Figure 2 - available via license: Creative Commons Attribution 4.0 International
Content may be subject to copyright.
Rotenone induced PD-like phenotypes in Drosophila. A and B, Rotenone at various concentration was added in fly food at 1 day post-eclosion through the lifetime. (a) Fly survival ratio were scored weekly. There were 60 flies in each group and 3 parallel experiments were carried out in total. Data were analyzed using Kaplan-Meier survival analysis (log-rank followed by Holm-Sidak method). Rotenone treatments at 25, 75, and 125 nM significantly reduced fly survival ratio compared with the group with no rotenone exposure (p < 0.05); (b) The locomotor activity was monitored weekly using climbing assays. There were 30 flies in each experimental group and this experiment was repeated three times in parallel. Rotenone treatment significantly reduced fly locomotor function compared with control with no rotenone exposure ( * p < 0.05 analyzed with ANOVA); (c) and (d) Rotenone caused dopaminergic neuronal degeneration. Rotenone at 25, 75 and 125 µM was added in fly food at 1 day post-eclosion for 14 days. The fly brains were dissected and subjected to whole mount brain immunoflouresence using anti-TH antibodies; (c) Quantification of anti-TH positive neurons in DA neuron clusters; (d) Representative images of PPM1/2 clusters of control and 75 µM Rotenone group. Arrows indicate examples of TH-positive neurons in both conditions.
Source publication
Parkinson’s disease (PD) is a progressive neurodegenerative movement disorder resulting from a selective loss of dopaminergic neurons. The pathogenesis of PD remains incompletely understood, but increasing evidence from human and animal studies has suggested that oxidative damage contributes to the neuronal loss in PD. In this study, we used roteno...
Contexts in source publication
Context 1
... assess whether rotenone induces degeneration in dopaminergic neurons, brains from flies with rotenone exposure were dissected and subjected to whole-mount brain immunostaining using specific anti-TH antibodies. We counted the TH-positive cellsin all the clusters (the details are described in the methods section) except the paired PAM cluster, be cause the density of the neurons in PAM was too high to allow precise quantification. In unexposed control flies, at 2 week of age, the DA clus- ters did not change significantly in number of TH-posi- tive cells or their morphology (Figures 2(c) and (d)). However, at 2 week of age, flies exposed to rotenone showed a striking decrease of anti-TH-positive staining in all the DA clusters, indicating that rotenone induced a loss of DA neurons in the brain. A and B, Rotenone at various concentration was added in fly food at 1 day post-eclosion through the lifetime. (a) Fly sur- vival ratio were scored weekly. There were 60 flies in each group and 3 parallel experiments were carried out in total. Data were analyzed using Kaplan-Meier survival analysis (log-rank followed by Holm-Sidak method). Rotenone treatments at 25, 75, and 125 nM significantly reduced fly survival ratio com- pared with the group with no rotenone exposure (p < 0.05); (b) The locomotor activity was monitored weekly using climbing assays. There were 30 flies in each experimental group and this experiment was repeated three times in parallel. Rotenone treatment significantly reduced fly locomotor function com- pared with control with no rotenone exposure ( * p < 0.05 ana- lyzed with ANOVA); (c) and (d) Rotenone caused dopaminer- gic neuronal degeneration. Rotenone at 25, 75 and 125 µM was added in fly food at 1 day post-eclosion for 14 days. The fly brains were dissected and subjected to whole mount brain im- munoflouresence using anti-TH antibodies; (c) Quantification of anti-TH positive neurons in DA neuron clusters; (d) Repre- sentative images of PPM1/2 clusters of control and 75 µM Rotenone group. Arrows indicate examples of TH-positive neurons in both ...
