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Article: Glycogen synthase kinase 3β (GSK3β) and presenilin (PS) are key regulators of kinesin-1-mediated cargo motility within axons.

Banerjee, Rupkatha / Gunawardena, Shermali

Frontiers in cell and developmental biology

2023 Volume 11, Page(s) 1202307

Abstract: It has been a quarter century since the discovery that molecular motors are phosphorylated, but fundamental questions still remain as to how specific kinases contribute to particular motor functions, ... ...

Abstract	It has been a quarter century since the discovery that molecular motors are phosphorylated, but fundamental questions still remain as to how specific kinases contribute to particular motor functions, particularly
Language	English
Publishing date	2023-06-09
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2737824-X
ISSN	2296-634X
ISSN	2296-634X
DOI	10.3389/fcell.2023.1202307
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Article ; Online: Glycogen synthase kinase 3β (GSK3β) and presenilin (PS) are key regulators of kinesin-1-mediated cargo motility within axons

Rupkatha Banerjee / Shermali Gunawardena

Frontiers in Cell and Developmental Biology, Vol

2023 Volume 11

Abstract	It has been a quarter century since the discovery that molecular motors are phosphorylated, but fundamental questions still remain as to how specific kinases contribute to particular motor functions, particularly in vivo, and to what extent these processes have been evolutionarily conserved. Such questions remain largely unanswered because there is no cohesive strategy to unravel the likely complex spatial and temporal mechanisms that control motility in vivo. Since diverse cargoes are transported simultaneously within cells and along narrow long neurons to maintain intracellular processes and cell viability, and disruptions in these processes can lead to cancer and neurodegeneration, there is a critical need to better understand how kinases regulate molecular motors. Here, we review our current understanding of how phosphorylation can control kinesin-1 motility and provide evidence for a novel regulatory mechanism that is governed by a specific kinase, glycogen synthase kinase 3β (GSK3β), and a scaffolding protein presenilin (PS).
Keywords	kinesin-1 ; GSK3β ; presenilin ; axonal transport ; phosphorylation ; Biology (General) ; QH301-705.5
Subject code	572
Language	English
Publishing date	2023-06-01T00:00:00Z
Publisher	Frontiers Media S.A.
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: PolyQ-Expansion Causes Mitochondria Fragmentation Independent of Huntingtin and Is Distinct from Traumatic Brain Injury (TBI)/Mechanical Stress-Mediated Fragmentation Which Results from Cell Death.

Swinter, Kelsey / Salah, Dania / Rathnayake, Rasika / Gunawardena, Shermali

Cells

2023 Volume 12, Issue 19

Abstract: Mitochondrial dysfunction has been reported in many Huntington's disease (HD) models; however, it is unclear how these defects occur. Here, we test the hypothesis that excess pathogenic huntingtin (HTT) impairs mitochondrial homeostasis, ... ...

Abstract	Mitochondrial dysfunction has been reported in many Huntington's disease (HD) models; however, it is unclear how these defects occur. Here, we test the hypothesis that excess pathogenic huntingtin (HTT) impairs mitochondrial homeostasis, using
MeSH term(s)	Animals ; Stress, Mechanical ; Cell Death ; Drosophila ; Huntington Disease/metabolism ; Mitochondria/metabolism ; Brain Injuries, Traumatic/pathology
Chemical Substances	polyglutamine (26700-71-0)
Language	English
Publishing date	2023-10-05
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells12192406
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Article ; Online: Nanoparticles in the brain: a potential therapeutic system targeted to an early defect observed in many neurodegenerative diseases.

Gunawardena, Shermali

Pharmaceutical research

2013 Volume 30, Issue 10, Page(s) 2459–2474

Abstract: Currently, there are no effective treatments or cures for many neurodegenerative diseases affecting an aging baby-boomer generation. The ongoing problem with many of the current therapeutic treatments is that most are aimed at dissolving or dissociating ... ...

