Animal Cap Assay
Animal Cap Assay - Related Videos
11:00
Tissue Determination Using the Animal Cap Transplant (ACT) Assay in Xenopus laevis
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Cited by 8 •
2010
Animal caps overexpressing gene product(s) are transplanted to the flank of developing Xenopus laevis embryos in order to establish whether tissue is determined.
06:58
Animal Behavior - Student Protocol
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2019
Making Choice Chambers ExpandBefore beginning the experiments, use one of the Sharpies from the lab table to trace a line around the circumference of the bottom of both bottles. One line should be 1 cm from the base of the bottle and the other should be 1.5 cm from the base of the bottle. Use a razor blade to make a small cut in each bottle at the mark. Then, insert the scissors and carefully cut along the line until the bottoms of the bottles are removed completely. Fit one bottle bottom into...
14:21
Dissection of Organizer and Animal Pole Explants from Xenopus laevis Embryos and Assembly of a Cell Adhesion Assay
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Cited by 2 •
2007
This video demonstrates the technique used for preparation of organizer and animal pole explants from Xenopus laevis embryos, including the use of the eyebrow knife - a specialized dissection tool made of one's eyebrow. The protocol for assembling an adhesion assay is also given, which probes for the presence of key adhesion molecules present on the surface organizer or animal pole cells that are critical for proper development.
08:55
In vitro Transcription and Capping of Gaussia Luciferase mRNA Followed by HeLa Cell Transfection
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Cited by 13 •
2012
This method describes high yield in vitro synthesis of both capped and uncapped mRNA from a linearized plasmid containing the Gaussia luciferase (GLuc) gene. The RNA is purified and a fraction of the uncapped RNA is enzymatically capped using the Vaccinia virus capping enzyme. In the final step, the mRNA is transfected into HeLa cells and cell culture supernatants are assayed for luciferase activity.
09:52
An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics
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Cited by 3 •
2020
Tracking individual translation events allows for high-resolution kinetic studies of cap-dependent translation mechanisms. Here we demonstrate an in vitro single-molecule assay based on imaging interactions between fluorescently labeled antibodies and epitope-tagged nascent peptides. This method enables single-molecule characterization of initiation and peptide elongation kinetics during active in vitro cap-dependent translation.
FAQs
Isolation removes the animal cap from many of the surrounding embryonic influences present in the intact embryo. Researchers can then observe how the explant develops on its own and compare that behavior with responses produced after exposure to candidate signals or tissues. This separation helps identify which properties arise within the cells and which depend on external cues.
Candidate signals or tissues can reveal whether embryonic ectoderm is responsive to a particular inductive cue. Changes in the cultured explant, including altered gene expression or cell differentiation, provide evidence that the added signal influences cell fate. The assay therefore connects an external developmental cue with a measurable response in isolated embryonic tissue.
Activin, BMP, and FGF signaling are examined because the animal cap assay has been central to studying how these pathways contribute to embryonic induction and patterning. Testing tissue responses in isolation allows investigators to relate pathway activity to changes in gene expression and differentiation, helping clarify how signaling influences the developmental potential of ectoderm.
Two major outcomes are changes in gene expression and cell differentiation. Gene-expression changes indicate that the cue has altered the developmental program of the explant, while differentiation shows a resulting change in cell identity or tissue behavior. Considering both outcomes gives a more informative picture of how surrounding signals regulate embryonic cell fate.
The procedure begins with a blastula-stage amphibian embryo, from which researchers excise the animal pole region. They culture the resulting explant, often in the presence of a candidate signaling molecule or tissue, and then assess changes in gene expression or differentiation. These steps connect a defined exposure with a developmental response in the isolated tissue.
This assay is useful when researchers need to test how embryonic ectoderm responds to inductive information without relying only on observations within the whole embryo. It supports studies of mesoderm induction, embryonic patterning, and signaling pathways. By linking experimental cues to tissue responses, the method helps analyze mechanisms that regulate early developmental decisions.