Learning and memory depend on long-term synaptic plasticity including long-term potentiation (LTP) and depression (LTD). Activity-regulated cytoskeleton-associated protein (Arc) plays versatile roles in synaptic plasticity mainly through inducing F-actin formation, underlying consolidation of LTP, and promoting AMPA receptor (AMPAR) endocytosis, underlying LTD. Insulin can also induce LTD by facilitating the internalization of AMPARs. In neuroblastoma cells, insulin induced a dramatic increase in Arc mRNA and Arc protein levels, which may underlie the memory-enhancing action of insulin. Thus, a hypothesis was made that, in response to insulin, increased AMPAR endocytosis leads to enhanced Arc expression, and vice versa. Primary cultures of neonatal Sprague-Dawley rat cortical neurons were used. Using Western-blot analysis and immunofluorescent staining, our results reveal that inhibiting AMPAR-mediated responses with AMPAR antagonists significantly enhanced whereas blocking AMPAR endocytosis with various reagents significantly prevented insulin (200 nM, 2 h)-induced Arc expression. Furthermore, via surface biotinylation assay, we demonstrate that acute blockade of new Arc synthesis after insulin stimulation using Arc antisense oligodeoxynucleotide prevented insulin-stimulated AMPAR endocytosis. These findings suggest for the first time that an interaction exists between insulin-stimulated AMPAR endocytosis and insulin-induced Arc expression.

Chronic high-fat-diet (HFD) consumption can lead to the development of brain insulin resistance, which then exerts deleterious effects on learning and memory. Activityregulated cytoskeleton-associated protein (Arc) is a memory-related protein, and its expression can be induced by insulin stimulation. In HFD-fed animals, their basal Arc protein levels in cerebral cortex and hippocampus are reduced. However, the effects of HFD on novelty-induced Arc protein expression that is important for cognitive function is still unknown. In the present study, after feeding HFD (60% kcal from fat) for 5 weeks, mice developed brain insulin resistance and had a significant reduction in the novelty-induced but not the basal Arc protein levels in their hippocampi. Further experiments were performed in primary rat hippocampal neurons. The results show that, under the condition of neuronal insulin resistance, acute insulin stimulation induced less activation of the phosphatidylinositol 3-kinase/protein kinase B/p70 ribosomal S6 kinase (PI3K/Akt/p70S6K) pathway, resulting in reduced induction of Arc protein expression. Accordingly, it is suggested that following HFD feeding, the reduction in novelty-induced Arc protein expression in animal's hippocampus is probably related to a suppressed activation of the PI3K/Akt/p70S6K pathway due to the existence of brain insulin resistance. K E Y W O R D SArc, high-fat diet, insulin resistance, PI3K

Protein kinase M zeta (PKMζ) and the kidney and brain protein (KIBRA) play important roles in various forms of memories. However, whether they are involved in performing the T-maze task is still unknown. In this study, the delayed nonmatch-to-sample (DNMS) task in a T-maze was given to rats. The percentage of correct choices denoting the performance accuracy was calculated and the protein levels of PKMζ and KIBRA in rat's prefrontal cortex were measured. The results showed significantly increased performance accuracy after the training phase, which was maintained on the next day in groups with a delay of 10 s but not 30 s, indicating that 30 s is too long for rats to maintain working memory. As for the expressions of PKMζ and KIBRA, significant increases were observed 1 day after the training phase, indicating that the formation of reference memory accompanies an increase in PKMζ and KIBRA. No significant difference was found among groups with various delay intervals, indicating that the expressions of PKMζ and KIBRA exert no effects on the performance of working memory. These results provide the first evidence that KIBRA as well as PKMζ is closely related to reference memory but not working memory in rats.

