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Whole-mount fluorescent in situ hybridization staining of the colonial tunicate Botryllus schlosseri.

ABSTRACT: Botryllus schlosseri is a colonial ascidian with characteristics that make it an attractive model for studying immunology, stem cell biology, evolutionary biology, and regeneration. Transcriptome sequencing and the recent completion of a draft genome sequence for B. schlosseri have revealed a large number of genes, both with and without vertebrate homologs, but analyzing the spatial and temporal expression of these genes in situ has remained a challenge. Here, we report a robust protocol for in situ hybridization that enables the simultaneous detection of multiple transcripts in whole adult B. schlosseri using Tyramide Signal Amplification in conjunction with digoxigenin- and dinitrophenol-labeled RNA probes. Using this protocol, we have identified a number of genes that can serve as markers for developing and mature structures in B. schlosseri, permitting analysis of phenotypes induced in loss-of-function experiments.

SUBMITTER: Langenbacher AD 

PROVIDER: S-EPMC4308483 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature

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Whole-mount fluorescent in situ hybridization staining of the colonial tunicate Botryllus schlosseri.

Langenbacher Adam D AD   Rodriguez Delany D   Di Maio Alessandro A   De Tomaso Anthony W AW  

Genesis (New York, N.Y. : 2000) 20140917 1

Botryllus schlosseri is a colonial ascidian with characteristics that make it an attractive model for studying immunology, stem cell biology, evolutionary biology, and regeneration. Transcriptome sequencing and the recent completion of a draft genome sequence for B. schlosseri have revealed a large number of genes, both with and without vertebrate homologs, but analyzing the spatial and temporal expression of these genes in situ has remained a challenge. Here, we report a robust protocol for in  ...[more]

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