Expanded dynamic range of fluorescent indicators for Ca(2+) by circularly permuted yellow fluorescent proteins.
AuthorsNagai T, Yamada S, Tominaga T, Ichikawa M, Miyawaki A
JournalProc Natl Acad Sci U S A
PubMed ID15247428
'Fluorescence resonance energy transfer (FRET) technology has been used to develop genetically encoded fluorescent indicators for various cellular functions. Although most indicators have cyan- and yellow-emitting fluorescent proteins (CFP and YFP) as FRET donor and acceptor, their poor dynamic range often prevents detection of subtle but significant signals. Here, we ... More
New statistical methods enhance imaging of cameleon fluorescence resonance energy transfer in cultured zebrafish spinal neurons.
AuthorsFan X, Majumder A, Reagin SS, Porter EL, Sornborger AT, Keith CH, Lauderdale JD
JournalJ Biomed Opt
PubMed ID17614725
Cameleons are genetically encoded fluorescence resonance energy transfer (FRET)-based Ca(2+) indicators. Attempts to use cameleons to detect neural activity in vertebrate systems have been largely frustrated by the small FRET signal, in contradistinction to the higher signals seen in Drosophila and Caenorhabditis elegans. We have developed a statistical optimization method ... More
Involvement of transient receptor potential vanilloid subtype 1 in analgesic action of methylsalicylate.
AuthorsOhta T, Imagawa T, Ito S,
JournalMol Pharmacol
PubMed ID18987162
Methylsalicylate (MS) is a naturally occurring compound that is used as a major active ingredient of balms and liniments supplied as topical analgesics. Despite the common use of MS as a pain reliever, the underlying molecular mechanism is not fully understood. Here we characterize the action of MS on transient ... More
Novel Gating and Sensitizing Mechanism of Capsaicin Receptor (TRPV1): TONIC INHIBITORY REGULATION OF EXTRACELLULAR SODIUM THROUGH THE EXTERNAL PROTONATION SITES ON TRPV1.
AuthorsOhta T, Imagawa T, Ito S,
JournalJ Biol Chem
PubMed ID18230619
Transient receptor potential V1 (TRPV1) is a nonselective cation channel expressed in nociceptors and activated by capsaicin. TRPV1 detects diverse stimuli, including acid, heat, and endogenous vanilloids, and functions as a molecular integrator of pain perception. Herein we demonstrate a novel regulatory role of extracellular Na(+) ([Na(+)](o)) on TRPV1 function. ... More
An engineered GM-CSF-CCL2 fusokine is a potent inhibitor of CCR2-driven inflammation as demonstrated in a murine model of inflammatory arthritis.
AuthorsRafei M, Berchiche YA, Birman E, Boivin MN, Young YK, Wu JH, Heveker N, Galipeau J,
JournalJ Immunol
PubMed ID19592643
CCR2 is a chemokine receptor widely expressed by lymphomyeloid cells involved in maladaptive autoimmune ailments. Therefore CCR2 is of great interest as a biological target for immune suppression due to its direct implication in autoimmune diseases such as rheumatoid arthritis. We have generated a novel fusion protein using GM-CSF and ... More
Measuring calcium signaling using genetically targetable fluorescent indicators.
AuthorsPalmer AE, Tsien RY
JournalNat Protoc
PubMed ID17406387
Genetically encoded Ca2+ indicators allow researchers to quantitatively measure Ca2+ dynamics in a variety of experimental systems. This protocol summarizes the indicators that are available, and highlights those that are most appropriate for a number of experimental conditions, such as measuring Ca2+ in specific organelles and localizations in mammalian tissue-culture ... More