Context 2
... generated a rotenone-based fly model using W1118 normal wild type Drosophila. We added rotenone in fly food at a concentration gradient of 5, 25, 75 and 125 μM from day 1 post-eclosion and keep the same drug level in the food throughout the lifetime. We found that rotenone caused early death in a dose-dependent manner and is significant when compared with unex- posed flies (Figure 2(a)). To measure the behavioral change resulted from rotenone exposure, we employed a climbing assay (negative geotactic test) at multiple time points in their lifetime. When flies were tapped to the bottom of a vial, nearly all untreated control flies rapidly climbed to the top of the vial on day 7 and day 14. How- ever, flies exposed to rotenone exhibited a significant dose-dependent motor deficit in performance, indicating locomotor dysfunction (Figure 2(b)). There was a pro- gressive, dose dependent decline in climbing ability in flies with rotenone exposure. Rotenone at 75 μM and 125 μM caused significant locomotor impairment and early mortality compared to the untreated group. Thus, we used these two concentrations to conduct further ...
Context 3
... generated a rotenone-based fly model using W1118 normal wild type Drosophila. We added rotenone in fly food at a concentration gradient of 5, 25, 75 and 125 μM from day 1 post-eclosion and keep the same drug level in the food throughout the lifetime. We found that rotenone caused early death in a dose-dependent manner and is significant when compared with unex- posed flies (Figure 2(a)). To measure the behavioral change resulted from rotenone exposure, we employed a climbing assay (negative geotactic test) at multiple time points in their lifetime. When flies were tapped to the bottom of a vial, nearly all untreated control flies rapidly climbed to the top of the vial on day 7 and day 14. How- ever, flies exposed to rotenone exhibited a significant dose-dependent motor deficit in performance, indicating locomotor dysfunction (Figure 2(b)). There was a pro- gressive, dose dependent decline in climbing ability in flies with rotenone exposure. Rotenone at 75 μM and 125 μM caused significant locomotor impairment and early mortality compared to the untreated group. Thus, we used these two concentrations to conduct further ...
Similar publications
Tianma Gouteng Yin (TGY) is a traditional Chinese medicine (TCM) decoction widely used to treat symptoms associated with typical Parkinson's disease (PD). In this study, the neuroprotective effects of water extract of TGY were tested on rotenone-intoxicated and human α-synuclein transgenic Drosophila PD models. In addition, the neuroprotective effe...
Citations
... Rotenone exposure induced the loss of MMP via mitochondrial complex I inhibition, leading to the opening of permeability transition pores and increasing intracellular ROS. With the increase of ROS in the dopaminergic neurons, redox homeostasis is lost, leading to antioxidant enzyme dysfunction, lipid peroxidation, neuro-inflammation, and neural cell damage, all of which contribute to the progress of neurodegeneration [24,29]. A promising approach that focuses on the inhibition of brain oxidative damage is the activation of the Nrf2 signaling pathway [30]. ...
Diacetylcurcumin manganese complex (DiAc-Cp-Mn) is a diacetylcurcumin (DiAc-Cp) derivative synthesized with Mn (II) to mimic superoxide dismutase (SOD). It exhibited superior reactive oxygen species (ROS) scavenging efficacy, particularly for the superoxide radical. The present study investigated the ROS scavenging activity, neuroprotective effects, and underlying mechanism of action of DiAc-Cp-Mn in a cellular model of Parkinson’s disease. This study utilized rotenone-induced neurotoxicity in SH-SY5Y cells to assess the activities of DiAc-Cp-Mn by measuring cell viability, intracellular ROS, mitochondrial membrane potential (MMP), SOD, and catalase (CAT) activities. The mRNA expression of the nuclear factor erythroid 2 p45-related factor (Nrf2), Kelch-like ECH-associated protein 1 (Keap1), inducible nitric oxide synthase (iNOS), and Interleukin 1β (IL-1β), which are oxidative and inflammatory genes, were also evaluated to clarify the molecular mechanism. The results of the in vitro assays showed that DiAc-Cp-Mn exhibited greater scavenging activity against superoxide radicals, hydrogen peroxide, and hydroxyl radicals compared to DiAc-Cp. In cell-based assays, DiAc-Cp-Mn demonstrated greater neuroprotective effects against rotenone-induced neurotoxicity when compared to its parent compound, DiAc-Cp. DiAc-Cp-Mn maintained MMP levels, reduced intracellular ROS levels, and increased the activities of SOD and CAT by activating the Nrf2-Keap1 signaling pathway. In addition, DiAc-Cp-Mn exerted its anti-inflammatory impact by down-regulating the mRNA expression of iNOS and IL-1β that provoked neuro-inflammation. The current study indicates that DiAc-Cp-Mn protects against rotenone-induced neuronal damage by reducing oxidative stress and inflammation.
