One of the most feared disabilities in survivors of cardiac arrest is neurological impairment. Global cerebral ischemia, as seen with cardiac arrest, causes delayed loss of CA1 pyramidal neurons in the hippocampus while the nearby dentate gyrus (DG) is relatively resistant. Despite much work on hippocampal neuronal injury, little is known of the effects of ischemia on hippocampal astrocytes. This grant seeks to understand the contribution of astrocytes to hippocampal selective vulnerability. Astrocytes are essential to many brain functions including synaptic transmission, metabolic and ionic homeostasis, antioxidant defense, inflammation, the blood brain barrier, and trophic support. This proposal will study three aspects of hippocampal astrocyte survival and response to ischemia and ischemia-like stress.
In aim 1, mitochondrial function will be assessed with ischemia and reperfusion. Mitochondria are important both for energy production and for the regulation of cell death. Hippocampal organotypic slice cultures, which reproduce selective CA1 vulnerability, and astrocytes isolated from these two subregions of the hippocampus will be used for electrophysiological studies and real time imaging. The timecourse of changes in mitochondrial membrane potential, reactive oxygen species, and intracellular calcium during reperfusion in astrocytes in CA1 and DG will be assessed. Comparing the vulnerable and resistant areas will allow discrimination of important changes. Respiratory function and mitochondrial complex activity will also be measured.
Aim 2 will focus on changes in astrocyte glutamate uptake with ischemia and reperfusion. Early impairment of glutamate uptake by astrocytes endangers neighboring neurons, and we have found early loss of GLT-1 in CA1 astrocytes. Detailed studies of glutamate uptake and transporter expression will be performed. ATP stores required for maintaining membrane potential and glutamate uptake will be monitored.
Aim 3 will use overexpression of the chaperone Hsp70 selectively in astrocytes and induction of GLT-1 as protective strategies and assess changes in astrocyte function as well as changes in neuronal survival. Together these studies will provide a picture of hippocampal astrocyte ischemic response and link astrocyte dysfunction to selective neuronal injury in hippocampus. Maintaining and improving astrocyte function provides a novel target for developing therapies to improve neurological function following cardiac arrest.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS053898-04
Application #
7773514
Study Section
Special Emphasis Panel (ZRG1-NOMD-A (01))
Program Officer
Bosetti, Francesca
Project Start
2007-03-01
Project End
2012-02-29
Budget Start
2010-03-01
Budget End
2011-02-28
Support Year
4
Fiscal Year
2010
Total Cost
$342,559
Indirect Cost
Name
Stanford University
Department
Anesthesiology
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
Stary, Creed M; Xu, Lijun; Li, Le et al. (2017) Inhibition of miR-181a protects female mice from transient focal cerebral ischemia by targeting astrocyte estrogen receptor-?. Mol Cell Neurosci 82:118-125
Bell, Josh D; Cho, Jang-Eun; Giffard, Rona G (2017) MicroRNA Changes in Preconditioning-Induced Neuroprotection. Transl Stroke Res 8:585-596
Stary, Creed M; Sun, Xiaoyun; Ouyang, YiBing et al. (2016) miR-29a differentially regulates cell survival in astrocytes from cornu ammonis 1 and dentate gyrus by targeting VDAC1. Mitochondrion 30:248-54
Xu, Li-Jun; Ouyang, Yi-Bing; Xiong, Xiaoxing et al. (2015) Post-stroke treatment with miR-181 antagomir reduces injury and improves long-term behavioral recovery in mice after focal cerebral ischemia. Exp Neurol 264:1-7
Ouyang, Yi-Bing; Stary, Creed M; White, Robin E et al. (2015) The use of microRNAs to modulate redox and immune response to stroke. Antioxid Redox Signal 22:187-202
Stary, Creed M; Giffard, Rona G (2015) Advances in astrocyte-targeted approaches for stroke therapy: an emerging role for mitochondria and microRNAS. Neurochem Res 40:301-7
Stary, Creed M; Sun, Xiaoyun; Giffard, Rona G (2015) Astrocytes Protect against Isoflurane Neurotoxicity by Buffering pro-brain-derived Neurotrophic Factor. Anesthesiology 123:810-9
Ouyang, Yi-Bing; Xu, Lijun; Liu, Siwei et al. (2014) Role of Astrocytes in Delayed Neuronal Death: GLT-1 and its Novel Regulation by MicroRNAs. Adv Neurobiol 11:171-88
Ouyang, Yi-Bing; Giffard, Rona G (2014) MicroRNAs affect BCL-2 family proteins in the setting of cerebral ischemia. Neurochem Int 77:2-8
Ouyang, Yi-Bing; Xu, Lijun; Yue, Sibiao et al. (2014) Neuroprotection by astrocytes in brain ischemia: importance of microRNAs. Neurosci Lett 565:53-8

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