The binding of advanced glycation end products (AGE) to the receptor for AGE (RAGE) is known to deteriorate various cell functions and is implicated in the pathogenesis of diabetic vascular complications. Here we show that AGE, tumor necrosis factor-␣ (TNF-␣), and 17␤-estradiol (E 2 ) up-regulated RAGE mRNA and protein levels in human microvascular endothelial cells and ECV304 cells, with the mRNA stability being essentially invariant. Transient transfection experiments with human RAGE promoter-luciferase chimeras revealed that the region from nucleotide number ؊751 to ؊629 and the region from ؊239 to ؊89 in the RAGE 5-flanking sequence exhibited the AGE/TNF-␣ and E 2 responsiveness, respectively. Site-directed mutation of an nuclear factor-B (NF-B) site at ؊671 or of Sp-1 sites at ؊189 and ؊172 residing in those regions resulted in an abrogation of the AGE/TNF-␣-or E 2 -mediated transcriptional activation. Electrophoretic mobility shift assays revealed that ECV304 cell nuclear extracts contained factors which retarded the NF-B and Sp-1 elements, and that the DNA-protein complexes were supershifted by antip65/p50 NF-B and anti-Sp-1/estrogen receptor ␣ antibodies, respectively. These results suggest that AGE, TNF-␣, and E 2 can activate the RAGE gene through NF-B and Sp-1, causing enhanced AGE-RAGE interactions, which would lead to an exacerbation of diabetic microvasculopathy.

Rod/cone photoreceptors of the outer retina and the melanopsin-expressing retinal ganglion cells (mRGCs) of the inner retina mediate non-image forming visual responses including entrainment of the circadian clock to the ambient light, the pupillary light reflex (PLR), and light modulation of activity. Targeted deletion of the melanopsin gene attenuates these adaptive responses with no apparent change in the development and morphology of the mRGCs. Comprehensive identification of mRGCs and knowledge of their specific roles in image-forming and non-image forming photoresponses are currently lacking. We used a Cre-dependent GFP expression strategy in mice to genetically label the mRGCs. This revealed that only a subset of mRGCs express enough immunocytochemically detectable levels of melanopsin. We also used a Cre-inducible diphtheria toxin receptor (iDTR) expression approach to express the DTR in mRGCs. mRGCs develop normally, but can be acutely ablated upon diphtheria toxin administration. The mRGC-ablated mice exhibited normal outer retinal function. However, they completely lacked non-image forming visual responses such as circadian photoentrainment, light modulation of activity, and PLR. These results point to the mRGCs as the site of functional integration of the rod/cone and melanopsin phototransduction pathways and as the primary anatomical site for the divergence of image-forming and non-image forming photoresponses in mammals.

No abstract

The relationship between interindividual differences in nicotine metabolism and CYP2A6 genetic polymorphism in humans was proved.