Aluminum (Al) is a neurotoxicant and is implicated in several neurodegenerative diseases, including Alzheimer's disease (AD). In AD brains, one of the pathological hallmarks is the extracellular deposition of senile plaques, which are mainly composed of aggregated amyloid-beta (Abeta). Endoproteolysis of the amyloid-beta precursor protein (AbetaPP) by the beta-secretase and the gamma-secretase generates Abeta. AbetaPP can also be cleaved by the alpha-secretase within the Abeta region, which releases a soluble fragment sAPPalpha and precludes the formation of Abeta. Al has been reported to increase the level of Abeta, promote Abeta aggregation, and increase Abeta neurotoxicity. In contrast, small G protein Rho and its effector, Rho-associated kinase (ROCK), are known to negatively regulate the amount of Abeta. Inhibition of the Rho-ROCK pathway may underlie the ability of nonsteroidal anti-inflammatory drugs and statins to reduce Abeta production. Whether the Rho-ROCK pathway is involved in Al-induced elevation and aggregation of Abeta is unknown. In the present study, cultured rat cortical neurons were treated with Al(malt)(3) in the absence or presence of ROCK inhibitor Y-27632. After the treatment of Al(malt)(3), the cell viability and the level of sAPPalpha were reduced, whereas the amyloid fibrils in the conditioned media were increased. Treatment with Y-27632 prevented these adverse effects of Al(malt)(3) and thus maintained neuronal survival. These results reveal that the activation of the Rho-ROCK signaling pathway was involved in Al-induced effects in terms of the cell viability, the production of sAPPalpha, and the formation of amyloid fibril, which provides a novel mechanism underlying Al-induced neurotoxicity.

INTRODUCTIONDi-(2-ethylhexyl)-phthalate (DEHP) is a widely used plasticizer found in a variety of polyvinyl chloride products. With time and use, DEHP is leached from plastic products into the external environment (Heudorf et al., 2007). DEHP is identified as an endocrine disruptor (Marsee et al., 2006;Parks et al., 2000). Compared to adults, exposure of DEHP in fetus and children is considered disproportionately high, for DEHP can transfer from the mother to her offspring through the placenta and lactation (Latini et al., 2003;Main et al., 2006). Animal studies have demonstrated that gestational and lactational exposure to DEHP in rats disrupts the testicular function and the androgen-dependent development in their male offspring (Lyche et al., 2009;Parks et al., 2000). In addition to reproductive toxicity, perinatal exposure to DEHP also induces developmental disturbances in the offspring. Evidence indicates that gestational exposure to DEHP reduces the body weight at birth in the offspring (Tanida et al., 2009). In addition, gestational and lactational DEHP exposure at low level (6.25 or 10 mg/kg/day) results in a growth restriction during development, even after maturation, the body weight of DEHP-exposed rats is still lower than that of non-exposed rats (Chen et al., 2010; Lin et al., 2011; Wei et al., 2012). Because the muscle mass contributes the most parts of body weight, the reduced body weight caused by perinatal exposure to DEHP may be the consequence of impaired myogenesis. Until recently, there is no investigation studying the effect of DEHP ABSTRACT -The purpose of this study was to investigate the effects of di-(2-ethylhexyl) phthalate (DEHP) treatment on MyoD and myogenin expression and myotube formation in the murine C2C12 cells. Myogenic differentiation is principally regulated by activities of myogenic regulatory factors, such as MyoD and myogenin, leading the elongation and fusion of mononucleated myoblasts into multinucleated myotubes. In the present study, myogenic differentiation of C2C12 cells was induced by serum depri--ylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) assay clearly demonstrated cell viability was not -ated myotubes, and the percent of nuclei incorporated into myosin heavy chain (MyHC)-stained myo--er MyHC, as well as myogenic regulatory factors MyoD and myogenin, were reduced in DEHP-treated myotubes during myogenic differentiation. Taken together, the results showed that DEHP may impair myogenic differentiation through repression of myogenic regulatory factors, such as MyoD and myogenin, resulting in a reduction of MyHC expression. This in vitro study suggests that DEHP may be an environmental risk factor for myogenesis.