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?(Fig.5A).5A). Furthermore, hypothalamic expression of mRNA encoding myeloid cell-specific markers Cd68 and Emr1 (which encodes F4/80) was also reduced compared to controls, suggesting an effect of CL on microglial accumulation in this brain area (Fig.?(Fig.5A).5A). Astrogliosis with advancing age is correlated with increased expression of glial fibrillary acidic protein (GFAP) (Nichols et?al. 1995). There is an age-related increase in levels of GFAP in 12- and 22-month-old controls and CL mice (compared to 6-week-old mice, data not shown), but the intensity of GFAP staining GS-7340 chemical structure and the number of immunostained astrocytes in the ARC of CL mice is reduced by Adenylyl cyclase approximately 20�C30% compared to CL8 control female mice at both 12 and 22?months of age (Fig.?(Fig.5B5B and C). Figure 5 Hypothalamic inflammatory gene expression and astrogliosis during aging in crowded litter (CL) female mice. (A) Quantification of mRNA encoding proinflammatory cytokines (Il6, Tnfa), NF-��B pathway genes (Nfkbia, Ikbkb, Ikbke), and microglia-specific ... Next, we evaluated the numbers of microglia in the hypothalamus of aged CL mice. Using immunostaining for the microglia-specific Iba1 marker, we found that numbers of microglial cells in the mediobasal hypothalamus (MBH) increase in an age-dependent manner (Fig.?(Fig.6A).6A). About 80�C100% of these Iba1+ cells produced tumor necrosis factor-�� (TNF-��) in control CL8 mice at 12 and 22?months of age, indicating that they are inflammatory (Fig.?(Fig.6A).6A). In CL12 and CL15 mice, however, the proportion BIBF 1120 of Iba1?+? microglia that produce TNF-�� is significantly reduced (P?