Techniques and probes for the study ofXenopus tropicalisdevelopment

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Abstract
The frogXenopus laevishas provided significant insights into developmental and cellular processes. However,X. laevishas an allotetraploid genome precluding its use in forward genetic analysis. Genetic analysis may be applicable toXenopus (Silurana) tropicalis, which has a diploid genome and a shorter generation time. Here, we show that many tools for the study ofX. laevisdevelopment can be applied toX. tropicalis. By using the developmental staging system of Nieuwkoop and Faber, we find thatX. tropicalisembryos develop at similar rates toX. laevis, although they tolerate a narrower range of temperatures. We also show that many of the analytical reagents available forX. laeviscan be effectively transferred toX. tropicalis. TheX. laevisprotocol for whole‐mount in situ hybridization to mRNA transcripts can be successfully applied toX. tropicaliswithout alteration. Additionally,X. laevisprobes often work inX. tropicalis—alleviating the immediate need to clone theX. tropicalisorthologs before initiating developmental studies. Antibodies that react againstX. laevisproteins can effectively detect theX. tropicalisprotein by using established immunohistochemistry procedures. Antisense morpholino oligonucleotides (MOs) offer a new alternative to study loss of gene activity during development. We show that MOs function inX. tropicalis. Finally,X. tropicalisoffers the possibility for forward genetics and genomic analysis.