Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping Platform
In recent years, zebrafish have become commonly used as a model for studying human traits and disorders. Their small size, high fecundity, and rapid development allow for more high-throughput experiments compared to other vertebrate models. Given that zebrafish share >70% gene homologs with h...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2020-11-01
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| Series: | Frontiers in Cell and Developmental Biology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2020.586296/full |
| _version_ | 1818935334166069248 |
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| author | Alexandra Colón-Rodríguez José M. Uribe-Salazar José M. Uribe-Salazar KaeChandra B. Weyenberg Aditya Sriram Alejandra Quezada Alejandra Quezada Gulhan Kaya Emily Jao Brittany Radke Pamela J. Lein Pamela J. Lein Megan Y. Dennis Megan Y. Dennis Megan Y. Dennis |
| author_facet | Alexandra Colón-Rodríguez José M. Uribe-Salazar José M. Uribe-Salazar KaeChandra B. Weyenberg Aditya Sriram Alejandra Quezada Alejandra Quezada Gulhan Kaya Emily Jao Brittany Radke Pamela J. Lein Pamela J. Lein Megan Y. Dennis Megan Y. Dennis Megan Y. Dennis |
| author_sort | Alexandra Colón-Rodríguez |
| collection | DOAJ |
| description | In recent years, zebrafish have become commonly used as a model for studying human traits and disorders. Their small size, high fecundity, and rapid development allow for more high-throughput experiments compared to other vertebrate models. Given that zebrafish share >70% gene homologs with humans and their genomes can be readily edited using highly efficient CRISPR methods, we are now able to rapidly generate mutations impacting practically any gene of interest. Unfortunately, our ability to phenotype mutant larvae has not kept pace. To address this challenge, we have developed a protocol that obtains multiple phenotypic measurements from individual zebrafish larvae in an automated and parallel fashion, including morphological features (i.e., body length, eye area, and head size) and movement/behavior. By assaying wild-type zebrafish in a variety of conditions, we determined optimal parameters that avoid significant developmental defects or physical damage; these include morphological imaging of larvae at two time points [3 days post fertilization (dpf) and 5 dpf] coupled with motion tracking of behavior at 5 dpf. As a proof-of-principle, we tested our approach on two novel CRISPR-generated mutant zebrafish lines carrying predicted null-alleles of syngap1b and slc7a5, orthologs to two human genes implicated in autism-spectrum disorder, intellectual disability, and epilepsy. Using our optimized high-throughput phenotyping protocol, we recapitulated previously published results from mouse and zebrafish models of these candidate genes. In summary, we describe a rapid parallel pipeline to characterize morphological and behavioral features of individual larvae in a robust and consistent fashion, thereby improving our ability to better identify genes important in human traits and disorders. |
| first_indexed | 2024-12-20T05:18:31Z |
| format | Article |
| id | doaj.art-f6bb783921c541c3978a11e0544dc4fb |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-12-20T05:18:31Z |
| publishDate | 2020-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-f6bb783921c541c3978a11e0544dc4fb2022-12-21T19:52:06ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2020-11-01810.3389/fcell.2020.586296586296Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping PlatformAlexandra Colón-Rodríguez0José M. Uribe-Salazar1José M. Uribe-Salazar2KaeChandra B. Weyenberg3Aditya Sriram4Alejandra Quezada5Alejandra Quezada6Gulhan Kaya7Emily Jao8Brittany Radke9Pamela J. Lein10Pamela J. Lein11Megan Y. Dennis12Megan Y. Dennis13Megan Y. Dennis14Genome Center, Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, Davis, CA, United StatesGenome Center, Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, Davis, CA, United StatesIntegrative Genetics and Genomics Graduate Group, University of California, Davis, Davis, CA, United StatesGenome Center, Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, Davis, CA, United StatesGenome Center, Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, Davis, CA, United StatesGenome Center, Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, Davis, CA, United StatesSacramento State RISE Program, California State University, Sacramento, Sacramento, CA, United StatesGenome Center, Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, Davis, CA, United StatesGenome Center, Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, Davis, CA, United StatesGenome Center, Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, Davis, CA, United StatesDepartment of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, Davis, CA, United StatesMIND Institute, School of Medicine, University of California, Davis, Davis, CA, United StatesGenome Center, Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, Davis, CA, United StatesIntegrative Genetics and Genomics Graduate Group, University of California, Davis, Davis, CA, United StatesMIND Institute, School of Medicine, University of California, Davis, Davis, CA, United StatesIn recent years, zebrafish have become commonly used as a model for studying human traits and disorders. Their small size, high fecundity, and rapid development allow for more high-throughput experiments compared to other vertebrate models. Given that zebrafish share >70% gene homologs with humans and their genomes can be readily edited using highly efficient CRISPR methods, we are now able to rapidly generate mutations impacting practically any gene of interest. Unfortunately, our ability to phenotype mutant larvae has not kept pace. To address this challenge, we have developed a protocol that obtains multiple phenotypic measurements from individual zebrafish larvae in an automated and parallel fashion, including morphological features (i.e., body length, eye area, and head size) and movement/behavior. By assaying wild-type zebrafish in a variety of conditions, we determined optimal parameters that avoid significant developmental defects or physical damage; these include morphological imaging of larvae at two time points [3 days post fertilization (dpf) and 5 dpf] coupled with motion tracking of behavior at 5 dpf. As a proof-of-principle, we tested our approach on two novel CRISPR-generated mutant zebrafish lines carrying predicted null-alleles of syngap1b and slc7a5, orthologs to two human genes implicated in autism-spectrum disorder, intellectual disability, and epilepsy. Using our optimized high-throughput phenotyping protocol, we recapitulated previously published results from mouse and zebrafish models of these candidate genes. In summary, we describe a rapid parallel pipeline to characterize morphological and behavioral features of individual larvae in a robust and consistent fashion, thereby improving our ability to better identify genes important in human traits and disorders.https://www.frontiersin.org/articles/10.3389/fcell.2020.586296/fullDanio reriophenotypingCRISPRdevelopmental disordersseizureVAST |
| spellingShingle | Alexandra Colón-Rodríguez José M. Uribe-Salazar José M. Uribe-Salazar KaeChandra B. Weyenberg Aditya Sriram Alejandra Quezada Alejandra Quezada Gulhan Kaya Emily Jao Brittany Radke Pamela J. Lein Pamela J. Lein Megan Y. Dennis Megan Y. Dennis Megan Y. Dennis Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping Platform Frontiers in Cell and Developmental Biology Danio rerio phenotyping CRISPR developmental disorders seizure VAST |
| title | Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping Platform |
| title_full | Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping Platform |
| title_fullStr | Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping Platform |
| title_full_unstemmed | Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping Platform |
| title_short | Assessment of Autism Zebrafish Mutant Models Using a High-Throughput Larval Phenotyping Platform |
| title_sort | assessment of autism zebrafish mutant models using a high throughput larval phenotyping platform |
| topic | Danio rerio phenotyping CRISPR developmental disorders seizure VAST |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2020.586296/full |
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