Transcriptome analysis for Notch3 target genes identifies Grip2 as a novel regulator of myogenic response in the cerebrovasculature

Arterioscler Thromb Vasc Biol. 2013 Jan;33(1):76-86. doi: 10.1161/ATVBAHA.112.251736. Epub 2012 Nov 1.


Objective: Notch3 is critically important for the structure and myogenic response of distal arteries, particularly of cerebral arteries. However, signaling pathways acting downstream of Notch3 remain largely unknown.

Methods and results: Transcriptome analysis using tail arteries of Notch3-null mice identified a core set of 17 novel Notch3-regulated genes confirmed in tail or brain arteries. Postnatal deletion of RBP-Jκ in smooth muscle cells recapitulated the structural, functional, and molecular defects of brain arteries induced by Notch3 deficiency. Transient in vivo blockade of the Notch pathway with γ-secretase inhibitors uncovered, in addition to Notch3, 6 immediate responders, including the voltage-gated potassium channel Kv1.5, which opposes to myogenic constriction of brain arteries, and the glutamate receptor-interacting protein 2 (Grip2) with no previously established role in the cerebrovasculature. We identified a vascular smooth muscle cell isoform of Grip2. We showed that Notch3-RBP-Jκ specifically regulates this isoform. Finally, we found that cerebral arteries of Grip2 mutant mice, which express an N-terminally truncated Grip2 protein, exhibited selective attenuation of pressure-induced contraction.

Conclusions: Our data provide insight into how Notch3 signals in the brain arteries, establishing the postnatal requirement of smooth muscle RBP-Jκ in this context. Notch3-regulated transcriptome provides potential for modulating myogenic response in the cerebrovasculature.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alanine / analogs & derivatives
  • Alanine / pharmacology
  • Amyloid Precursor Protein Secretases / antagonists & inhibitors
  • Amyloid Precursor Protein Secretases / metabolism
  • Animals
  • Azepines / pharmacology
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Cerebral Arteries / metabolism
  • Enzyme Inhibitors / pharmacology
  • Gene Expression Profiling
  • Gene Expression Regulation
  • Immunoglobulin J Recombination Signal Sequence-Binding Protein / genetics
  • Immunoglobulin J Recombination Signal Sequence-Binding Protein / metabolism
  • Intracellular Signaling Peptides and Proteins
  • Kv1.5 Potassium Channel / genetics
  • Kv1.5 Potassium Channel / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Muscle, Smooth, Vascular / drug effects
  • Muscle, Smooth, Vascular / growth & development
  • Muscle, Smooth, Vascular / metabolism*
  • Myocytes, Smooth Muscle / drug effects
  • Myocytes, Smooth Muscle / metabolism*
  • Nerve Tissue Proteins / deficiency
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Receptor, Notch3
  • Receptors, Notch / deficiency
  • Receptors, Notch / genetics
  • Receptors, Notch / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction
  • Vasoconstriction* / drug effects
  • Vasoconstrictor Agents / pharmacology
  • Vasodilation
  • Vasodilator Agents / pharmacology


  • Azepines
  • Carrier Proteins
  • Enzyme Inhibitors
  • Grip2 protein, mouse
  • Immunoglobulin J Recombination Signal Sequence-Binding Protein
  • Intracellular Signaling Peptides and Proteins
  • Kcna5 protein, mouse
  • Kv1.5 Potassium Channel
  • N2-((2S)-2-(3,5-difluorophenyl)-2-hydroxyethanoyl)-N1-((7S)-5-methyl-6-oxo-6,7-dihydro-5H-dibenzo(b,d)azepin-7-yl)-L-alaninamide
  • Nerve Tissue Proteins
  • Notch3 protein, mouse
  • Rbpj protein, mouse
  • Receptor, Notch3
  • Receptors, Notch
  • Vasoconstrictor Agents
  • Vasodilator Agents
  • Amyloid Precursor Protein Secretases
  • Alanine