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001-es BibID:BIBFORM031849
035-os BibID:PMID:19193035 WOS:000264535100005
Első szerző:Bannwarth, Michael
Cím:Indo-1 derivatives for local calcium sensing / Michael Bannwarth, Ivan R. Corrêa, Monika Sztretye, Sandrine Pouvreau, Cindy Fellay, Annina Aebischer, Leandro Royer, Eduardo Ríos, Kai Johnsson
Dátum:2009
ISSN:1554-8929
Megjegyzések:The role of calcium in signal transduction relies on the precise spatial and temporal control of its concentration. The existing means to detect fluctuations in Ca2+ concentrations with adequate temporal and spatial resolution are limited. We introduce here a method to measure Ca2+ concentrations in defined locations in living cells that is based on linking the Ca2+-sensitive dye Indo-1 to SNAP-tag fusion proteins. Fluorescence spectroscopy of SNAP-Indo-1 conjugates in vitro showed that the conjugates retained the Ca2+-sensing ability of Indo-1. In a proof-of-principle experiment, local Ca2+ sensing was demonstrated in single cells dissociated from muscle of adult mice expressing a nucleus-localized SNAP-tag fusion. Ca2+ concentrations inside nuclei of resting cells were measured by shifted excitation and emission ratioing of confocal microscopic images of fluorescence. After permeabilizing the plasma membrane, changes in the bathing solution induced corresponding changes in nuclear [Ca2+] that were readily detected and used for a preliminary calibration of the technique. This work thus demonstrates the synthesis and application of SNAP-tag-based Ca2+ indicators that combine the spatial specificity of genetically encoded calcium indicators with the advantageous spectroscopic properties of synthetic indicators.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
külföldön készült közlemény
Megjelenés:Acs Chemical Biology 4 : 3 (2009), p. 179-190. -
További szerzők:Corrêa, Ivan R. Sztretye Mónika (1981-) (élettanász, elektrofiziológus) Pouvreau, Sandrine Fellay, Cindy Aebischer, Annina Royer, Leandro (1975-) (kutató) Ríos, Eduardo (1945-) (kutató) Johnsson, Kai
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001-es BibID:BIBFORM031848
035-os BibID:PMID:20713548 WOS:000284828600009
Első szerző:Royer, Leandro (kutató)
Cím:Paradoxical buffering of calcium by calsequestrin demonstrated for the calcium store of skeletal muscle / Leandro Royer, Monika Sztretye, Carlo Manno, Sandrine Pouvreau, Jingsong Zhou, Bjorn C. Knollmann, Feliciano Protasi, Paul D. Allen, Eduardo Ríos
Dátum:2010
ISSN:0022-1295
Megjegyzések:Contractile activation in striated muscles requires a Ca(2+) reservoir of large capacity inside the sarcoplasmic reticulum (SR), presumably the protein calsequestrin. The buffering power of calsequestrin in vitro has a paradoxical dependence on [Ca(2+)] that should be valuable for function. Here, we demonstrate that this dependence is present in living cells. Ca(2+) signals elicited by membrane depolarization under voltage clamp were compared in single skeletal fibers of wild-type (WT) and double (d) Casq-null mice, which lack both calsequestrin isoforms. In nulls, Ca(2+) release started normally, but the store depleted much more rapidly than in the WT. This deficit was reflected in the evolution of SR evacuability, E, which is directly proportional to SR Ca(2+) permeability and inversely to its Ca(2+) buffering power, B. In WT mice E starts low and increases progressively as the SR is depleted. In dCasq-nulls, E started high and decreased upon Ca(2+) depletion. An elevated E in nulls is consistent with the decrease in B expected upon deletion of calsequestrin. The different value and time course of E in cells without calsequestrin indicate that the normal evolution of E reflects loss of B upon SR Ca(2+) depletion. Decrement of B upon SR depletion was supported further. When SR calcium was reduced by exposure to low extracellular [Ca(2+)], release kinetics in the WT became similar to that in the dCasq-null. E became much higher, similar to that of null cells. These results indicate that calsequestrin not only stores Ca(2+), but also varies its affinity in ways that progressively increase the ability of the store to deliver Ca(2+) as it becomes depleted, a novel feedback mechanism of potentially valuable functional implications. The study revealed a surprisingly modest loss of Ca(2+) storage capacity in null cells, which may reflect concurrent changes, rather than detract from the physiological importance of calsequestrin.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
külföldön készült közlemény
Megjelenés:Journal of General Physiology 136 : 3 (2010), p. 325-338. -
További szerzők:Sztretye Mónika (1981-) (élettanász, elektrofiziológus) Manno, Carlo (1978-) (kutató) Pouvreau, Sandrine Zhou, Jingsong (1968-) (kutató) Knollmann, Bjorn C. Protasi, Feliciano Allen, Paul D. (1960-) (kuató) Ríos, Eduardo (1945-) (kutató)
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3.

001-es BibID:BIBFORM072527
Első szerző:Sztretye Mónika (élettanász, elektrofiziológus)
Cím:Indo-1 Hybrid Biosensors For Calcium Monitoring In Cellular Organelles / Monika Sztretye, Sandrine Pouvreau, Michael Bannwarth, Ivan R. Corrêa, Cindy Fellay, Annina Aebischer, Leandro Royer, Jianxun Yi, Jingsong Zhou, Kai Johnsson, Eduardo Ríos
Dátum:2009
ISSN:0006-3495 1542-0086
Tárgyszavak:Orvostudományok Elméleti orvostudományok idézhető absztrakt
folyóiratcikk
skeletal muscle
Megjelenés:Biopysical Journal. - 96 : 3 Suppl. 1. (2009), p. 541a. -
További szerzők:Pouvreau, Sandrine Bannwarth, Michael Corrêa, Ivan R. Fellay, Cindy Aebischer, Annina Royer, Leandro (1975-) (kutató) Yi, Jianxun (1970-) (kutató) Zhou, Jingsong (1968-) (kutató) Johnsson, Kai Ríos, Eduardo (1945-) (kutató)
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