Abstract	Currently, there are no effective treatments or cures for many neurodegenerative diseases affecting an aging baby-boomer generation. The ongoing problem with many of the current therapeutic treatments is that most are aimed at dissolving or dissociating aggregates and preventing cell death, common neuropathology often seen towards the end stage of disease. Often such treatments have secondary effects that are more devastating than the disease itself. Thus, effective therapeutics must be focused on directly targeting early events such that global deleterious effects of drugs are minimized while beneficial therapeutic effects are maximized. Recent work indicates that in many neurodegenerative diseases long distance axonal transport is perturbed, leading to axonal blockages. Axonal blockages are observed before pathological or behavioral phenotypes are seen indicating that this pathway is perturbed early in disease. Thus, developing novel therapeutic treatments to an early defect is critical in curing disease. Here I review neurodegenerative disease and current treatment strategies, and discuss a novel nanotechnology based approach that is aimed at targeting an early pathway, with the rationale that restoring an early problem will prevent deleterious downstream effects. To accomplish this, knowledge exchange between biologists, chemists, and engineers will be required to manufacture effective novel biomaterials for medical use.
MeSH term(s)	Axonal Transport ; Brain/drug effects ; Brain/metabolism ; Brain/pathology ; Cell Survival/drug effects ; Drug Carriers/chemistry ; Drug Delivery Systems/methods ; Early Diagnosis ; Humans ; Nanoparticles/chemistry ; Neurodegenerative Diseases/drug therapy ; Neurodegenerative Diseases/metabolism ; Neurodegenerative Diseases/pathology ; Neuroprotective Agents/administration & dosage ; Neuroprotective Agents/pharmacokinetics ; Neuroprotective Agents/therapeutic use
Chemical Substances	Drug Carriers ; Neuroprotective Agents
Language	English
Publishing date	2013-04-27
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	843063-9
ISSN	1573-904X ; 0724-8741 ; 0739-0742
ISSN (online)	1573-904X
ISSN	0724-8741 ; 0739-0742
DOI	10.1007/s11095-013-1037-0
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Article: Developing nanotherapies for neurodegenerative diseases: ORMOSIL and its potential in axonal transport.

Gunawardena, Shermali

Therapeutic delivery

2012 Volume 3, Issue 10, Page(s) 1189–1198

Abstract: In neurons, essential components packaged into vesicles are transported down microtubules to the ends of axons (synapses) where they are utilized. Components are also transported from the synapse to the cell body. This transport pathway is crucial for ... ...

Abstract	In neurons, essential components packaged into vesicles are transported down microtubules to the ends of axons (synapses) where they are utilized. Components are also transported from the synapse to the cell body. This transport pathway is crucial for normal development, cell survival and plasticity. Recent work has established that defects in transport can contribute to the initiation of neurodegenerative disease, culminating in cell death and degeneration. Thus, delivering therapeutic treatments to an early defect is critical since many current strategies target pathology that occurs at later stages in the disease. Current treatments also affect the entire organism, causing side-effects that are often more deleterious than the disease. This article discusses how engineered synthetic structures can be used to directly target axonal transport--a pathway that is affected during the early stages of disease. Studies in this area will require the exchange of fundamental knowledge between biologists, chemists and engineers to effectively manufacture novel biomaterials for medical use.
MeSH term(s)	Animals ; Axonal Transport ; Drug Delivery Systems ; Drug Design ; Humans ; Microtubules/metabolism ; Nanoparticles ; Neurodegenerative Diseases/drug therapy ; Neurodegenerative Diseases/physiopathology ; Siloxanes/chemistry
Chemical Substances	Siloxanes ; ormosil
Language	English
Publishing date	2012-10-30
Publishing country	England
Document type	Journal Article ; Review
ISSN	2041-5990
ISSN	2041-5990
DOI	10.4155/tde.12.101
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Article ; Online: In Vivo Visualization of Moving Synaptic Cargo Complexes within Drosophila Larval Segmental Axons.

