PRDC regulates placode neurogenesis in chick by modulating BMP signalling

The epibranchial placodes generate the neurons of the geniculate, petrosal, and nodose cranial sensory ganglia. Previously, it has been shown that bone morphogenetic proteins (BMPs) are involved in the formation of these structures. However, it has been unclear as to whether BMP signalling has an ongoing function in directing the later development of the epibranchial placodes, and how this signalling is regulated. Here, we demonstrate that BMPs maintain placodal neurogenesis and that their activity is modulated by a member of the Cerberus/Dan family of BMP antagonists, Protein Related to Dan and Cerberus (PRDC). We find that Bmp4 is expressed in the epibranchial placodes while Bmp7 and PRDC are expressed in the pharyngeal pouches. The timing and regional expression of these three genes suggest that BMP7 is involved in inducing placode neurogenesis and BMP4 in maintaining it and that BMP activity is modulated by PRDC. To investigate this hypothesis, we have performed both gain- and loss- of-function experiments with PRDC and find that it can modulate the BMP signals that induce epibranchial neurogenesis: a gain of PRDC function results in a loss of Bmp4 and hence placode neurogenesis is inhibited; conversely, a loss of PRDC function induces ectopic Bmp4 and an expansion of placode neurogenesis. This modulation is therefore necessary for the number and positioning of the epibranchial neurons.     

Response of recruitment to light availability across a tropical lowland rain forest community

1.  Many hypotheses about species coexistence involve differential resource use and trade-offs in species' life-history traits. Quantifying resource use across most species in diverse communities, although, has seldom been attempted.
2.  We use a hierarchical Bayesian approach to quantify the light dependence of recruitment in 263 woody species in a 50-ha long-term forest census plot in Panama. Data on sapling recruitment were obtained using the 1985–1990 and 1990–1995 census intervals. Available light was estimated for each recruit from yearly censuses of canopy density.
3.  We use a power function (linear log–log relationship) to model the light effect on recruitment. Different responses of recruitment to light are expressed by the light effect parameter b . The distribution of b had a central mode at 0.8, suggesting that recruitment of many species responds nearly linearly to increasing light.
4.  Nearly every species showed increases in recruitment with increasing light. Just nine species (3%) had recruitment declining with light, while 198 species (75%) showed increasing recruitment in both census intervals. Most of the increases in recruitment were decelerating, i.e. the increase was less at higher light ( b  < 1). In the remaining species, the response to light varied between census intervals (24 species) or species did not have recruits in both intervals (41 species).
5.   Synthesis. Nearly all species regenerate better in higher light, and recruitment responses to light are spread along a continuum ranging from modest increase with light to a rather strict requirement for high light. These results support the hypothesis that spatio-temporal variation in light availability may contribute to the diversity of tropical tree species by providing opportunities for niche differentiation with respect to light requirements for regeneration.     

Comparative genomic evidence for a complete nuclear pore complex in the last eukaryotic common ancestor

Background

The Nuclear Pore Complex (NPC) facilitates molecular trafficking between nucleus and cytoplasm and is an integral feature of the eukaryote cell. It exhibits eight-fold rotational symmetry and is comprised of approximately 30 nucleoporins (Nups) in different stoichiometries. Nups are broadly conserved between yeast, vertebrates and plants, but few have been identified among other major eukaryotic groups.

Methodology/Principal Findings

We screened for Nups across 60 eukaryote genomes and report that 19 Nups (spanning all major protein subcomplexes) are found in all eukaryote supergroups represented in our study (Opisthokonts, Amoebozoa, Viridiplantae, Chromalveolates and Excavates). Based on parsimony, between 23 and 26 of 31 Nups can be placed in LECA. Notably, they include central components of the anchoring system (Ndc1 and Gp210) indicating that the anchoring system did not evolve by convergence, as has previously been suggested. These results significantly extend earlier results and, importantly, unambiguously place a fully-fledged NPC in LECA. We also test the proposal that transmembrane Pom proteins in vertebrates and yeasts may account for their variant forms of mitosis (open mitoses in vertebrates, closed among yeasts). The distribution of homologues of vertebrate Pom121 and yeast Pom152 is not consistent with this suggestion, but the distribution of fungal Pom34 fits a scenario wherein it was integral to the evolution of closed mitosis in ascomycetes. We also report an updated screen for vesicle coating complexes, which share a common evolutionary origin with Nups, and can be traced back to LECA. Surprisingly, we find only three supergroup-level differences (one gain and two losses) between the constituents of COPI, COPII and Clathrin complexes.

Conclusions/Significance

Our results indicate that all major protein subcomplexes in the Nuclear Pore Complex are traceable to the Last Eukaryotic Common Ancestor (LECA). In contrast to previous screens, we demonstrate that our conclusions hold regardless of the position of the root of the eukaryote tree.     

Obesity as an Obstetric Risk Factor: Does It Matter in a Perinatal Center?

Objectives: Obesity before and during pregnancy is associated with several obstetrics risk factors for both mother and fetus. The aim of this retrospective study was to analyze the influence of BMI before pregnancy on distinct perinatal parameters. Research Methods and Procedures: The study includes 5067 singleton pregnancies from 2001 to 2004 at the Department of Obstetrics and Gynecology, University of Leipzig. The study group was divided into BMI groups: <18.5, ≥18.5 to <25, ≥25 to <30, ≥30 to <35, ≥35 to <40, and ≥40 kg/m². Analysis of perinatal data included rate of intrauterine death, rate of cesarean section and shoulder dystocia, time of hospital stay for mother and newborn, and gestational age of delivery. Neonatal outcome variables included percentage of newborns weighing >4000 grams, rate of umbilical cord pH <7.10, and rate of 1‐, 5‐, and 10‐minute Apgar scores of <8. Results: There was no difference in the gestational age at delivery among the groups. In the group with BMI ≥30 kg/m², the cesarean section rate was significantly elevated to 25.1%, with a more dramatic increase up to 30.2% in the group with BMI ≥35 kg/m² and 43.1% in the group with BMI ≥40 kg/m², mainly because of a higher number of secondary cesarean sections. Although newborns of obese women showed worse initial neonatal adaptation, the 10‐minute Apgar values did not differ among the groups. The higher rate of operative deliveries and the trend to an increased rate of shoulder dystocia did not influence duration of the hospital stay for mothers and newborns or morbidity of both. Discussion: A high pre‐pregnancy BMI is clearly associated with a higher rate of cesarean section deliveries. However, under the compensating conditions of a tertiary perinatal center, overall morbidity of mothers and newborns seems not to be increased.     

Stonin 2 is an AP-2-dependent endocytic sorting adaptor for synaptotagmin internalization and recycling

Clathrin-mediated endocytosis is involved in the internalization, recycling, and degradation of cycling membrane receptors as well as in the biogenesis of synaptic vesicle proteins. While many constitutively internalized cargo proteins are recognized directly by the clathrin adaptor complex AP-2, stimulation-dependent endocytosis of membrane proteins is often facilitated by specialized sorting adaptors. Although clathrin-mediated endocytosis appears to be a major pathway for presynaptic vesicle cycling, no sorting adaptor dedicated to synaptic vesicle membrane protein endocytosis has been indentified in mammals. Here, we show that stonin 2, a mammalian ortholog of Drosophila stoned B, facilitates clathrin/AP-2-dependent internalization of synaptotagmin and targets it to a recycling vesicle pool in living neurons. The ability of stonin 2 to facilitate endocytosis of synaptotagmin is dependent on its association with AP-2, an intact mu-homology domain, and functional AP-2 heterotetramers. Our data identify stonin 2 as an AP-2-dependent endocytic sorting adaptor for synaptotagmin internalization and recycling.     

Asn- and Asp-mediated interactions between transmembrane helices during translocon-mediated membrane protein assembly

Inter-helix hydrogen bonding involving asparagine (Asn, N), glutamine (Gln, Q), aspartic acid (Asp, D) or glutamic acid (Glu, E) can drive efficient di- or trimerization of transmembrane helices in detergent micelles and lipid bilayers. Likewise, Asn-Asn and Asp-Asp pairs can promote the formation of helical hairpins during translocon-mediated membrane protein assembly in the endoplasmic reticulum. By in vitro translation of model integral membrane protein constructs in the presence of rough microsomes, we show that Asn- or Asp-mediated interactions with a neighbouring transmembrane helix can enhance the membrane insertion efficiency of a marginally hydrophobic transmembrane segment. Our observations suggest that inter-helix hydrogen bonds can form during Sec61 translocon-assisted insertion and thus could be important for membrane protein assembly.     

Attenuation of a vaccine strain of vaccinia virus via inactivation of interferon viroceptor

BACKGROUND: Interferons (IFNs) play an important role in host antiviral responses, but viruses, including vaccinia viruses (VV), employ mechanisms to disrupt IFN activities, and these viral mechanisms are often associated with their virulence. Here, we explore an attenuation strategy with a vaccine strain of VV lacking a virus-encoded IFN-gamma receptor homolog (viroceptor). METHODS: To facilitate the monitoring of virus properties, first we constructed a Lister vaccine strain derivative VV-RG expressing optical reporters. Further, we constructed a VV-RG derivative, VV-RG8, which lacks the IFN-gammaR viroceptor (B8R gene product). Replication, immunological and pathogenic properties of the constructed strains were compared. RESULTS: Viruses did not show significant differences in humoral and cellular immune responses of immune-competent mice. Replication of constructed viruses was efficient both in vitro and in vivo, but showed marked difference in kinetics of propagation. In cultured CV-1 epithelial cells, the VV-RG8 strain retained the propagation potential of the parental virus, while, in the C6 glial cells, significant delay was observed in the kinetics of the VV-RG8 replication cycle compared to VV-RG. The pathogenesis of the viruses was tested by survival assay and biodistribution in nude mice. High dose inoculation of nude mice with VV-RG8 caused less pronounced virus dissemination, improved weight gain, and increased survival rate, as compared with the VV-RG strain. CONCLUSIONS: The replication-competent virus VV-RG8 carrying a mutation at the B8R gene is less pathogenic for mice than the parental vaccine virus. We anticipate that step-wise inactivation of VV vaccine genes involved in evasion of host immune response may provide an alternative approach for generation of hyper-attenuated replication-competent vaccines.