Intracellular amyloidogenesis by human islet amyloid polypeptide induces apoptosis in COS-1 cells

H. Jay Hiddinga, Norman L. Eberhardt

Research output: Contribution to journalArticle

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Abstract

Human islet amyloid polypeptide (hIAPP) is co-secreted with insulin from pancreatic islet β cells. This peptide spontaneously aggregates in the form of fibrils, and amyloid deposits are associated with dead or degenerating β cells, a hallmark of noninsulin-dependent diabetes mellitus. We demonstrated that COS-1 cells transfected with vectors expressing hIAPP exhibited intracellular amyloid deposits that were associated with cell death (O'Brien, Butler, Kreutter, Kane, Eberhardt, Am J Pathol 1995, 147:609-616). To establish the mechanism of cell death, we transfected COS-1 cells with vectors expressing amyloidogenic hIAPP or nonamyloidogenic rat IAPP and mutant hIAPP constructs and assayed them for markers characteristic of apoptosis and necrosis by fluorescence-activated cell sorting analysis. Amyloidogenic hIAPP-transfected COS cells contained up to threefold more apoptotic cells present at 96 hours after transfection compared with the nonamyloidogenic vector controls. The hIAPP-induced apoptosis was negligible at 24 and 48 hours after transfection and was maximal at 96 hours which parallels the time course of amyloidogenesis. Immunohistochemical staining and confocal microscopy showed that hIAPP is localized with distinct clustering in the endoplasmic reticulum and Golgi apparatus with no discernable extracellular staining. These experiments provide direct evidence that intracellular hIAPP amyloid causes cell death by triggering apoptotic pathways.

Original languageEnglish (US)
Pages (from-to)1077-1088
Number of pages12
JournalAmerican Journal of Pathology
Volume154
Issue number4
StatePublished - Apr 1999

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Islet Amyloid Polypeptide
COS Cells
Apoptosis
Cell Death
Amyloid Plaques
Islets of Langerhans
Transfection
Staining and Labeling
Golgi Apparatus
Amyloid
Confocal Microscopy
Endoplasmic Reticulum
Type 2 Diabetes Mellitus
Cluster Analysis
Cause of Death
Flow Cytometry
Necrosis
Insulin
Peptides

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Intracellular amyloidogenesis by human islet amyloid polypeptide induces apoptosis in COS-1 cells. / Hiddinga, H. Jay; Eberhardt, Norman L.

In: American Journal of Pathology, Vol. 154, No. 4, 04.1999, p. 1077-1088.

Research output: Contribution to journalArticle

Hiddinga, H. Jay ; Eberhardt, Norman L. / Intracellular amyloidogenesis by human islet amyloid polypeptide induces apoptosis in COS-1 cells. In: American Journal of Pathology. 1999 ; Vol. 154, No. 4. pp. 1077-1088.
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