...Francisco J. Aulestia; María Teresa Alonso; Javier García-Sancho High Ca 2+ content in the Golgi apparatus (Go) is essential for protein processing and sorting. In addition, the Go can shape the cytosolic Ca 2+ signals by releasing or sequestering Ca 2+ . We generated two new aequorin-based Ca 2...

... aequorin-based probes Nat. Protoc. 2013 8 2105 2118 10.1038/nprot.2013.127 24113784 3 Demaurex N. Frieden M. Measurements of the free luminal ER Ca2+ concentration with targeted “cameleon” fluorescent proteins Cell Calcium 2003 34 109 119 10.1016/S0143-4160(03)00081-2...

... compared with the scRNA cells for [Ca 2+ ] values below 2.5 μM. 1 To whom correspondence should be addressed (email [email protected] ). aequorin Ca 2+ uniporter Ca 2+ uniporter protein mitochondrial (MCU) Ca 2+ uptake protein 1 mitochondrial (MICU1) knockdown mitochondrion...

... millimolar levels in a few seconds. Measuring the dynamics of [Ca 2+ ] M during prolonged stimulation has been previously precluded by the high Ca 2+ affinity of the probes available. We have now developed a mitochondrially targeted double-mutated form of the photoprotein aequorin which is able to measure...

... 40)] cells by using targeted aequorins for selective monitorization of Ca 2+ transport by organelles. We find that CALHM1 increases Ca 2+ leak from the ER and, more importantly, reduces ER Ca 2+ uptake by decreasing both the transport capacity and the Ca 2+ affinity of SERCA (sarcoplasmic/endoplasmic...

... Ca 2+ stores using targeted aequorins for selective measurements in different subcellular compartments. Both, HEK-293T [HEK (human embryonic kidney)-293 cells expressing the large T-antigen of SV40 (simian virus 40)] and HeLa cells accumulated Ca 2+ into the ER (endoplasmic reticulum) to near...

... be responsible for the degeneration of muscle occurring in DMD. In the present study, we used subsarcolemmal- and mitochondrial-targeted aequorin to study the effect of the antiapoptotic Bcl-2 protein overexpression on carbachol-induced near-plasma membrane and mitochondrial calcium responses in myotubes derived...

..., cytosolic-luciferase-expressing adenovirus ; DMEM, Dulbecco s modified Eagle s medium; GLP-1, glucagon-like peptide-1 ; hGH, human growth hormone; IP3, Ins(1,4,5)P3 ; KATP, ATP-sensitive K+ channel ; mitAeq, mitochondrial-aequorin-expressing adenovirus ; mitLuc, mitochondrial-luciferase-expressing...

...% of the mitochondrial space to several hundred micromolar in 2–3% of mitochondria. Intermediate levels were found in the rest of the mitochondrial space. Single-cell imaging experiments with aequorin showed that the heterogeneity had both an intercellular and a subcellular origin. Those mitochondria responding...

... the importance of this phenomenon for nutrient sensing in pancreatic islets and β-cells by imaging adenovirally expressed Ca 2+ and ATP sensors (aequorin and firefly luciferase). [Ca 2+ ] m increases provoked by KCl or tolbutamide evoked an immediate increase in cytosolic and mitochondrial free ATP concentration...

...Evelyn L. RIDGLEY; Zhao-hui XIONG; Larry RUBEN Here we examine a cell death process induced by reactive oxygen species (ROS) in the haemoflagellate Trypanosoma brucei brucei . Ca 2+ distribution in cellular compartments was measured with stable transformants expressing aequorin targeted...