Adenosine A3 receptor is involved in ADP-induced microglial process extension and migration

The extension of microglial processes toward injured sites in the brain is triggered by the stimulation of the purinergic receptor P2Y(12) by extracellular ATP. We recently showed that P2Y(12) stimulation by ATP induces microglial process extension in collagen gels. In the present study, we found that a P2Y(12) agonist, 2-methylthio-ADP (2MeSADP), failed to induce the process extension of microglia in collagen gels and that co-stimulation with adenosine, a phosphohydrolytic derivative of ATP, and 2MeSADP restored the chemotactic process extension. An adenosine A3 receptor (A3R)-selective agonist restored the chemotactic process extension, but other receptor subtype agonists did not. The removal of adenosine by adenosine deaminase and the blocking of A3R by an A3R-selective antagonist inhibited ADP-induced process extension. The A3R antagonist inhibited ADP-induced microglial migration, and an A3R agonist promoted 2MeSADP-stimulated migration. ADP and the A3R agonist activated Jun N-terminal kinase in microglia, and a Jun N-terminal kinase inhibitor inhibited the ADP-induced process extension. An RT-PCR analysis showed that A1R and A3R were expressed by microglia sorted from adult rat brains and that the A2AR expression level was very low. These results suggested that A3R signaling may be involved in the ADP-induced process extension and migration of microglia.     

Structural basis for extracellular interactions between calcitonin receptor-like receptor and receptor activity-modifying protein 2 for adrenomedullin-specific binding

The calcitonin receptor-like receptor (CRLR), a class B GPCR, forms a heterodimer with receptor activity-modifying protein 2 (RAMP2), and serves as the adrenomedullin (AM) receptor to control neovascularization, while CRLR and RAMP1 form the calcitonin gene-related peptide (CGRP) receptor. Here, we report the crystal structures of the RAMP2 extracellular domain alone and in the complex with the CRLR extracellular domain. The CRLR-RAMP2 complex exhibits several intermolecular interactions that were not observed in the previously reported CRLR-RAMP1 complex, and thus the shape of the putative ligand-binding pocket of CRLR-RAMP2 is distinct from that of CRLR-RAMP1. The CRLR-RAMP2 interactions were confirmed for the full-length proteins on the cell surface by site-specific photo-crosslinking. Mutagenesis revealed that AM binding requires RAMP2 residues that are not conserved in RAMP1. Therefore, the differences in both the shapes and the key residues of the binding pocket are essential for the ligand specificity.     

Accumulating microglia phagocytose injured neurons in hippocampal slice cultures: involvement of p38 MAP kinase

In this study, microglial migration and phagocytosis were examined in mouse organotypic hippocampal slice cultures, which were treated with N-methyl-D-aspartate (NMDA) to selectively injure neuronal cells. Microglial cells were visualized by the expression of enhanced green fluorescent protein. Daily observation revealed microglial accumulation in the pyramidal cell layer, which peaked 5 to 6 days after NMDA treatment. Time-lapse imaging showed that microglia migrated to the pyramidal cell layer from adjacent and/or remote areas. There was no difference in the number of proliferating microglia between control and NMDA-treated slices in both the pyramidal cell layer and stratum radiatum, suggesting that microglial accumulation in the injured areas is mainly due to microglial migration, not to proliferation. Time-lapse imaging also showed that the injured neurons, which were visualized by propidium iodide (PI), disappeared just after being surrounded by microglia. Daily observation revealed that the intensity of PI fluorescence gradually attenuated, and this attenuation was suppressed by pretreatment with clodronate, a microglia toxin. These findings suggest that accumulating microglia phagocytosed injured neurons, and that PI fluorescence could be a useful indicator for microglial phagocytosis. Using this advantage to examine microglial phagocytosis in living slice cultures, we investigated the involvements of mitogen-activated protein (MAP) kinases in microglial accumulation and phagocytosis. p38 MAP kinase inhibitor SB203580, but not MAP kinase/extracellular signal-regulated kinase inhibitor PD98059 or c-Jun N-terminal kinase inhibitor SP600125, suppressed the attenuation of PI fluorescence. On the other hand, microglial accumulation in the injured areas was not inhibited by any of these inhibitors. These data suggest that p38 MAP kinase plays an important role in microglial phagocytosis of injured neurons.     

