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Early, nonlethal ploidy and genome size quantification using confocal microscopy in zebrafish embryos
Author(s) -
Small Christopher D.,
Davis James P.,
Crawford Bryan D.,
Benfey Tillmann J.
Publication year - 2021
Publication title -
journal of experimental zoology part b: molecular and developmental evolution
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.823
H-Index - 63
eISSN - 1552-5015
pISSN - 1552-5007
DOI - 10.1002/jez.b.23069
Subject(s) - ploidy , biology , zebrafish , polyploid , genome , genome size , developmental biology , live cell imaging , embryo , confocal microscopy , cytoplasm , genetics , microbiology and biotechnology , cell , gene
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.