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The Arx transcription factor is expressed in the developing ventral telencephalon and subsets of its derivatives. Mutation of human ARX ortholog causes neurological disorders including epilepsy, lissencephaly, and mental retardation. We have isolated the mouse Arx endogenous enhancer modules that control its tightly compartmentalized forebrain expression. Interestingly, they are scattered downstream of its coding region and partially included within the introns of the downstream PolA1 gene. These enhancers are ultraconserved noncoding sequences that are highly conserved throughout the vertebrate phylum. Functional characterization of the Arx GABAergic enhancer element revealed its strict dependence on the activity of Dlx transcription factors. Dlx overexpression induces ectopic expression of endogenous Arx and its isolated enhancer, whereas loss of Dlx expression results in reduced Arx expression, suggesting that Arx is a key mediator of Dlx function. To further elucidate the mechanisms involved, a combination of gain-of-function studies in mutant Arx or Dlx tissues was pursued. This analysis provided evidence that, although Arx is necessary for the Dlx-dependent promotion of interneuron migration, it is not required for the GABAergic cell fate commitment mediated by Dlx factors. Although Arx has additional functions independent of the Dlx pathway, we have established a direct genetic relationship that controls critical steps in the development of telencephalic GABAergic neurons. These findings contribute elucidating the genetic hierarchy that likely underlies the etiology of a variety of human neurodevelopmental disorders.

Recent advances in the study of cerebral cortical early development are described in this chapter. The role of the anterior neural ridge in regulating telencephalon induction in the neural plate is discussed, followed by a review of the evidence for the roles of ventral, rostral and dorsal patterning centres in regulating regionalization of the telencephalon. The patterning centres produce secreted molecules (SHH, FGF, BMP, WNT) that regulate the expression of transcription factors which control regional identity, cell type specification, proliferation and differentiation. These intrinsic patterning mechanisms appear to be sufficient to generate much of the regional organization of the cerebral cortex present in newborn mice. While intrinsic mechanisms have a major role in cortical regionalization and in the production of cortical projection neurons, many cortical interneurons are derived from the basal ganglia and then migrate into the cerebral cortex. Furthermore, thalamic afferents appear to have an important role in maturation of the postnatal rodent cortex. Thus, both intrinsic and extrinsic mechanisms control development of the cerebral cortex.

The homeobox genes in the Dlx family are required for differentiation of basal forebrain neurons and craniofacial morphogenesis. Herein, we studied the expression of Dlx-1, Dlx-2, and Dlx-5 RNA and protein in the mouse forebrain from embryonic day 10.5 (E10.5) to E12.5. We provide evidence that Dlx-2 is expressed before Dlx-1, which is expressed before Dlx-5. We also demonstrate that these genes are expressed in the same cells, which may explain why single mutants of the Dlx genes have mild phenotypes. The DLX proteins are localized primarily to the nucleus, although DLX-5 also can be found in the cytoplasm. During development, the fraction of Dlx-positive cells increases in the ventricular zone. Analysis of the distribution of DLX-1 and DLX-2 in M-phase cells suggests that these proteins are distributed symmetrically to daughter cells during mitosis. We propose that DLX-negative cells in the ventricular zone are specified progressively to become DLX-2-expressing cells during neurogenesis; as these cells differentiate, they go on to express DLX-1, DLX-5, and DLX-6. This process appears to be largely the same in all regions of the forebrain that express the Dlx genes. In the basal telencephalon, these DLX-positive cells differentiate into projection neurons of the striatum and pallidum as well as interneurons, some of which migrate to the cerebral cortex and the olfactory bulb.

In mammals, some axons from each retina cross at the optic chiasm, whereas others do not. Although several loci have been identified within the chiasmatic region that appear to provide guidance cues to the retinal axons, the underlying molecular mechanisms that regulate this process are poorly understood. Here we investigate whether the earliest retinal axon trajectories and a cellular population (CD44 and stage-specific embryonic antigen 1 [SSEA] neurons), previously implicated in directing axon growth in the developing chiasm (reviewed in Mason and Sretavan [1997] Curr. Op. Neurobiol. 7:647-653), correlate with the expression patterns of several regulatory genes (BF-1, BF-2, Dlx-2, Nkx-2.1, Nkx-2.2, and Shh). These studies demonstrate that gene expression patterns in the chiasmatic region reflect the longitudinal subdivisions of the forebrain in that axon tracts in this region generally are aligned parallel to these subdivisions. Moreover, zones defined by overlapping domains of regulatory gene expression coincide with sites implicated in providing guidance information for retinal axon growth in the developing optic chiasm. Together, these data support the hypothesis that molecularly distinct, longitudinally aligned domains in the forebrain regulate the pattern of retinal axon projections in the developing hypothalamus.

International audience; This chapter describes retroviruses as the RNA viruses whose life cycle includes their replication by a DNA intermediate that integrates into the host genome. Transcription of the provirus leads to the production of virions without any apparent cytopathic effect on the cells. The demonstration that transfection of a cloned DNA copy of a retrovirus leads to the production of infectious virions indicated that retroviruses could be used as vectors for introducing DNA into eukaryotic cells. However, the definitive demonstration has been performed by inserting the herpes simplex thymidine kinase gene into a proviral copy of either the spleen necrosis virus or the Harvey murine sarcoma virus. One of the major advantages of retroviruses over DNA viruses is that, for most of them, integration into the host chromosome is an obligate step in their life cycle. The integration mechanism leads to the provirus having a specific structure in which the genomic organization is conserved