Downregulation of myosin II-B by siRNA alters the subcellular localization of the amyloid precursor protein and increases amyloid-beta deposition in N2a cells

Albert Einstein College of Medicine, New York, New York, United States
Biochemical and Biophysical Research Communications (Impact Factor: 2.28). 11/2007; 362(3):633-8. DOI: 10.1016/j.bbrc.2007.08.061
Source: PubMed

ABSTRACT The Alzheimer's disease (AD) brain pathology is characterized by extracellular deposits of amyloid-beta (Abeta) peptides and intraneuronal fibrillar structures. These pathological features may be functionally linked, but the mechanism by which Abeta accumulation relates to neuronal degeneration is still poorly understood. Abeta peptides are fragments cleaved from the amyloid precursor protein (APP), a transmembrane protein ubiquitously expressed in the nervous system. Although the proteolytic processing of APP has been implicated in AD, the physiological function of APP and the subcellular site of APP cleavages remain unknown. The overall structure of the protein and its fast anterograde transport along the axon support the idea that APP functions as a vesicular receptor for cytoskeletal motor proteins. In the current study, we test the hypothesis that myosin II, important contributor to the cytoskeleton of neuronal cells, may influence the trafficking and/or the processing of APP. Our results demonstrate that downregulation of myosin II-B, the major myosin isoform in neurons, is able to increase Abeta deposition, concomitantly altering the subcellular localization of APP. These new insights might be important for the understanding of the function of APP and provide a novel conceptual framework in which to analyze its pathological role.

  • [Show abstract] [Hide abstract]
    ABSTRACT: OBJECTIVE: Human genetic mutations that affect the N-terminal head-domain of the nonmuscle myosin-II (MyoII) molecule can result in nonsyndromic sensorineural hearing loss but the underlying mechanism is unknown. Ultimately, MyoII must be appropriately localized in order to execute endogenous functions. The aim of the current study is to determine whether the head-domain of MyoII regulates in vivo localization of the molecule in living and fixed preparations of the auditory organ. METHODS: A genetic/transgenic GAL4-UAS approach was used to selectively drive the expression of zip/MyoII (Drosophila homologue of human nonmuscle MyoII) in Drosophila melanogaster auditory (Johnston's organ) sensory neurons. To follow the distribution of the full-length transgene encoded by MyoII, the N-terminus was fused to green fluorescent protein. Additionally, headless zip/MyoII molecules with and without isoleucine-glutamine or IQ motifs were also expressed in Johnston's organ neurons. RESULTS: Removing the entire head domain of MyoII induced localization in neuronal dendrites while removing only a portion of the head but keeping the IQ motif induced localization in the soma and axons of the neurons. CONCLUSIONS: The findings suggest that the head domain regulates in vivo localization of MyoII in auditory neurons.
    International journal of pediatric otorhinolaryngology 03/2013; DOI:10.1016/j.ijporl.2013.02.011 · 0.85 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: P2X7 receptors trigger Ca(2+) -dependent exocytotic glutamate release, but also function as a route for non-exocytotic glutamate release from neurons or astrocytes. To gain an insight into the mechanisms involving the P2X7 receptor as a direct pathway for glutamate release, we compared the behavior of a full-length rat P2X7 receptor, a truncated rat P2X7 receptor in which the carboxyl tail had been deleted, a rat P2X7 receptor with the 18-amino acid cysteine-rich motif of the carboxyl tail deleted, and a rat P2X2 receptor, all of which are expressed in HEK293 cells. We found that the P2X7 receptor function as a route for glutamate release was antagonized in a non-competitive way by extracellular Mg(2+) , did not require the recruitment of pore-forming molecules, and was dependent on the carboxyl tail. Indeed, the truncated P2X7 receptor and the P2X7 receptor with the deleted cysteine-rich motif both lost their function as a pathway for glutamate release, while still evoking intracellular Ca(2+) elevation. No glutamate efflux was observed through the P2X2 receptor. Notably, HEK293 cells (lacking the machinery for Ca(2+) -dependent exocytosis), when transfected with P2X7 receptors, appear to be a suitable model for investigating the P2X7 receptor as a route for non-exocytotic glutamate efflux. © 2013 International Society for Neurochemistry, J. Neurochem. (2013) 10.1111/jnc.12143.
    Journal of Neurochemistry 01/2013; DOI:10.1111/jnc.12143 · 4.24 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: To examine the role of protein L-isoaspartyl O-methyltransferase (PIMT; EC on the secretion of Aβ peptides. HEK293 APPsw cells were treated with PIMT siRNA or adenosine dialdehyde (AdOX), a broad-spectrum methyltransferase inhibitor. Under the conditions, the level of Aβ secretion and regulatory mechanism by PIMT were examined. Knock-down of PIMT and treatment with AdOX significantly increased Aβ(40) secretion. Reductions in levels of PIMT decreased the secretion of soluble amyloid precursor protein alpha (sAPPα) without altering the total expression of APP or its membrane-bound C83 fragment. However, the levels of the C99 fragment generated by β-secretase were enhanced. Moreover, the decreased secretion of sAPPα resulting from PIMT knock-down seemed to be linked with the suppression of the expression of α-secretase gene products, α-disintegrin and metalloprotease 10 (ADAM10) and ADAM17, as indicated by Western blot analysis. In contrast, ADAM10 was not down-regulated in response to treatment with the protein arginine methyltransferase (PRMT) inhibitor, AMI-1. This study demonstrates a novel role for PIMT, but not PRMT, as a negative regulator of Aβ peptide formation and a potential protective factor in the pathogenesis of AD.
    Acta Pharmacologica Sinica 03/2011; 32(3):288-94. DOI:10.1038/aps.2010.228 · 2.35 Impact Factor
    This article is viewable in ResearchGate's enriched format