Proteomic Analysis of Human Cerebral Endothelial Cells Activated by Glutamate/MK-801: Significance in Ischemic Stroke Injury

Abstract

Glutamate is a major excitatory neurotransmitter in the central nervous system and plays a significant role in the pathophysiology of ischemic stroke. During acute ischemic cerebrovascular disease, glutamate efflux in the CNS produces excitotoxicity in neurons and may mediate forms of stress in other tissues expressing glutamate ionotropic (N-methyl-D-aspartate (NMDA)) receptors, e.g., cerebral endothelial cells. While endothelial cell stress in response to glutamate has been reported (oxidant stress, loss of barrier function), changes in protein expression produced by glutamate (an agonist of metabotropic and NMDA receptors) have not been documented. Here, we have examined how exposure of human cerebral endothelial cells to glutamate, in the presence and absence of the NMDA receptor antagonist MK-801, can alter the proteomic profile of cerebral endothelial cells. We found several important changes in the proteins expressed by cerebral endothelial cells in response to glutamate. Interestingly, MK-801 itself had some direct effects on cerebral endothelial cells. Taken together, our findings demonstrate that cerebral endothelial cells respond to glutamate by altering their protein expression profile. We assume that protein alterations found in the cerebral endothelial proteome, in response to glutamate and which were blocked by MK-801, may be important vascular targets in better understanding the pathogenesis of ischemic stroke.

Addition of NMDA-receptor antagonist MK801 during oxygen/glucose deprivation moderately attenuates the upregulation of glucose uptake after subsequent reoxygenation in brain endothelial cells

“…Recently, deleterious effects of homocysteine onto the BBB were reported to be mediated by NMDAR [2]. Furthermore, we and others were able to show that MK801 alone can affect the proteome and the intracellular Ca-level of BBB models [16,17]. Consequently, it should be taken into consideration that currently unknown serum components may act as NMDAR agonists or that MK801 can affect the intracellular Ca-level without prior NMDAR stimulation which can lead to decreased ROS formation and subsequent lower glucose transporter functionality.…”

Major remodeling of brain microvessels during neonatal period in the mouse: A proteomic and transcriptomic study

“…These effects involve MMP (ADAM) secretions. 8,39 Few studies have reported brain vascular proteome data, and most focus on BBB function after insults, 40 or are based on cultured brain microvascular ECs. 41,42 This present study focuses on the physiological patterns of at-risk stages for bleeding and constitutive elements involved in fMV cohesion.…”

Ionotropic glutamate receptors: Which ones, when, and where in the mammalian neocortex

“…In addition, activation of presumably GluN1/GluN2C NMDAR with glutamate in the presence of D‐serine and endothelial nitric oxide synthase leads to vasodilation (LeMaistre et al, 2012). Activation of endothelial NMDAR has been reported to impair blood–brain barrier function and to play a role in ischemia and edema (Minagar et al, ).…”