PUBLICATION
Early, nonlethal ploidy and genome size quantification using confocal microscopy in zebrafish embryos
- Authors
- Small, C.D., Davis, J.P., Crawford, B.D., Benfey, T.J.
- ID
- ZDB-PUB-210714-28
- Date
- 2021
- Source
- Journal of experimental zoology. Part B, Molecular and developmental evolution 336(6): 496-510 (Journal)
- Registered Authors
- Crawford, Bryan D., Small, Chris
- Keywords
- cell size, genome size, polyploid, triploid, whole genome duplication, zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Cell Size
- Genome Size
- Microscopy, Confocal
- Muscles/cytology
- Ploidies*
- Zebrafish/embryology*
- Zebrafish/genetics*
- PubMed
- 34254444 Full text @ J. Exp. Zool. B Mol. Dev. Evol.
Citation
Small, C.D., Davis, J.P., Crawford, B.D., Benfey, T.J. (2021) Early, nonlethal ploidy and genome size quantification using confocal microscopy in zebrafish embryos. Journal of experimental zoology. Part B, Molecular and developmental evolution. 336(6):496-510.
Abstract
Ploidy transitions through whole genome duplication have shaped evolution by allowing the sub- and neo-functionalization of redundant copies of highly conserved genes to express novel traits. The nuclear:cytoplasmic (n:c) ratio is maintained in polyploid vertebrates resulting in larger cells, but body size is maintained by a concomitant reduction in cell number. Ploidy can be manipulated easily in most teleosts, and the zebrafish, already well established as a model system for biomedical research, is therefore an excellent system in which to study the effects of increased cell size and reduced cell numbers in polyploids on development and physiology. Here we describe a novel technique using confocal microscopy to measure genome size and determine ploidy non-lethally at 48 h post-fertilization (hpf) in transgenic zebrafish expressing fluorescent histones. Volumetric analysis of myofiber nuclei using open-source software can reliably distinguish diploids and triploids from a mixed-ploidy pool of embryos for subsequent experimentation. We present an example of this by comparing heart rate between confirmed diploid and triploid embryos at 54 hpf.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping