Beta-actin is required for proper mouse neural crest ontogeny

The mouse genome consists of six functional actin genes of which the expression patterns are temporally and spatially regulated during development and in the adult organism. Deletion of beta-actin in mouse is lethal during embryonic development, although there is compensatory expression of other actin isoforms. This suggests different isoform specific functions and, more in particular, an important function for beta-actin during early mammalian development. We here report a role for beta-actin during neural crest ontogeny. Although beta-actin null neural crest cells show expression of neural crest markers, less cells delaminate and their migration arrests shortly after. These phenotypes were associated with elevated apoptosis levels in neural crest cells, whereas proliferation levels were unchanged. Specifically the pre-migratory neural crest cells displayed higher levels of apoptosis, suggesting increased apoptosis in the neural tube accounts for the decreased amount of migrating neural crest cells seen in the beta-actin null embryos. These cells additionally displayed a lack of membrane bound N-cadherin and dramatic decrease in cadherin-11 expression which was more pronounced in the pre-migratory neural crest population, potentially indicating linkage between the cadherin-11 expression and apoptosis. By inhibiting ROCK ex vivo, the knockout neural crest cells regained migratory capacity and cadherin-11 expression was upregulated. We conclude that the presence of beta-actin is vital for survival, specifically of pre-migratory neural crest cells, their proper emigration from the neural tube and their subsequent migration. Furthermore, the absence of beta-actin affects cadherin-11 and N-cadherin function, which could partly be alleviated by ROCK inhibition, situating the Rho-ROCK signaling in a feedback loop with cadherin-11

Mutational analysis of human profilin I reveals a second PI(4,5)-P2 binding site neighbouring the poly(L-proline) binding site

Background: Profilin is a small cytoskeletal protein which interacts with actin, proline-rich proteins and phosphatidylinositol 4,5-bisphosphate (PI(4,5)-P-2). Crystallography, NMR and mutagenesis of vertebrate profilins have revealed the amino acid residues that are responsible for the interactions with actin and poly(L-proline) peptides. Although Arg88 of human profilin I was shown to be involved in PI(4,5)-P-2-binding, it was suggested that carboxy terminal basic residues may be involved as well. Results : Using site directed mutagenesis we have refined the PI(4,5)-P-2 binding site of human profilin I. For each mutant we assessed the stability and studied the interactions with actin, a proline-rich peptide and PI(4,5)-P-2 micelles. We identified at least two PI(4,5)-P-2-binding regions in human profilin I. As expected, one region comprises Arg88 and overlaps with the actin binding site. The second region involves Arg136 in the carboxy terminal helix and neighbours the poly(L-proline) binding site. In addition, we show that adding a small protein tag to the carboxy terminus of profilin strongly reduces binding to poly(L-proline), suggesting local conformational changes of the carboxy terminal a-helix may have dramatic effects on ligand binding. Conclusions : The involvement of the two terminal a-helices of profilin in ligand binding imposes important structural constraints upon the functions of this region. Our data suggest a model in which the competitive interactions between PI(4,5)-P-2 and actin and PI(4,5)-P-2 and poly(L-proline) regulate profilin functions

Survey among Belgian pig producers about the introduction of group housing systems for gestating sows

There is a global move from individual to group housing of gestating sows. In the European Union, individual gestating stalls will be banned by 2013. Just like in other industrialized regions, these stalls have been the standard housing system for intensively kept sows from the 1960s onward in the Flemish region of Belgium. Because the socioeconomic consequences for the pig industry may be far-reaching and because farmer attitude may influence the realization of the hoped-for improvement in animal welfare in practice, we conducted a survey from 2003 until 2009 among representative samples of Flemish pig producers every 2 yr. The share of farms with group housing increased from 10.5% in 2003 to 29.8% in 2007, but then dropped to 24.6% in 2009. It appears that after 2005 users of old group housing systems in particular stopped farming. Because sow herd size increased more on farms with vs. without group housing and because the proportion of the herd that was group-housed also tended to increase between 2003 to 2009, the change to group housing took place faster when expressed at the level of the sow (from 9.1% in 2003 to 34.1% in 2009) instead of farm. The percentage of farmers planning to convert to group housing within 2 yr was 4.1% in 2003, and 6 to 7% thereafter. These were typically young farmers (P = 0.006) with a large sow herd (P < 0.001) and with a likely successor (P = 0.03). Free access stalls were the most common group housing system (31% of farms, 37% of sows). Their popularity is expected to increase further at the expense of electronic feeding stations, ad libitum feeding, and stalls/troughs with manual feed delivery. User satisfaction was generally high but depended on whether or not all gestating sows were kept in group (P < 0.001), the provisioning of environmental enrichment (P = 0.057), and the age (P = 0.012) and type (P = 0.016) of system. The main criteria for choosing a certain group housing system were the investment costs and sow health and welfare. The importance of economic reasons (P = 0.007) and type of labor (P = 0.043) decreased with the age of the system. In 2003 and 2005 the main reason for not having converted to group housing was that farmers would stop keeping sows by 2013. In 2007 and 2009 the reasons mainly concerned uncertainty about the future and maximally delaying the conversion. Belgium is one of the European Union countries where the pig industry is expected to undergo drastic changes during the few years remaining before the ban on individual housing

A transition in the Dutch wastewater system? : The struggle between discourses and with lock-ins

Recently, calls have increased for a paradigm shift or transition towards resource recovery and a circular economy in the Dutch wastewater system. However, we have observed diverging interpretations on the nature of the transition. This reflects the political environment of sustainability transitions: political struggle emerges over the definition of problems, futures and strategies to be used. In order to help clarify the emerging debate and identify political choices, we conducted a discourse analysis. We identified three discourses that reveal some of the political choices to be made. One discourse is becoming dominant and focusses on optimising the large-scale infrastructure, market development and legislative changes. The discourse draws on the existing infrastructure and current political-economic institutions, which gives it an advantage in becoming dominant. Our findings also suggest that this discourse shapes a transition pathway that is characterised by lock-in effects and, at most, incremental changes instead of a fundamental shift in the established Dutch wastewater system

Morphological characterization of humus forms in coastal dune systems: experience from the Flemish coast and northwest France

Humus profiles of dune soils on 46 sites along the Flemish coast and in northwest France were classified according to three classification systems (Delecour, 1980; Green et al., 1993 and Jabiol et al., 1995). These classification systems are not well adapted to young ecosystems on almost pure sand. Problems arose in the keys themselves and in the terminology and criteria which are used to classify the soils