The development of a new family of intracellular calcium probes

Iatridou, Helene; Foukaraki, Evangelia; Kuhn, Michael; Marcus, Elizabeth M.; Haugland, Richard P.; Katerinopoulos, Haralambos E.

doi:10.1016/0143-4160(94)90058-2

Cited by 55 publications

(54 citation statements)

References 13 publications

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“…Intracellular ionized magnesium contaminates the mag-fura-2 signal, and calibration of single excitation indicators like Calcium Green-5N is difficult (Brocard et al, 1993;Rajdev and Reynolds, 1993;Stout et al, 1996). BTC ratio values are not affected significantly by alterations in [Mg 2ϩ ] in the 0 -10 mM range (Iatridou et al, 1994;Hyrc et al, 1997).…”

Section: Resultsmentioning

confidence: 99%

“…C ytosolic calcium determination was performed with fluorescent calcium indicators: f ura-2/AM ester, f ura-2/ K ϩ salt, f ura-2 coupled to dextran [f ura-2/dextran; molecular weight (MW) 3000], and benzothiazole coumarin (BTC) (Iatridou et al, 1994;Konishi and Watanabe, 1995). All indicators were purchased from Molecular Probes (Eugene, OR).…”

Section: Methodsmentioning

confidence: 99%

“…We used f ura-2/AM ester, f ura-2/ K ϩ salt, f ura-2/dextran, or a benzothiazole coumarin-based indicator, BTC, to determine [Ca 2ϩ ] i in neurons subjected to lethal and nonlethal challenges. The low affinity of BTC for calcium (K D ϳ7-26 M) allows detection of [Ca 2ϩ ] i in the micromolar range (Iatridou et al, 1994;Zhao et al, 1996).…”

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confidence: 99%

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Ionized Intracellular Calcium Concentration Predicts Excitotoxic Neuronal Death: Observations with Low-Affinity Fluorescent Calcium Indicators

et al. 1997

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Cytosolic calcium ([Ca 2ϩ ] i ) is an important mediator of neuronal signal transduction, participating in diverse biochemical reactions that elicit changes in synaptic efficacy, metabolic rate, and gene transcription. Excessive [Ca 2ϩ ] i also has been implicated as a cause of acute neuronal injury, although measurement of [Ca 2ϩ (Rothman and Olney, 1986;Choi, 1992). Studies using primary neuronal culture systems have shown that cultured neurons are selectively vulnerable to brief applications of glutamate (Rothman, 1985; or to selective agonists of the NMDA class of glutamate receptors (Rothman, 1985;Choi et al., 1988;Hartley and Choi, 1989). In contrast, prolonged activation of the AM PA or kainate receptors is required to produce neurotoxicity (Koh et al., 1990).Schanne and colleagues (1979) originally described the dependence of extracellular calcium in hepatocyte toxicity, which provided a potential mechanism for neuronal toxicity with the subsequent discovery that glutamate receptor activation produced neuronal calcium influx (Connor et al., 1987(Connor et al., , 1988Murphy et al., 1987). Additional studies by other investigators led to the hypothesis that calcium entry was responsible for excitotoxic neuronal injury (Choi, 1987) (for review, see Dubinsky, 1993a). A correlation between intracellular calcium levels and glutamate toxicity was suggested by the dependence of the latter on extracellular calcium Rothman et al., 1987 influx during nonlethal AMPA treatment (Marcoux et al., 1988;Eimerl and Schramm, 1994; Lu et al., 1996). The study by Hartley and coworkers (1993) suggested that a direct relationship existed between calcium accumulation and subsequent death in neurons exposed to NMDA. Although previous studies using fluorescent intracellular calcium indicators have noted that lethal excitotoxic insults may be followed by prolonged or delayed [Ca 2ϩ ] i elevation (Ogura et al., 1988;de Erausquin et al., 1990;Randall and Thayer, 1992;Dubinsky, 1993b; Tymianski et al., 1993a;Limbrick et al., 1995), such indicators have failed to demonstrate a consistent difference between intracellular free calcium concentration ([Ca 2ϩ ] i ) in cells during lethal and nonlethal challenges (de Erausquin et al., 1990;Michaels and Rothman, 1990; Tymianski et al., 1993a).One potential interpretation of these results is that there is not a direct relationship between the magnitude of acute [Ca 2ϩ ] i elevation and the extent of subsequent neuronal death. Another possibility is that current methods of [Ca 2ϩ ] i measurement fail to distinguish toxic levels of [Ca 2ϩ ] i . In the present study we examine the hypothesis that conventional, high-affinity calcium indicators (e.g., fura-2) are unable to measure accurately the [Ca 2ϩ ] i in the micromolar range and therefore fail to detect differences in [Ca 2ϩ ] i between lethal and nonlethal excitotoxic exposures. We Received April 18, 1997; revised June 2, 1997; accepted June 12, 1997. This work was supported by National Institutes o...

