Identification and Characterization of Rvs162/Rvs167-3, a Novel N-BAR Heterodimer in the Human Fungal Pathogen Candida albicans

Membrane reshaping resides at the core of many important cellular processes, and among its mediators are the BAR (Bin, Amphiphysin, Rvs) domain-containing proteins. We have explored the diversity and function of the Rvs BAR proteins in Candida albicans and identified a novel family member, Rvs167-3 (orf19.1861). We show that Rvs167-3 specifically interacts with Rvs162 to form a stable BAR heterodimer able to bind liposomes in vitro. A second, distinct heterodimer is formed by the canonical BAR proteins Rvs161 and Rvs167. Purified Rvs161/Rvs167 complex also binds liposomes, indicating that C. albicans expresses two functional BAR heterodimers. We used live-cell imaging to localize green fluorescent protein (GFP)-tagged Rvs167-3 and Rvs167 and show that both proteins concentrate in small cortical spots. However, while Rvs167 strictly colocalizes with the endocytic marker protein Abp1, we do not observe any colocalization of Rvs167-3 with sites of endocytosis marked by Abp1. Furthermore, the rvs167-3Δ/Δ mutant is not defective in endocytosis and strains lacking Rvs167-3 or its partner Rvs162 do not display increased sensitivity to high salt concentrations or decreased cell wall integrity, phenotypes which have been observed for rvs167Δ/Δ and rvs161Δ/Δ strains and which are linked to endocytosis defects. Taken together, our results indicate different roles for the two BAR heterodimers in C. albicans: the canonical Rvs161/Rvs167 heterodimer functions in endocytosis, whereas the novel Rvs162/Rvs167-3 heterodimer seems not to be involved in this process. Nevertheless, despite their different roles, our phenotypic analysis revealed a genetic interaction between the two BAR heterodimers, suggesting that they may have related but distinct membrane-associated functions.     

Effect of competition intensity on recruitment of palatable and unpalatable grasses

In this study, we made an attempt to reveal how competition intensity from established plants impacts on palatable and unpalatable grass seedlings recruitment, in a natural mesic grassland of central Argentina. Our objective was to assess the seedling recruitment of a palatable species (Chascolytrum subaristatum) and an unpalatable species (Nassella trichotoma) in microsites differing in competition intensity from established plants. Identity (C. subaristatum and N. trichotoma) and defoliation severity were used as surrogate for competition intensity. In March 2017, we permanently marked established individuals of N. trichotoma and C. subaristatum and placed two circular plots adjacent to each individual. In one plot we added seeds of N. trichotoma and in the other seeds of C. subaristatum. After seeding, established plants were randomly assigned to one of three level of defoliation: without defoliation, low defoliation severity and high defoliation severity. From April to November 2017 (i.e. over a complete annual growing cycle), we measured seedling density, recruitment and growth. Our results supported the hypothesis that seedlings of palatable grasses are more competitive than seedlings of unpalatable grasses. Seedling of the palatable grass C. subaristatum recruited successfully regardless the intensity of competition from established plants, whereas seedlings of the unpalatable grass N. trichotoma recruited better under low competitive pressure from established plants. Our results suggest that the availability of microsites with low competitive pressure from the established vegetation, created by selective grazing of palatable grasses, promotes the recruitment of unpalatable grass seedlings. This mechanism may contribute to the species replacement process commonly observed in heavy grazed grasslands.     

Saikosaponin-d,a novel SERCA inhibitor,induces autophagic cell death in apoptosis-defective cells

Autophagy is an important cellular process that controls cells in a normal homeostatic state by recycling nutrients to maintain cellular energy levels for cell survival via the turnover of proteins and damaged organelles. However, persistent activation of autophagy can lead to excessive depletion of cellular organelles and essential proteins, leading to caspase-independent autophagic cell death. As such, inducing cell death through this autophagic mechanism could be an alternative approach to the treatment of cancers. Recently, we have identified a novel autophagic inducer, saikosaponin-d (Ssd), from a medicinal plant that induces autophagy in various types of cancer cells through the formation of autophagosomes as measured by GFP-LC3 puncta formation. By computational virtual docking analysis, biochemical assays and advanced live-cell imaging techniques, Ssd was shown to increase cytosolic calcium level via direct inhibition of sarcoplasmic/endoplasmic reticulum Ca²⁺ ATPase pump, leading to autophagy induction through the activation of the Ca²⁺/calmodulin-dependent kinase kinase–AMP-activated protein kinase–mammalian target of rapamycin pathway. In addition, Ssd treatment causes the disruption of calcium homeostasis, which induces endoplasmic reticulum stress as well as the unfolded protein responses pathway. Ssd also proved to be a potent cytotoxic agent in apoptosis-defective or apoptosis-resistant mouse embryonic fibroblast cells, which either lack caspases 3, 7 or 8 or had the Bax-Bak double knockout. These results provide a detailed understanding of the mechanism of action of Ssd, as a novel autophagic inducer, which has the potential of being developed into an anti-cancer agent for targeting apoptosis-resistant cancer cells.     

