ArticleBMC developmental biology2013
In situ hybridization protocol for enhanced detection of gene expression in the planarian Schmidtea mediterranea.
Article in BMC developmental biology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 205 papers.
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Who cites it
205 citing papers in PubMed, 343 citations in OpenAlex.
- Combinatorial mechanisms specify cellular location and neurotransmitter identity during planarian neurogenesis.Nature communications · 2026Article
- Multiplexed gene expression analysis in dissociated and intact planarian flatworm tissues using in situ hybridization chain reaction.Biology open · 2026Article
- Ephrin Signaling Patterns Sensory Neurons During Tissue Homeostasis in Planarians.bioRxiv : the preprint server for biology · 2026Article
- Lack of specificity of planarian progenitor responses to injury in regeneration.bioRxiv : the preprint server for biology · 2026Article
- TIP60 acts as a wound-induced factor essential for efficient regeneration in planaria.The Journal of biological chemistry · 2026Article
- Protocol for purifying and sequencing cell subpopulations based on RNA FISH signal in planaria.STAR protocols · 2026Article
- Redox-modulated ERK dynamics support wound-dependent tissue formation during early planarian regeneration.iScience · 2026Article
- Host-transposon mutualism supports regeneration in planarians.Developmental cell · 2026Article
- Drift of positional identity drives reproductive aging in a long-lived regenerative animal.Current biology : CB · 2026Article
- Conserved innate immunity components limit transgene expression in adult planarians.Development (Cambridge, England) · 2026Article
- SoxB1-mediated chromatin remodeling promotes sensory neuron differentiation in planarians.Genetics · 2026Article
- Serotonin signaling modulates growth and motility in juvenile Fasciola hepatica.PLoS neglected tropical diseases · 2026Article
- Schistosome esophageal gland factor MEG-8.2 drives host cell lysis and interacts with host immune proteins.PLoS pathogens · 2026Article
- Single-nucleotide m⁶A mapping uncovers redundant YTHDF function in planarian progenitor fate selection.The EMBO journal · 2026Article
- RoboA reinforces planarian stem cell fate through FoxA and Anosmin1a.Nature communications · 2026Article
- 4D single-cell spatial transcriptomics reveals dynamic morphogenetic gradients and regenerative domains in planarians.GigaScience · 2026Article
- Metabolites produced by agat+ cells support regeneration in the planarian Schmidtea mediterranea.Developmental biology · 2026Article
- A Tousled-like kinase (TLK) homolog required for male germline stem cell homeostasis in the planarian Schmidtea mediterranea.Developmental biology · 2026Article
- Immunolocalization in Xenopus.Methods in molecular biology (Clifton, N.J.) · 2026Article
- SoxB1-Mediated Chromatin Remodeling Promotes Sensory Neuron Differentiation in Planarians.bioRxiv : the preprint server for biology · 2025Article
145 more citing papers are in PubMed but not listed here.
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe freshwater planarian Schmidtea mediterranea has emerged as a powerful model for studies of regenerative, stem cell, and germ cell biology. Whole-mount in situ hybridization (WISH) and whole-mount fluorescent in situ hybridization (FISH) are critical methods for determining gene expression patterns in planarians. While expression patterns for a number of genes have been elucidated using established protocols, determining the expression patterns for particularly low-abundance transcripts remains a challenge.
resultsWe show here that a short bleaching step in formamide dramatically enhances signal intensity of WISH and FISH. To further improve signal sensitivity we optimized blocking conditions for multiple anti-hapten antibodies, developed a copper sulfate quenching step that virtually eliminates autofluorescence, and enhanced signal intensity through iterative rounds of tyramide signal amplification. For FISH on regenerating planarians, we employed a heat-induced antigen retrieval step that provides a better balance between permeabilization of mature tissues and preservation of regenerating tissues. We also show that azide most effectively quenches peroxidase activity between rounds of development for multicolor FISH experiments. Finally, we apply these modifications to elucidate the expression patterns of a few low-abundance transcripts.
conclusionThe modifications we present here provide significant improvements in signal intensity and signal sensitivity for WISH and FISH in planarians. Additionally, these modifications might be of widespread utility for whole-mount FISH in other model organisms.
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