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  • 1990-1994  (2)
  • 1880-1889
  • 293 cells  (1)
  • Adenosine triphosphate  (1)
  • 1
    ISSN: 1432-2013
    Keywords: Adenosine triphosphate ; Caged-adenosine triphosphate ; Potassium channel ; Metabolism ; Heart ; Cardiac ventricle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We have used ‘caged-ATP’ to investigate the kinetic behavior of KATP channels in ventricular cells from rat heart. In whole cells, loaded with ‘caged-ATP’, an increase of intracellular [ATP] following a UV light flash produced a decrease of KATP channel current that was too slow (τ ≈ 300 ms) to be explained by the expected timecourse of ATP release (τ ≈ 3 ms) and the time-course of channel blockade by ATP (τ ≈ 20 ms). In isolated membrane patches, caged-ATP itself caused partial blockade of KATP channels. Under these conditions, photorelease of ATP caused channel activity to decline further. The results suggest that caged-ATP can bind to the KATP channel but, on binding, decreases the open probability to a lesser extent than does ATP. Additionally, the observations indicate that for photolytically-generated ATP to bind to the channel, caged-ATP must first unbind (slowly) from the channel. We conclude that ‘caged-ATP’ is not fully caged with respect to its allosteric action on the KATP channel.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2013
    Keywords: Indo-1 ; Flash photolysis ; DM-nitrophen ; Nd: YAG laser ; Cardiac cells ; 293 cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We have constructed a modular instrument to measure intracellular [Ca2+] ([Ca2+]i) in single isolated cells while simultaneously imposing step changes in [Ca2+]i using “caged Ca2+”. By combining the outputs of a xenon arc lamp with a frequency-tripled (Nd: YAG) laser, the instrument can operate with low maintained illumination to measure [Ca2+]i using a ratiometric Ca2+-sensitive fluorophore and also activate the release of Ca2+ from a caged-Ca2+ compound with a high energy pulse of ultraviolet light. This instrument is simple to assemble, introduces little electrical noise, provides a wide range of illumination power, produces only moderate photobleaching of the Ca2+ indicator and can be readily adapted to diverse cellular preparations. We demonstrate the use of this system to measure step changes in [Ca2+]i in adult rat ventricular myocytes and a human embryonic kidney cell line (293 cells) in culture.
    Type of Medium: Electronic Resource
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