Automated quantification of the schooling behaviour of sticklebacks

Ardekani, Reza; Greenwood, Anna K.; Peichel, Catherine L.; Tavaré, Simon

doi:10.1186/1687-5281-2013-61

Cited by 6 publications

(3 citation statements)

References 18 publications

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“…The school-only trials were scored by the use of two methods. The time spent with the school, defined by a fish swimming within one body length of the school ( Wark et al 2011 ), was determined with custom-designed tracking software ( Ardekani et al 2013 ). The latency to approach the school was determined manually by recording the frame number at which fish first joined the school.…”

Section: Methodsmentioning

confidence: 99%

Genetic Mapping of Natural Variation in Schooling Tendency in the Threespine Stickleback

Greenwood

Ardekani

McCann

et al. 2015

G3 Genes|Genomes|Genetics

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Although there is a heritable basis for many animal behaviors, the genetic architecture of behavioral variation in natural populations remains mostly unknown, particularly in vertebrates. We sought to identify the genetic basis for social affiliation in two populations of threespine sticklebacks (Gasterosteus aculeatus) that differ in their propensity to school. Marine sticklebacks from Japan school strongly whereas benthic sticklebacks from a lake in Canada are more solitary. Here, we expanded on our previous efforts to identify quantitative trait loci (QTL) for differences in schooling tendency. We tested fish multiple times in two assays that test different aspects of schooling tendency: 1) the model school assay, which presents fish with a school of eight model sticklebacks; and 2) the choice assay, in which fish are given a choice between the model school and a stationary artificial plant. We found low-to-moderate levels of repeatability, ranging from 0.1 to 0.5, in schooling phenotypes. To identify the genomic regions that contribute to differences in schooling tendency, we used QTL mapping in two types of crosses: benthic × marine backcrosses and an F2 intercross. We found two QTL for time spent with the school in the model school assay, and one QTL for number of approaches to the school in the choice assay. These QTL were on three different linkage groups, not previously linked to behavioral differences in sticklebacks. Our results highlight the importance of using multiple crosses and robust behavioral assays to uncover the genetic basis of behavioral variation in natural populations.

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Section: Methodsmentioning

confidence: 99%

Genetic Mapping of Natural Variation in Schooling Tendency in the Threespine Stickleback

Greenwood

Ardekani

McCann

et al. 2015

G3 Genes|Genomes|Genetics

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“…Indeed, there are already hundreds of previously identified candidate genes for social behavior waiting to be characterized [21][22][23][24][25][26][27]. Sticklebacks enjoy well-established behavioral assays [28,29] that are amenable to automation [15,30].…”

Section: Introductionmentioning

confidence: 99%

Minimally invasive brain injections for viral-mediated transgenesis: New tools for behavioral genetics in sticklebacks

James

Bell

2021

PLoS ONE

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Behavioral genetics in non-model organisms is currently gated by technological limitations. However, with the growing availability of genome editing and functional genomic tools, complex behavioral traits such as social behavior can now be explored in diverse organisms. Here we present a minimally invasive neurosurgical procedure for a classic behavioral, ecological and evolutionary system: threespine stickleback (Gasterosteus aculeatus). Direct brain injection enables viral-mediated transgenesis and pharmaceutical delivery which bypasses the blood-brain barrier. This method is flexible, fast, and amenable to statistically powerful within-subject experimental designs, making it well-suited for use in genetically diverse animals such as those collected from natural populations. Developing this minimally invasive neurosurgical protocol required 1) refining the anesthesia process, 2) building a custom surgical rig, and 3) determining the normal recovery pattern allowing us to clearly identify warning signs of failure to thrive. Our custom-built surgical rig (publicly available) and optimized anesthetization methods resulted in high (90%) survival rates and quick behavioral recovery. Using this method, we detected changes in aggression from the overexpression of either of two different genes, arginine vasopressin (AVP) and monoamine oxidase (MAOA), in outbred animals in less than one month. We successfully used multiple promoters to drive expression, allowing for tailored expression profiles through time. In addition, we demonstrate that widely available mammalian plasmids work with this method, lowering the barrier of entry to the technique. By using repeated measures of behavior on the same fish before and after transfection, we were able to drastically reduce the necessary sample size needed to detect significant changes in behavior, making this a viable approach for examining genetic mechanisms underlying complex social behaviors.

