Inhibition of NAADP signalling on reperfusion protects the heart by preventing lethal calcium oscillations via two-pore channel 1 and opening of the mitochondrial permeability transition pore

Aims In the heart, a period of ischaemia followed by reperfusion evokes powerful cytosolic Ca2+ oscillations that can cause lethal cell injury. These signals represent attractive cardioprotective targets, but the underlying mechanisms of genesis are ill-defined. Here, we investigated the role of the second messenger nicotinic acid adenine dinucleotide phosphate (NAADP), which is known in several cell types to induce Ca2+ oscillations that initiate from acidic stores such as lysosomes, likely via two-pore channels (TPCs, TPC1 and 2). Methods and results An NAADP antagonist called Ned-K was developed by rational design based on a previously existing scaffold. Ned-K suppressed Ca2+ oscillations and dramatically protected cardiomyocytes from cell death in vitro after ischaemia and reoxygenation, preventing opening of the mitochondrial permeability transition pore. Ned-K profoundly decreased infarct size in mice in vivo. Transgenic mice lacking the endo-lysosomal TPC1 were also protected from injury. Conclusion NAADP signalling plays a major role in reperfusion-induced cell death and represents a potent pathway for protection against reperfusion injury

Defining murine organogenesis at single-cell resolution reveals a role for the leukotriene pathway in regulating blood progenitor formation.

During gastrulation, cell types from all three germ layers are specified and the basic body plan is established 1 . However, molecular analysis of this key developmental stage has been hampered by limited cell numbers and a paucity of markers. Single-cell RNA sequencing circumvents these problems, but has so far been limited to specific organ systems 2 . Here, we report single-cell transcriptomic characterization of >20,000 cells immediately following gastrulation at E8.25 of mouse development. We identify 20 major cell types, which frequently contain substructure, including three distinct signatures in early foregut cells. Pseudo-space ordering of somitic progenitor cells identifies dynamic waves of transcription and candidate regulators, which are validated by molecular characterization of spatially resolved regions of the embryo. Within the endothelial population, cells that transition from haemogenic endothelial to erythro-myeloid progenitors specifically express Alox5 and its co-factor Alox5ap, which control leukotriene production. Functional assays using mouse embryonic stem cells demonstrate that leukotrienes promote haematopoietic progenitor cell generation. Thus, this comprehensive single-cell map can be exploited to reveal previously unrecognized pathways that contribute to tissue development

Mid-Spring Burning Reduces Spotted Knapweed and Increases Native Grasses during a Michigan Experimental Grassland Establishment

The definitive version is available at www.blackwell-synergy.com Infestations of the exotic perennial Spotted knapweed (Centaurea maculosa Lam.) hinder the restoration and management of native ecosystems on droughty, infertile sites throughout the Midwestern United States. We studied the effects of annual burning on knapweed persistence on degraded, knapweed-infested gravel-mine spoils in western Michigan. Our experiment included 48, 4-m2 plots seeded to native warm-season grasses in 1999 using a factorial arrangement of initial herbicide and fertility treatments. Beginning in 2003, we incorporated fire as an additional factor and burned half of the plots in late April or May for three years (2003-2005). Burning increased the dominance of warm-season grasses and decreased both biomass and dominance of knapweed in most years. Burning reduced adult knapweed densities in all three years of the study, reduced seedling densities in the first two years, and reduced juvenile densities in the last two years. Knapweed density and biomass also declined on the unburned plots through time, suggesting that warm-season grasses may effectively compete with knapweed even in the absence of fire. By the end of the study, mean adult knapweed densities on both burned (0.4-m2) and unburned plots (1.3-m2) were reduced to levels where the seeded grasses should persist with normal management, including the use of prescribed fire. These results support the use of carefully timed burning to help establish and maintain fire-adapted native plant communities on knapweed-infested sites in the Midwest by substantially reducing knapweed density, biomass, and seedling recruitment, and by further shifting the competitive balance toward native warm-season grasses