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  • 1
    ISSN: 1432-2013
    Keywords: pH ; Fluorescent indicator ; Intracellular pH ; Calibration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In this paper we present methods to measure intracellular pH (pHi) with fluorescent indicators. These methods are based on the change in intracellular pH following the addition of weak acids and weak bases to the extracellular medium. The first method requires that the fluorescence of the indicator is proportional to the change in pHi that follows the addition of a weak acid or weak base to the extracellular medium. The second is a null method which uses a mixture of weak acid and weak base that does not change the fluorescent signal. This null method can be used in situations in which the fluorescent signal is a monotonic but non-linear function of pH. The first method depends upon four assumptions. (i) That only the uncharged forms of the weak acids and bases cross the surface membrane. (ii) That the pKa is the same inside and outside the cell. (iii) That the buffering power is constant. (iv) That there is no significant pH regulation on the time scale of the change in pHi. The null method only requires the first two assumptions. We have made estimates of pHi in four different cell types and compared the results obtained with these methods with those obtained from other methods of pHi calibration.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2013
    Keywords: Key words Sodium ; Action potential ; Endocardial ; Epicardial
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  In this study we report measurements of the intracellular sodium concentration ([Na+]i) in cardiac myocytes isolated from sub-endocardial and sub-epicardial regions of the rabbit left ventricle. These measurements show that [Na+]i is significantly higher in sub-epicardial than in sub-endocardial myocytes both at rest and during steady-state stimulation at 0.5 Hz. During a 10-min rest the rate of fall of [Na+]i was identical in cells from the two regions but during post-rest recovery [Na+]i rose significantly faster in sub-endocardial cells. The effect that this difference in sodium regulation may have on the rate of recovery of the calcium transient post-rest is discussed. The steady-state differences in [Na+]i do not appear to have the effect on contraction we would expect i.e. steady-state contraction is larger in sub-endocardial cells where [Na+]i is lower. Changes in [Na+]i brought about by altering the frequency of stimulation are associated with the expected changes of contraction. Action potentials were found to be significantly longer in sub-endocardial cells and following rest action potential duration was shortened although regional differences were maintained. No regional differences in this effect of rest or in recovery of the action potential post-rest were found.
    Type of Medium: Electronic Resource
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