Neural crest requires Impdh 2 for development of the enteric nervous system, great vessels, and craniofacial skeleton

Mutations that impair the proliferation of enteric neural crest-derived cells (ENCDC) cause Hirschsprung disease, a potentially lethal birth defect where the enteric nervous system (ENS) is absent from distal bowel. Inosine 5′ monophosphate dehydrogenase (IMPDH) activity is essential for de novo GMP synthesis, and chemical inhibition of IMPDH induces Hirschsprung disease-like pathology in mouse models by reducing ENCDC proliferation. Two IMPDH isoforms are ubiquitously expressed in the embryo, but only IMPDH2 is required for life. To further understand the role of IMPDH2 in ENS and neural crest development, we characterized a conditional Impdh2 mutant mouse. Deletion of Impdh2 in the early neural crest using the Wnt1-Cre transgene produced defects in multiple neural crest derivatives including highly penetrant intestinal aganglionosis, agenesis of the craniofacial skeleton, and cardiac outflow tract and great vessel malformations. Analysis using a Rosa26 reporter mouse suggested that some or all of the remaining ENS in Impdh2 conditional-knockout animals was derived from cells that escaped Wnt1-Cre mediated DNA recombination. These data suggest that IMPDH2 mediated guanine nucleotide synthesis is essential for normal development of the ENS and other neural crest derivatives

Exposing the role of coparenting and parenting for adolescent personal identity processes

In line with a family systems perspective, this study examined the association between two aspects of family climate, that of coparenting^(cooperation, triangulation) and parenting (autonomy-support, dependency-oriented and achievement-oriented psychological control) and their relation to adolescent personal identity formation (commitment making, identification with commitment, exploration in breadth, exploration in depth, ruminative exploration, reconsideration of commitment). Using structural equation modeling, we tested the hypothesis that coparenting would be associated with adolescent identity formation via parenting. Cross-sectional self-report data were collected from 1,105 Swiss adolescents (aged 13-18 years; 51% female). Structural equation modeling revealed associations between coparental cooperation and more adaptive identity formation via parental autonomy support. Conversely, coparental triangulation was associated with maladaptive identity dimensions via parental dependency-oriented psychological control. These associations were not moderated by age, gender, or family structure

Further insight into adolescent personal identity statuses: Differences based on self-esteem, family climate, and family communication.

During adolescence, youngsters are faced with the challenging task of forming an identity. This process can be either supported or hindered by adolescents' family context. The present study used a six-process model of personal identity including the five identity processes described by the dual-cycle model of identity (exploration in breadth, commitment making, exploration in depth, identification with commitment, and ruminative exploration) as well as a sixth identity process of reconsideration of commitment, commonly described in the three-factor model of identity. In the current investigation, we sought to evaluate how adolescents in identity statuses derived from this six-process model differed based on psychological adjustment, perceived family climate, and family communication. A total of 1105 Swiss adolescents (Mage = 15.08; 51% female) completed self-report questionnaires at one time point. Using a person-centered approach, identity statuses were empirically derived and unique profiles for each identity status were identified. We identified six identity statuses: Achievement, Foreclosure, Ruminative Moratorium, Reconsidering Achievement, Troubled Diffusion, and Carefree Diffusion. Statuses with the highest degree of commitment showed the most optimal profiles of psychological adjustment and perceived family climate, whereas those with the lowest levels of commitment demonstrated the least optimal profiles. Adolescents in the Reconsidering Achievement status, however, reported high levels of both parental support and psychological control. The use of the six-process model of identity allowed for the derivation of six identity statuses and provided further insight into how adolescents in different identity statuses confront identity-related issues in the context of their family

Tissue-specific Regulation of the Ecto-5′-nucleotidase Promoter: ROLE OF THE cAMP RESPONSE ELEMENT SITE IN MEDIATING REPRESSION BY THE UPSTREAM REGULATORY REGION

