The experiments in this project use innovative gene delivery techniques and a multipronged approach to assess the fundamental role of astrocyte activation in neurologic function in an intact mouse model of Alzheimer's disease (AD). Studies use adeno-associated virus vectors (AAV) bearing the astrocyte-specific promoter Gfa2 to target the protein phosphatase calcineurin (CN) and NFAT transcription factors in astrocytes of wild-type and APP/PS1 mice. AAV-Gfa2 vectors are bilaterally delivered to the hippocampus at different ages/disease stages and mice are assessed on different AD biomarkers.
In Aim 1, cognitive status is assessed using the active avoidance task, while synaptic function is evaluated using hippocampal slice electrophysiology and Western blot measures of synaptic proteins.
In Aim 2, hippocampal glutamate regulation is investigated using ceramic enzyme-based microelectrode arrays and measures of glutamate transporter levels.
In Aim 3, levels of glial activation and neuroinflammation are determined with immunohistochemical (IHC) analyses and assessment of cytokine levels using Multiplex ELISAs.
In Aim 4, IHC is used to determine the extent of A? deposition, while ELISAs are used to quantify levels of A?40 and A?42 in soluble and insoluble hippocampal tissue fractions, and Westerns used to assess BACE protein expression. AAV-Gfa2 vectors encode either potent inhibitors or activators of CN/NFAT signaling and therefore will determine the necessity and sufficiency of this astrocytic pathway in driving and/or maintaining neurologic dysfunction in AD mice. These studies provide a highly novel approach to the study of activated astrocytes and could have a major impact on the development of treatment strategies for AD and other neurodegenerative conditions.

Public Health Relevance

Increasing evidence implicates activated astrocytes in a variety of neurodegenerative conditions, including Alzheimer's disease (AD). However, these cells are difficult to target selectively with therapeutics. In this project, we use cutting-edge adeno associated virus vectors to selectively prevent astrocyte activation and improve neurologic function in intact AD model mice. This approach could emerge as a new treatment strategy for AD and other neurodegenerative disorders.

National Institute of Health (NIH)
National Institute on Aging (NIA)
Research Project (R01)
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Cell Death in Neurodegeneration Study Section (CDIN)
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Wise, Bradley C
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University of Kentucky
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Schools of Medicine
United States
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Niedowicz, Dana M; Reeves, Valerie L; Platt, Thomas L et al. (2014) Obesity and diabetes cause cognitive dysfunction in the absence of accelerated ?-amyloid deposition in a novel murine model of mixed or vascular dementia. Acta Neuropathol Commun 2:64
Wang, Wang-Xia; Danaher, Robert J; Miller, Craig S et al. (2014) Expression of miR-15/107 family microRNAs in human tissues and cultured rat brain cells. Genomics Proteomics Bioinformatics 12:19-30
Furman, Jennifer L; Norris, Christopher M (2014) Calcineurin and glial signaling: neuroinflammation and beyond. J Neuroinflammation 11:158
Sama, Diana M; Norris, Christopher M (2013) Calcium dysregulation and neuroinflammation: discrete and integrated mechanisms for age-related synaptic dysfunction. Ageing Res Rev 12:982-95
Thibault, Olivier; Pancani, Tristano; Landfield, Philip W et al. (2012) Reduction in neuronal L-type calcium channel activity in a double knock-in mouse model of Alzheimer's disease. Biochim Biophys Acta 1822:546-9
Mohmmad Abdul, Hafiz; Baig, Irfan; Levine 3rd, Harry et al. (2011) Proteolysis of calcineurin is increased in human hippocampus during mild cognitive impairment and is stimulated by oligomeric Abeta in primary cell culture. Aging Cell 10:103-13
Mathis, Diana M; Furman, Jennifer L; Norris, Christopher M (2011) Preparation of acute hippocampal slices from rats and transgenic mice for the study of synaptic alterations during aging and amyloid pathology. J Vis Exp :
Norris, Christopher M; Blalock, Eric M; Chen, Kuey-Chu et al. (2010) Hippocampal 'zipper' slice studies reveal a necessary role for calcineurin in the increased activity of L-type Ca(2+) channels with aging. Neurobiol Aging 31:328-38
Furman, Jennifer L; Artiushin, Irina A; Norris, Christopher M (2010) Disparate effects of serum on basal and evoked NFAT activity in primary astrocyte cultures. Neurosci Lett 469:365-9
Abdul, Hafiz Mohmmad; Furman, Jennifer L; Sama, Michelle A et al. (2010) NFATs and Alzheimer's Disease. Mol Cell Pharmacol 2:7-14

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