Recent evidence supports the notion that transformation of undifferentiated neural stem cell (NSC) precursors may contribute to the development of glioblastoma multiforme (GBM). The over-expression and mutation of the epidermal growth factor receptor (EGFR), along with other cellular pathway mutations, plays a significant role in GBM maintenance progression. Though EGFR signaling is important in determining neural cell fate and conferring astrocyte differentiation, there is a limited understanding of its role in NSC and tumor stem cell (TSC) biology. We hypothesized that EGFR expression and mutation in post-natal NSCs may contribute to cellular aggressiveness including enhanced cellular proliferation, survival and migration. Stable subclones of C17.2 murine NSCs were transfected to over-express either the wild-type EGFR (wtEGFR) or its most common mutated variant EGFRvIII. Activated EGFR signaling in these cells induced behaviors characteristic of GBM TSCs, including enhanced proliferation, survival and migration, even in the absence of EGF ligand. wtEGFR activation was also found to block neuronal differentiation and was associated with a dramatic increase in chemotaxis in the presence of EGF. EGFRvIII expression lead to an increase in NSC proliferation and survival, while it simultaneously blocked neuronal differentiation and promoted glial fate. Our findings suggest that activated EGFR signaling enhances the aggressiveness of NSCs. Understanding the regulatory mechanisms of NSCs may lend insight into deregulated mechanisms of GBM TSC invasion, proliferation, survival and resistance to current treatment modalities.

Fragile X Syndrome (FXS) is a debilitating neurodevelopmental disorder thought to arise from disrupted synaptic communication in several key brain regions including the amygdala - a central processing center for information with emotional and social relevance. Recent studies reveal defects in both excitatory and inhibitory neurotransmission in mature amygdala circuits in Fmr1-/y mutants, the animal model of FXS. However, whether these defects are the result of altered synaptic development or simply faulty mature circuits remains unknown. Using a combination of electrophysiological and genetic approaches, we show the development of both pre- and postsynaptic components of inhibitory neurotransmission in the FXS amygdala is dynamically altered during critical stages of neural circuit formation. Surprisingly, we observe that there is a homeostatic correction of defective inhibition, which, despite transiently restoring inhibitory synaptic efficacy to levels at or beyond those of control, ultimately fails to be maintained. Using inhibitory interneuron-specific conditional knockout and rescue mice, we further reveal that Fragile X Mental Retardation Protein (FMRP) function in amygdala inhibitory microcircuits can be segregated into distinct pre- and postsynaptic components. Collectively, these studies reveal a previously unrecognized complexity of disrupted neuronal development in FXS and therefore have direct implications for establishing novel temporal and region-specific targeted therapies to ameliorate core amygdala-based behavioral symptoms.

The expression of major histocompatibility class II genes is necessary for proper antigen presentation and induction of an immune response. This expression is initiated by the class II transactivator, CIITA. The establishment of the active form of CIITA is controlled by a series of post-translational events, including GTP binding, ubiquitination, and dimerization. However, the role of phosphorylation is less clearly defined as are the consequences of phosphorylation on CIITA activity and the identity of the kinases involved. In this study we show that the extracellular signal-regulated kinases 1 and 2 (ERK1/2) interact directly with CIITA, targeting serine residues in the amino terminus of the protein, including serine 288. Inhibition of this phosphorylation by dominant-negative forms of ERK or by treatment of cells with the ERK inhibitor PD98059 resulted in the increase in CIITA-mediated gene expression from a class II promoter, enhanced the nuclear concentration of CIITA, and impaired its ability to bind to the nuclear export factor, CRM1. In contrast, inhibition of ERK1/2 activity had little effect on serine-to-alanine mutant forms of CIITA. These data suggest a model whereby ERK1/2-mediated phosphorylation of CIITA down-regulates CIITA activity by priming it for nuclear export, thus providing a means for cells to tightly regulate the extent of antigen presentation.The class II transactivator CIITA 2 plays a critical role in initiating an immune response by activating the expression of major histocompatibility (MHC) class II genes and associated molecules (2-6). MHC II glycoproteins are necessary for the presentation of antigenic peptides to CD4 ϩ T lymphocytes and the subsequent initiation and propagation of CD4 ϩ T cell-mediated immune responses and are involved in the development and maintenance of homeostasis of the CD4 ϩ T cell population. Although constitutive expression of MHC class II genes is primarily restricted to a specific subset of antigen-presenting cells that include B cells and dendritic cells, expression is inducible in a variety of other cell types and tissues by cytokines such as interferon-␥ and tumor necrosis factor-␣ (1). Both constitutive and inducible expression of MHC II and other related genes are contingent upon the activation of CIITA (7,8). Loss of a functional CIITA protein results in an immunodeficiency called bare lymphocyte syndrome, which is characterized by a complete absence of MHC class II-mediated antigen presentation. Patients with this disease suffer from recurrent infections due to opportunistic infections and, ultimately, death in early childhood (9 -11).CIITA is a protein of 1130 amino acids that does not bind directly to DNA. Instead, it regulates gene expression by interacting with other transcription factors and chromatin-remodeling proteins at the W/X/Y regulatory elements in the promoter regions of class II genes (for review, see Refs. 12-17). Nuclear factors binding to CIITA include the regulatory factor X (RFX) complex (RFX5, RFXAP, RFXANK/RFX-B) (18) and nuc...

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