We have isolated the 5' region of the ecto-5'-nucleotidase (low K(m) 5'-NT) gene and established that a 969-base pair (bp) fragment confers cell-specific expression of a CAT reporter gene that correlates with the expression of endogenous ecto-5'-NT mRNA and enzymatic activity. A 768-bp upstream negative regulatory region has been identified that conferred lymphocyte-specific negative regulation in a heterologous system with a 244-bp deoxycytidine kinase core promoter. DNase I footprinting identified several protected areas including Sp1, Sp1/AP-2, and cAMP response element (CRE) binding sites within the 201-bp core promoter region and Sp1, NRE-2a, TCF-1/LEF-1, and Sp1/NF-AT binding sites in the upstream regulatory region. Whereas the CRE site was essential in mediating the negative activity of the upstream regulatory region in Jurkat but not in HeLa cells, mutation of the Sp1/AP-2 site decreased promoter activity in both cell lines. Electrophoretic mobility shift assay analysis of proteins binding to the CRE site identified both ATF-1 and ATF-2 in Jurkat cells. Finally, phorbol 12-myristate 13-acetate increased the activity of both the core and the 969-bp promoter fragments, and this increase was abrogated by mutations at the CRE site. In summary, we have identified a tissue-specific regulatory region 5' of the ecto-5'-NT core promoter that requires the presence of a functional CRE site within the basal promoter for its suppressive activity

Characterization of the Human Inosine-5′-monophosphate Dehydrogenase Type II Gene

Inosine-5'-monophosphate dehydrogenase (IMPDH) activity and mRNA levels are induced up to 15-fold upon mitogenic or antigenic stimulation of human peripheral blood T lymphocytes. This increase in IMPDH activity is required for cellular proliferation and has been associated with malignant transformation. We have cloned the human IMPDH type II gene and show that it contains 14 exons and is approximately 5.8 kilobases in length. Exons vary in size from 49 to 207 base pairs and introns from 73 to 1065 base pairs. The transcription start site was mapped to a position 50 nucleotides upstream of the translation initiation site. The 5'-flanking region consisting of 463 base pairs upstream of the translation initiation site confers induced transcription and differential regulation upon a chloramphenicol acetyltransferase reporter gene when transfected into Jurkat T cells and human peripheral blood T lymphocytes, respectively. DNase I footprinting analysis using Jurkat T cell nuclear extract identified four protected regions in the promoter which coincide with consensus transcription factor binding sites for the nuclear factors AP2, ATF, CREB, Egr-1, Nm23, and Sp1. These findings suggest that several of these nuclear factors may play a critical role in the regulation of IMPDH type II gene expression during T lymphocyte activation

Regulation of Inosine-5′-monophosphate Dehydrogenase Type II Gene Expression in Human T Cells: ROLE FOR A NOVEL 5′ PALINDROMIC OCTAMER SEQUENCE

Expression of the gene encoding human inosine- 5'-monophosphate dehydrogenase (IMPDH) type II, an enzyme catalyzing the rate-limiting step in the generation of guanine nucleotides, is increased more than 10-fold in activated peripheral blood T lymphocytes and is required for T cell activation. We have examined the 5'-regulatory sequences that are important for the transcriptional regulation of this gene in T cells. DNase I mapping of genomic DNA identified a hypersensitive element near the transcription initiation site. Fine mapping by in vivo footprinting demonstrated five transcription factor binding sites that are occupied in both resting and activated peripheral blood T lymphocytes; these are tandem CRE motifs, a Sp1 site, an overlapping Egr-1/Sp1 site, and a novel palindromic octamer sequence (POS). The tandem CRE and POS sites are of major functional importance as judged by mutational and electrophoretic mobility shift analyses. These data provide evidence that expression of the human IMPDH type II gene is predominantly regulated by the nuclear factors ATF-2 and an as yet unidentified POS-binding protein. Additional major protein-DNA interactions do not occur within the promoter region after T lymphocyte activation, indicating a requirement for additional protein-protein interactions and/or post-translational modifications of pre-bound transcription factors to account for the observed increase in IMPDH type II gene expression

Dissecting the molecular organization of the translocon-associated protein complex

