Focal adhesion kinase is essential for cardiac looping and multichamber heart formation

Focal adhesion kinase (FAK) is a critical mediator of matrix‐ and growth factor‐induced signaling during development. Myocyte‐restricted FAK deletion in mid‐gestation mice results in impaired ventricular septation and cardiac compaction. However, whether FAK regulates early cardiogenic steps remains unknown. To explore a role for FAK in multi‐chambered heart formation, we utilized anti‐sense morpholinos to deplete FAK in Xenopus laevis. Xenopus FAK morphants exhibited impaired cardiogenesis, pronounced pericardial edema, and lethality by tadpole stages. Spatial‐temporal assessment of cardiac marker gene expression revealed that FAK was not necessary for midline migration, differentiation, fusion of cardiac precursors, or linear heart tube formation. However, myocyte proliferation was significantly reduced in FAK morphant heart tubes and these tubes failed to undergo proper looping morphogenesis. Collectively our data imply that FAK plays an essential role in chamber outgrowth and looping morphogenesis likely stimulated by fibroblast growth factors (and possibly other) cardiotrophic factors. genesis 48:492–504, 2010. © 2010 Wiley‐Liss, Inc.     

Photosynthetic and carbohydrate status of easy-and difficult-to-acclimatize sea oats (Uniola paniculata L.) genotypes during In vitro culture and Ex vitro acclimatization

Summary The photosynthetic and carbohydrate status of an easy-to-acclimatize (EK 16-3) and a difficult-to-acclimatize (EK 11-1) genotype of Uniola paniculata L. (sea oats), a native dune species of the southeastern US, were evaluated during in vitro culture and ex vitro acclimatization. Net photosynthetic rate was eight times greater for EK 16-3 than EK 11-1 plantlets after ex vitro transfer. In vitro-produced leaves were morphologically similar to ex vitro-produced leaves and exhibited similar photosynthetic competence. EK 11-1 plantlets exhibited greater transpiration rates at the time of ex vitro transfer than EK 16-3 plantlets. However, the small magnitude of this difference, although significant, indicated that control of water loss was probably not the main cause for poor acclimatization of EK 11-1 plantlets. Carbohydrate analysis in vitro revealed that EK 16-3 plantlets utilized leaf starch reserves more rapidly than EK 11-1 plantlets. Starch utilization correlated with the development of leaves with expanded leaf blades during in vitro rooting in EK 16-3 plantlets. After ex vitro transfer, both genotypes exhibited significant decreases of starch and soluble sugar content in shoots and roots. However, the higher photosynthetic ability of shoots in EK 16-3 resulted in greater accumulation of shoot soluble sugars than EK 11-1 after 2-wk ex vitro culture. After 6-wk in vitro rooting, there were significantly higher chlorophyll and soluble protein contents, ribulose 1,5-bisphosphate carboxylase (rubisco) and phosphoenolpyruvate carboxylase activities in EK 16-3 than EK 11-1 shoots. These differences also correlated with the development of anatomical and morphological leaf features in EK 16-3 similar to those of greenhouse-produced leaves.     

Evolution of life‐history traits and mating strategy in males: a case study on two populations of a Drosophila parasitoid

Abiotic and biotic factors affect life‐history traits and lead populations to exhibit different behavioural strategies. Due to the direct link between their behaviour and fitness, parasitoid females have often been used to test the theories explaining these differences. In male parasitoids, however, such investigations are vastly understudied, although their mating strategy directly determines their fitness. In this study, we compared the pattern of life history traits and the mating strategy of males in two populations of the Drosophila parasitoid Asobara tabida, exposed to different biotic and abiotic conditions, with the major difference being that one of them was recently exposed to strong competition with the dominant competitor Leptopilina boulardi after recent climate change, the other population being settled in a location where L. boulardi has not been recorded. The results showed that individuals of both populations have a different reproductive strategy: in one population, females produced a more female‐biased sex ratio, while males accumulated more lipids during their larval development, invested more energy in reproduction and decreased their locomotor activity, suggesting a higher proportion of matings on their emergence patch, all of these factors being possibly linked to the new competition pressure. In both populations, mating without sperm transfer may persist for several days after males become sperm‐depleted, and may be more frequent than mating with sperm transfer over their whole lifespan. This point is discussed from an evolutionary point of view.     

