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Zheng, Lin
Publications (7 of 7) Show all publications
Zheng, L., Cedazo-Minguez, A., Hallbeck, M., Jerhammar, F., Marcusson, J. & Terman, A. (2012). Intracellular distribution of amyloid beta peptide and its relationship to the lysosomal system.. Translational Neurodegeneration, 1(1), 19
Open this publication in new window or tab >>Intracellular distribution of amyloid beta peptide and its relationship to the lysosomal system.
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2012 (English)In: Translational Neurodegeneration, ISSN 2047-9158, Vol. 1, no 1, p. 19-Article in journal (Refereed) Published
Abstract [en]

Background

Amyloid beta peptide (Aβ) is the main component of extraneuronal senile plaques typical of Alzheimer’s disease (AD) brains. Although Aβ is produced by normal neurons, it is shown to accumulate in large amounts within neuronal lysosomes in AD. We have recently shown that under normal conditions the majority of Aβ is localized extralysosomally, while oxidative stress significantly increases intralysosomal Aβ content through activation of macroautophagy. It is also suggested that impaired Aβ secretion and resulting intraneuronal increase of Aβ can contribute to AD pathology. However, it is not clear how Aβ is distributed inside normal neurons, and how this distribution is effected when Aβ secretion is inhibited.

Methods

Using retinoic acid differentiated neuroblastoma cells and neonatal rat cortical neurons, we studied intracellular distribution of Aβ by double immunofluorescence microscopy for Aβ40 or Aβ42 and different organelle markers. In addition, we analysed the effect of tetanus toxin-induced exocytosis inhibition on the intracellular distribution of Aβ.

Results

Under normal conditions, Aβ was found in the small cytoplasmic granules in both neurites and perikarya. Only minor portion of Aβ was colocalized with trans-Golgi network, Golgi-derived vesicles, early and late endosomes, lysosomes, and synaptic vesicles, while the majority of Aβ granules were not colocalized with any of these structures. Furthermore, treatment of cells with tetanus toxin significantly increased the amount of intracellular Aβ in both perikarya and neurites. Finally, we found that tetanus toxin increased the levels of intralysosomal Aβ although the majority of Aβ still remained extralysosomally.

Conclusion

Our results indicate that most Aβ is not localized to Golgi-related structures, endosomes, lysosomes secretory vesicles or other organelles, while the suppression of Aβ secretion increases intracellular intra- and extralysosomal Aβ.

Place, publisher, year, edition, pages
BioMed Central, 2012
Keywords
Alzheimer disease; Amyloid β-protein; Colocalization; Exocytosis; Immunocytochemistry; Lysosomes
National Category
Neurosciences
Identifiers
urn:nbn:se:liu:diva-99373 (URN)10.1186/2047-9158-1-19 (DOI)23210724 (PubMedID)2-s2.0-84875525734 (Scopus ID)
Available from: 2013-10-16 Created: 2013-10-16 Last updated: 2019-10-14Bibliographically approved
Zheng, L. (2012). Lysosomal Involvement in the Pathogenesis of Alzheimer's Disease. (Doctoral dissertation). Linköping: Linköping University Electronic Press
Open this publication in new window or tab >>Lysosomal Involvement in the Pathogenesis of Alzheimer's Disease
2012 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Alzheimer’s disease (AD), the major cause of senile dementia, is associated with progressive formation of neurofibrillary tangles and extraneuronal plaques composed of amyloid beta peptide (Aβ). Aβ has been also found within Alzheimer neurons in association with the lysosomal system, an acidic vacuolar compartment possessing numerous hydrolytic enzymes. Lysosomes have been shown to be involved in both the formation of Aβ and its toxicity to neurons. Another line of evidence implicates oxidative stress as an important factor in the development of AD. It is reported that oxidative damage is one of the earliest changes in AD and plays an important role in the development of the disease. Although both the lysosomal system and reactive oxygen species are involved in AD, the mechanisms of this involvement are not well understood.

To gain insight into the relationship between oxidative stress and the lysosomal system in AD pathogenesis, we focused our study on: 1) The effect of oxidative stress on intracellular distribution of Aβ; 2) the role of endogenous Aβ in oxidant-induced apoptosis; 3) the role of autophagy and APP processing in oxidant induced damage; and, 4) the intraneuronal localization of Aβ and its relationship to the lysosomal system.

