Screening for Arabidopsis mutants with altered Ca2+ signal response using aequorin-based Ca2+ reporter system
نویسندگان
چکیده
منابع مشابه
Ca2+ Measurements in Mammalian Cells with Aequorin-based Probes.
Aequorin is a Ca2+ sensitive photoprotein suitable to measure intracellular Ca2+ transients in mammalian cells. Thanks to recombinant cDNAs expression, aequorin can be specifically targeted to various subcellular compartments, thus allowing an accurate measurement of Ca2+ uptake and release of different intracellular organelles. Here, we describe how to use this probe to measure cytosolic Ca2+ ...
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Calcium ion is a versatile second messenger for diverse cell signaling in response to developmental and environmental cues. The specificity of Ca(2+)-mediated signaling is defined by stimulus-elicited Ca(2+) signature and downstream decoding processes. Here, an Aequorin-based luminescence recording system was developed for monitoring Ca(2+) in response to various stimuli in Arabidopsis. With th...
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The presence and physiological role of Ca2+-induced Ca2+ release (CICR) in nonmuscle excitable cells has been investigated only indirectly through measurements of cytosolic [Ca2+] ([Ca2+]c). Using targeted aequorin, we have directly monitored [Ca2+] changes inside the ER ([Ca2+]ER) in bovine adrenal chromaffin cells. Ca2+ entry induced by cell depolarization triggered a transient Ca2+ release f...
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photoproteins are excellent reporter systems because they don’t have virtually background signal. aequorin is the most well-known photoprotein. three improved engineered photoproteins photina, i-photina and c-photina, were also recently developed and optimized for generation of ca2+ mobilization assays precisely. the total light emission is greater than aequorin and their reaction kinetics is a...
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The photoprotein aequorin was the first probe used to measure specifically the [Ca(2+)] inside the lumen of the endoplasmic reticulum ([Ca(2+)](ER)) of intact cells and it provides values for the steady-state [Ca(2+)](ER), around 500 microM, that closely match those obtained now by other procedures. Aequorin-based methods to measure [Ca(2+)](ER) offer several advantages: (i) targeting of the pr...
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ژورنال
عنوان ژورنال: STAR Protocols
سال: 2021
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2021.100558