TY - JOUR
T1 - Amyloid-β induced membrane damage instigates tunneling nanotube-like conduits by p21-activated kinase dependent actin remodulation
AU - Dilna, Aysha
AU - Deepak, K. V.
AU - Damodaran, Nandini
AU - Kielkopf, Claudia S.
AU - Kagedal, Katarina
AU - Ollinger, Karin
AU - Nath, Sangeeta
N1 - Funding Information:
SN thanks to Manipal Academy of Higher Education intramural grants (India; 2019-2022, #MAHE/CDS/PHD/MIFR/2019), Magnus Bergvalls (Sweden, 2015-2016, #2014-00192), Gun och Bertil Stohnes research grants (Sweden, 2014-2015) and Alzheimer fonden (Sweden; 2012-2013). We acknowledge Dr. TMA Pai Scholarship of Manipal Academy of Higher Education (for DKV). We thank our long-term collaborator Prof. Martin Hallbeck of Linkoping University, Sweden, for the valuable suggestions on designing of cell-to-cell transfer experiments. We thank Dr. Ravi Manjithaya of JNCASR, Bangalore, India, for giving us SH-SY5Y cell lines and giving us access to the SIM-superresolution microscope. Thanks a lot to Likhesh Sharma, PhD, Product Manager GE Healthcare for assisting with imaging by SIM-superresolution microscope. We thank Prof. Satyajit Mayor of the National center for biological sciences, India, for his kind gift of GFP-GPI construct, CAAX-mCherry plasmid, sharing of some reagents and for his valuable inputs. The CAAX-mCherry plasmid was a kind gift to Prof. Satyajit Mayor by Prof. Jacco van Rheenen of Hubrecht Institute for Developmental Biology and Stem Cell Research. Thanks to Prof. Subba Rao Gangi Setty of the Indian Institute of Science and Prof. Gopal Pande of Manipal institute of higher education, India, for their valuable inputs.
Funding Information:
SN thanks to Manipal Academy of Higher Education intramural grants (India; 2019-2022 , # MAHE/CDS/PHD/MIFR/2019 ), Magnus Bergvalls (Sweden, 2015-2016 , # 2014-00192 ), Gun och Bertil Stohnes research grants (Sweden, 2014-2015 ) and Alzheimer fonden (Sweden; 2012-2013 ). We acknowledge Dr. TMA Pai Scholarship of Manipal Academy of Higher Education (for DKV). We thank our long-term collaborator Prof. Martin Hallbeck of Linkoping University, Sweden, for the valuable suggestions on designing of cell-to-cell transfer experiments. We thank Dr. Ravi Manjithaya of JNCASR, Bangalore, India, for giving us SH-SY5Y cell lines and giving us access to the SIM-superresolution microscope. Thanks a lot to Likhesh Sharma, PhD, Product Manager GE Healthcare for assisting with imaging by SIM-superresolution microscope. We thank Prof. Satyajit Mayor of the National center for biological sciences, India, for his kind gift of GFP-GPI construct, CAAX-mCherry plasmid, sharing of some reagents and for his valuable inputs. The CAAX-mCherry plasmid was a kind gift to Prof. Satyajit Mayor by Prof. Jacco van Rheenen of Hubrecht Institute for Developmental Biology and Stem Cell Research. Thanks to Prof. Subba Rao Gangi Setty of the Indian Institute of Science and Prof. Gopal Pande of Manipal institute of higher education, India, for their valuable inputs.
Publisher Copyright:
© 2021 Elsevier B.V.
PY - 2021/12/1
Y1 - 2021/12/1
N2 - Alzheimer's disease (AD) pathology progresses gradually via anatomically connected brain regions. Direct transfer of amyloid-β1–42 oligomers (oAβ) between connected neurons has been shown, however, the mechanism is not fully revealed. We observed formation of oAβ induced tunneling nanotubes (TNTs)-like nanoscaled f-actin containing membrane conduits, in differentially differentiated SH-SY5Y neuronal models. Time-lapse images showed that oAβ propagate from one cell to another via TNT-like structures. Preceding the formation of TNT-like conduits, we detected oAβ−induced plasma membrane (PM) damage and calcium-dependent repair through lysosomal-exocytosis, followed by massive endocytosis to re-establish the PM. Massive endocytosis was monitored by an influx of the membrane-staining dye TMA-DPH and PM damage was quantified by propidium iodide influx in the absence of Ca2+. The massive endocytosis eventually caused accumulation of internalized oAβ in Lamp1 positive multivesicular bodies/lysosomes via the actin cytoskeleton remodulating p21-activated kinase1 (PAK1) dependent endocytic pathway. Three-dimensional quantitative confocal imaging, structured illumination superresolution microscopy, and flowcytometry quantifications revealed that oAβ induces activation of phospho-PAK1, which modulates the formation of long stretched f-actin extensions between cells. Moreover, the formation of TNT-like conduits was inhibited by preventing PAK1-dependent internalization of oAβ using the small-molecule inhibitor IPA-3, a highly selective cell-permeable auto-regulatory inhibitor of PAK1. The present study reveals that the TNT-like conduits are probably instigated as a consequence of oAβ induced PM damage and repair process, followed by PAK1 dependent endocytosis and actin remodeling, probably to maintain cell surface expansion and/or membrane tension in equilibrium.
AB - Alzheimer's disease (AD) pathology progresses gradually via anatomically connected brain regions. Direct transfer of amyloid-β1–42 oligomers (oAβ) between connected neurons has been shown, however, the mechanism is not fully revealed. We observed formation of oAβ induced tunneling nanotubes (TNTs)-like nanoscaled f-actin containing membrane conduits, in differentially differentiated SH-SY5Y neuronal models. Time-lapse images showed that oAβ propagate from one cell to another via TNT-like structures. Preceding the formation of TNT-like conduits, we detected oAβ−induced plasma membrane (PM) damage and calcium-dependent repair through lysosomal-exocytosis, followed by massive endocytosis to re-establish the PM. Massive endocytosis was monitored by an influx of the membrane-staining dye TMA-DPH and PM damage was quantified by propidium iodide influx in the absence of Ca2+. The massive endocytosis eventually caused accumulation of internalized oAβ in Lamp1 positive multivesicular bodies/lysosomes via the actin cytoskeleton remodulating p21-activated kinase1 (PAK1) dependent endocytic pathway. Three-dimensional quantitative confocal imaging, structured illumination superresolution microscopy, and flowcytometry quantifications revealed that oAβ induces activation of phospho-PAK1, which modulates the formation of long stretched f-actin extensions between cells. Moreover, the formation of TNT-like conduits was inhibited by preventing PAK1-dependent internalization of oAβ using the small-molecule inhibitor IPA-3, a highly selective cell-permeable auto-regulatory inhibitor of PAK1. The present study reveals that the TNT-like conduits are probably instigated as a consequence of oAβ induced PM damage and repair process, followed by PAK1 dependent endocytosis and actin remodeling, probably to maintain cell surface expansion and/or membrane tension in equilibrium.
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U2 - 10.1016/j.bbadis.2021.166246
DO - 10.1016/j.bbadis.2021.166246
M3 - Article
C2 - 34403739
AN - SCOPUS:85113167406
SN - 0925-4439
VL - 1867
JO - Biochimica et Biophysica Acta - Molecular Basis of Disease
JF - Biochimica et Biophysica Acta - Molecular Basis of Disease
IS - 12
M1 - 166246
ER -