Changes in mitochondrial dynamics during amyloid β-induced PC12 cell apoptosis
- 246 Downloads
- 7 Citations
Abstract
Changes in mitochondrial morphology and dynamics influence mitochondrial function and ultimately damage neurons in Alzheimer’s disease (AD). Amyloid β (Aβ) is a major factor in the pathogenesis of AD. Although it has been proved that Aβ can affect the dynamics of mitochondria, there is little known on the precise dynamic process. Thus, MTT, Hoechst 33342, and Annexin V/PI analysis were used to study Aβ25–35 neurotoxity on PC12 cells, live cell station and image processing were applied to study the moving parameters and characters of mitochondria. We also studied changes of mitochondrial membrane potential and reactive oxygen species production. The results showed that long-term exposure of PC12 cells to Aβ25–35 resulted in increase of mitochondrial number and decrease of mitochondrial length and size, which presented fluctuated during early time and dramatic changes occurred after 6 h. Low concentration exposure caused little mitochondrial changes before 24 h while short time exposure induced mitochondrial fragmentation that could be recovered to normal. Mitochondrial membrane potential dissipation and reactive oxygen species production were observed, as well as apparent cell apoptosis with significant morphological changes. These data suggest that mitochondrial fission can be reversed during Aβ25–35-induced PC12 cell apoptosis, depending on the concentration and exposure time of Aβ25–35, which may be helpful in AD prevention and therapy.
Keywords
Amyloid β Mitochondrial dynamics Live cell station Apoptosis PC12 cellsNotes
Acknowledgments
This study was supported by the Key Laboratory for Biomedical Engineering of the Ministry of China; the Economic and Trade Commission of Zhejiang Province; and the Key Laboratory of Chinese Medicine Screening, Exploitation and Medicinal Effectiveness Appraise for Cardio-cerebral Vascular and Nervous System of Zhejiang Province. We thank Kurtis Feng for critical reading of the manuscript.
Supplementary material
Supplementary material 1 (MPG 11162 kb)
Supplementary material 2 (MPG 15078 kb)
Supplementary material 3 (MPG 10376 kb)
Supplementary material 4 (MPG 8978 kb)
References
- 1.Samanta MK, Wilson B, Santhi K et al (2006) Alzheimer disease and its management: a review. Am J Ther 13:516–526CrossRefPubMedGoogle Scholar
- 2.Su B, Wang X, Zheng L et al (2010) Abnormal mitochondrial dynamics and neurodegenerative diseases. Biochim Biophys Acta 1802:135–142PubMedGoogle Scholar
- 3.Small DH, Mok SS, Bornstein JC (2001) Alzheimer’s disease and Abeta toxicity: from top to bottom. Nat Rev Neurosci 2:595–598CrossRefPubMedGoogle Scholar
- 4.Yankner BA, Lu T (2009) Amyloid beta-protein toxicity and the pathogenesis of Alzheimer disease. J Biol Chem 284:4755–4759CrossRefPubMedGoogle Scholar
- 5.Chan DC (2006) Mitochondria: dynamic organelles in disease, aging, and development. Cell 125:1241–1252CrossRefPubMedGoogle Scholar
- 6.Suen DF, Norris KL, Youle RJ (2008) Mitochondrial dynamics and apoptosis. Genes Dev 22:1577–1590CrossRefPubMedGoogle Scholar
- 7.Lu B (2009) Mitochondrial dynamics and neurodegeneration. Curr Neurol Neurosci Rep 9:212–219CrossRefPubMedGoogle Scholar
- 8.Loo DT, Copani A, Pike CJ et al (1993) Apoptosis is induced by beta-amyloid in cultured central nervous system neurons. Proc Natl Acad Sci USA 90:7951–7955CrossRefPubMedGoogle Scholar
- 9.Barsoum MJ, Yuan H, Gerencser AA et al (2006) Nitric oxide-induced mitochondrial fission is regulated by dynamin-related GTPases in neurons. EMBO J 25:3900–3911CrossRefPubMedGoogle Scholar
- 10.Rui Y, Tiwari P, Xie Z et al (2006) Acute impairment of mitochondrial trafficking by beta-amyloid peptides in hippocampal neurons. J Neurosci 26:10480–10487CrossRefPubMedGoogle Scholar