Optimal-foraging predator favors commensalistic Batesian mimicry

Background

Mimicry, in which one prey species (the Mimic) imitates the aposematic signals of another prey (the Model) to deceive their predators, has attracted the general interest of evolutionary biologists. Predator psychology, especially how the predator learns and forgets, has recently been recognized as an important factor in a predator–prey system. This idea is supported by both theoretical and experimental evidence, but is also the source of a good deal of controversy because of its novel prediction that in a Model/Mimic relationship even a moderately unpalatable Mimic increases the risk of the Model (quasi-Batesian mimicry).

Methodology/Principal Findings

We developed a psychology-based Monte Carlo model simulation of mimicry that incorporates a “Pavlovian” predator that practices an optimal foraging strategy, and examined how various ecological and psychological factors affect the relationships between a Model prey species and its Mimic. The behavior of the predator in our model is consistent with that reported by experimental studies, but our simulation''s predictions differed markedly from those of previous models of mimicry because a more abundant Mimic did not increase the predation risk of the Model when alternative prey were abundant. Moreover, a quasi-Batesian relationship emerges only when no or very few alternative prey items were available. Therefore, the availability of alternative prey rather than the precise method of predator learning critically determines the relationship between Model and Mimic. Moreover, the predation risk to the Model and Mimic is determined by the absolute density of the Model rather than by its density relative to that of the Mimic.

Conclusions/Significance

Although these predictions are counterintuitive, they can explain various kinds of data that have been offered in support of competitive theories. Our model results suggest that to understand mimicry in nature it is important to consider the likely presence of alternative prey and the possibility that predation pressure is not constant.     

Impact of altitude and topography on the genetic diversity of Quercus serrata populations in the Chichibu Mountains, central Japan

In order to elucidate the factors affecting the genetic diversity of Quercus serrata in secondary forests in mountainous regions, we evaluated the level and distribution of genetic variation within and between 15 populations using seven microsatellite markers. The populations were at altitudes ranging from 140 to 1200 m in and around the Chichibu Mountains, central Japan.The expected heterozygosity (H_E) ranged from 0.766 to 0.837. The two populations that exhibited the highest and the second highest values of H_E are located beside a river and a lake, respectively. The two populations exhibiting the lowest and the second lowest values of H_E are, in contrast, located on a summit and a ridge. The observed heterozygosity (H_O) varied between 0.638 and 0.844, and the value of this variable was also higher for the populations beside water than those on summits or ridges. The soils at the waterside are wet, in contrast to those on ridges and summits, which tend to be shallow and subject to rapid desiccation. These results suggest that a lack of soil moisture is likely to inhibit the development and regeneration of Q. serrata, and that genetic diversity is reduced in arid areas. The genetic differentiation was low (F_ST=0.013) among the investigated populations, although all five populations in Yamanashi prefecture clustered together in an UPGMA tree. According to a multiple regression analysis, there was no significant isolation by distance among the populations along either the horizontal or vertical axes. Therefore, genetic variation within populations is affected by topography, but variation between populations is hardly affected by geographical factors. Furthermore, the results of this study suggest two conclusions. First, that altitude is not always a useful variable when estimating the genetic diversity of plant populations in mountainous regions. Second, that genetic diversity can vary even among the undifferentiated plant populations in small areas like the Chichibu Mountains.     

The nutritional contribution of males affects the feeding behavior and spatial distribution of females in a bruchid beetle,<Emphasis Type="Italic"> Bruchidius dorsalis</Emphasis>

The relationship between the quality/quantity of male investment and the feeding behavior of females was investigated in a bruchid weevil, Bruchidius dorsalis (Fahraeus), whose males donate nutrition via seminal fluid to females. Experiments on the effect of feeding regimes of both sexes on the mating frequency of females showed that females mated at a higher frequency if given low-quality food or poor male investment. On the other hand, the experiment that examined the effect of male investment quality on female feeding behavior showed that females receiving the high-quality investment exhibited feeding behavior less often. These results suggest that male investment and feeding behavior play the same role for B. dorsalis females. These experiments also showed that there are sex-related asymmetries in mating and feeding behaviors: females mated more often but males fed more often. Moreover, a field census suggested that only males visited non-host flowers to feed on the pollen and nectar during the non-flowering period of the host plants; females always stayed on the host plants irrespective of the flowering phenology. These results suggest that in B. dorsalis courtship role reversal and sex-specific feeding modes are fairly fixed and obligatory, and that male investment, derived from sexual selection, could affect the feeding behavior and spatial distribution of both sexes, which may have far-reaching impact in various ecological contexts.     

