Techniques and probes for the study of Xenopus tropicalis development.
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.
Khokha
Mustafa K
MK
Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, California, USA.
Chung
Christina
C
Bustamante
Erika L
EL
Gaw
Lisa W K
LW
Trott
Kristin A
KA
Yeh
Joanna
J
Lim
Nancy
N
Lin
Jennifer C Y
JC
Taverner
Nicola
N
Amaya
Enrique
E
Papalopulu
Nancy
N
Smith
James C
JC
Zorn
Aaron M
AM
Harland
Richard M
RM
Grammer
Timothy C
TC
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
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