2007 Annual Report 1a.Objectives (from AD-416) Aim 1: Continue to determine the effects of BB supplementation on behavioral aging using paradigms sensitive to: motor (probes sensitive to balance, strength, coordination) and cognitive (short- and long-term memories) behaviors. Aim 2: Determine the effects of BB supplementation on signaling and neurogenesis and correlate these with alterations in behavioral parameters determined in SA1. Aim 3: Since there is compelling evidence to suggest that controlling inflammation may be the key mitigating the neurodegenerative diseases, we will determine the mechanisms involved in the anti-flammatory/antioxidant activity of BB in cells. 1b.Approach (from AD-416) While the mechanism(s) linking aging, inflammation and oxidative stress to cognitive and motor dysfunction are not fully understood, it is acknowledged that a complex pattern of sequential signal transduction and gene expression is involved. Thus, in addition to the other mechanistic parameters given above, we will also characterize the age-induced neurodegenerative processes in response to BB supplementation at the level of gene transcription. Additionally, we will determine the effect of BB extract on the generality and magnitude of the behavioral effects by further molecular behavioral analyses. Importantly, to accomplish this, we will utilize a “top-down” approach by continuing to assess the specificity of the reduction in motor and cognitive behavioral deficits on various tasks and then to more reductionistic analyses to determine the mechanisms involved in the beneficial effects of BBS. Recently, it appears that inflammation may be the primary culprit in the continued neurodegeneration seen in diseases such as Alzheimer’s and Parkinson’s disease. 3.Progress Report This report serves to document research conducted under a Trust Fund Cooperative Agreement between ARS and the U.S. Highbush Blueberry Council. Previously, we found that one of the major effects of blueberry (BB) application to cells may be to alter oxidative stress/inflammatory signaling. Our findings suggest that BB may antagonize oxidative stress effects by enhancing phospo (p) mitogen activated protein kinase (MAPK) and lowering activation of p cyclic AMP response element binding protein (CREB). pCREB is important in acetylating Forkhead proteins, which prevents their interaction with sirtuins. Sirtuins are very important in mediating cellular health and functioning. Since BB supplementation appears to lower pCREB activation the Forkhead proteins are not deactivated. We also showed that in M1muscarinic receptor chimerics (M1 receptors containing an M3 muscarinic receptor i3 loop) the dopamine-induced increases in pMAPK were reduced, as were the increases in pCREB, suggesting that at least part of the protection against oxidative stress-induced decrements in calcium buffering seen in the chimerics may be mediated through the i3 loop of the receptor and enhancement of pCREB. Although it appears that oxidative stress induced deficits in calcium buffering in the M3 truncated - and chimeric-transfected COS 7 cells, it is clear that these changes are less dependent upon alterations MAPK or CREB. It appears that antioxidants might be targeting additional sites on these chimerics to decrease oxidative stress sensitivity. Last year we found that BBs also interact with genes that regulate oxidative and inflammatory stressors (e.g., IL-1beta). These genes are downregulated in BV-2 mouse microglial cells treated with the inflammatory agent lipopolysaccharide (LPS). Treatment with BB extracts significantly and dose-dependently reduced the lipopolysaccharide (LPS)-induced NO production in conditioned media from these cells, along with the reduction of reactive oxygen species and the attenuation of inducible NO synthase (iNOS), cyclo-oxygenases 2 (COX-2), and the pre-processed form of IL-1beta in the LPS-activated BV2 cells. Additional findings this year showed that the post C18 fraction (post C-18 contains no sugars and organic acids) was effective in reducing nuclear factor kappa B (an inflammatory marker), as well as p38 mitogen activated protein kinase, iNOS and COX-2 activation. The results from this study suggest that BB polyphenols attenuate inflammatory responses of the brain microglial cells and could be used to modulate inflammatory conditions in the central nervous system. Additional details of this research can be found in the report for the parent CRIS 1950-51000-063-00D Nutritional Modulation of Brain Aging and Cognitive Decline.