title: Silencing Estrogen Receptor-α with siRNA in the Intact Rodent Brain.
abstract: The ability to silence the expression of gene products in a chemically, spatially, and temporally specific manner in the brains of animals has enabled key breakthroughs in the field of behavioral neuroscience. Using this technique, estrogen receptor alpha (ERα) has been specifically implicated in a multitude of behaviors in mice, including sexual, aggressive, locomotor, and maternal behaviors. ERα has been identified in a variety of brain regions, including the medial preoptic area, ventromedial hypothalamus, and amygdala. In this chapter we describe the techniques involved in the generation of the small hairpin RNAs (shRNAs) specifically designed to silence ERα, the construction of the adeno-associated viral (AAV) vector for delivery of the shRNA, the procedures to confirm the silencing of ERα (in vitro and in vivo) and in vivo delivery of the shRNAs to the brains of animals.
authors.affiliation: Division of Natural Sciences, College of Mount Saint Vincent, Bronx, NY, USA. ana.ribeiro@mountsaintvincent.edu.|Laboratory of Neurobiology and Behavior, The Rockefeller University, New York, NY, USA. ana.ribeiro@mountsaintvincent.edu.
authors.forename: Ana C
authors.initials: AC
authors.lastname: Ribeiro
authors.affiliation: Department of Psychology, University of Tromsø, Tromsø, Norway.
authors.forename: Anders
authors.initials: A
authors.lastname: Ågmo
authors.affiliation: Laboratory of Molecular Neurosurgery, Weill Medical College of Cornell University, New York, NY, USA.
authors.forename: Sergei
authors.initials: S
authors.lastname: Musatov
authors.affiliation: Laboratory of Neurobiology and Behavior, The Rockefeller University, New York, NY, USA.
authors.forename: Donald W
authors.initials: DW
authors.lastname: Pfaff
mesh_terms: D000818:Animals; D001522:Behavior, Animal; D001921:Brain / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics; D047628:Estrogen Receptor alpha / Q000235:genetics* / Q000378:metabolism; D017930:Genes, Reporter; D005822:Genetic Vectors; D007150:Immunohistochemistry; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D014161:Transduction, Genetic; D057188:Workflow
chemical_list: D047628:Estrogen Receptor alpha; D034741:RNA, Small Interfering; D008156:Luciferases
keywords: Adeno-associated virus; Behavior; Estrogen receptor alpha; Mice; RNA interference; shRNA