In our study, hyperoxia (40% versus 8% ambient oxygen) was used as a model of mild oxidative stress in vitro, while transfected cells producing different amounts of Aβ were used to assess toxicity due to endogenous Aβ. It was found that: 1) oxidative stress induces autophagic uptake of Aβ, resulting in its partial accumulation within lysosomes; 2) oxidative stress can induce neuronal death through macroautophagy of Aβ and consequent lysosomal membrane permeabilization; 3) increased cellular Aβ production is associated with enhanced oxidative stress and enhanced macroautophagy, resulting in increased intralysosomal Aβ accumulation and consequent apoptosis; and, 4) in normal conditions, intracellular Aβ shows primarily cytosolic distribution, not related to lysosomes and other acidic vacuoles, endoplasmic reticulum, Golgi complexes, synaptic vesicles or mitochondria. Only a minor portion of Aβ shows partial colocalization with cellular organelles. Inhibition of secretion significantly increased Aβ colocalization with endoplasmic reticulum, Golgi complexes, synaptic vesicles and lysosomes, as well as the amount of mitochondrial and cytosolic Aβ.

Oxidative stress induces intralysosomal autophagy-generated Aβ accumulation, consequently causing lysosomal membrane permeabilization and apoptosis. Our findings provide a possible explanation of the interactive role of oxidative stress and lysosomal system in AD pathogenesis, and may be helpful for a future therapeutic strategy against AD.

Place, publisher, year, edition, pages
Linköping: Linköping University Electronic Press, 2012. p. 55
Series
Linköping University Medical Dissertations, ISSN 0345-0082 ; 1282
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-73412 (URN)978-91-7393-005-5 (ISBN)
Public defence
2012-02-03, Berzeliussalen, ingång 65, Campus US, Linköpings universitet, Linköping, 13:00 (English)
Opponent
Supervisors
Available from: 2012-01-03 Created: 2012-01-03 Last updated: 2019-12-10Bibliographically approved
Zheng, L., Terman, A., Hallbeck, M., Dehvari, N., Cowburn, R. F., Benedikz, E., . . . Marcusson, J. (2011). Macroautophagy-generated increase of lysosomal amyloid β-protein mediates oxidant-induced apoptosis of cultured neuroblastoma cells. Autophagy, 7(12), 1528-1545
Open this publication in new window or tab >>Macroautophagy-generated increase of lysosomal amyloid β-protein mediates oxidant-induced apoptosis of cultured neuroblastoma cells
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2011 (English)In: Autophagy, ISSN 1554-8627, E-ISSN 1554-8635, Vol. 7, no 12, p. 1528-1545Article in journal (Refereed) Published
Abstract [en]

Increasing evidence suggests the toxicity of intracellular amyloid β-protein (Aβ) to neurons, as well as the involvement of oxidative stress in Alzheimer disease (AD). Here we show that normobaric hyperoxia (exposure of cells to 40% oxygen for five days, and consequent activation of macroautophagy and accumulation of Aβ within lysosomes, induced apoptosis in differentiated SH-SY5Y neuroblastoma cells. Cells under hyperoxia showed: (1) increased numbers of autophagic vacuoles that contained amyloid precursor protein (APP) as well as Aβ monomers and oligomers, (2) increased reactive oxygen species production, and (3) enhanced apoptosis. Oxidant-induced apoptosis positively correlated with cellular Aβ production, being the highest in cells that were stably transfected with APP Swedish KM670/671NL double mutation. Inhibition of γ-secretase, prior and/or in parallel to hyperoxia, suggested that the increase of lysosomal Aβ resulted mainly from its autophagic uptake, but also from APP processing within autophagic vacuoles. The oxidative stress-mediated effects were prevented by macroautophagy inhibition using 3-methyladenine or ATG5 downregulation. Our results suggest that upregulation of macroautophagy and resulting lysosomal Aβ accumulation are essential for oxidant-induced apoptosis in cultured neuroblastoma cells and provide aditional support for the interactive role of oxidative stress and the lysosomal system in AD-related neurodegeneration.

