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.
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In some cases, reporting the developmental stage is more informative than chronological age-for example, in juvenile Xenopus, in which rate of development can be manipulated by incubation temperature.
“In some cases, however, reporting the developmental stage is more informative than chronological age—for example, in juvenile Xenopus, in which rate of development can be manipulated by incubation temperature [109].”
From Reporting animal research: Explanation and elaboration for the ARRIVE guidelines 2.0 (2020) · cites this paper as [109]
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References 53 · Cited by 241
- Authors
- Khokha, M. K., Chung, C., Bustamante, E. L., Gaw, L. W. K., Trott, K. A., Yeh, J., Lim, N., Lin, J. C. Y., Taverner, N., Amaya, E., Papalopulu, N., Smith, J. C., Zorn, A. M., Harland, R. M., & Grammer, T. C.
- Affiliations
- University of California, Berkeley (US)
- Wellcome Trust (GB)
- Cincinnati Children's Hospital Medical Center (US)
- Corresponding author
- Harland, Richard M.
- ORCID
- Venue
- Developmental Dynamics
- Publisher
- Wiley
- Published
- 2002-11-25
- Volume
- 225
- Issue
- 4
- Pages
- 499-510
- ISSN
- 1058-8388 (ISSN-L)
- 1097-0177
- OpenAlex
- W2149642765
- Cited by
- 241
- References
- 53
- Citations by year
- 2026: 7 · 2025: 11 · 2024: 8 · 2023: 9 · 2022: 13 · 2021: 11 · 2020: 15 · 2019: 11 · 2018: 20 · 2017: 8 · 2016: 6 · 2015: 17 · 2014: 10 · 2013: 4 · 2012: 13
- FWCI
- 3.13
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Developmental Dynamics (publishedVersion)
- PubMed (publishedVersion)
- Research Explorer (The University of Manchester) (submittedVersion)
- Funders
- University of Virginia
- National Institute of General Medical Sciences (GM66684, GM 42341, R01GM042341, R01GM066684)
- Eunice Kennedy Shriver National Institute of Child Health and Human Development (K-12-HD 00850, K12HD000850)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Zebrafish Biomedical Research Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
- Keywords
- Xenopus tropicalis, Xenopus laevis, amphibian embryonic development, developmental staging, whole-mount in situ hybridization, antibody cross-reactivity, morpholino antisense oligonucleotides, forward genetics, diploid genome, heat tolerance
- MeSH terms
- Animals, Base Sequence, Cytoskeletal Proteins, Ectoderm, Endoderm, Immunohistochemistry, Mesoderm, Molecular Sequence Data, RNA, Messenger, Time Factors, Xenopus, Developmental Biology, Trans-Activators, Gene Library, Oligonucleotides, Antisense, In Situ Hybridization, DNA, Complementary, Gene Expression Regulation, Developmental, Xenopus Proteins, beta Catenin
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