In eukaryotic cells, one-third of all proteins must be transported across or inserted into the endoplasmic reticulum (ER) membrane by the ER protein translocon. The translocon-associated protein (TRAP) complex is an integral component of the translocon, assisting the Sec61 protein-conducting channel by regulating signal sequence and transmembrane helix insertion in a substrate-dependent manner. Here we use cryo-electron tomography (CET) to study the structure of the native translocon in evolutionarily divergent organisms and disease-linked TRAP mutant fibroblasts from human patients. The structural differences detected by subtomogram analysis form a basis for dissecting the molecular organization of the TRAP complex. We assign positions to the four TRAP subunits within the complex, providing insights into their individual functions. The revealed molecular architecture of a central translocon component advances our understanding of membrane protein biogenesis and sheds light on the role of TRAP in human congenital disorders of glycosylation

Coevolution of Glauber-like Ising dynamics on typical networks

We consider coevolution of site status and link structures from two different initial networks: a one dimensional Ising chain and a scale free network. The dynamics is governed by a preassigned stability parameter $S$, and a rewiring factor $\phi$, that determines whether the Ising spin at the chosen site flips or whether the node gets rewired to another node in the system. This dynamics has also been studied with Ising spins distributed randomly among nodes which lie on a network with preferential attachment. We have observed the steady state average stability and magnetisation for both kinds of systems to have an idea about the effect of initial network topology. Although the average stability shows almost similar behaviour, the magnetisation depends on the initial condition we start from. Apart from the local dynamics, the global effect on the dynamics has also been studied. These parameters show interesting variations for different values of $S$ and $\phi$, which helps in determining the steady-state condition for a given substrate.Comment: 8 pages, 10 figure

Plexin-D1 Is a Novel Regulator of Germinal Centers and Humoral Immune Responses

Long-lived humoral immune responses depend upon the generation of memory B cells and long-lived plasma cells during the germinal center (GC) reaction. These memory compartments, characterized by class-switched IgG and high-affinity Abs, are the basis for successful vaccination. We report that a new member of the plexin family of molecules, plexin-D1, controls the GC reaction and is required for secondary humoral immune responses. Plexin-D1 was not required for B cell maturation, marginal zone precursor development, dark and light zone formation, Igλ+ and Igκ+ B cell skewing, B1/B2 development, and the initial extrafollicular response. Plexin-D1 expression was increased following B cell activation, and PlxnD1−/− mice exhibited defective GC reactions during T-dependent immune activation. PlxnD1−/− B cells showed a defect in migration toward the GC chemokines, CXCL12, CXCL13, and CCL19. Accordingly, PlxnD1−/− mice exhibited defective production of IgG1 and IgG2b, but not IgG3 serum Ab, accompanied by reductions in long-lived bone marrow plasmacytes and recall humoral memory responses. These data show a new role for immune plexins in the GC reaction and generation of immunologic memory

Different reactions to adverse neighborhoods in games of cooperation

In social dilemmas, cooperation among randomly interacting individuals is often difficult to achieve. The situation changes if interactions take place in a network where the network structure jointly evolves with the behavioral strategies of the interacting individuals. In particular, cooperation can be stabilized if individuals tend to cut interaction links when facing adverse neighborhoods. Here we consider two different types of reaction to adverse neighborhoods, and all possible mixtures between these reactions. When faced with a gloomy outlook, players can either choose to cut and rewire some of their links to other individuals, or they can migrate to another location and establish new links in the new local neighborhood. We find that in general local rewiring is more favorable for the evolution of cooperation than emigration from adverse neighborhoods. Rewiring helps to maintain the diversity in the degree distribution of players and favors the spontaneous emergence of cooperative clusters. Both properties are known to favor the evolution of cooperation on networks. Interestingly, a mixture of migration and rewiring is even more favorable for the evolution of cooperation than rewiring on its own. While most models only consider a single type of reaction to adverse neighborhoods, the coexistence of several such reactions may actually be an optimal setting for the evolution of cooperation.Comment: 12 pages, 5 figures; accepted for publication in PLoS ON