Place, publisher, year, edition, pages
Landes Bioscience, 2011
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-72778 (URN)10.4161/auto.7.12.18051 (DOI)000298182600012 ()
Note
Funding agencies|Gustav V and Queen Victoria Foundation||County Council of Ostergotland||Stiftelsen Olle Engkvist Byggmastare||Stifielsen for Gamla Tjanarinnor||Gunoch Bertil Stohnes Stiftelse||Lions forskningsfond||Svenska Lundbeckstiftelsen||Karolinska Institute Fund for Geriatric Research||Alice och Knut Wallenberg Stiftelse||Swedish Alzheimer Foundation||Swedish Brain Power||Available from: 2011-12-07 Created: 2011-12-07 Last updated: 2019-10-14Bibliographically approved
Zheng, L., Kågedal, K., Dehvari, N., Benedikz, E., Cowburn, R., Marcusson, J. & Terman, A. (2009). Oxidative stress induces macroautophagy of amyloid beta-protein and ensuing apoptosis. Free Radical Biology & Medicine, 46(3), 422-429
Open this publication in new window or tab >>Oxidative stress induces macroautophagy of amyloid beta-protein and ensuing apoptosis
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2009 (English)In: Free Radical Biology & Medicine, ISSN 0891-5849, E-ISSN 1873-4596, Vol. 46, no 3, p. 422-429Article in journal (Refereed) Published
Abstract [en]

There is increasing evidence for the toxicity of intracellular amyloid beta-protein (A beta) to neurons and the involvement of lysosomes in this process in Alzheimer disease (AD). We have recently shown that oxidative stress, a recognized determinant of AD. enhances macroautophagy and leads to intralysosomal accumulation of A beta in Cultured neuroblastoma cells. We hypothesized that oxidative stress promotes AD by stimulating macroautophagy of A that further may induce cell death by destabilizing lysosomal membranes. To investigate such possibility, we compared the effects of hyperoxia (40% ambient oxygen) in cultured HEK293 cells that were transfected with an empty vector (Vector), wild-type APP (APPwt), or Swedish mutant APP (APPswe). Exposure to hyperoxia for 5 days increased the number of cells with A beta-containing lysosomes, as well as the number of apoptotic cells, compared to normoxic conditions. The rate of apoptosis in all three cell lines demonstrated dependence on intralysosomal A beta content (Vector<APPwt<APPswe). Furthermore, the degree of apoptosis was positively correlated with lysosomal membrane permeabilization, whereas inhibitors Of macroautophagy and lysosomal function decreased oxidant-induced apoptosis and diminished the differences in apoptotic response between different cell lines. These results suggest that oxidative stress can induce neuronal death through macroautophagy of A beta and consequent lysosomal membrane permeabilization, which may help explain the mechanisms behind neuronal loss in AD.

Keywords
Alzheimer disease, Amyloid beta-protein, Amyloid precursor protein, Apoptosis, Autophagy, Lysosomes, Oxidative stress, Free radicals
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-16625 (URN)10.1016/j.freeradbiomed.2008.10.043 (DOI)
Available from: 2009-02-08 Created: 2009-02-06 Last updated: 2017-12-14Bibliographically approved
Zheng, L., Roberg, K., Jerhammar, F., Marcusson, J. & Terman, A. (2006). Autophagy of amyloid beta-protein in differentiated neuroblastoma cells exposed to oxidative stress. Neuroscience Letters, 394(3), 184-189
Open this publication in new window or tab >>Autophagy of amyloid beta-protein in differentiated neuroblastoma cells exposed to oxidative stress
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2006 (English)In: Neuroscience Letters, ISSN 0304-3940, E-ISSN 1872-7972, Vol. 394, no 3, p. 184-189Article in journal (Refereed) Published
Abstract [en]

