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XB-ART-6124
Dev Dyn. December 1, 2002; 225 (4): 499-510.

Techniques and probes for the study of Xenopus tropicalis development.

Khokha MK , Chung C , Bustamante EL , Gaw LW , Trott KA , Yeh J , Lim N , Lin JC , Taverner N , Amaya E , Papalopulu N , Smith JC , Zorn AM , Harland RM , Grammer TC .


Abstract
The frog Xenopus laevis has provided significant 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--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.

PubMed ID: 12454926
Article link: Dev Dyn.
Grant support: GM42341 NIGMS NIH HHS , GM66684 NIGMS NIH HHS , K12-HD00850 NICHD NIH HHS

Genes referenced: a2m ag1 egr2 en2 epha4 foxg1 gsc hhex hoxb9 hoxd10 meis3 myod1 nkx2-5 nrp1 otx2 pax2 pax3 pax8 shh snai2 sox15 sox3 t tubb2b twist1 vegt ventx2.1 ventx2.2 zic2

Antibodies referenced: Notochord Ab1 Somite Ab1
Morpholinos referenced: ctnnb1 MO1 ctnnb1 MO2


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