title: Techniques and probes for the study of Xenopus tropicalis development.
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.
authors.affiliation: Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, California, USA.
authors.forename: Mustafa K
authors.initials: MK
authors.lastname: Khokha
authors.forename: Christina
authors.initials: C
authors.lastname: Chung
authors.forename: Erika L
authors.initials: EL
authors.lastname: Bustamante
authors.forename: Lisa W K
authors.initials: LW
authors.lastname: Gaw
authors.forename: Kristin A
authors.initials: KA
authors.lastname: Trott
authors.forename: Joanna
authors.initials: J
authors.lastname: Yeh
authors.forename: Nancy
authors.initials: N
authors.lastname: Lim
authors.forename: Jennifer C Y
authors.initials: JC
authors.lastname: Lin
authors.forename: Nicola
authors.initials: N
authors.lastname: Taverner
authors.forename: Enrique
authors.initials: E
authors.lastname: Amaya
authors.forename: Nancy
authors.initials: N
authors.lastname: Papalopulu
authors.forename: James C
authors.initials: JC
authors.lastname: Smith
authors.forename: Aaron M
authors.initials: AM
authors.lastname: Zorn
authors.forename: Richard M
authors.initials: RM
authors.lastname: Harland
authors.forename: Timothy C
authors.initials: TC
authors.lastname: Grammer
mesh_terms: D000818:Animals; D001483:Base Sequence; D003598:Cytoskeletal Proteins / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D015509:Developmental Biology / Q000379:methods*; D004475:Ectoderm / Q000378:metabolism; D004707:Endoderm / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008648:Mesoderm / Q000378:metabolism; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism; D014981:Xenopus / Q000196:embryology*; D029867:Xenopus Proteins; D051176:beta Catenin
chemical_list: C495266:CTNNB1 protein, Xenopus; D003598:Cytoskeletal Proteins; D018076:DNA, Complementary; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D015534:Trans-Activators; D029867:Xenopus Proteins; D051176:beta Catenin