A novel reporter allele for monitoring Dll4 expression within the embryonic and adult mouse

Biology Open

卷:7

Joshua D. Wythe¹  Alexander M. Rhyner¹  Alexander M. Herman¹  Sean P. Marrelli²  W. Patrick Devine³  Benoit G. Bruneau⁴ 

[1]Cardiovascular Research Institute, Baylor College of Medicine, Houston, TX 77030, USA

[2]|Department of Neurology, McGovern Medical School at UT Health, Houston, TX 77005, USA

[3]|Department of Pathology, University of California San Francisco, San Francisco, CA 94113, USA

[4]|Gladstone Institute of Cardiovascular Disease, University of California San Francisco, San Francisco, CA 94110, USA

关键词: Delta;    Notch;    Vascular;    Angiogenesis;   

DOI  :  10.1242/bio.026799

来源: DOAJ

【 摘 要 】

Canonical Notch signaling requires the presence of a membrane bound ligand and a corresponding transmembrane Notch receptor. Receptor engagement induces multiple proteolytic cleavage events culminating in the nuclear accumulation of the Notch intracellular domain and its binding to a transcriptional co-factor to mediate gene expression. Notch signaling networks are essential regulators of vascular patterning and angiogenesis, as well as myriad other biological processes. Delta-like 4 (Dll4) encodes the earliest Notch ligand detected in arterial cells, and is enriched in sprouting endothelial tip cells. Dll4 expression has often been inferred by proxy using a lacZ knockin reporter allele. This is problematic, as a single copy of Dll4 is haploinsufficient. Additionally, Notch activity regulates Dll4 transcription, making it unclear whether these reporter lines accurately reflect Dll4 expression. Accordingly, precisely defining Dll4 expression is essential for determining its role in development and disease. To address these limitations, we generated a novel BAC transgenic allele with a nuclear-localized β-galactosidase reporter (Dll4-BAC-nlacZ). Through a comparative analysis, we show the BAC line overcomes previous issues of haploinsufficiency, it recapitulates Dll4 expression in vivo, and allows superior visualization and imaging. As such, this novel Dll4 reporter is an important addition to the growing Notch toolkit.

【 授权许可】

Unknown