Oxidative stress is considered important for the pathogenesis of Alzheimer disease (AD), which is characterized by the formation of senile plaques rich in amyloid beta-protein (Aβ). Aβ cytotoxicity has been found dependent on lysosomes, which are abundant in AD neurons and are shown to partially co-localize with Aβ. To determine whether oxidative stress has any influence on the relationship between lysosomes and Aβ1-42 (the most toxic form of Aβ), we studied the effect of hyperoxia (40% versus 8% ambient oxygen) on the intracellular localization of Aβ1-42 (assessed by immunocytochemistry) in retinoic acid differentiated SH-SY5Y neuroblastoma cells maintained in serum-free OptiMEM medium. In control cells, Aβ1-42 was mainly localized to small non-lysosomal cytoplasmic granules. Only occasionally Aβ1-42 was found in large (over 1 μm) lysosomal-associated membrane protein 2 positive vacuoles, devoid of the early endosomal marker rab5. These large Aβ1-42-containing lysosomes were not detectable in the presence of serum (known to suppress autophagy), while their number increased dramatically (up to 24-fold) after exposure of cells to hyperoxia during 5 days. Activation of autophagy by hyperoxia was confirmed by transmission electron microscopy. Furthermore, an inhibitor of autophagic sequestration 3-methyladenine prevented the accumulation of Aβ1-42-positive lysosomes due to hyperoxia. In parallel experiments, intralysosomal accumulation of Aβ1-40 following oxidative stress has been found as well. The results suggest that Aβ can be autophagocytosed and its accumulation within neuronal lysosomes is enhanced by oxidative stress. © 2005 Elsevier Ireland Ltd. All rights reserved.

Keywords
Alzheimer disease; Amyloid beta-protein; Autophagy; Lysosomes; Reactive oxygen species
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-35894 (URN)10.1016/j.neulet.2005.10.035 (DOI)28972 (Local ID)28972 (Archive number)28972 (OAI)
Available from: 2009-10-10 Created: 2009-10-10 Last updated: 2017-12-13Bibliographically approved
Zheng, L., Marcusson, J. & Terman, A. (2006). Oxidative stress and Alzheimer disease - The autophagy connection?. AUTOPHAGY, 2(2), 143-145
Open this publication in new window or tab >>Oxidative stress and Alzheimer disease - The autophagy connection?
2006 (English)In: AUTOPHAGY, ISSN 1554-8627, Vol. 2, no 2, p. 143-145Article in journal (Refereed) Published
Abstract [en]

Intraneuronal accumulation of amyloid beta-protein (A beta) is believed to be responsible for degeneration and apoptosis of neurons and consequent senile plaque formation in Alzheimer disease (AD), the main cause of senile dementia. Oxidative stress, an early determinant of AD, has been recently found to induce intralysosomal A beta accumulation in cultured differentiated neuroblastoma cells through activation of macroautophogy. Because A beta is known to destabilize lysosomal membranes, potentially resulting in apoptotic cell death, this finding suggests the involvement of oxidative stress-induced macroautophagy in the pathogenesis of AD.

Keywords
Alzheimer disease, amyloid beta-protein, autophagy, lysosomes, oxidative stress
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-48087 (URN)
Available from: 2009-10-11 Created: 2009-10-11 Last updated: 2011-01-11
Zheng, L., Roberg, K., Jerhammar, F., Marcusson, J. & Terman, A. (2006). Oxidative Stress Induces Intralysosomal Accumulation of Alzheimer Amyloid β-Protein in Cultured Neuroblastoma Cells. Annals of the New York Academy of Sciences, 1067, 248-251
Open this publication in new window or tab >>Oxidative Stress Induces Intralysosomal Accumulation of Alzheimer Amyloid β-Protein in Cultured Neuroblastoma Cells
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2006 (English)In: Annals of the New York Academy of Sciences, ISSN 0077-8923, E-ISSN 1749-6632, Vol. 1067, p. 248-251Article in journal (Refereed) Published
Abstract [en]

Oxidative stress is considered important for the pathogenesis of Alzheimer's disease (AD), which is characterized by the formation of extracellular senile plaques, mainly composed of amyloid β-protein (Aβ). Aβ also accumulates within AD neurons and is believed to exert cellular toxicity through lysosomal labilization. We report that the exposure of human neuroblastoma cells to hyperoxia (40% vs. 8% ambient oxygen) induced the accumulation of large (over 1 μM) Aβ-containing lysosomes, which were not typical of control cells, showing a distinct localization of Aβ and lysosomal markers. An inhibitor of autophagy, 3-methyladenine, suppressed the effect of hyperoxia. The results suggest a link between the involvement of oxidative stress and lysosomes in AD.

Keywords
Alzheimer disease; amyloid beta-protein; autophagy; lysosomes; oxidative stress
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-35998 (URN)10.1196/annals.1354.032 (DOI)29306 (Local ID)29306 (Archive number)29306 (OAI)
Available from: 2009-10-10 Created: 2009-10-10 Last updated: 2017-12-13Bibliographically approved
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