Fighting by Kittens and Piglets during Suckling: What Does it Mean?

Newborn kittens and piglets show some notable similarities in suckling‐associated behaviours. Both develop a pattern of teat use in which each member of the litter predominantly occupies one or two particular nipples or teats, both show agonistic behaviour towards littermates during suckling, and both are born with ‘weapons’–kittens with sharp claws and piglets with sharp needle teeth. Traditionally, the differential use of nipples or teats has been thought to be due to differential quality of the mammary glands. However, neither our findings in kittens nor an examination of the literature on piglets support this view. As mammary glands usually are of equal quality and as there are normally more glands than young, there seems little reason for littermates to fight. Furthermore, there is also little evidence that individual performance in suckling contests or use of ‘weapons’ is related to weight gain or to dominance relations. Given the present lack of satisfying adaptive explanations for the seemingly aggressive behaviour and use of weapons by kittens and piglets, we suggest these to be epiphenomena of developmental processes anticipating the use of these behaviours and structures in later adaptive contexts.     

Discrete Notch signaling requirements in the specification of hematopoietic stem cells

Hematopoietic stem cells (HSCs) require multiple molecular inputs for proper specification, including activity of the Notch signaling pathway. A requirement for the Notch1 and dispensability of the Notch2 receptor has been demonstrated in mice, but the role of the remaining Notch receptors has not been investigated. Here, we demonstrate that three of the four Notch receptors are independently required for the specification of HSCs in the zebrafish. The orthologues of the murine Notch1 receptor, Notch1a and Notch1b, are each required intrinsically to fate HSCs, just prior to their emergence from aortic hemogenic endothelium. By contrast, the Notch3 receptor is required earlier within the developing somite to regulate HSC emergence in a non-cell-autonomous manner. Epistatic analyses demonstrate that Notch3 function lies downstream of Wnt16, which is required for HSC specification through its regulation of two Notch ligands, dlc and dld. Collectively, these findings demonstrate for the first time that multiple Notch signaling inputs are required to specify HSCs and that Notch3 performs a novel role within the somite to regulate the neighboring precursors of hemogenic endothelium.     

Postural control mechanisms in the upside-down catfish (Synodontis nigriventris)

The catfishSynodontis nigriventris normally swims upside-down but can assume any posture in response to a substrate it swims close to. Postural reflexes of the body and the eyes, labyrinthine anatomy and passively maintained posture were investigated to obtain indications for possible mechanisms controlling the peculiar postural behavior of this fish. Saccade-like resetting movements of the eyes during counter-roll to body tilt about the longitudinal axis, and maintained tilted swimming positions in blinded fish suggest that these animals reset their vestibular CNS circuits to zero when in tilted positions.Synodontis nigriventris is thus able to maintain any posture without interference from tilt-counteracting vestibular reflexes. The normal upside-down swimming apparently results from a central bias for this position and a supporting ventral light response.We conclude that if the reafference principle applies to the phenomena investigated, the efference copy may be fed through an integrator before reaching vestibular reflex circuits.     

The impact of livestock grazing on the spatial pattern of vegetation in north-eastern Patagonia,Argentina

Background: The occurrence of shrub patches, alternating with either bare soil or low herbaceous cover, is a common feature in arid and semi-arid shrublands throughout the world. This patchy pattern of vegetation may result from water limitation, modulated by plant interactions; grazing (offtake and tramping) by livestock may cause further patchiness vegetation structure.

Aims: We hypothesised that vegetation patchiness in the semi-arid shrublands of north-eastern Patagonia would be increased by livestock grazing, but not by positive interactions between adult plants of shrubs and grasses.

Methods: We compared vegetation cover and pattern at three grazing intensities (exclosure, light and heavy grazing) and measured the growth of a representative shrub and grass in the presence and absence of the other to quantify the role of plant-to-plant interactions and its interaction with grazing for vegetation structure.

Results: In the grazing exclosure and in moderately grazed areas, vegetation cover among shrub patches was larger, whereas the top cover of shrubs was lower than in the heavily grazed areas. We did not find any evidence of positive interactions between shrub and grass life forms.

Conclusions: Our results were consistent with the hypothesis that livestock grazing increased the formation of patchy vegetation cover in arid and semi-arid shrublands.