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Ionized Intracellular Calcium Concentration Predicts Excitotoxic Neuronal Death: Observations with Low-Affinity Fluorescent Calcium Indicators

et al. 1997

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“…is also a dualexcitation ratiometric Ca 2ϩ indicator (165). The excitation maximum shifts from 464 to 400 nm upon binding Ca 2ϩ .…”

Section: Btcmentioning

confidence: 99%

Measurement of Intracellular Calcium

Takahashi

Camacho

Lechleiter

et al. 1999

Physiological Reviews

674

511

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To a certain extent, all cellular, physiological, and pathological phenomena that occur in cells are accompanied by ionic changes. The development of techniques allowing the measurement of such ion activities has contributed substantially to our understanding of normal and abnormal cellular function. Digital video microscopy, confocal laser scanning microscopy, and more recently multiphoton microscopy have allowed the precise spatial analysis of intracellular ion activity at the subcellular level in addition to measurement of its concentration. It is well known that Ca2+ regulates numerous physiological cellular phenomena as a second messenger as well as triggering pathological events such as cell injury and death. A number of methods have been developed to measure intracellular Ca2+. In this review, we summarize the advantages and pitfalls of a variety of Ca2+ indicators used in both optical and nonoptical techniques employed for measuring intracellular Ca2+ concentration.

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“…The BTC signal was calibrated by obtaining the minimum (before the first flash) and maximum (after complete photolysis of all caged Ca2+) fluorescence ratios. The BTC fluorescence signal was then converted to [Ca2+]i using the equation of Grynkiewicz et al (1985) and a Kd of 7 jM (Iatridou et al, 1994).…”

Section: Solutionsmentioning

confidence: 99%

Co-localization of L-type Ca2+ channels and insulin-containing secretory granules and its significance for the initiation of exocytosis in mouse pancreatic B-cells.

et al. 1995

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We have monitored L-type Ca2+ channel activity, local cytoplasmic Ca2+ transients, the distribution of insulincontaining secretory granules and exocytosis in individual mouse pancreatic B-cells. Subsequent to the opening of the Ca2+ channels, exocytosis is initiated with a latency <100 ms. The entry of Ca2+ that precedes exocytosis is unevenly distributed over the cell and is concentrated to the region with the highest density of secretory granules. In this region, the cytoplasmic Ca2+ concentration is 5-to 10-fold higher than in the remainder of the cell reaching concentrations of several micromolar. Single-channel recordings confirm that the L-type Ca2+ channels are clustered in the part of the cell containing the secretory granules. This arrangement, which is obviously reminiscent of the 'active zones' in nerve terminals, can be envisaged as being favourable to the B-cell as it ensures that the Ca2+ transient is maximal and restricted to the part of the cell where it is required to rapidly initiate exocytosis whilst at the same time minimizing the expenditure of metabolic energy to subsequently restore the resting Ca2+ concentration.

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The development of a new family of intracellular calcium probes

Cited by 55 publications

References 13 publications

Ionized Intracellular Calcium Concentration Predicts Excitotoxic Neuronal Death: Observations with Low-Affinity Fluorescent Calcium Indicators

Ionized Intracellular Calcium Concentration Predicts Excitotoxic Neuronal Death: Observations with Low-Affinity Fluorescent Calcium Indicators

Measurement of Intracellular Calcium

Co-localization of L-type Ca2+ channels and insulin-containing secretory granules and its significance for the initiation of exocytosis in mouse pancreatic B-cells.

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