title: Comprehensive analysis of transcription dynamics from brain samples following behavioral experience.
abstract: The encoding of experiences in the brain and the consolidation of long-term memories depend on gene transcription. Identifying the function of specific genes in encoding experience is one of the main objectives of molecular neuroscience. Furthermore, the functional association of defined genes with specific behaviors has implications for understanding the basis of neuropsychiatric disorders. Induction of robust transcription programs has been observed in the brains of mice following various behavioral manipulations. While some genetic elements are utilized recurrently following different behavioral manipulations and in different brain nuclei, transcriptional programs are overall unique to the inducing stimuli and the structure in which they are studied(1,2). In this publication, a protocol is described for robust and comprehensive transcriptional profiling from brain nuclei of mice in response to behavioral manipulation. The protocol is demonstrated in the context of analysis of gene expression dynamics in the nucleus accumbens following acute cocaine experience. Subsequent to a defined in vivo experience, the target neural tissue is dissected; followed by RNA purification, reverse transcription and utilization of microfluidic arrays for comprehensive qPCR analysis of multiple target genes. This protocol is geared towards comprehensive analysis (addressing 50-500 genes) of limiting quantities of starting material, such as small brain samples or even single cells. The protocol is most advantageous for parallel analysis of multiple samples (e.g. single cells, dynamic analysis following pharmaceutical, viral or behavioral perturbations). However, the protocol could also serve for the characterization and quality assurance of samples prior to whole-genome studies by microarrays or RNAseq, as well as validation of data obtained from whole-genome studies.
authors.affiliation: The Alexander Silberman Institute of Life Sciences & Edmond and Lily Safra Center for Brain Sciences, The Hebrew University of Jerusalem.
authors.forename: Hagit
authors.initials: H
authors.lastname: Turm
authors.affiliation: The Alexander Silberman Institute of Life Sciences & Edmond and Lily Safra Center for Brain Sciences, The Hebrew University of Jerusalem.
authors.forename: Diptendu
authors.initials: D
authors.lastname: Mukherjee
authors.affiliation: The Alexander Silberman Institute of Life Sciences & Edmond and Lily Safra Center for Brain Sciences, The Hebrew University of Jerusalem.
authors.forename: Doron
authors.initials: D
authors.lastname: Haritan
authors.affiliation: The Alexander Silberman Institute of Life Sciences & Edmond and Lily Safra Center for Brain Sciences, The Hebrew University of Jerusalem.
authors.forename: Maayan
authors.initials: M
authors.lastname: Tahor
authors.affiliation: The Alexander Silberman Institute of Life Sciences & Edmond and Lily Safra Center for Brain Sciences, The Hebrew University of Jerusalem; ami.citri@mail.huji.ac.il.
authors.forename: Ami
authors.initials: A
authors.lastname: Citri
mesh_terms: D000818:Animals; D001522:Behavior, Animal / Q000187:drug effects / Q000502:physiology*; D003042:Cocaine / Q000494:pharmacology; D020869:Gene Expression Profiling; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D042282:Microdissection; D046210:Microfluidic Analytical Techniques / Q000379:methods; D009714:Nucleus Accumbens / Q000187:drug effects / Q000502:physiology* / Q000601:surgery; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D014158:Transcription, Genetic
chemical_list: D012333:RNA, Messenger; D003042:Cocaine