Measuring G Protein Activation with Spectrally Resolved Fluorescence Fluctuation Spectroscopy

G protein-coupled receptor signaling has been posited to occur through either collision coupling or pre-assembled complexes with G protein transducers. To investigate the dynamics of G protein signaling, we introduce fluorescence covariance matrix analysis (FCMA), a novel implementation of fluorescence cumulant analysis applied to spectrally resolved fluorescence images. We labeled the GPCR, Gα, and Gβγ units with distinct fluorescent protein labels and we applied FCMA to measure directly the complex formation during stimulation of dopamine and adrenergic receptors. To determine the prevalence of hetero-oligomers, we compared the GPCR data to those from control samples expressing three fluorescent protein labels with known stoichiometries. Interactions between Gα and Gβγ subunits determined by FCMA were sensitive to stimulation with GPCR ligands. However, GPCR/G protein interactions were too weak to be distinguished from background. These findings support a collision coupling mechanism rather than pre-assembled complexes for the two GPCRs studied.

The formation of multiple layers of ice particles in the polar summer mesopause region

This paper presents a two-dimensional theoretical model to study the formation process of multiple layers of small ice particles in the polar summer mesosphere as measured by rockets and associated with polar mesosphere summer echoes (PMSE). The proposed mechanism primarily takes into account the transport processes induced by gravity waves through collision coupling between the neutral atmosphere and the ice particles. Numerical solutions of the model indicate that the dynamic influence of wind variation induced by gravity waves can make a significant contribution to the vertical and horizontal transport of ice particles and ultimately transform them into thin multiple layers. Additionally, the pattern of the multiple layers at least partially depends on the vertical wavelength of the gravity wave, the ice particle size and the wind velocity. The results presented in this paper will be helpful to better understand the occurrence of multiple layers of PMSE as well as its variation process.