Gene expression analysis on sections of zebrafish regenerating fins reveals limitations in the whole-mount in situ hybridization method.
The caudal fin of adult zebrafish is used to study the molecular mechanisms that govern regeneration processes. Most reports of gene expression in regenerating caudal fins rely on in situ hybridization (ISH) on whole-mount samples followed by sectioning of the samples. In such reports, expression is mostly confined to cells other than those located between the dense collagenous structures that are the actinotrichia and lepidotrichia. Here, we re-examined the expression of genes by performing ISH directly on cryo-sections of regenerates. We detected expression of some of these genes in cell types that appeared to be non-expressing when ISH was performed on whole-mount samples. These results demonstrate that ISH reagents have a limited capacity to penetrate between the regenerating skeletal matrices and suggest that ISH performed directly on fin sections is a preferable method to study gene expression in fin regenerates.
Smith
A
A
Ottawa Health Research Institute, Ottawa, ON, Canada.
Zhang
J
J
Guay
D
D
Quint
E
E
Johnson
A
A
Akimenko
M A
MA
D000818:Animals; D001391:Azo Compounds; D015925:Cryopreservation; D020869:Gene Expression Profiling / Q000379:methods*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D017403:In Situ Hybridization / Q000379:methods*; D012038:Regeneration / Q000502:physiology*; D013623:Tail / Q000502:physiology*; D015027:Zebrafish / Q000502:physiology*
D001391:Azo Compounds; C009798:C.I. direct red 80