title: Forward Genetic Screening Using Behavioral Tests in Zebrafish: A Proof of Concept Analysis of Mutants.
abstract: The zebrafish enjoys several advantages over other model organisms. It is small, easy to maintain, prolific, and numerous genetic tools are available for it. For example, forward genetic screens have allowed investigators to identify important genes potentially involved in a variety of functions from embryogenesis to cancer. However, despite its sophisticated behavioral repertoire, behavioral methods have rarely been utilized in forward genetic screens. Here, we employ a two-tiered strategy, a proof of concept study, to explore the feasibility of behavioral screens. We generated mutant lines using transposon-based insertional mutagenesis, allowing us to bias mutant selection with target genes expressed within the brain. Furthermore, we employed an efficient and fast behavioral pre-selection in which we investigated the locomotory response of 5-day post-fertilization old larval fish to hyperosmotic shock. Based on this assay, we selected five lines for our lower throughput secondary adult behavioral screen. The latter screen utilized tests in which computer animated image presentation and video-tracking-based automated quantification of behavior allowed us to compare heterozygous zebrafish with their wild-type siblings on their responses to a variety of stimuli. We found significant mutation induced adult behavioral alterations in 4 out of the 5 lines analyzed, including changes in response to social or fear inducing stimuli, to handling and novelty, or in habituation to novelty. We discuss the pros and cons of behavioral phenotyping and of the use of different forward genetic methods in biomedical research with zebrafish.
authors.affiliation: Department of Psychology, University of Toronto Mississauga, 3359 Mississauga Road North, Rm CCT4004, Mississauga, ON, L5L 1C6, Canada. robert_gerlai@yahoo.com.
authors.forename: Robert
authors.initials: R
authors.lastname: Gerlai
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic, Guggenheim 13, 221 4th Ave. SW, Rochester, MN, 55902, USA.
authors.forename: Tanya L
authors.initials: TL
authors.lastname: Poshusta
authors.affiliation: Department of Psychology, University of Toronto Mississauga, 3359 Mississauga Road North, Rm CCT4004, Mississauga, ON, L5L 1C6, Canada.
authors.forename: Mindy
authors.initials: M
authors.lastname: Rampersad
authors.affiliation: Department of Psychology, University of Toronto Mississauga, 3359 Mississauga Road North, Rm CCT4004, Mississauga, ON, L5L 1C6, Canada.
authors.forename: Yohaan
authors.initials: Y
authors.lastname: Fernandes
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic, Guggenheim 13, 221 4th Ave. SW, Rochester, MN, 55902, USA.
authors.forename: Tammy M
authors.initials: TM
authors.lastname: Greenwood
authors.affiliation: Department of Psychology, University of Toronto Mississauga, 3359 Mississauga Road North, Rm CCT4004, Mississauga, ON, L5L 1C6, Canada.|Department of Biochemistry and Molecular Biology, Mayo Clinic, Guggenheim 13, 221 4th Ave. SW, Rochester, MN, 55902, USA.
authors.forename: Margot A
authors.initials: MA
authors.lastname: Cousin
authors.affiliation: Department of Psychology, University of Toronto Mississauga, 3359 Mississauga Road North, Rm CCT4004, Mississauga, ON, L5L 1C6, Canada.|Department of Biochemistry and Molecular Biology, Mayo Clinic, Guggenheim 13, 221 4th Ave. SW, Rochester, MN, 55902, USA.
authors.forename: Eric W
authors.initials: EW
authors.lastname: Klee
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic, Guggenheim 13, 221 4th Ave. SW, Rochester, MN, 55902, USA. clark.karl@mayo.edu.
authors.forename: Karl J
authors.initials: KJ
authors.lastname: Clark
mesh_terms: D000818:Animals; D001522:Behavior, Animal; D005786:Gene Expression Regulation; D005820:Genetic Testing; D006185:Habituation, Psychophysiologic; D007814:Larva / Q000235:genetics; D009043:Motor Activity; D009154:Mutation / Q000235:genetics*; D009995:Osmosis; D013997:Time Factors; D015027:Zebrafish / Q000235:genetics*
keywords: Behavioral phenotyping; Fear; Forward and reverse genetics; Habituation; Social behavior; Tol2 transposon