Characterization of Chilling Sensitivity of Tropical and Temperate Grasses

C16-C18 fatty acid composition of some lipids in several species of temperate and tropical grasses were measured. The fatty acid composition of phosphatidylglycerol (PG) and sulfoquinovosyl diglyceride (SQDG) indicated significant differences between temperate and tropical grasses. Especially the differences of the fatty acid composition of PG were remarkable. The unsaturated fatty acid content of PG in tropical grasses was lower than the contents in temperate grasses. These differences constantly appeared through the year. These differences were caused by the low content of polyunsaturated fatty acids, or the low contents of C18 fatty acids

Measurement of inositol 1,4,5-trisphosphate in living cells using an improved set of resonance energy transfer-based biosensors

Improved versions of inositol-1,4,5-trisphosphate (InsP3) sensors were created to follow intracellular InsP3 changes in single living cells and in cell populations. Similar to previous InsP3 sensors the new sensors are based on the ligand binding domain of the human type-I InsP3 receptor (InsP3R-LBD), but contain a mutation of either R265K or R269K to lower their InsP3 binding affinity. Tagging the InsP3R-LBD with N-terminal Cerulean and C-terminal Venus allowed measurement of Ins P3 in single-cell FRET experiments. Replacing Cerulean with a Luciferase enzyme allowed experiments in multi-cell format by measuring the change in the BRET signal upon stimulation. These sensors faithfully followed the agonist-induced increase in InsP3 concentration in HEK 293T cells expressing the Gq-coupled AT1 angiotensin receptor detecting a response to agonist concentration as low as 10 pmol/L. Compared to the wild type InsP3 sensor, the mutant sensors showed an improved off-rate, enabling a more rapid and complete return of the signal to the resting value of InsP3 after termination of M3 muscarinic receptor stimulation by atropine. For parallel measurements of intracellular InsP3 and Ca2+ levels in BRET experiments, the Cameleon D3 Ca2+ sensor was modified by replacing its CFP with luciferase. In these experiments depletion of plasma membrane PtdIns(4,5)P2 resulted in the fall of InsP3 level, followed by the decrease of the Ca2+-signal evoked by the stimulation of the AT1 receptor. In contrast, when type-III PI 4-kinases were inhibited with a high concentration of wortmannin or a more specific inhibitor, A1, the decrease of the Ca2+-signal preceded the fall of InsP3 level indicating an InsP3-, independent, direct regulation of capacitative Ca2+ influx by plasma membrane inositol lipids. Taken together, our results indicate that the improved InsP3 sensor can be used to monitor both the increase and decrease of InsP3 levels in live cells suitable for high-throughput BRET applications. © 2015, Public Library of Science. All rights reserved

Subaru Studies of the Cosmic Dawn

An overview on the current status of the census of the early universe population is given. Observational surveys of high redshift objects provide direct opportunities to study the early epoch of the Universe. The target population included are Lyman Alpha Emitters (LAE), Lyman Break Galaxies (LBG), gravitationally lensed galaxies, quasars and gamma-ray bursts (GRB). The basic properties of these objects and the methods used to study them are reviewed. The present paper highlights the fact that the Subaru Telescope group made significant contributions in this field of science to elucidate the epoch of the cosmic dawn and to improve the understanding of how and when infant galaxies evolve into mature ones.Comment: 14 pages, 11 figures, accepted for publication in the Proceedings of the Japan Academy, Series