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Ultrastructural evidence that apoptosis is the mechanism by which human amylin evokes death in RINm5F pancreatic islet beta-cells
Authors:Saafi E L  Konarkowska B  Zhang S  Kistler J  Cooper G J
Institution:The Biochemistry and Molecular Biology Group, University of Auckland, Auckland, New Zealand.
Abstract:A view is emerging that human amylin (HA) kills pancreatic islet beta-cells by apoptosis. This study strengthens this view by documenting time-dependent morphological and ultrastructural changes in 10 microm HA-treated cultured RINm5F islet beta-cells. Membrane blebbing and microvilli loss were the earliest detectable apoptosis-related phenomena, already evident 1 h after HA exposure. Following 6-12 h of HA-treatment, chromatin margination became evident, consistent with detecting DNA laddering about the same time. Nuclear shrinkage, nuclear membrane convolution and prominent cytoplasmic vacuolization were clearly recognized at 22 h post-treatment. Together, these cellular changes constitute a strong case for HA-induced apoptosis, and further demonstrates that electron microscopy is a more sensitive tool for early apoptosis detection in cultured cells than classical biochemical assays like visualizing DNA laddering. The ultrastructural changes reported here contribute further evidence to be included in the ongoing dissection of molecular mechanisms underlying HA-induced apoptosis, as may occur in type-2 diabetes mellitus.
Keywords:amylin  islet amyloid  apoptosis  RINm5F islet β‐cells  type‐2 diabetes  cellular morphology  ultrastructure  scanning electron microscopy  transmission electron microscopy
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