NDLI: Understanding early organogenesis using a simplified in situ hybridization protocol in Xenopus.

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Understanding early organogenesis using a simplified in situ hybridization protocol in Xenopus.

Understanding early organogenesis using a simplified in situ hybridization protocol in Xenopus.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Deimling, Steven J. Halabi, Rami R. Grover, Stephanie A. Wang, Jean H. Drysdale, Thomas A.
Description	Author Affiliation: Deimling SJ ( Developmental and Stem Cell Biology, Hospital for Sick Children.); Halabi RR ( Children's Health Research Institute, University of Western Ontario); Grover SA ( Neurosciences and Mental Health, Hospital for Sick Children.); Wang JH ( Children's Health Research Institute, University of Western Ontario); Drysdale TA ( Children's Health Research Institute, University of Western Ontario)
Abstract	Organogenesis is the study of how organs are specified and then acquire their specific shape and functions during development. The Xenopuslaevis embryo is very useful for studying organogenesis because their large size makes them very suitable for identifying organs at the earliest steps in organogenesis. At this time, the primary method used for identifying a specific organ or primordium is whole mount in situ hybridization with labeled antisense RNA probes specific to a gene that is expressed in the organ of interest. In addition, it is relatively easy to manipulate genes or signaling pathways in Xenopus and in situ hybridization allows one to then assay for changes in the presence or morphology of a target organ. Whole mount in situ hybridization is a multi-day protocol with many steps involved. Here we provide a simplified protocol with reduced numbers of steps and reagents used that works well for routine assays. In situ hybridization robots have greatly facilitated the process and we detail how and when we utilize that technology in the process. Once an in situ hybridization is complete, capturing the best image of the result can be frustrating. We provide advice on how to optimize imaging of in situ hybridization results. Although the protocol describes assessing organogenesis in Xenopus laevis, the same basic protocol can almost certainly be adapted to Xenopus tropicalis and other model systems.
File Format	HTM / HTML
e-ISSN	1940087X
Journal	Journal of Visualized Experiments
Issue Number	95
Language	English
Publisher	MyJove Corp.
Publisher Date	2015-01-12
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine In Situ Hybridization Organogenesis Physiology Xenopus Laevis Embryology Animals Models, Animal Research Support, Non-u.s. Gov't Video-audio Media
Content Type	Text
Resource Type	Article

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