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Pitx2, a homeodomain transcription factor, is essential for normal development of the pituitary gland, craniofacial region, eyes, heart, abdominal viscera, and limbs. Complete loss of Pitx2 in mice (Pitx2(-/-)) results in embryonic lethality by approximately e15 due to cardiac defects, whereas embryos with partial loss of function (Pitx2(neo/-) or Pitx2(neo/neo)) survive until later in development (e17-e19). Pitx2 is expressed in discrete populations of postmitotic neurons in the mouse brain, but its role in mammalian central nervous system (CNS) development is not known. We undertook an analysis of Pitx2-deficient embryos to determine whether loss of Pitx2 affects CNS development. The CNS is normal in hypomorphic e16.5 Pitx2(neo/-) and e18.5 Pitx2(neo/neo) embryos, with no evidence of midline or other defects. Midgestation (e10.5) Pitx2(-/-) embryos have normally formed neural tube structures and cerebral vesicles, whereas older (e14.5) Pitx2(-/-) embryos exhibit loss of gene expression and axonal projections in the subthalamic nucleus (a group of cells in the ventrolateral thalamus) and in the developing superior colliculus of dorsal midbrain. Our results suggest a role for Pitx2 in regulating regionally specific terminal neuronal differentiation in the developing ventrolateral thalamus and midbrain.

Patterning of the early neural tube is achieved in part by the inductive signals, which arise from neuroepithelial signaling centers. The zona limitans intrathalamica (ZLI) is a neuroepithelial domain in the alar plate of the diencephalon which separates the prethalamus from the thalamus. The ZLI has recently been considered to be a possible secondary organizer, effecting its inductions via sonic hedgehog (Shh), a signaling molecule which drives morphogenetic information for the thalamus. Using experimental embryological techniques involving the generation of chimeric embryos, we show that the formation of the ZLI in the diencephalic alar plate is due to an interaction between the prechordal and epichordal plate neuroepithelia. We also provide evidence that Shh expression in the ZLI underlies the morphogenetic activity of this putative diencephalic organizer. Ectopic Shh led to the auto-induction of its own gene expression in host cells, as well as to the expression of other genes involved in diencephalic regionalization and histogenesis. Analysis of long-term surviving embryos after Shh ectopic expression demonstrated that Shh was able to induce thalamic structures and local overgrowth. Overall, these results indicate that Shh expressed in the ZLI plays an important role in diencephalic growth and in the development of the thalamus.

The Wiskott-Aldrich syndrome protein (WASp) plays a major role in coupling T cell antigen receptor (TCR) stimulation to induction of actin cytoskeletal changes required for T cell activation. Here, we report that WASp inducibly binds the sorting nexin 9 (SNX9) in T cells and that WASp, SNX9, p85, and CD28 colocalize within clathrin-containing endocytic vesicles after TCR/CD28 costimulation. SNX9, implicated in clathrin-mediated endocytosis, binds WASp via its SH3 domain and uses its PX domain to interact with the phosphoinositol 3-kinase regulatory subunit p85 and product, phosphoinositol (3,4,5)P 3. The data reveal ligationinduced CD28 endocytosis to be clathrin-and phosphoinositol 3-kinase-dependent and TCR/CD28-evoked CD28 internalization and NFAT activation to be markedly enhanced by SNX9 overexpression, but severely impaired by expression of an SNX9 mutant (SNX9⌬PX) lacking p85-binding capacity. CD28 endocytosis and CD28-evoked actin polymerization also are impaired in WASpdeficient T cells. These findings suggest that SNX9 couples WASp to p85 and CD28 so as to link CD28 engagement to its internalization and to WASp-mediated actin remodeling required for CD28 cosignaling. Thus, the WASp/SNX9/p85/CD28 complex enables a unique interface of endocytic, actin polymerizing, and signal transduction pathways required for CD28-mediated T cell costimulation.actin cytoskeleton ͉ costimulation ͉ lymphocyte activation ͉ signaling

The diencephalon is a central area of the vertebrate developing brain, where the thalamic nuclear complex, the pretectum and the anterior tegmental structures are generated. It has been subdivided into prosomeres, which are transversal domains defined by morphological and molecular criteria. The zona limitans intrathalamica is a central boundary in the diencephalon that separates the posterior diencephalon (prosomeres 1 and 2), from the anterior diencephalon (prosomere 3). This intrathalamic limit appears early on in neural tube development, and the molecular pattern that it reveals suggests an important role in the diencephalic histogenesis. We hereby present a fate map of the presumptive territories in the diencephalon of a chick embryo at the 10-11 somite stages (HH9-10), by homotopic and isochronic quail-chick grafts. The anatomical interpretation of chimeric brains was aided by correlative whole-mount in situ hybridization with RNA probes for chicken genes expressed in specific diencephalic territories. The resulting fate map describes the distribution of the presumptive diencephalic prosomeres in the neural tube, and demonstrates their topologically conserved relationships throughout the neural development. Moreover, we show that the presumptive epithelium of ZLI can be localized at early developmental stages in the diencephalic alar plate at the anterior limit of the Wnt8b gene expression domain.