Banerjee, Rupkatha / White, Joseph A / Gunawardena, Shermali

Methods in molecular biology (Clifton, N.J.)

2020 Volume 2143, Page(s) 293–300

Abstract: Identifying moving synaptic vesicle complexes and isolating specific proteins present within such complexes in vivo is challenging. Here we detail a protocol that we have developed that is designed to simultaneously visualize the axonal transport of two ... ...

Abstract	Identifying moving synaptic vesicle complexes and isolating specific proteins present within such complexes in vivo is challenging. Here we detail a protocol that we have developed that is designed to simultaneously visualize the axonal transport of two fluorescently tagged synaptic vesicle proteins in living Drosophila larval segmental nerves in real time. Using a beam-splitter and split view software, larvae expressing GFP-tagged Synaptobrevin (Syb) and mRFP-tagged Rab4-GTPase or YFP-tagged Amyloid Precursor protein (APP) and mRFP-tagged Rab4-GTPase are imaged simultaneously using separate wavelengths. Merged kymographs from the two wavelengths are evaluated for colocalization analysis. Vesicle velocity analysis can also be done. Such analysis enables us to visualize the motility behaviors of two synaptic proteins present on a single vesicle complex and identify candidate proteins moving on synaptic vesicles in vivo, under physiological conditions.
MeSH term(s)	Amyloid beta-Protein Precursor/analysis ; Amyloid beta-Protein Precursor/genetics ; Animals ; Axonal Transport ; Axons/metabolism ; Computer Systems ; Drosophila Proteins/analysis ; Drosophila Proteins/genetics ; Drosophila melanogaster/growth & development ; Drosophila melanogaster/metabolism ; Fluorescent Dyes/analysis ; GTP Phosphohydrolases/analysis ; GTP Phosphohydrolases/genetics ; Intravital Microscopy/methods ; Kymography ; Larva ; Luminescent Proteins/analysis ; Luminescent Proteins/genetics ; Microscopy, Fluorescence/methods ; R-SNARE Proteins/analysis ; R-SNARE Proteins/genetics ; Software ; Synaptic Vesicles/physiology ; Synaptic Vesicles/ultrastructure
Chemical Substances	Amyloid beta-Protein Precursor ; Drosophila Proteins ; Fluorescent Dyes ; Luminescent Proteins ; R-SNARE Proteins ; GTP Phosphohydrolases (EC 3.6.1.-) ; Rab4 protein, Drosophila (EC 3.6.1.-)
Language	English
Publishing date	2020-06-02
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-0585-1_21
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Article ; Online: A stop or go switch: glycogen synthase kinase 3β phosphorylation of the kinesin 1 motor domain at Ser314 halts motility without detaching from microtubules.

Banerjee, Rupkatha / Chakraborty, Piyali / Yu, Michael C / Gunawardena, Shermali

Development (Cambridge, England)

2021 Volume 148, Issue 24

Abstract: It is more than 25 years since the discovery that kinesin 1 is phosphorylated by several protein kinases. However, fundamental questions still remain as to how specific protein kinase(s) contribute to particular motor functions under physiological ... ...

