Signaling adaptor protein v-Crk activates Rho and regulates cell motility in 3Y1 rat fibroblast cell line.

The adaptor protein Crk has been reported to associate with focal adhesions and is thought to be involved in integrin-mediated signaling pathway. However, the precise mechanism of Crk-dependent regulation of cytoskeleton still remains under investigation. In this study, we have established a v-Crk-inducible cell line in rat fibroblasts 3Y1 cells and found that v-Crk activated Rho and induced actin stress fiber formation. In addition to the induction of tyrosine-phosphorylation of p130(Cas) and paxillin, we demonstrated that v-Crk induced threonine-phosphorylated bands sized at 72/78 kDa found specifically in 3Y1 cells. Both of the inhibitors of Rho and Rho-associated kinase, C3 and Y27632, respectively, inhibited these v-Crk-induced biochemical effects. Although v-Crk-induced cells exhibited a decrease of cell motility, integrin stimulation recovered the suppression of motility. Furthermore, v-Crk enhanced motility in chemotactic assay toward fibronectin with additional activation of Rho and the increase of levels of CD44 cleavage. These results suggest that v-Crk activated Rho and induced actin stress fiber formation and CD44 cleavage leading to the regulation of cell motility.     

LIF-Free Embryonic Stem Cell Culture in Simulated Microgravity

Background

Leukemia inhibitory factor (LIF) is an indispensable factor for maintaining mouse embryonic stem (ES) cell pluripotency. A feeder layer and serum are also needed to maintain an undifferentiated state, however, such animal derived materials need to be eliminated for clinical applications. Therefore, a more reliable ES cell culture technique is required.

Methodology/Principal Findings

We cultured mouse ES cells in simulated microgravity using a 3D-clinostat. We used feeder-free and serum-free media without LIF.

Conclusions/Significance

Here we show that simulated microgravity allows novel LIF-free and animal derived material-free culture methods for mouse ES cells.     

Candida nonsorbophila sp. nov., a new ascomycetous yeast species isolated in Thailand

Four yeast strains, RS42, SSK10, ST-520 and ST-521, isolated from water in a mangrove forest, bark of a tree and fruit of Ficus sp., respectively, were found to represent a hitherto undescribed anamorphic species. The four strains are related to Candida sinolaborantium in the D1/D2 domain of the large subunit rRNA gene, but differed by eight nucleotide substitutions and two indels, and for this reason are regarded as members of a separate species. Because ascospore formation was not detected, it is described as a new species of Candida, Candida nonsorbophila sp. nov. The type strain is RS42^T (BCC 25963^T=NBRC 103860^T=CBS 10862^T). This species is distinguished from C. sinolaborantium by the inability to assimilate l -sorbose, l -rhamnose and galactitol, and a higher maximum growth temperature.     

Life history of <Emphasis Type="Italic">Stenopsylla nigricornis</Emphasis> (Hemiptera: Psylloidea: Triozidae) and phytohormones involved in its gall induction

Many phytophagous insects have an ability to manipulate plant tissue and induce galls, but the mechanism is not yet fully understood. Some insects have multivoltine life cycles, and each generation induces galls on different plant species or different organs in the same host. Such host-use patterns are interesting study subjects to clarify the gall-inducing mechanisms of insects. We focused on a multivoltine and gall-inducing psyllid Stenopsylla nigricornis Kuwayama (Hemiptera: Psylloidea: Triozidae), which is associated with Symplocos lucida Sieb. (Symplocaceae). Based on periodic field surveys in Kyushu, Japan, S. nigricornis is revealed to have a bivoltine life history. Then, we revealed that the spring generation induces galls on leaves, while the autumn generation does so on flower buds and overwintering leaf buds. We also analyzed phytohormones in normal plant tissue, S. nigricornis nymphs, and their galls. As a result, nymphs were discovered to contain much higher concentrations of isopentenyladenosine and its possible precursor, isopentenyladenosine riboside than plant tissues, strongly suggesting that the phytohormone is involved in gall induction by S. nigricornis. Because flower bud galls contained significantly lower concentrations of abscisic acid (ABA) than normal flower bud, the autumn generation nymphs are considered to regulate the ABA level and to promote the earlier opening of host flower buds.     

