Mapping the consequence of Notch1 proteolysis in vivo with NIP-CRE.
The four highly conserved Notch receptors receive short-range signals that control many biological processes during development and in adult vertebrate tissues. The involvement of Notch1 signaling in tissue self-renewal is less clear, however. We developed a novel genetic approach N(1)IP-CRE (Notch1 Intramembrane Proteolysis) to follow, at high resolution, the descendents of cells experiencing Notch1 activation in the mouse. By combining N(1)IP-CRE with loss-of-function analysis, Notch activation patterns were correlated with function during development, self-renewal and malignancy in selected tissues. Identification of many known functions of Notch1 throughout development validated the utility of this approach. Importantly, novel roles for Notch1 signaling were identified in heart, vasculature, retina and in the stem cell compartments of self-renewing epithelia. We find that the probability of Notch1 activation in different tissues does not always indicate a requirement for this receptor and that gradients of Notch1 activation are evident within one organ. These findings highlight an underappreciated layer of complexity of Notch signaling in vivo. Moreover, NIP-CRE represents a general strategy applicable for monitoring proteolysis-dependent signaling in vivo.
Vooijs
Marc
M
Department of Molecular Biology and Pharmacology and the Department of Medicine, Division of Dermatology, Washington University Medical School, St Louis, MO 63110, USA. m.vooijs@umcutrecht.nl
Ong
Chin-Tong
CT
Hadland
Brandon
B
Huppert
Stacey
S
Liu
Zhenyi
Z
Korving
Jeroen
J
van den Born
Maaike
M
Stappenbeck
Thaddeus
T
Wu
Yumei
Y
Clevers
Hans
H
Kopan
Raphael
R
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D004475:Ectoderm / Q000378:metabolism; D004707:Endoderm / Q000378:metabolism; D004817:Epidermis / Q000196:embryology / Q000378:metabolism; D005318:Fetal Heart / Q000196:embryology / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D008648:Mesoderm / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008954:Models, Biological; D009474:Neurons / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D017434:Protein Structure, Tertiary; D051881:Receptor, Notch1 / Q000737:chemistry / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
C497762:Notch1 protein, mouse; D051881:Receptor, Notch1; D004247:DNA; D010447:Peptide Hydrolases