title: Whole-mount single molecule FISH method for zebrafish embryo.
abstract: Noise in gene expression renders cells more adaptable to changing environment by imposing phenotypic and functional heterogeneity on genetically identical individual cells. Hence, quantitative measurement of noise in gene expression is essential for the study of biological processes in cells. Currently, there are two complementary methods for quantitatively measuring noise in gene expression at the single cell level: single molecule FISH (smFISH) and single cell qRT-PCR (or single cell RNA-seq). While smFISH has been developed for culture cells, tissue sections and whole-mount invertebrate organisms, the method has not been reported for whole-mount vertebrate organisms. Here, we report an smFISH method that is suitable for whole-mount zebrafish embryo, a popular vertebrate model organism for the studies of development, physiology and disease. We show the detection of individual transcripts for several cell-type specific and ubiquitously expressed genes at the single cell level in whole-mount zebrafish embryo. We also demonstrate that the method can be adapted to detect two different genes in individual cells simultaneously. The whole-mount smFISH method described in this report is expected to facilitate the study of noise in gene expression and its role in zebrafish, a vertebrate animal model relevant to human biology.
authors.affiliation: 1] Kyoto University, Graduate School of Biostudies, Kyoto, Japan [2] ERATO Sato Live Bio-Forecasting Project, Japan Science and Technology Agency (JST), Kyoto, Japan [3] Advanced Telecommunications Research Institute International (ATR), The Thomas N. Sato BioMEC-X Laboratories, Kyoto, Japan.
authors.forename: Yuma
authors.initials: Y
authors.lastname: Oka
authors.affiliation: 1] ERATO Sato Live Bio-Forecasting Project, Japan Science and Technology Agency (JST), Kyoto, Japan [2] Advanced Telecommunications Research Institute International (ATR), The Thomas N. Sato BioMEC-X Laboratories, Kyoto, Japan [3] Nara Institute of Science and Technology, Graduate School of Biological Sciences, Nara, Japan [4] Cornell University, Department of Biomedical Science, NY, USA [5] Centenary Institute, Sydney, Australia.
authors.forename: Thomas N
authors.initials: TN
authors.lastname: Sato
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism; D017930:Genes, Reporter; D017404:In Situ Hybridization, Fluorescence; D018613:Microscopy, Confocal; D060888:Real-Time Polymerase Chain Reaction; D015027:Zebrafish / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism