Abstract P622: Regulation of Neuroprotective Myeloid Cell Leukemia 1 by Rapamycin and AT2R Agonists in Dopaminergic Neuronal Cell-line

Hypertension ◽  
2015 ◽  
Vol 66 (suppl_1) ◽  
Author(s):  
Jamal Bajwa ◽  
Lakshmi Pulakat
Keyword(s):  
Oxidative Stress ◽  
Cell Death ◽  
Cell Line ◽  
Neuronal Damage ◽  
Neuronal Cells ◽  
Neuronal Cell ◽  
Myeloid Cell ◽  
Serum Starvation ◽  
Cell Leukemia ◽  
Neuronal Cell Line

Myeloid Cell Leukemia I (MCL-1) is a critical protein for neuronal cell survival. MCL-1 is one of the anti-apoptotic proteins in the Bcl2 family. In neurons, MCL-1 regulates the rate of programmed cell death during development and after neuronal damage. It is now well established that without sufficient MCL-1 dopaminergic neuronal cells succumb to cell death under conditions of oxidative stress that result in neurodegenerative diseases such as Parkinsonism. Therefore, identifying drugs that can up-regulate the expression of MCL-1 in neuronal cells is critical for enhancing neuronal resistance to oxidative stress and improving neuronal survival. Aim of this study was to evaluate the effects of treatments with Rapamycin and a novel AT2R peptide agonist NP-6A4 on the MCL-1 expression in SH-SY5Y neuronal cell line. SH-SY5Y cells are a human-derived in vitro model of neuronal function and differentiation, expressing both adrenergic and dopaminergic markers. This cell line is a highly translational model for Parkinson's disease. Cells were maintained in a 1:1 mixture of DMEM and Ham's F-12 with 10% FBS. Cells were subjected to serum starvation and were treated with Rap (10nM), NP-6A4 (300nM) or their combination for 6 hours. MCL-1 protein expression was assessed by immunofluorescence using anti-MCL-1 antibody and a fluorophore-conjugated secondary antibody. Cells were imaged using a confocal microscope and fluorescence was quantified using Leica LAS AF software. It was observed that Rap treatment significantly suppressed MCL-1 expression in SH-SY5Y cells (~40% suppression, p<0.001), whereas Rap+NP-6A4 treatment reversed the Rap-mediated suppression of MCL-1 (p<0.0002). This data indicates that Rapamycin suppresses MCL-1 in dopaminergic neuronal cells and AT2R agonist, NP-6A4 is capable of reversing this effect.

scholarly journals A Bifunctional Anti-Amyloid Blocks Oxidative Stress and the Accumulation of Intraneuronal Amyloid-Beta

Molecules ◽  
2018 ◽  
Vol 23 (8) ◽  
pp. 2010 ◽  
Author(s):  
Silvia Hilt ◽  
Robin Altman ◽  
Tamás Kálai ◽  
Izumi Maezawa ◽  
Qizhi Gong ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Line ◽  
Small Molecule ◽  
Amyloid Beta ◽  
Disease Process ◽  
Neuronal Cell ◽  
Super Resolution ◽  
Microscopy Imaging ◽  
Neuronal Cell Line ◽  
Intimate Link

There is growing recognition regarding the role of intracellular amyloid beta (Aβ) in the Alzheimer’s disease process, which has been linked with aberrant signaling and the disruption of protein degradation mechanisms. Most notably, intraneuronal Aβ likely underlies the oxidative stress and mitochondrial dysfunction that have been identified as key elements of disease progression. In this study, we employed fluorescence imaging to explore the ability of a bifunctional small molecule to reduce aggregates of intracellular Aβ and attenuate oxidative stress. Structurally, this small molecule is comprised of a nitroxide spin label linked to an amyloidophilic fluorene and is known as spin-labeled fluorene (SLF). The effect of the SLF on intracellular Aβ accumulation and oxidative stress was measured in MC65 cells, a human neuronal cell line with inducible expression of the amyloid precursor protein and in the N2a neuronal cell line treated with exogenous Aβ. Super-resolution microscopy imaging showed SLF decreases the accumulation of intracellular Aβ. Confocal microscopy imaging of MC65 cells treated with a reactive oxygen species (ROS)-sensitive dye demonstrated SLF significantly reduces the intracellular Aβ-induced ROS signal. In order to determine the contributions of the separate SLF moieties to these protective activities, experiments were also carried out on cells with nitroxides lacking the Aβ targeting domain or fluorene derivatives lacking the nitroxide functionality. The findings support a synergistic effect of SLF in counteracting both the conformational toxicity of both endogenous and exogenous Aβ, its promotion of ROS, and Aβ metabolism. Furthermore, these studies demonstrate an intimate link between ROS production and Aβ oligomer formation.

scholarly journals Deletions in mitochondrial DNA (mt-DNA) during hypoxic cell death in the human NT2-N neuronal cell line

1997 ◽  
Vol 42 (3) ◽  
pp. 387-387
Author(s):  
Terje Rootwelt ◽  
Usha Reddy ◽  
Bernard Fromenty ◽  
David Pleasure
Keyword(s):  
Cell Death ◽  
Mitochondrial Dna ◽  
Cell Line ◽  
Neuronal Cell ◽  
Hypoxic Cell ◽  
Mt Dna ◽  
Neuronal Cell Line

Angiotensin II Triggers Apoptosis Via Enhancement of NADPH Oxidase-Dependent Oxidative Stress in a Dopaminergic Neuronal Cell Line

2015 ◽  
Vol 40 (4) ◽  
pp. 854-863 ◽  
Author(s):  
Hong-Rui Zhao ◽  
Teng Jiang ◽  
You-Yong Tian ◽  
Qing Gao ◽  
Zhang Li ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Angiotensin Ii ◽  
Nadph Oxidase ◽  
Cell Line ◽  
Neuronal Cell ◽  
Neuronal Cell Line ◽  
Dopaminergic Neuronal Cell

scholarly journals Hypoxic cell death in the human NT2-N neuronal cell line

1997 ◽  
Vol 42 (3) ◽  
pp. 387-387
Author(s):  
Terje Rootwelt ◽  
Michelle Dunn ◽  
Marc Yudkoff ◽  
Takayuki Itoh ◽  
David Pleasure
Keyword(s):  
Cell Death ◽  
Cell Line ◽  
Neuronal Cell ◽  
Hypoxic Cell ◽  
Neuronal Cell Line

scholarly journals Rhes, a striatal-enriched protein, promotes mitophagy via Nix

2019 ◽  
Vol 116 (47) ◽  
pp. 23760-23771 ◽  
Author(s):  
Manish Sharma ◽  
Uri Nimrod Ramírez-Jarquín ◽  
Oscar Rivera ◽  
Melissa Kazantzis ◽  
Mehdi Eshraghi ◽  
...  
Keyword(s):  
Cell Death ◽  
Cell Line ◽  
Neuronal Cell ◽  
Dependent Manner ◽  
Systemic Injection ◽  
Neuronal Cell Line ◽  
In The Wild ◽  
Striatal Lesion ◽  
The Brain

Elimination of dysfunctional mitochondria via mitophagy is essential for cell survival and neuronal functions. But, how impaired mitophagy participates in tissue-specific vulnerability in the brain remains unclear. Here, we find that striatal-enriched protein, Rhes, is a critical regulator of mitophagy and striatal vulnerability in brain. In vivo interactome and density fractionation reveal that Rhes coimmunoprecipitates and cosediments with mitochondrial and lysosomal proteins. Live-cell imaging of cultured striatal neuronal cell line shows Rhes surrounds globular mitochondria, recruits lysosomes, and ultimately degrades mitochondria. In the presence of 3-nitropropionic acid (3-NP), an inhibitor of succinate dehydrogenase, Rhes disrupts mitochondrial membrane potential (ΔΨm) and promotes excessive mitophagy and cell death. Ultrastructural analysis reveals that systemic injection of 3-NP in mice promotes globular mitochondria, accumulation of mitophagosomes, and striatal lesion only in the wild-type (WT), but not in the Rhes knockout (KO), striatum, suggesting that Rhes is critical for mitophagy and neuronal death in vivo. Mechanistically, Rhes requires Nix (BNIP3L), a known receptor of mitophagy, to disrupt ΔΨm and promote mitophagy and cell death. Rhes interacts with Nix via SUMO E3-ligase domain, and Nix depletion totally abrogates Rhes-mediated mitophagy and cell death in the cultured striatal neuronal cell line. Finally, we find that Rhes, which travels from cell to cell via tunneling nanotube (TNT)-like cellular protrusions, interacts with dysfunctional mitochondria in the neighboring cell in a Nix-dependent manner. Collectively, Rhes is a major regulator of mitophagy via Nix, which may determine striatal vulnerability in the brain.

ELEVATED CYTOSOLIC Ca2+DOES NOT MEDIATE CELL DEATH FROM CHEMICAL ANOXIA IN A NEURONAL CELL LINE

1991 ◽  
Vol 75 (3) ◽  
pp. A612-A612
Author(s):  
M. E. Johnson ◽  
C. B. Uhl ◽  
GJ. Gores ◽  
J. C. Sill
Keyword(s):  
Cell Death ◽  
Cell Line ◽  
Neuronal Cell ◽  
Neuronal Cell Line ◽  
Mediate Cell ◽  
Chemical Anoxia
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