Kate S Heffernan1, Kazi Rahman1, Yoland Smith2, Adriana Galvan3. 1. Division of Neuropharmacology and Neurological Disorders, Yerkes National Primate Research Center, Emory University, Atlanta, GA, USA. 2. Division of Neuropharmacology and Neurological Disorders, Yerkes National Primate Research Center, Emory University, Atlanta, GA, USA; Udall Center of Excellence for Parkinson's Disease Research, Emory University, Atlanta, GA, USA; Department of Neurology, School of Medicine, Emory University, Atlanta, GA, USA. 3. Division of Neuropharmacology and Neurological Disorders, Yerkes National Primate Research Center, Emory University, Atlanta, GA, USA; Udall Center of Excellence for Parkinson's Disease Research, Emory University, Atlanta, GA, USA; Department of Neurology, School of Medicine, Emory University, Atlanta, GA, USA. Electronic address: agalvan@emory.edu.
Abstract
BACKGROUND: The study of astrocytic functions in non-human primates (NHPs) has been hampered by the lack of genetic tools to selectively target astrocytes. Viral vectors with selective and efficient transduction of astrocytes could be a potent tool to express marker proteins, modulators, or sensors in NHP astrocytes, but the availability of thoroughly characterized astrocytic selective promoter sequences to use in these species remains extremely limited. NEW METHOD: We describe the specificity and efficiency of an astrocyte-specific promoter, GfaABC1D in the brain of the rhesus macaque, with emphasis in basal ganglia regions. AAV5-pZac2.1-GfaABC1D-tdTomato was locally injected into the globus pallidus external segment (GPe) and putamen. The extent, efficiency, and specificity of transduction was analyzed with immunohistochemistry at the light and electron microscope levels. RESULTS: The GfaABC1D promoter directed the expression of tdTomato in an astrocyte-specific manner in directly or indirectly targeted regions (including both segments of the globus pallidus, putamen, subthalamic nucleus and cortex). COMPARISON WITH EXISTING METHODS: Due to its small size, the GfaABC1D promoter is advantageous over other previously used glial fibrillary acidic protein-based promoter sequences, facilitating its use to drive expression of various transgenes in adeno-associated viruses (AAV) or other viral vectors. CONCLUSION: GfaABC1D is an efficient promoter that selectively targets astrocytes in the monkey basal ganglia and expands the viral vector toolbox to study astrocytic functions in non-human primates.
BACKGROUND: The study of astrocytic functions in non-human primates (NHPs) has been hampered by the lack of genetic tools to selectively target astrocytes. Viral vectors with selective and efficient transduction of astrocytes could be a potent tool to express marker proteins, modulators, or sensors in NHP astrocytes, but the availability of thoroughly characterized astrocytic selective promoter sequences to use in these species remains extremely limited. NEW METHOD: We describe the specificity and efficiency of an astrocyte-specific promoter, GfaABC1D in the brain of the rhesus macaque, with emphasis in basal ganglia regions. AAV5-pZac2.1-GfaABC1D-tdTomato was locally injected into the globus pallidus external segment (GPe) and putamen. The extent, efficiency, and specificity of transduction was analyzed with immunohistochemistry at the light and electron microscope levels. RESULTS: The GfaABC1D promoter directed the expression of tdTomato in an astrocyte-specific manner in directly or indirectly targeted regions (including both segments of the globus pallidus, putamen, subthalamic nucleus and cortex). COMPARISON WITH EXISTING METHODS: Due to its small size, the GfaABC1D promoter is advantageous over other previously used glial fibrillary acidic protein-based promoter sequences, facilitating its use to drive expression of various transgenes in adeno-associated viruses (AAV) or other viral vectors. CONCLUSION: GfaABC1D is an efficient promoter that selectively targets astrocytes in the monkey basal ganglia and expands the viral vector toolbox to study astrocytic functions in non-human primates.
Authors: Ye Zhang; Steven A Sloan; Laura E Clarke; Christine Caneda; Colton A Plaza; Paul D Blumenthal; Hannes Vogel; Gary K Steinberg; Michael S B Edwards; Gordon Li; John A Duncan; Samuel H Cheshier; Lawrence M Shuer; Edward F Chang; Gerald A Grant; Melanie G Hayden Gephart; Ben A Barres Journal: Neuron Date: 2015-12-10 Impact factor: 17.173
Authors: John D Cahoy; Ben Emery; Amit Kaushal; Lynette C Foo; Jennifer L Zamanian; Karen S Christopherson; Yi Xing; Jane L Lubischer; Paul A Krieg; Sergey A Krupenko; Wesley J Thompson; Ben A Barres Journal: J Neurosci Date: 2008-01-02 Impact factor: 6.167