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Effects of K + ‐induced depolarization and purinergic receptor activation on elemental content in insulin‐producing RINm5F‐cells.
Author(s) -
Pålsgård Eva,
Lindh Ulf,
JunttiBerggren Lisa,
Berggren PerOlof,
Roomans Godfried M.
Publication year - 1995
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1006/cbir.1995.1004
Subject(s) - stimulation , depolarization , chemistry , insulin , incubation , microanalysis , biophysics , calcium , analytical chemistry (journal) , biochemistry , chromatography , biology , endocrinology , organic chemistry
X‐ray microanalysis was used to detect elemental changes in the insulin‐producing tumor cell‐line RINm5F. To improve discrimination between mobile ions and ions bound to macromolecules a new approach was employed, consisting of multivariate statistical analysis of correlations between the concentrations of Na, Mg, P, S, Cl, K, and Ca. RINm5F cells, cultured on Formvar‐coated titanium grids, were stimulated with high K + or ATP, that are both known to stimulate insulin release. The buffers used contained Ca 2+ or one of the Ca 2+ ‐analogues Sr 2+ and Ba 2+ , to represent Ca 2+ uptake in response to stimulation. After stimulation the cells were shock‐frozen and freeze‐dried overnight. Incubation for 10‐20 seconds in a Ca 2+ ‐containing buffer did not significantly affect elemental composition, whereas cellular Mg, P and K decreased in a Sr 2+ ‐containing buffer. Depolarization with high K + concentration caused an increase in the cellular Na content, both in Ca 2+ ‐ and Sr 2+ ‐containing buffers, but not in the buffer where Ca 2+ had been replaced by Ba 2+ . X‐ray microanalysis is useful for detection of elemental changes subsequent to stimulation of cultured cells. Moreover, multivariate statistical analysis strengthens the idea that stimulation of RINm5F cells causes redistribution of ions possibly due to changes in the state of binding of some elements to cellular proteins.