Proceedings of the National Academy of Sciences of the United States of America
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Proc. Natl. Acad. Sci. U.S.A. · Apr 2015
Expanded palette of Nano-lanterns for real-time multicolor luminescence imaging.
Fluorescence live imaging has become an essential methodology in modern cell biology. However, fluorescence requires excitation light, which can sometimes cause potential problems, such as autofluorescence, phototoxicity, and photobleaching. Furthermore, combined with recent optogenetic tools, the light illumination can trigger their unintended activation. ⋯ The brightness of these cyan and orange Nano-lanterns was ∼20 times brighter than wild-type Renilla luciferase, which allowed us to perform multicolor live imaging of intracellular submicron structures. The rapid dynamics of endosomes and peroxisomes were visualized at around 1-s temporal resolution, and the slow dynamics of focal adhesions were continuously imaged for longer than a few hours without photobleaching or photodamage. In addition, we extended the application of these multicolor Nano-lanterns to simultaneous monitoring of multiple gene expression or Ca(2+) dynamics in different cellular compartments in a single cell.
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Proc. Natl. Acad. Sci. U.S.A. · Mar 2015
CFTR and sphingolipids mediate hypoxic pulmonary vasoconstriction.
Hypoxic pulmonary vasoconstriction (HPV) optimizes pulmonary ventilation-perfusion matching in regional hypoxia, but promotes pulmonary hypertension in global hypoxia. Ventilation-perfusion mismatch is a major cause of hypoxemia in cystic fibrosis. We hypothesized that cystic fibrosis transmembrane conductance regulator (CFTR) may be critical in HPV, potentially by modulating the response to sphingolipids as mediators of HPV. ⋯ Our findings demonstrate a central role of CFTR and sphingolipids in HPV. Upon hypoxia, nSMase triggers TRPC6 translocation, which requires its interaction with CFTR. Concomitant SphK1-dependent formation of S1P and activation of S1P2/4 result in phospholipase C-mediated TRPC6 and rho kinase activation, which conjointly trigger vasoconstriction.
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Proc. Natl. Acad. Sci. U.S.A. · Mar 2015
Novel Epac fluorescent ligand reveals distinct Epac1 vs. Epac2 distribution and function in cardiomyocytes.
Exchange proteins directly activated by cAMP (Epac1 and Epac2) have been recently recognized as key players in β-adrenergic-dependent cardiac arrhythmias. Whereas Epac1 overexpression can lead to cardiac hypertrophy and Epac2 activation can be arrhythmogenic, it is unknown whether distinct subcellular distribution of Epac1 vs. Epac2 contributes to differential functional effects. ⋯ Epac2 and Epac1 were differentially concentrated along T tubules and around the nucleus, respectively. Epac1-KO abolished OMe-CPT-induced nuclear CaMKII activation and export of transcriptional regulator histone deacetylase 5. In conclusion, Epac1 is localized and functionally involved in nuclear signaling, whereas Epac2 is located at the T tubules and regulates arrhythmogenic sarcoplasmic reticulum Ca leak.
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Proc. Natl. Acad. Sci. U.S.A. · Mar 2015
Ocean-driven thinning enhances iceberg calving and retreat of Antarctic ice shelves.
Iceberg calving from all Antarctic ice shelves has never been directly measured, despite playing a crucial role in ice sheet mass balance. Rapid changes to iceberg calving naturally arise from the sporadic detachment of large tabular bergs but can also be triggered by climate forcing. Here we provide a direct empirical estimate of mass loss due to iceberg calving and melting from Antarctic ice shelves. ⋯ Net mass loss due to iceberg calving for these ice shelves (302 ± 27 Gt/y) is comparable in magnitude to net mass loss due to basal melt (312 ± 14 Gt/y). Moreover, we find that iceberg calving from these decaying ice shelves is dominated by frequent calving events, which are distinct from the less frequent detachment of isolated tabular icebergs associated with ice shelves in neutral or positive mass balance regimes. Our results suggest that thinning associated with ocean-driven increased basal melt can trigger increased iceberg calving, implying that iceberg calving may play an overlooked role in the demise of shrinking ice shelves, and is more sensitive to ocean forcing than expected from steady state calving estimates.
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Proc. Natl. Acad. Sci. U.S.A. · Mar 2015
Epigenetic modification of the oxytocin receptor gene influences the perception of anger and fear in the human brain.
In humans, the neuropeptide oxytocin plays a critical role in social and emotional behavior. The actions of this molecule are dependent on a protein that acts as its receptor, which is encoded by the oxytocin receptor gene (OXTR). DNA methylation of OXTR, an epigenetic modification, directly influences gene transcription and is variable in humans. ⋯ These data indicate that the human endogenous oxytocin system is involved in attenuation of the fear response, corroborating research implicating intranasal oxytocin in the same processes. Our findings highlight the importance of including epigenetic mechanisms in the description of the endogenous oxytocin system and further support a central role for oxytocin in social cognition. This approach linking epigenetic variability with neural endophenotypes may broadly explain individual differences in phenotype including susceptibility or resilience to disease.