Novel antibody to human BASP1 labels apoptotic cells post-caspase activation

Apoptosis is associated with morphological changes, including membrane blebbing, cell shrinkage, and chromatin condensation. However, the molecular mechanisms of the dynamic changes in cellular components during apoptosis are largely unknown. Here we developed a new rat monoclonal antibody, 9B1, that specifically immunolabeled dying cells in tissues and in cell cultures. The 9B1 antibody labeled the cytoplasm of apoptotic cells in a caspase-dependent manner. We identified human brain abundant membrane attached signal protein 1 (hBASP1) as the 9B1 antigen using the liquid chromatography with tandem mass spectrometry (LC/MS/MS) method. hBASP1 was present in the nucleus of HeLa cells, but relocated from the nucleus to the cytoplasm after the caspase activation step of apoptosis. Immunostaining analysis revealed that 9B1 preferentially labeled this cytoplasmic form of hBASP1. Labeling by 9B1 to distinguish apoptotic changes could be a novel criterion for determining whether cells with activated caspases are fated for survival or death.     

Coexistence mechanisms of evergreen,deciduous and coniferous trees in a mid-montane mixed forest on Mt. Emei,Sichuan, China

Structure and regeneration of a mid-montane (2200 m a.s.l.)mixed forest codominated by evergreen (Lithocarpuscleistocarpus), deciduous (Acer flabellatum)andconiferous (Tsuga chinensis, Abies fabri, andTaxus chinensis) trees were analyzed in a 40m× 60 m plot on Mt. Emei, Sichuan, China. Plant communitystructure and composition varied depending on topographic micro-habitat withinthe plot. Four topographic communities (topo-communities) were distinguishedwith dominant species corresponding to topography: (1)Abies – valley bank, (2) Acer– lower steep slope, (3) Lithocarpus – uppergentle slope, and (4) Tsuga – ridge. The coexistencemechanisms of the evergreen, deciduous, and coniferous trees were determined byidentifying the regeneration process characteristic of each dominant species,asrepresented by their seedling dispersion patterns and seedling establishmentalong the topographic gradients. The saplings and seedlings of the dominantswere distributed differently according to the topography:Lithocarpus under the canopy of parent trees and in gapsofthe upper gentle slope and ridge, Acer mainly in bothwell-lit and shady sites on the lower steep slope and on fallen logs,Abies on the valley bank but only in well-lit sites,Tsuga on the ridge, Taxus mostly onrock and well-lit sites on the slopes. Distribution of surviving saplings andseedlings was also related to the species of nearby canopy trees. We suggestthat Lithocarpus, Tsuga and Acer tendto be self-replacing in their own topographic habitats, andAbies survives as a fugitive by occupying occasionalsuitable gaps. The variation in soil conditions, particularly nutrients, withtopography affected seedling establishment and the growth of trees. Thesuccessional change of quantitative species composition, as predicted by theMarkovian model, shows the mixed forest to be in a sustained climax stage.     

Bioconversion of dethiobiotin to biotin by a cell-free system of a bioYB transformant of Bacillus sphaericus

Abstract Potassium-limited cultures of Candida utilis grown with ammonium chloride as the sole nitrogen source attained a higher dry weight than similar cultures grown with sodium nitrate as the sole nitrogen source. This increase depended on the extracellular ammonia concentration and was not due to accumulation of storage polymers. We conclude that in this yeast ammonium ions can fulfill to some extent the physiological role of potassium ions and that the intracellular concentration of ammonium ions is predominantly determined by the ammonia concentration in the culture medium.     

Expression of tumor necrosis factor at a specific developmental stage of mouse embryos

We investigated the expression of the tumor necrosis factor (TNF) gene during development of mouse embryos, and observed its transient expression on Days 9 and 10 of gestation. We also detected a 25-kDa protein showing immunological cross-reactivity with mouse TNF antibody in an extract of 10-day embryos. These results suggest that TNF plays a role in mammalian ontogenesis.