Microinjection for Transgenesis and Genome Editing in Threespine Sticklebacks

Erickson, Priscilla A.; Ellis, Nicholas; Miller, Craig T.

doi:10.3791/54055

Cited by 18 publications

(22 citation statements)

References 43 publications

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“…CRISPR/cas9 systems (Erickson et al . ) and TALEN systems (Ishikawa A & Kitano J unpublished data) have been established in sticklebacks. It is now possible to investigate the functions of these candidate genes in vivo.…”

Section: Discussionmentioning

confidence: 99%

Genetic basis for variation in salinity tolerance between stickleback ecotypes

et al. 2016

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Adaptation to different salinities can drive and maintain divergence between populations of aquatic organisms. Anadromous and stream ecotypes of threespine stickleback (Gasterosteus aculeatus) are an excellent model to explore the genetic mechanisms underlying osmoregulation divergence. Using a parapatric pair of anadromous and stream stickleback ecotypes, we employed an integrated genomic approach to identify candidate genes important for adaptation to different salinity environments. Quantitative trait loci (QTL) mapping of plasma sodium concentrations under a seawater challenge experiment identified a significant QTL on chromosome 16. To identify candidate genes within this QTL, we first conducted RNA-seq and microarray analysis on gill tissue to find ecotypic differences in gene expression that were associated with plasma Na levels. This resulted in the identification of ten candidate genes. Quantitative PCR analysis on gill tissue of additional Japanese stickleback populations revealed that the majority of the candidate genes showed parallel divergence in expression levels. Second, we conducted whole-genome sequencing and found five genes that are predicted to have functionally important amino acid substitutions. Finally, we conducted genome scan analysis and found that eight of these candidate genes were located in genomic islands of high differentiation, suggesting that they may be under divergent selection. The candidate genes included those involved in ATP synthesis and hormonal signalling, whose expression or amino acid changes may underlie the variation in salinity tolerance. Further functional molecular analysis of these genes will reveal the causative genetic and genomic changes underlying divergent adaptation.

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Section: Discussionmentioning

confidence: 99%

Genetic basis for variation in salinity tolerance between stickleback ecotypes

et al. 2016

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“…A major challenge for the future is to identify the genes and mutations that underlie additional phenotypic traits in sticklebacks. The development of new genome-editing tools like TALENs and CRISPR/Cas9 for use in sticklebacks [22] will greatly facilitate this research, enabling us to discern whether there are general patterns in the genetic and molecular architecture of phenotypic evolution, at least in sticklebacks. It will also be important to compare results in sticklebacks to those in other systems, like those highlighted in this special issue [77][78][79][80][81], to determine whether the genetic and molecular basis of phenotypic evolution is contingent on the study system or whether general evolutionary patterns will emerge.…”

Section: Perspectives and Future Directionsmentioning

confidence: 99%

The genetic and molecular architecture of phenotypic diversity in sticklebacks

Peichel

Marques

2017

Phil. Trans. R. Soc. B

162

201

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One contribution of 17 to a theme issue 'Evodevo in the genomics era, and the origins of morphological diversity'. A major goal of evolutionary biology is to identify the genotypes and phenotypes that underlie adaptation to divergent environments. Stickleback fish, including the threespine stickleback (Gasterosteus aculeatus) and the ninespine stickleback (Pungitius pungitius), have been at the forefront of research to uncover the genetic and molecular architecture that underlies phenotypic diversity and adaptation. A wealth of quantitative trait locus (QTL) mapping studies in sticklebacks have provided insight into longstanding questions about the distribution of effect sizes during adaptation as well as the role of genetic linkage in facilitating adaptation. These QTL mapping studies have also provided a basis for the identification of the genes that underlie phenotypic diversity. These data have revealed that mutations in regulatory elements play an important role in the evolution of phenotypic diversity in sticklebacks. Genetic and molecular studies in sticklebacks have also led to new insights on the genetic basis of repeated evolution and suggest that the same loci are involved about half of the time when the same phenotypes evolve independently. When the same locus is involved, selection on standing variation and repeated mutation of the same genes have both contributed to the evolution of similar phenotypes in independent populations. This article is part of the themed issue 'Evo-devo in the genomics era, and the origins of morphological diversity'.

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“…Although GWAS, QTL mapping and other -omics approaches can provide us with a list of candidate genes or mutations, gene manipulation is necessary to confirm causal association with phenotype. In the three-spined stickleback, genome editing and transgenic techniques have already been established (Chan et al, 2010;Colosimo et al, 2005;Erickson et al, 2016;Hart and Miller, 2017;Hosemann et al, 2004;Howes et al, 2017;Wucherpfennig et al, 2019). These tools allow us to overexpress and knock out genes to directly test the function of candidate genes in vivo.…”

Section: The Future Of Physiological Genomics Of Seasonal Reproductiomentioning

confidence: 99%

Diversity in reproductive seasonality in the three-spined stickleback,Gasterosteus aculeatus

Ishikawa

Kitano

2020

Journal of Experimental Biology

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The annual timing of reproduction is a key life history trait with a large effect on fitness. Populations often vary in the timing and duration of reproduction to adapt to different seasonality of ecological and environmental variables between habitats. However, little is known about the molecular genetic mechanisms underlying interpopulation variation in reproductive seasonality. Here, we demonstrate that the three-spined stickleback (Gasterosteus aculeatus) is a good model for molecular genetic analysis of variations in reproductive seasonality. We first compiled data on reproductive seasons of diverse ecotypes, covering marine-anadromous, lake and stream ecotypes, of three-spined stickleback inhabiting a wide range of latitudes. Our analysis showed that both ecotype and latitude significantly contribute to variation in reproductive seasons. Stream ecotypes tend to start breeding earlier and end later than other ecotypes. Populations from lower latitudes tend to start breeding earlier than those from higher latitudes in all three ecotypes. Additionally, stream ecotypes tend to have extended breeding seasons at lower latitudes than at higher latitudes, leading to nearly year-round reproduction in the most southern stream populations. A review of recent progress in our understanding of the physiological mechanisms underlying seasonal reproduction in the three-spined stickleback indicates that photoperiod is an important external cue that stimulates and/or suppresses reproduction in this species. Taking advantage of genomic tools available for this species, the three-spined stickleback will be a good model to investigate what kinds of genes and mutations underlie variations in the physiological signalling pathways that regulate reproduction in response to photoperiod.

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Microinjection for Transgenesis and Genome Editing in Threespine Sticklebacks

Cited by 18 publications

References 43 publications

Genetic basis for variation in salinity tolerance between stickleback ecotypes

Genetic basis for variation in salinity tolerance between stickleback ecotypes

The genetic and molecular architecture of phenotypic diversity in sticklebacks

Diversity in reproductive seasonality in the three-spined stickleback,Gasterosteus aculeatus

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