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“…In addition to having a fully sequenced genome, they have been used in comparative cross-taxa studies looking for a conservation in the molecular underpinnings of social behavior (Rittschof et al 2014; Saul et al 2019) with both emerging and classic model systems. Indeed, there are already hundreds of previously identified candidate genes for social behavior waiting to be characterized (Sanogo et al 2011; Laine et al 2012; Greenwood et al 2013; Mommer & Bell 2014; Greenwood & Peichel 2015; Bell et al 2016; Bukhari et al 2017) and well-established behavioral assays (van Iersel 1953; Rowland 1982) that are amenable to automation (Ardekani et al 2013; Norton & Gutiérrez 2019).…”

Section: Introductionmentioning

confidence: 99%

Minimally invasive brain injections for viral-mediated transgenesis: New tools for behavioral genetics in sticklebacks

James

Bell

2020

Preprint

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24Major Findings 25 Forced expression of MAOA or AVP resulted in more attacks, showing a causal link between genes and 26 behavior.Abstract 35 Establishing a causal relationship between genes and social behavior is challenging. To enable 36 direct manipulation of candidate genes and thereby examine how their expression contributes to 37 behavior, we developed a neurosurgical method to deliver pharmacological agents or transgenic 38 elements directly into the threespine stickleback (Gasterosteus aculeatus) brain. Threespine sticklebacks 39 are a classic system for the study of behavior, ecology, and evolution. Male sticklebacks defending 40 nesting territories are highly aggressive toward intruders. Previous studies in stickleback have shown 41 that aggression is heritable, and that hundreds of genes are differentially expressed in the brain following 42 territorial intrusion. 43 We use viral-mediated transgenesis to test the effects on territorial aggression of overexpression 44 of candidate genes, monoamine oxidase (MAOA) and arginine vasopressin (AVP), in the stickleback 45 brain. Male sticklebacks received transcranial injections of mammalian homolog cDNA packaged in a 46 replication-deficient Herpes Simplex Virus 1 carrier. Animals transfected with either AVP or MAOA 47 constructs were more aggressive in response to a territorial intruder, unlike control animals transfected 48 with a fluorescent protein. 49 Viral-mediated transgenesis is a promising method to examine genetic underpinnings of 50 behaviors. Our success demonstrates that widely available mammalian plasmids work with this method, 51 lowering the barrier of entry to the technique. This method is flexible, fast, and amenable to statistically 52 powerful within-subject experimental designs, making it practical for use in natural populations. It 53 further enhances the growing molecular toolkit in threespine stickleback, a classic ethological system, 54 and is the first step toward using chemogenetics and optogenetics.55 56 57 Complex behaviors have been repeatedly shown to be heritable (reviewed in Dochtermann et al., 58 2019), yet establishing a causal relationship between genes and social behavior remains challenging. 59Partially, this difficulty arises from limitations of the primarily correlative methods for examining the 60 interplay between genes and behavior (Charney 2017), such as QTL, GWAS, and RNAseq studies. 61These types of studies are certainly a necessary step generating many candidate genes. However, to fully 62 characterize how a gene contributes to behavior, it is necessary to consider not just sequence differences, 63 but also regulatory and epigenetic influences. Therefore, to demonstrate and fully characterize a causal 64 relationship between a gene and behavior, it is crucial to have a method for manipulating gene 65 expression at a specific time and location. We developed one such method, viral-mediated transgenesis, 66 for the classic ethological system of threespined stickleback (Gasterosteus aculeatus).

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Automated quantification of the schooling behaviour of sticklebacks

Cited by 6 publications

References 18 publications

Genetic Mapping of Natural Variation in Schooling Tendency in the Threespine Stickleback

Genetic Mapping of Natural Variation in Schooling Tendency in the Threespine Stickleback

Minimally invasive brain injections for viral-mediated transgenesis: New tools for behavioral genetics in sticklebacks

Minimally invasive brain injections for viral-mediated transgenesis: New tools for behavioral genetics in sticklebacks

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