Panayiotis Zagouras

Intracellular Cleavage of Notch Leads to a Heterodimeric Receptor on the Plasma Membrane

Previous models for signal transduction via the Notch pathway have depicted the full-length Notch receptor expressed at the cell surface. We present evidence demonstrating that the Notch receptor on the plasma membrane is cleaved. This cleavage is an evolutionarily conserved, general property of Notch and occurs in the trans-Golgi network as the receptor traffics toward the plasma membrane. Although full-length Notch is detectable in the cell, it does not reach the surface. Cleavage results in a C-terminal fragment, N(TM), that appears to be cleaved N-terminal to the transmembrane domain, and an N-terminal fragment, N(EC), that contains most of the extracellular region. We provide evidence that these fragments are tethered together on the plasma membrane by a link that is sensitive to reducing conditions, forming a heterodimeric receptor.

Involvement of Notch-1 in mammalian retinal neurogenesis: association of Notch-1 activity with both immature and terminally differentiated cells

The Notch pathway is thought to define an evolutionarily conserved signaling mechanism that regulates the differentiation of immature cells through cells interactions. We have examined the expression of the Notch-1 receptor, the central element of this pathway, in the developing rat retina, where cell-fate choices depend upon a series of local cell interactions. Notch-1 immunoreactivity is associated with differentiating cells at different stages of retinal neurogenesis, suggesting that Notch-1 may play a role in the successive cell-fate determination which governs retinal development. In addition, the Notch-1 immunoreactivity is detected in nuclei of postmitotic, differentiated neurons of the adult retina. Our observations raise the possibility that besides its role in the differentiation of immature cell populations Notch-1 activity may also be involved in the maintenance of the differentiated state.

Molecular Characterization of Novel and Selective Peroxisome Proliferator-Activated Receptor α Agonists with Robust Hypolipidemic Activity in Vivo

The nuclear receptor peroxisome proliferator-activated receptor α (PPARα) is recognized as the primary target of the fibrate class of hypolipidemic drugs and mediates lipid lowering in part by activating a transcriptional cascade that induces genes involved in the catabolism of lipids. We report here the characterization of three novel PPARα agonists with therapeutic potential for treating dyslipidemia. These structurally related compounds display potent and selective binding to human PPARα and support robust recruitment of coactivator peptides in vitro. These compounds markedly potentiate chimeric transcription systems in cell-based assays and strikingly lower serum triglycerides in vivo. The transcription networks induced by these selective PPARα agonists were assessed by transcriptional profiling of mouse liver after short- and long-term treatment. The induction of several known PPARα target genes involved with fatty acid metabolism were observed, reflecting the expected pharmacology associated with PPARα activation. We also noted the down-regulation of a number of genes related to immune cell function, the acute phase response, and glucose metabolism, suggesting that these compounds may have anti-inflammatory action in the mammalian liver. Whereas these compounds are efficacious in acute preclinical models, extended safety studies and further clinical testing will be required before the full therapeutic promise of a selective PPARα agonist is realized.