Several laboratories have developed genetic methods to monitor Notch activity in developing and adult mice. These approaches have been useful in identifying Notch signaling with high temporal and spatial resolution. This research has contributed substantially to our understanding of the role of Notch in cell specification and cellular physiology. Here, we present two protocols to monitor Notch activity in the mouse brain: (1) by intraventricular electroporation and (2) by intracranial viral injections of Notch reporter constructs. These methods allow monitoring of Notch signaling in specific brain regions from development to adulthood. In addition, using the appropriate modifications, the Notch reporter systems can also be used to monitor Notch activity in other organs of the mouse such as retina, skin, skeletal muscle, and cancer cells.
This is a preview of subscription content, log in to check access.
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
We would like to thank Nicholas Gaiano and Kenichi Mizutani for allowing us to reprint one of their results and for helping with the intraventricular electroporation technique. We would also like to thank Mauro Giacca and Lorena Zentilin for packaging the AAVs. This work is supported by the Swiss National Foundation, the Synapsis Foundation for Alzheimer’s Research, and Swiss Heart Association.
Ishibashi M, Moriyoshi K, Sasai Y et al (1994) Persistent expression of helix-loop-helix factor HES-1 prevents mammalian neural differentiation in the central nervous system. EMBO J 13:1799–1805PubMedCentralPubMedGoogle Scholar
Shimizu K, Chiba S, Saito T et al (2002) Functional diversity among Notch1, Notch2, and Notch3 receptors. Biochem Biophys Res Commun 291:775–779PubMedCrossRefGoogle Scholar
Ohtsuka T, Imayoshi I, Shimojo H et al (2006) Visualization of embryonic neural stem cells using Hes promoters in transgenic mice. Mol Cell Neurosci 31:109–122PubMedCrossRefGoogle Scholar
Masamizu Y, Ohtsuka T, Takashima Y et al (2006) Real-time imaging of the somite segmentation clock: Revelation of unstable oscillators in the individual presomitic mesoderm cells. Proc Natl Acad Sci U S A 103:1313–1318PubMedCentralPubMedCrossRefGoogle Scholar
Vilas-Boas F, Fior R, Swedlow JR et al (2011) A novel reporter of notch signalling indicates regulated and random Notch activation during vertebrate neurogenesis. BMC Biol 9:58PubMedCentralPubMedCrossRefGoogle Scholar
Smith E, Claudinot S, Lehal R et al (2012) Generation and characterization of a Notch1 signaling-specific reporter mouse line. Genesis 50:700–710PubMedGoogle Scholar
Ilagan MX, Lim S, Fulbright M et al (2011) Real-time imaging of Notch activation using a luciferase complementation-based reporter. Sci Signal 4:rs7PubMedCentralPubMedGoogle Scholar
Nagy A (2000) Cre recombinase: the universal reagent for genome tailoring. Genesis 26:99–109PubMedGoogle Scholar
Gradinaru V, Thompson KR, Deisseroth K (2008) eNpHR: a Natronomonas halorhodopsin enhanced for optogenetic applications. Brain Cell Biol 36:129–139PubMedCentralPubMedGoogle Scholar
Maillard I, Weng AP, Carpenter AC et al (2004) Mastermind critically regulates Notch-mediated lymphoid cell fate decisions. Blood 104:1696–1702PubMedGoogle Scholar
Aschauer DF, Kreuz S, Rumpel S (2013) Analysis of transduction efficiency, tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One 8:e76310PubMedCentralPubMedGoogle Scholar
Fernández ME, Croce S, Boutin C et al (2011) Targeted electroporation of defined lateral ventricular walls: a novel and rapid method to study fate specification during postnatal forebrain neurogenesis. Neural Dev 6:13PubMedCentralPubMedGoogle Scholar
Tanigaki K, Han H, Yamamoto N et al (2002) Notch-RBP-J signaling is involved in cell fate determination of marginal zone B cells. Nat Immunol 3:443–450PubMedGoogle Scholar