oligomeric forms of insulin amyloid aggregation disrupt outgrowth and complexity of neuron-like pc12 cells低聚物的形式的淀粉样蛋白聚合干扰胰岛素的产物和复杂性neuron-like pc12细胞.pdfVIP

Oligomeric Forms of Insulin Amyloid Aggregation Disrupt Outgrowth and Complexity of Neuron-Like PC12 Cells 1 1 2 1 Ehsan Kachooei , Ali Akbar Moosavi-Movahedi *, Fariba Khodagholi , Hassan Ramshini , Fatemeh Shaerzadeh2, Nader Sheibani3 1 Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran, 2 Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran, 3 Department of Ophthalmology and Visual Sciences, and Pharmacology, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, United States of America Abstract Formation of protein amyloid fibrils consists of a series of intermediates including oligomeric aggregates, proto-fibrillar structures, and finally mature fibrils. Recent studies show higher toxicity for oligomeric and proto-fibrillar intermediates of protein relative to their mature fibrils. Here the kinetic of the insulin amyloid fibrillation was evaluated using a variety of techniques including ThT fluorescence, Congo red absorbance, circular dichroism, and atomic force microscopy (AFM). The solution surface tension changes were attributed to hydrophobic changes in insulin structure and were detected by Du ¨ Nouy Ring method. Determination of the surface tension of insulin oligomeric, proto-fibrillar and fibrillar forms indicated that the hydrophobicity of solution is enhanced by the formation of the oligomeric forms of insulin compared to other forms. In order to investigate the toxicity of the different forms of insulin we monitored morphological alterations of the differentiated neuron-like PC12 cells following incubation with native, oligomeric aggregates, proto-fibrillar, and fibrillar forms of insulin. The cell body area,