Abstract	It is more than 25 years since the discovery that kinesin 1 is phosphorylated by several protein kinases. However, fundamental questions still remain as to how specific protein kinase(s) contribute to particular motor functions under physiological conditions. Because, within an whole organism, kinase cascades display considerable crosstalk and play multiple roles in cell homeostasis, deciphering which kinase(s) is/are involved in a particular process has been challenging. Previously, we found that GSK3β plays a role in motor function. Here, we report that a particular site on kinesin 1 motor domain (KHC), S314, is phosphorylated by GSK3β in vivo. The GSK3β-phosphomimetic-KHCS314D stalled kinesin 1 motility without dissociating from microtubules, indicating that constitutive GSK3β phosphorylation of the motor domain acts as a STOP. In contrast, uncoordinated mitochondrial motility was observed in CRISPR/Cas9-GSK3β non-phosphorylatable-KHCS314A Drosophila larval axons, owing to decreased kinesin 1 attachment to microtubules and/or membranes, and reduced ATPase activity. Together, we propose that GSK3β phosphorylation fine-tunes kinesin 1 movement in vivo via differential phosphorylation, unraveling the complex in vivo regulatory mechanisms that exist during axonal motility of cargos attached to multiple kinesin 1 and dynein motors.
MeSH term(s)	Adenosine Triphosphatases/genetics ; Animals ; Axonal Transport/genetics ; Axons/metabolism ; CRISPR-Cas Systems/genetics ; Cell Movement/genetics ; Cell Movement/physiology ; Drosophila Proteins/genetics ; Drosophila melanogaster/genetics ; Dyneins/genetics ; Glycogen Synthase Kinase 3 beta/genetics ; Kinesins/genetics ; Larva/genetics ; Microtubules/genetics ; Neurons/metabolism ; Phosphorylation/genetics ; Protein Domains/genetics
Chemical Substances	Drosophila Proteins ; Glycogen Synthase Kinase 3 beta (EC 2.7.11.1) ; Adenosine Triphosphatases (EC 3.6.1.-) ; Khc protein, Drosophila (EC 3.6.1.-) ; Dyneins (EC 3.6.4.2) ; Kinesins (EC 3.6.4.4)
Language	English
Publishing date	2021-12-23
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	90607-4
ISSN	1477-9129 ; 0950-1991
ISSN (online)	1477-9129
ISSN	0950-1991
DOI	10.1242/dev.199866
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Article ; Online: HTT (huntingtin) and RAB7 co-migrate retrogradely on a signaling LAMP1-containing late endosome during axonal injury.

Krzystek, Thomas J / White, Joseph A / Rathnayake, Rasika / Thurston, Layne / Hoffmar-Glennon, Hayley / Li, Yichen / Gunawardena, Shermali

Autophagy

2022 Volume 19, Issue 4, Page(s) 1199–1220

Abstract: Abbreviations: Atg5: Autophagy-related 5; Atg8a: Autophagy-related 8a; AL: autolysosome; AP: autophagosome; BAF1: bafilomycin ... ...

Abstract	Abbreviations: Atg5: Autophagy-related 5; Atg8a: Autophagy-related 8a; AL: autolysosome; AP: autophagosome; BAF1: bafilomycin A
MeSH term(s)	Humans ; Kinesins/metabolism ; Autophagy ; Mitogen-Activated Protein Kinase 8/metabolism ; Axons/metabolism ; Transcription Factors/metabolism ; Carrier Proteins ; Endosomes/metabolism ; Huntingtin Protein/metabolism ; Lysosomal Membrane Proteins/metabolism
Chemical Substances	Kinesins (EC 3.6.4.4) ; Mitogen-Activated Protein Kinase 8 (EC 2.7.11.24) ; Transcription Factors ; Carrier Proteins ; HTT protein, human ; Huntingtin Protein ; LAMP1 protein, human ; Lysosomal Membrane Proteins ; KIF5C protein, human (EC 3.6.1.-)
Language	English
Publishing date	2022-09-09
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2454135-7
ISSN	1554-8635 ; 1554-8627
ISSN (online)	1554-8635
ISSN	1554-8627
DOI	10.1080/15548627.2022.2119351
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Article ; Online: Ethanol stimulates the in vivo axonal movement of neuropeptide dense-core vesicles in Drosophila motor neurons.

Iacobucci, Gary J / Gunawardena, Shermali

Journal of neurochemistry

2017 Volume 144, Issue 4, Page(s) 466–482

Abstract: Proper neuronal function requires essential biological cargoes to be packaged within membranous vesicles and transported, intracellularly, through the extensive outgrowth of axonal and dendritic fibers. The precise spatiotemporal movement of these ... ...

