RESUMEN
We revise the calibration of scintillating screens commonly used to detect relativistic electron beams with low average current, e.g., from laser-plasma accelerators, based on new and expanded measurements that include higher charge density and different types of screens than previous work [Buck et al., Rev. Sci. Instrum. 81, 033301 (2010)]. Electron peak charge densities up to 10 nC/mm2 were provided by focused picosecond-long electron beams delivered by the Electron Linac for beams with high Brilliance and low Emittance (ELBE) at the Helmholtz-Zentrum Dresden-Rossendorf. At low charge densities, a linear scintillation response was found, followed by the onset of saturation in the range of nC/mm2. The absolute calibration factor (photons/sr/pC) in this linear regime was measured to be almost a factor of 2 lower than that reported by Buck et al. retrospectively implying a higher charge in the charge measurements performed with the former calibration. A good agreement was found with the results provided by Glinec et al. [Rev. Sci. Instrum. 77, 103301 (2006)]. Furthermore long-term irradiation tests with an integrated dose of approximately 50 nC/mm2 indicate a significant decrease of the scintillation efficiency over time. Finally, in order to enable the transfer of the absolute calibration between laboratories, a new constant reference light source has been developed.
RESUMEN
The transient receptor potential (TRP) family of ion channels comprises receptors that are activated by a vast variety of physical as well as chemical stimuli. TRP channels interact in a complex manner with several intracellular signaling cascades, both up- and downstream of receptor activation. Investigating cascades stimulated downstream of the cold and menthol receptor TRPM8, we found evidence for both, functional and structural interaction of TRPM8 with Gαq. We demonstrated menthol-evoked increase in intracellular Ca(2+) under extracellular Ca(2+)-free conditions, which was blocked by the PLC inhibitors U73122 or edelfosine. This metabotropic Ca(2+) signal could be observed also in cells expressing a channel-dead (i.e. non-conducting) or a chloride-conducting TRPM8 pore mutant. However, this intracellular metabotropic Ca(2+) signal could not be detected in Gαq deficient cells or in the presence of dominant-negative GαqX. Evidence for a close spatial proximity necessary for physical interaction of TRPM8 and Gαq was provided by acceptor bleaching experiments demonstrating FRET between TRPM8-CFP and Gαq-YFP. A Gαq-YFP mobility assay (FRAP) revealed a restricted diffusion of Gαq-YFP under conditions when TRPM8 is immobilized in the plasma membrane. Moreover, a menthol-induced and TRPM8-mediated G protein activation could be demonstrated by FRET experiments monitoring the dissociation of Gαq-YFP from a Gß/Gγ-CFP complex, and by the exchange of radioactive [(35)S]GTPγS for GDP. Our observations lead to a view that extends the operational range of the TRPM8 receptor from its function as a pure ion channel to a molecular switch with additional metabotropic capacity.