Techniques and Probes for the Study of Xenopus tropicalis Development.

Khokha, M.K., Chung, C., Bustamante, E., Gaw, L., Trott, K., Yeh, J., Lim, N., Lin, J., Taverner, N.,

Amaya, E., Papalopulu, N., Smith, J.C., Zorn, A.M., Harland, R.M. and Grammer, T.C.

Developmental Dynamics 225:499-510 (2002). Full Article (1700K)

Abstract

The frog Xenopus laevis has provided signicant insights into developmental and cellular processes. However,

X. laevis has an allotetraploid genome precluding its use in forward genetic analysis. Genetic analysis may be

applicable to Xenopus (Silurana) tropicalis, which has a diploid genome and a shorter generation time. Here,

we show that many tools for the study of X.laevis development can be applied to X. tropicalis. By using the

developmental staging system of Nieuwkoop and Faber,we find that X. tropicalis embryos develop at similar

rates to X. laevis, although they tolerate a narrower range of temperatures. We also show that many of the

analytical reagents available for X.laevis can be effectively transferred to X. tropicalis. The X. laevis protocol

for whole-mount in situ hybridization to mRNA transcripts can be successfully applied to X. tropicalis without

alteration. Additionally, X. laevis probes often work in X. tropicalis &emdash;alleviating the immediate need to

clone the X. tropicalis orthologs before initiating developmental studies.Antibodies that react against X. laevis

proteins can effectively detect the X. tropicalis protein 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 in X. tropicalis. Finally, X. tropicalis offers the

possibility for forward genetics and genomic analysis.

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