Cytosolic Ca2+ gradients in pancreatic islet-cells stimulated by glucose and carbachol.
Digital image analysis was employed to resolve the spatial differences in distribution of cytosolic free Ca2+ concentrations ([Ca2+]i) in mouse pancreatic islet-cells stimulated with glucose and carbachol. Using Indo-1 loaded mouse islet-cells, we have demonstrated that glucose induces steep spatial gradients of [Ca2+]i in isolated mouse islet-cells. Furthermore, the largest [Ca2+]i increase was always spatially restricted to a region just beneath the plasma membrane. Low concentrations of carbachol (0.6 microM) induced steep spatial gradients of [Ca2+]i which originated from the center of the cells. However, 10 microM carbachol increased [Ca2+]i to high levels collapsing the [Ca2+]i gradients in the center of the cells. Different patterns of [Ca2+]i oscillations were observed between dissociated pancreatic islet-cells and mouse pancreatic islets when challenged with 11 mM glucose. Under these conditions we could identify cells within the islet which oscillate with the same pattern as the whole islet. We postulate that "initiators" of insulin release, as glucose, induce greater [Ca2+]i increases at exocytotic sites than those induced by "potentiators", as carbachol.
Martín
F
F
Center for Bioengineering, Department of Physiology, School of Medicine, UHM, Alicante, Spain.
Ribas
J
J
Soria
B
B
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002217:Carbachol / Q000494:pharmacology*; D002478:Cells, Cultured; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D005456:Fluorescent Dyes; D005947:Glucose / Q000494:pharmacology*; D007211:Indoles; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods
D005456:Fluorescent Dyes; D007211:Indoles; D002217:Carbachol; D005947:Glucose; C048960:indo-1; D002118:Calcium