A single incidental dark pulse during daytime attenuated food anticipatory behavior.

Using an open-source operant feeding device (FED3), we measured food-seeking nose poking behavior in mice. When the mice were exposed to 4 h restricted feeding at night, all mice exhibited robust food anticipatory nose poking starting ~4 h before scheduled mealtime. When the light-dark cycle was advanced by 6 h, mice exhibited two distinct bouts of anticipatory poking, one corresponding to actual mealtime which continued at the same time of day, and one corresponding to predicted mealtime which shifted parallel with the light-dark cycle. Likewise, two similar bouts of food-seeking behavior appeared when the light-dark cycle was delayed for 9 h. These data suggest that food anticipatory behavior is encoded to a circadian oscillator that entrains to the light-dark cycle. Two weeks after advancing the light-dark cycle, mice incidentally received a 3.5 h dark pulse in the middle of the day. This single dark pulse had a negligible effect on running wheel behavior but caused a temporary attenuation of both food anticipatory poking and pellet intake. These results suggest that the circadian oscillator controlling food anticipatory poking is sensitive to light disruption and that proper food anticipation is critical for sufficient food intake during temporally restricted feeding.

Protocol to study circadian food-anticipatory poking in mice using the feeding experimentation device version 3.

Food-anticipatory nose poking is a unique food-seeking behavior driven by the food-entrainable oscillator. Here, we present a protocol to record a novel food-seeking nose poking behavior in mice under temporally restricted feeding followed by food deprivation using the open-source feeding experimentation device version 3 (FED3). We describe steps for setting up the FED3 and cage, training, and habituation. We then detail procedures for setting up the schedule for time-restricted feeding and food deprivation and for generating ethograms from FED3 data. For complete details on the use and execution of this protocol, please refer to Ehichioya et al.1.

Ambient ultrafine particles reduce endothelial nitric oxide production via S-glutathionylation of eNOS.

Exposure to airborne particulate pollutants is intimately linked to vascular oxidative stress and inflammatory responses with clinical relevance to atherosclerosis. Particulate matter (PM) has been reported to induce endothelial dysfunction and atherosclerosis. Here, we tested whether ambient ultrafine particles (UFP, diameter <200 nm) modulate eNOS activity in terms of nitric oxide (NO) production via protein S-glutathionylation. Treatment of human aortic endothelial cells (HAEC) with UFP significantly reduced NO production. UFP-mediated reduction in NO production was restored in the presence of JNK inhibitor (SP600125), NADPH oxidase inhibitor (Apocynin), anti-oxidant (N-acetyl cysteine), and superoxide dismutase mimetics (Tempol and MnTMPyP). UFP exposure increased the GSSG/GSH ratio and eNOS S-glutathionylation, whereas over-expression of Glutaredoxin-1 (to inhibit S-glutathionylation) restored UFP-mediated reduction in NO production by nearly 80%. Thus, our findings suggest that eNOS S-glutathionylation is a potential mechanism underlying ambient UFP-induced reduction of NO production.