Effects of logging rotation in a lowland dipterocarp forest on mating system and gene flow in <Emphasis Type="Italic">Shorea parvifolia</Emphasis>

Selective logging is one of several silvicultural practices used in sustainable forest management in the lowland dipterocarp forest in Indonesia. Selecting only trees with diameters >50 cm for logging can reduce the density of reproductive trees, thereby affecting pollen dispersal and influencing the mating system among remaining trees. We evaluated the effect of logging rotations on the mating system, gene flow and genetic diversity in populations of Shorea parvifolia in primary forest, and in first and second rotation forest. Our results revealed that multiple (or at least two) selective logging events with a 30-year logging rotation had a significant impact on the genetic diversity of pollen clouds. However, the average pollen dispersal distance did not differ significantly among the multiple selective logging rotations. The multiple rotations reduced the outcrossing rate and the number effective of pollen donors in the logged forest. Moreover, the number of pollen donors in a plot was affected by the basal area of reproductive trees present. These results suggest that reducing the number of reproductive trees by logging with multiple rotations might increase the bi-parental inbreeding rate due to the reduction in density of reproductive trees in a selectively logged forest. We conclude that multiple rotations with a 30-year cycle of selective logging as currently practiced would reduce the density of reproductive trees, and would not be sustainable in terms of maintaining genetic diversity in tropical forests of Southeast Asia.     

Fighting and Stinging Responses are Affected by a Dopamine Receptor Blocker Flupenthixol in Honey Bee Virgin Queens

Fighting and aggression are important tasks for self-preservation in animals. In honey bees, virgin queens fight against each other for survival in a monogynous colony. Because the virgin queens have higher levels of dopamine (DA) in the brain than do mated queens with low aggressiveness, DA may promote fighting and aggression behaviours of virgin queens. In the present study, we investigated the effect of DA on the fighting and stinging response of honey bee virgin queens. We injected two concentrations (10^?3 M and 10^?2 M) of DA and the DA receptor blocker flupenthixol into the abdomen of one-day-old virgin queens and observed fighting and stinging responses. DA injection did not affect fighting and stinging. Injections of 10^?3 M flupenthixol decreased the winning rate significantly, whereas 10^?2 M flupenthixol increased the winning rate, indicating the opposite effects on fighting responses depending on the degrees of blockade of DA signalling. In terms of the stinging response, 10^?2 M flupenthixol-injected virgin queens stung significantly more often than control and 10^?3 M flupenthixol-injected virgin queens. These results suggest an involvement of DA signalling in the regulation of fighting and aggression in virgin queens, although a blockade of DA does not always inhibit these behaviours.     

Return of Drones: Flight Experience Improves Returning Performance in Honeybee Drones

The effect of experience on the behavior of worker bees has been extensively investigated; however, few such studies have been conducted on male bees. Honeybee (Apis mellifera) males (drones), unlike the males of other social hymenopterans, return to their nest after performing a mating flight and have, therefore, an opportunity to learn from their experiences. This provides a chance to understand the significance of experience in social hymenopteran males. Here, we investigated whether experience improves the returning performance in drones (rate and time of return to the hive). We compared the returning performance of “Experienced” drones that were allowed to fly freely and thus had an opportunity to learn the position of the hive before the experiment with “Naive” drones that were not allowed to fly and therefore, had no opportunity to learn. We found that Experienced drones returned to the hive after a displacement, whereas Naive drones did not. Furthermore, time to return decreased with the age of drones. These results suggest that flight experience improves the returning performance, which should increase the possibility of mating success and overall colony fitness.