- 11.Pike CJ, Walencewicz-Wasserman AJ, Kosmoski J et al (1995) Structure-activity analyses of beta-amyloid peptides: contributions of the beta 25–35 region to aggregation and neurotoxicity. J Neurochem 64:253–265CrossRefPubMedGoogle Scholar
- 12.Xu YK, Xu KD, Li JY et al (2007) Bi-directional transport of GLUT4 vesicles near the plasma membrane of primary rat adipocytes. Biochem Biophys Res Commun 359:121–128CrossRefPubMedGoogle Scholar
- 13.Xu KD, Yan M, Wang YP et al (2009) Free cholesterol overloading induced smooth muscle cells death and activated both ER- and mitochondrial-dependent death pathway. Atherosclerosis 207:123–130CrossRefGoogle Scholar
- 14.Gogvadze V, Orrenius S, Zhivotovsky B (2006) Multiple pathways of cytochrome c release from mitochondria in apoptosis. Biochim Biophys Acta 1757:639–647CrossRefPubMedGoogle Scholar
- 15.Lin YH, Liu AH, Wu HL et al (2006) Salvianolic acid B, an antioxidant from Salvia miltiorrhiza, prevents Abeta(25–35)-induced reduction in BPRP in PC12 cells. Biochem Biophys Res Commun 348:593–599CrossRefPubMedGoogle Scholar
- 16.Tang XQ, Yang CT, Chen J et al (2008) Effect of hydrogen sulphide on beta-amyloid-induced damage in PC12 cells. Clin Exp Pharmacol Physiol 35:180–186PubMedGoogle Scholar
- 17.Vaisid T, Kosower NS, Elkind E et al (2008) Amyloid beta peptide toxicity in differentiated PC12 cells: calpain–calpastatin, caspase, and membrane damage. J Neurosci Res 86:2314–2325CrossRefPubMedGoogle Scholar
- 18.Esposito G, De Filippis D, Carnuccio R et al (2006) The marijuana component cannabidiol inhibits beta-amyloid-induced tau protein hyperphosphorylation through Wnt/beta-catenin pathway rescue in PC12 cells. J Mol Med 84:253–258CrossRefPubMedGoogle Scholar
- 19.Iuvone T, De Filippis D, Esposito G et al (2006) The spice sage and its active ingredient rosmarinic acid protect PC12 cells from amyloid-beta peptide-induced neurotoxicity. J Pharmacol Exp Ther 317:1143–1149CrossRefPubMedGoogle Scholar
- 20.Knott AB, Perkins G, Schwarzenbacher R et al (2008) Mitochondrial fragmentation in neurodegeneration. Nat Rev Neurosci 9:505–518CrossRefPubMedGoogle Scholar
- 21.Wang X, Su B, Siedlak SL et al (2008) Amyloid-beta overproduction causes abnormal mitochondrial dynamics via differential modulation of mitochondrial fission/fusion proteins. Proc Natl Acad Sci USA 105:19318–19323CrossRefPubMedGoogle Scholar
- 22.Pletjushkina OY, Lyamzaev KG, Popova EN et al (2006) Effect of oxidative stress on dynamics of mitochondrial reticulum. Biochim Biophys Acta 1757:518–524CrossRefPubMedGoogle Scholar
- 23.Szabadkai G, Simoni AM, Bianchi K et al (2006) Mitochondrial dynamics and Ca2+ signaling. Biochim Biophys Acta 1763:442–449CrossRefPubMedGoogle Scholar
- 24.Irrcher I, Adhihetty PJ, Joseph AM et al (2003) Regulation of mitochondrial biogenesis in muscle by endurance exercise. Sports Med 33:783–793CrossRefPubMedGoogle Scholar
- 25.Kann O, Kovacs R (2007) Mitochondria and neuronal activity. Am J Physiol Cell Physiol 292:C641–C657CrossRefPubMedGoogle Scholar
- 26.Han XJ, Lu YF, Li SA et al (2007) Involvement of calcineurin in glutamate-induced mitochondrial dynamics in neurons. Neurosci Res 60:114–119CrossRefPubMedGoogle Scholar
- 27.Kim BH, Lee HG, Choi JK et al (2004) The cellular prion protein (PrPC) prevents apoptotic neuronal cell death and mitochondrial dysfunction induced by serum deprivation. Brain Res Mol Brain Res 124:40–50CrossRefPubMedGoogle Scholar
- 28.Chen H, McCaffery JM, Chan DC (2007) Mitochondrial fusion protects against neurodegeneration in the cerebellum. Cell 130:548–562CrossRefPubMedGoogle Scholar
- 29.Baloh RH, Schmidt RE, Pestronk A et al (2007) Altered axonal mitochondrial transport in the pathogenesis of Charcot–Marie–Tooth disease from mitofusin 2 mutations. J Neurosci 27:422–430CrossRefPubMedGoogle Scholar