Dual-FRET imaging of IP3 and Ca2+ revealed Ca2+-induced IP3 production maintains long lasting Ca2+ oscillations in fertilized mouse eggs

In most species, fertilization induces Ca(2+) transients in the egg. In mammals, the Ca(2+) rises are triggered by phospholipase Czeta (PLCzeta) released from the sperm; IP3 generated by PLCzeta induces Ca(2+) release from the intracellular Ca(2+) store through IP3 receptor, termed IP3-induced Ca(2+) release. Here, we developed new fluorescent IP3 sensors (IRIS-2s) with the wider dynamic range and higher sensitivity (Kd = 0.047-1.7 muM) than that we developed previously. IRIS-2s employed green fluorescent protein and Halo-protein conjugated with the tetramethylrhodamine ligand as fluorescence resonance energy transfer (FRET) donor and acceptor, respectively. For simultaneous imaging of Ca(2+) and IP3, using IRIS-2s as the IP3 sensor, we developed a new single fluorophore Ca(2+) sensor protein, DYC3.60. With IRIS-2s and DYC3.60, we found that, right after fertilization, IP3 concentration ([IP3]) starts to increase before the onset of the first Ca(2+) wave. [IP3] stayed at the elevated level with small peaks followed after Ca(2+) spikes through Ca(2+) oscillations. We detected delays in the peak of [IP3] compared to the peak of each Ca(2+) spike, suggesting that Ca(2+)-induced regenerative IP3 production through PLC produces small [IP3] rises to maintain [IP3] over the basal level, which results in long lasting Ca(2+) oscillations in fertilized eggs

B cell-derived GABA elicits IL-10⁺ macrophages to limit anti-tumour immunity

GABAを標的とする抗腫瘍免疫機構 --代謝産物を介した免疫細胞間制御の一端を解明--. 京都大学プレスリリース. 2021-11-10.Small, soluble metabolites not only are essential intermediates in intracellular biochemical processes, but can also influence neighbouring cells when released into the extracellular milieu1-3. Here we identify the metabolite and neurotransmitter GABA as a candidate signalling molecule synthesized and secreted by activated B cells and plasma cells. We show that B cell-derived GABA promotes monocyte differentiation into anti-inflammatory macrophages that secrete interleukin-10 and inhibit CD8⁺ T cell killer function. In mice, B cell deficiency or B cell-specific inactivation of the GABA-generating enzyme GAD67 enhances anti-tumour responses. Our study reveals that, in addition to cytokines and membrane proteins, small metabolites derived from B-lineage cells have immunoregulatory functions, which may be pharmaceutical targets allowing fine-tuning of immune responses

Latitudinal gradient of spruce forest understory and tundra phenology in Alaska as observed from satellite and ground-based data

The latitudinal gradient of the start of the growing season (SOS) and the end of the growing season (EOS) were quantified in Alaska (61°N to 71°N) using satellite-based and ground-based datasets. The Alaskan evergreen needleleaf forests are sparse and the understory vegetation has a substantial impact on the satellite signal. We evaluated SOS and EOS of understory and tundra vegetation using time-lapse camera images. From the comparison of three SOS algorithms for determining SOS from two satellite datasets (SPOT-VEGETATION and Terra-MODIS), we found that the satellite-based SOS timing was consistent with the leaf emergence of the forest understory and tundra vegetation. The ensemble average of SOS over all satellite algorithms can be used as a measure of spring leaf emergence for understory and tundra vegetation. In contrast, the relationship between the ground-based and satellite-based EOSs was not as strong as that of SOS both for boreal forest and tundra sites because of the large biases between those two EOSs (19 to 26 days). The satellite-based EOS was more relevant to snowfall events than the senescence of understory or tundra. The plant canopy radiative transfer simulation suggested that 84–86% of the NDVI seasonal amplitude could be a reasonable threshold for the EOS determination. The latitudinal gradients of SOS and EOS evaluated by the satellite and ground data were consistent and the satellite-derived SOS and EOS were 3.5 to 5.7 days degree− 1 and − 2.3 to − 2.7 days degree− 1, which corresponded to the spring (May) temperature sensitivity of − 2.5 to − 3.9 days °C− 1 in SOS and the autumn (August and September) temperature sensitivity of 3.0 to 4.6 days °C− 1 in EOS. This demonstrates the possible impact of phenology in spruce forest understory and tundra ecosystems in response to climate change in the warming Artic and sub-Arctic regions