Abstract	Proper neuronal function requires essential biological cargoes to be packaged within membranous vesicles and transported, intracellularly, through the extensive outgrowth of axonal and dendritic fibers. The precise spatiotemporal movement of these cargoes is vital for neuronal survival and, thus, is highly regulated. In this study we test how the axonal movement of a neuropeptide-containing dense-core vesicle (DCV) responds to alcohol stressors. We found that ethanol induces a strong anterograde bias in vesicle movement. Low doses of ethanol stimulate the anterograde movement of neuropeptide-DCV while high doses inhibit bi-directional movement. This process required the presence of functional kinesin-1 motors as reduction in kinesin prevented the ethanol-induced stimulation of the anterograde movement of neuropeptide-DCV. Furthermore, expression of inactive glycogen synthase kinase 3 (GSK-3β) also prevented ethanol-induced stimulation of neuropeptide-DCV movement, similar to pharmacological inhibition of GSK-3β with lithium. Conversely, inhibition of PI3K/AKT signaling with wortmannin led to a partial prevention of ethanol-stimulated transport of neuropeptide-DCV. Taken together, we conclude that GSK-3β signaling mediates the stimulatory effects of ethanol. Therefore, our study provides new insight into the physiological response of the axonal movement of neuropeptide-DCV to exogenous stressors. Cover Image for this Issue: doi: 10.1111/jnc.14165.
MeSH term(s)	Animals ; Axonal Transport/drug effects ; Axons/drug effects ; Axons/metabolism ; Central Nervous System Depressants/pharmacology ; Drosophila/physiology ; Ethanol/pharmacology ; Glycogen Synthase Kinase 3 beta/antagonists & inhibitors ; Glycogen Synthase Kinase 3 beta/metabolism ; Immunohistochemistry ; Kinesin/physiology ; Larva ; Lithium/pharmacology ; Motor Neurons/drug effects ; Motor Neurons/metabolism ; Neuropeptides/metabolism ; Phosphatidylinositol 3-Kinases/antagonists & inhibitors ; Signal Transduction/drug effects ; Stimulation, Chemical ; Synaptic Vesicles/drug effects ; Synaptic Vesicles/metabolism ; Wortmannin/pharmacology
Chemical Substances	Central Nervous System Depressants ; Neuropeptides ; Ethanol (3K9958V90M) ; Lithium (9FN79X2M3F) ; Phosphatidylinositol 3-Kinases (EC 2.7.1.-) ; Glycogen Synthase Kinase 3 beta (EC 2.7.11.1) ; Kinesin (EC 3.6.4.4) ; Wortmannin (XVA4O219QW)
Language	English
Publishing date	2017-10-18
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	80158-6
ISSN	1471-4159 ; 0022-3042 ; 1474-1644
ISSN (online)	1471-4159
ISSN	0022-3042 ; 1474-1644
DOI	10.1111/jnc.14230
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Article: The Non-amyloidal Component Region of α-Synuclein Is Important for α-Synuclein Transport Within Axons.

Anderson, Eric N / Hirpa, Delnessaw / Zheng, Kan Hong / Banerjee, Rupkatha / Gunawardena, Shermali

Frontiers in cellular neuroscience

2020 Volume 13, Page(s) 540

Abstract: Proper transport of the Parkinson's disease (PD) protein, α-synuclein (α-syn), is thought to be crucial for its localization and function at the synapse. Previous work has shown that defects in long distance transport within narrow caliber axons occur ... ...

Abstract	Proper transport of the Parkinson's disease (PD) protein, α-synuclein (α-syn), is thought to be crucial for its localization and function at the synapse. Previous work has shown that defects in long distance transport within narrow caliber axons occur early in PD, but how such defects contribute to PD is unknown. Here we test the hypothesis that the NAC region is involved in facilitating proper transport of α-syn within axons
Language	English
Publishing date	2020-01-10
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2452963-1
ISSN	1662-5102
ISSN	1662-5102
DOI	10.3389/fncel.2019.00540
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