. In this review, we will describe the principle aspects of CNS development in birds and mammals, starting from early stages of embryogenesis (gastrulation and neurulation) and culminating with the formation of a variety of different regions which contribute to the structural complexity of the brain (regionalization and morphogenesis). We will pay special attention to the cellular and molecular mechanisms involved in neural tube regionalization and the key role played by localized secondary organizers in the patterning of neural primordia. KEY WORDS: patterning, neural plate, neural tube, gastrulation, neurulation, secondary organizer, anterior neural ridge, zona limitans intrathalamica, isthmic organizer Neural plate and neural tube formationA fundamental early step in neural development is the allocation of a group of ectodermal cells as precursors of the entire nervous system (Hemmati-Brivanlou and Melton, 1997). This process involves an inductive interaction first demonstrated in amphibian embryos by Spemann and Mangold in the 1920's (see Spemann and Mangold, 2001). Their experiments which involved the grafting of differently pigmented species of newt established the concept of neural induction as an instructive interaction between the dorsal lip of the blastopore (the "organizer") and the neighboring ectoderm. The discovery of a neural organization center for the amphibian gastrula initiated a search for homologous structures in other vertebrates. Soon thereafter, the equivalent region was discovered in most vertebrate species, including the shield of teleosts. In birds and mammals, the region was named "Hensen's node" and "the node", respectively. When C.H. Waddington transplanted the Hensen node of a chick embryo, he observed the induction of Int. J. Dev. Biol. 54: 7-20 (2010) Abbreviations used in this paper: ANR, anterior neural ridge; AP, anteroposterior; BMP, bone morphogenetic protein; DV, dorso-ventral; FGF, fibroblast growth factor; IsO, Isthmic organizer; ML, medio-lateral; TGF, transforming growth factor; ZLI, zona limitans intrathalamica.an ectopic neural plate or the formation of a partial new embryonic axis containing neural tube, notochord and somites (Waddington, 1933;Waddington, 1936). This demonstration provided the first evidence that in chick embryos, the nervous system is induced by signals from non-neural cells. Recent works demonstrated that the capacity of ectodermal cells to undergo neural differentiation represents their default state. In fact, neural differentiation must be suppressed in the lateral ectoderm by signals transmitted between neighboring cells, in order to develop as epidermis. These molecular signals are members of the bone morphogenetic protein (BMP) subclass of transforming growth factor β (TGF-β)-related proteins (for review see Wittler and Kessel, 2004). C. Vieira et al.Recent studies using chick embryos have shown that neural induction really begins prior to the formation of the organizer region and thus must be initiated by signals derived from other cellul...

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BackgroundImproved access to skilled health personnel for childbirth is a priority strategy to improve maternal health. This study investigates interventions to achieve this where traditional birth attendants were providers of childbirth care and asks what has been done and what has worked?Methods and FindingsWe systematically reviewed published and unpublished literature, searching 26 databases and contacting experts to find relevant studies. We included references from all time periods and locations. 132 items from 41 countries met our inclusion criteria and are included in an inventory; six were intervention evaluations of high or moderate quality which we further analysed. Four studies report on interventions to deploy midwives closer to communities: two studies in Indonesia reported an increase in use of skilled health personnel; another Indonesian study showed increased uptake of caesarean sections as midwives per population increased; one study in Bangladesh reported decreased risk of maternal death. Two studies report on interventions to address financial barriers: one in Bangladesh reported an increase in use of skilled health personnel where financial barriers for users were addressed and incentives were given to skilled care providers; another in Peru reported that use of emergency obstetric care increased by subsidies for preventive and maternity care, but not by improved quality of care.ConclusionsThe interventions had positive outcomes for relevant maternal health indicators. However, three of the studies evaluate the village midwife programme in Indonesia, which limits the generalizability of conclusions. Most studies report on a main intervention, despite other activities, such as community mobilization or partnerships with traditional birth attendants. Many authors note that multiple factors including distance, transport, family preferences/support also need to be addressed. Case studies of interventions in the inventory illustrate how different countries attempted to address these complexities. Few high quality studies that measure effectiveness of interventions exist.