ISSN:
1432-2013
Keywords:
Key words Ca2+ entry
;
Kinetics
;
Thapsigargin
;
Parotid cells
;
Fura-2
Source:
Springer Online Journal Archives 1860-2000
Topics:
Medicine
Notes:
Abstract The molecular mechanism(s) involved in mediating Ca2+ entry into rat parotid acinar and other non-excitable cells is not known. In this study we have examined the kinetics of Ca2+ entry in fura-2-loaded parotid acinar cells, which were treated with thapsigargin to deplete internal Ca2+ pools (Ca2+-pool-depleted cells). The rate of Ca2+ entry was determined by measuring the initial increase in free cytosolic [Ca2+] ([Ca2+]i) in Ca2+-pool-depleted, and control (untreated), cells upon addition of various [Ca2+] to the medium. In untreated cells, a low-affinity component was detected with K Ca = 3.4 ± 0.7 mM (where K Ca denotes affinity for Ca2+) and V max = 9.8 ± 0.4 nM [Ca2+]i /s. In thapsigargin-treated cells, two Ca2+ influx components were detected with K Ca values of 152 ± 79 μM (V max = 5.1 ± 1.9 nM [Ca2+]i/s) and 2.4 ± 0.9 mM (V max = 37.6 ± 13.6 nM [Ca2+]i/s), respectively. We have also examined the effect of Ca2+ and depolarization on these two putative Ca2+ influx components. When cells were treated with thapsigargin in a Ca2+-free medium, Ca2+ influx was higher than into cells treated in a Ca2+-containing medium and, while there was a 46% increase in the V max of the low-affinity component (no change in K Ca), the high-affinity component was not clearly detected. In depolarized Ca2+-pool-depleted cells (with 50 mM KCl in the medium) the high-affinity component was considerably decreased while there was an apparent increase in the K Ca of the low-affinity component, without any change in the V max. These results demonstrate that Ca2+ influx into parotid acinar cells (1) is increased (four- to five-fold) upon internal Ca2+ pool depletion, and (2) is mediated via at least two components, with low and high affinities for Ca2+.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/s004240050111
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