONCLUSIONS:

These results support the idea that lipid raft integrity, ensured by seladin-1/DHCR24, plays a crucial protective role in the ischemic brain by guaranteeing EAAT2-mediated uptake of glutamate excess.

• Authors: Macarena Hernández-Jiménez, Diego Martínez-López, Enrique Gabandé-Rodríguez, Adrian Martín-Segura, Ignacio Lizasoain, María D Ledesma, Carlos G Dotti, María A Moro
• Journal: Stroke (Stroke) Vol. 47 Issue 1 Pg. 206-13 (Jan 2016) ISSN: 1524-4628 [Electronic] United States
• PMID: 26628388 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2015 American Heart Association, Inc.

Chemical References

• Androstenes
• Excitatory Amino Acid Transporter 2
• Nerve Tissue Proteins
• Slc1a2 protein, mouse
• 3-beta-(2-(diethylamino)ethoxy)androst-5-en-17-one
• Glutamic Acid
• Oxidoreductases Acting on CH-CH Group Donors
• Dhcr24 protein, mouse

Topics

• Androstenes (pharmacology)
• Animals
• Animals, Newborn
• Cells, Cultured
• Excitatory Amino Acid Transporter 2 (genetics, metabolism)
• Glutamic Acid (metabolism)
• Male
• Membrane Microdomains (drug effects, genetics, metabolism)
• Mice
• Mice, Inbred C57BL
• Mice, Knockout
• Nerve Tissue Proteins (antagonists & inhibitors, deficiency, genetics)
• Oxidoreductases Acting on CH-CH Group Donors (antagonists & inhibitors, deficiency, genetics)
• Stroke (genetics, metabolism, prevention & control)