The Ras-related Protein,Rap1A,Mediates Thrombin-stimulated,Integrin-dependent Glioblastoma Cell Proliferation and Tumor Growth

Rap1 is a Ras family GTPase with a well documented role in ERK/MAP kinase signaling and integrin activation. Stimulation of the G-protein-coupled receptor PAR-1 with thrombin in human 1321N1 glioblastoma cells led to a robust increase in Rap1 activation. This response was sustained for up to 6 h and mediated through RhoA and phospholipase D (PLD). Thrombin treatment also induced a 5-fold increase in cell adhesion to fibronectin, which was blocked by down-regulating PLD or Rap1A or by treatment with a β₁ integrin neutralizing antibody. In addition, thrombin treatment led to increases in phospho-focal adhesion kinase (tyrosine 397), ERK1/2 phosphorylation and cell proliferation, which were significantly inhibited in cells treated with β₁ integrin antibody or Rap1A siRNA. To assess the role of Rap1A in tumor formation in vivo, we compared growth of 1321N1 cells stably expressing control, Rap1A or Rap1B shRNA in a mouse xenograft model. Deletion of Rap1A, but not of Rap1B, reduced tumor mass by >70% relative to control. Similar observations were made with U373MG glioblastoma cells in which Rap1A was down-regulated. Collectively, these findings implicate a Rap1A/β₁ integrin pathway, activated downstream of G-protein-coupled receptor stimulation and RhoA, in glioblastoma cell proliferation. Moreover, our data demonstrate a critical role for Rap1A in glioblastoma tumor growth in vivo.     

Reproductive biology and reproductive output assessment in natural and introduced subpopulations of Apium bermejoi,a ‘Critically Endangered’ endemic plant from Menorca (western Mediterranean)

Apium bermejoi is a critically endangered species that is native to Menorca, an island of 702 km² located in the western Mediterranean basin. We investigated the reproductive biology of this species and evaluated flowering/fruiting phenology and reproductive output in wild subpopulations over four years (2007–2010). We used ex situ cultivated plants to study floral biology and performed bagging experiments to elucidate the breeding system. We addressed the role of ants in seed dispersal/predation in wild subpopulations by analysing the presence of A. bermejoi seeds in the waste accumulated on the refuse piles of all anthills. Our observations indicated that flowers are protandrous but that there is no dichogamy between flowers within and among umbels of the same individual. Moreover, individuals are mixed in the field because the close germination of several seeds and the plant’s caespitose habit promote cross‐pollination. Bagging experiments demonstrated that the species is autogamous, and seeds from selfing (bagged with muslin) showed high final germination (>80%). However, the fruit set of selfed umbels (0.3–0.4 per 1) was lower than that of open‐pollinated umbels (0.6–0.8 per 1), and the high pollen/ovule (P/O) ratio (3641±133, n= 30) was also indicative of xenogamy. Plants bagged with muslin, bagged with tulle or barred from contact from ants exhibited similar fruit sets, lower than those observed in isolated and control groups, suggesting that anemophily and pollination by flying insects were less effective than ant‐based pollination. We identified eight species of floral visitors, and we noted that ants were the most frequent visitors. Although we found some A. bermejoi seeds in the refuse piles of Messor bouvieri anthills, most of the seeds were not damaged (99.1%), and the number of seeds found represents only a small proportion of the overall fruit production. Because sexual reproduction is the main reproductive mechanism of these plants, despite their high potential for space colonisation and vegetative propagation, the reproductive output for each year determines the subpopulation configuration in the following year.     

Ecology and morphological variability of Aulacoseira granulata (Bacillariophyceae) in Spanish reservoirs

The ecology of Aulacoseira (formerly Melosira) granulata andits different forms in     

Y enriched and Y specific DNA sequences from the genome of the mediterranean fruit fly,Ceratitis capitata

DNA sequences that are enriched or specific to the genome of the male medfly, Ceratitis capitata, have been isolated using a differential hybridization approach. Twelve phage clones from a genomic library have been identified that consistently display more intense hybridization with a genomic DNA probe from males as opposed to one from females. Southern DNA blot analysis reveals that these recombinant clones contain at least one EcoRI fragment that is either specific to the male genome, or more highly represented in it, as compared with the female genome. These EcoRI fragments, when used as probes, all generate a similar pattern of intense multiple bands in genomic DNA of males. This suggests the presence of repetitive sequences that are at least partially homologous in these regions of the genome that are specific to or enriched in males. In situ hybridization to mitotic chromosomes confirms a Y chromosomal origin for the male specific repetitive sequences. Data on the genomic organization, representation and evolutionary conservation of these sequences that are specific to or enriched in males are presented. Studies of the genomic organization and representation of flanking sequences that are not male specific are presented as well.by R. Appels     

Gene transfer of H-2 class II genes: antigen presentation by mouse fibroblast and hamster B-cell lines

We have transferred the mouse Ak alpha and Ak beta genes, which encode the class II I-Ak molecule, into mouse L-cell fibroblasts and hamster B cells. I-Ak molecules are expressed on the surface of both cell types. The L-cell and hamster B-cell I-Ak molecules appear normal by serological analyses and two-dimensional gel electrophoresis. Furthermore, the I-Ak molecules on L cells can act as targets for the allogenic T-cell killing of the transformed L cells. The I-Ak molecules in both mouse fibroblasts and hamster B cells can present certain antigens to T-cell helper hybridomas. Thus only class II molecules are required to convert the nonantigen-presenting cell. Accordingly, it will be possible to dissect the structure-function relationships existing between Ia molecules, foreign antigen, and T-cell receptor molecules by in vitro site-directed mutagenesis and gene transfer.