Techniques and probes for the study of<i>Xenopus tropicalis</i>development

Khokha, Mustafa K.; Chung, Christina Z.; Bustamante, Erika L.; Gaw, Lisa W.K.; Trott, Kristin A.; Yeh, Joanna; Lim, Nancy; Lin, Jennifer; Taverner, Nicola; Amaya, Enrique; Papalopulu, Nancy; Smith, James C.; Zorn, Aaron M.; Harland, Richard M.; Grammer, Timothy C.

doi:10.1002/dvdy.10184

Cited by 227 publications

(207 citation statements)

References 78 publications

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“…The intense development of genomic resources and the concerted effort put into sequencing the genome of X. tropicalis argue strongly in favor of using this species to develop genetic approaches in the future. The observation that more than 85% of the coding sequence of caspase 9 is conserved between the two species is a strong argument in favor of using data from either in functional studies (Khokha et al, 2002).…”

Section: Discussionmentioning

confidence: 99%

Apoptosis of tail muscle during amphibian metamorphosis involves a caspase 9-dependent mechanism

et al. 2005

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The climax of amphibian metamorphosis is marked by thyroid hormone-dependent tadpole tail resorption, implicating apoptosis of multiple cell types, including epidermal cells, fibroblasts, nerve cells, and muscles. The molecular cascades leading to and coordinating the death of different cell types are not fully elucidated. It is known that the mitochondrial pathway, and in particular the Bax and XR11 genes, regulates the balance between apoptosis and survival in muscle. However, the down-stream factors modulated by changes in mitochondrial permeability have not been studied in a functional context. To investigate further the mitochondrial-dependent pathway, we analyzed the regulation and the role of caspase 9 in Xenopus tadpoles. We report that caspase 9 mRNA is expressed in the tail before metamorphosis and increases before and during climax. Similarly, at the protein level, the production of active forms of caspase 9 increases in muscle tissue as metamorphosis progresses. To assess the functional role of caspase 9, we designed a dominant-negative protein. Overexpression of this dominant-negative abrogates both Bax-induced cell death in vitro and muscle apoptosis in vivo during natural metamorphosis. These findings consolidate a model of metamorphic muscle death that directly implicates the mitochondrial pathway and the apoptosome. Developmental Dynamics 233:76 -87, 2005.

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Section: Discussionmentioning

confidence: 99%

Apoptosis of tail muscle during amphibian metamorphosis involves a caspase 9-dependent mechanism

et al. 2005

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“…The genome of Xenopus tropicalis was sequenced recently to construct a genetic linkage map (Kochan et al, 2003). Previous studies have compared the developmental processes in Xenopus laevis and Xenopus tropicalis (Khokha et al, 2002, Sargent and Mohun, 2005, Shook et al, 2004. Fortunately, the same methods established to investigate early development in Xenopus laevis, including mRNA injection, whole-mount in situ hybridization and transplantation can also be used in the study of Xenopus tropicalis development (Khokha et al, 2002, Offield et al, 2000.…”

Section: Introductionmentioning

confidence: 99%

“…Previous studies have compared the developmental processes in Xenopus laevis and Xenopus tropicalis (Khokha et al, 2002, Sargent and Mohun, 2005, Shook et al, 2004. Fortunately, the same methods established to investigate early development in Xenopus laevis, including mRNA injection, whole-mount in situ hybridization and transplantation can also be used in the study of Xenopus tropicalis development (Khokha et al, 2002, Offield et al, 2000. In addition, many of the genes involved in early development in Xenopus laevis are expressed in the same regions and have the same functions in Xenopus tropicalis (D'Souza et al, 2003, Fisher et al, 2003, Fletcher et al, 2004, Knochel et al, 2001.…”

Section: Introductionmentioning

confidence: 99%

Comparison of induction during development between Xenopus tropicalis and Xenopus laevis

Sedohara¹,

Suzawa²,

Asashima³

2006

Int. J. Dev. Biol.

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Several in vitro systems exist for the induction of animal caps using growth factors such as activin. In this paper, we compared the competence of activin-treated animal cap cells dissected from the late blastulae of Xenopus tropicalis and Xenopus laevis. The resultant tissue explants from both species differentiated into mesodermal and endodermal tissues in a dosedependent manner. In addition, RT-PCR analysis revealed that organizer and mesoderm markers were expressed in a similar temporal and dose-dependent manner in tissues from both organisms. These results indicate that animal cap cells from Xenopus tropicalis have the same competence in response to activin as those from Xenopus laevis.

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“…To synthesize RNA sense probes we used the same clones and the Maxiscript SP6 (AM1308) from Ambion, using digoxigenin labeled dUTP. The wholemount in situ hybridization procedure was done according to the standard protocol (Khokha et al, 2002). Embryos were stained with BM Purple at room temperature or 4 o C depending on ideal conditions for each probe.…”

Section: Whole-mount In Situ Hybridizationmentioning

confidence: 99%

Nucleoporin gene expression in Xenopus tropicalis embryonic development

Reza

Khokha²,

Viso³

2016

Int. J. Dev. Biol.

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Nucleoporins (nups) compose the structure of the nuclear pore complex (NPC) of all cells, but several studies have illuminated nucleoporins' additional roles in development and the cell cycle. However, a comprehensive study of nup expression in embryonic development has not yet been reported. We synthesized antisense probes for all nup genes and used whole-mount in situ hybridization techniques to determine the expression pattern of all members of the nup family of genes at three different developmental stages in Xenopus tropicalis. We found that the expression of nups was not ubiquitous in embryos, but was localized to specific and distinguishable anatomical structures at all three stages tested. We also found that the expression patterns for nups within the same subcomplexes were not necessarily identical. Thus, nup expression is subject to a significant level of regulation during development. These results provide new information for functional studies of nups to unravel their roles in embryonic development. KEY WORDS: nups, NPC, nuclear pore complex, in situ hybridizationThe nuclear membrane forms an essential boundary for the nucleus and is a major site of macromolecular traffic in the cell, necessitating a mechanism for selective transport (Lusk et al., 2007). The nuclear pore complex (NPC) is a cylindrical array of proteins, embedded in the nuclear envelope, which creates a portal for macromolecular transport. The structure of the NPC has been analyzed and categorized into a cytoplasmic ring, a central spoke ring, and a nuclear ring (Grossman et al., 2012). Each of these subunits is composed of proteins called nucleoporins (nups), which form different subcomplexes: transmembrane, scaffold (outer and inner ring subcomplexes), cytoplasmic (ring/ linker), nuclear basket, and channel nups. Transmembrane nups anchor the NPC to the nuclear envelope, while scaffold and central channel nups make up the central structure of the transport complex (Fig. 1A). Several cytoplasmic, channel and nuclear basket nups have phenylalanine-glycine (FG) repeats that assist in trafficking molecules. For instance, central FG-repeat nups arrayed in the central pore create the permeability barrier of the NPC (Onischenko and Weis, 2011). A new study has recently elucidated the underlying protein-protein interaction network within the NPC. Analyzing the X-rays crystallographics of scaffold nucleoporins enabled the construction of full-length atomic structures, ultimately leading to a massive composite structure of the NPC (Lin et al., 2016).

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Techniques and probes for the study ofXenopus tropicalisdevelopment

Cited by 227 publications

References 78 publications

Apoptosis of tail muscle during amphibian metamorphosis involves a caspase 9-dependent mechanism

Apoptosis of tail muscle during amphibian metamorphosis involves a caspase 9-dependent mechanism

Comparison of induction during development between Xenopus tropicalis and Xenopus laevis

Nucleoporin gene expression in Xenopus tropicalis embryonic development

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