Immunohistochemistry and RNA In Situ Hybridization in Mouse Brain Development
During development, the mouse brain is progressively divided into functionally distinct compartments. Numerous neuronal and glial cell types are subsequently generated in response to various inductive signals. Each cell expresses a unique combination of genes encoding proteins from transcription factors to neurotransmitters that define its role in brain function. To understand these important and highly sophisticated processes, it is critical to accurately locate the various proteins and cells that produce them. In this chapter, we introduce the techniques of Immunohistochemistry, which detects the localization of specific proteins, and RNA in situ hybridization, which enables the visualization of specific mRNAs.
KeywordsImmunohistochemistry RNA in situ hybridization Cryosection Antibody Digoxigenin Fluorescent
We would like to thank Dr. Simeone for providing the RNA in situ probe for Otx2. The monoclonal antibody against Pax6 developed by Dr. Jessell was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by Department of Biological Sciences, The University of Iowa, Iowa City, IA 52242, USA. Research in the Liu lab has been supported by US NSF (IOS-0949877 and IOS-1257540), US NIH (HD083625) and a Penn State University new faculty start-up fund.
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