... Finally, PLG's role in the autophagy-apoptosis balance is facilitated by the increase of BCL2 phosphorylation at Ser70 via MAPK8 signaling. The PD-relevant neuroprotective and senolytic properties of PLG have been studied in SCNA transgenic SK-N-SH cells, senescent WI-38 human fibroblasts, 6-OHDA treated PC12 cells, rotenonetreated SK-N-SH cells, rotenone-treated mice, MPTP treated rats, 6-OHDA treated rats, and in lipopolysaccharide treated BV2 microglial cells (192,199,202,(204)(205)(206). Notably, it has not yet been studied in D. melanogaster. ...
Drosophila melanogaster is a valuable model organism for a wide range of biological exploration. The well-known advantages of D. melanogaster include its relatively simple biology, the ease with which it is genetically modified, the relatively low financial and time costs associated with their short gestation and life cycles, and the large number of offspring they produce per generation. D. melanogaster has facilitated the discovery of many significant insights into the pathology of Parkinson’s disease (PD) and has served as an excellent preclinical model of PD-related therapeutic discovery. In this review, we provide an overview of the major D. melanogaster models of PD, each of which provide unique insights into PD-relevant pathology and therapeutic targets. These models are discussed in the context of their past, current, and future potential use for studying the utility of secondary metabolites as therapeutic agents in PD. Over the last decade, senolytics have garnered an exponential interest in their ability to mitigate a broad spectrum of diseases, including PD. Therefore, an emphasis is placed on the senolytic and senomorphic properties of secondary metabolites. It is expected that D. melanogaster will continue to be critical in the effort to understand and improve treatment of PD, including their involvement in translational studies focused on secondary metabolites.
... 50 Several rotenone-based toxin cell, rodent, and Drosophila models have been used to study the molecular mechanisms of cell death and potential therapeutics for PD. 51 However, little is known about the precise mechanistic action of these agents in Drosophila; the involvement of mitochondrial complex I is well supported by several experimental studies. 52,53 In our study, exposure to rotenone in Drosophila reproduces key aspects of PD, for instance, deficits in locomotion, death, and loss of dopaminergic neuronal cells. ...
Parkinson's disease (PD) is the second most common neurodegenerative disorder; however, its etiology remains elusive. Antioxidants are considered to be a promising approach for decelerating neurodegenerative disease progression owing to extensive examination of the relationship between oxidative stress and neurodegenerative diseases. In this study, we investigated the therapeutic effect of melatonin against rotenone-induced toxicity in the Drosophila model of PD. The 3-5 day old flies were divided into four groups: control, melatonin alone, melatonin and rotenone, and rotenone alone groups. According to their respective groups, flies were exposed to a diet containing rotenone and melatonin for 7 days. We found that melatonin significantly reduced the mortality and climbing ability of Drosophila because of its antioxidative potency. It alleviated the expression of Bcl 2, tyrosine hydroxylase (TH), NADH dehydrogenase, mitochondrial membrane potential, and mitochondrial bioenergetics and decreased caspase 3 expression in the Drosophila model of rotenone-induced PD-like symptoms. These results indicate the neuromodulatory effect of melatonin, and that it is likely modulated against rotenone-induced neurotoxicity by suppressing oxidative stress and mitochondrial dysfunctions.
... Overall, the cell viability and mitochondrial hydroxyl radical assay results were consistent with previous studies which have shown protective effects of curcumin, NAC and DFO against rotenone in in vitro models of PD [18,21,38,39]. However, unlike the results reported by Mursaleen et al. [38] where the P68 + DQA curcumin nanoformulations were assessed in SH-SY5Y cells alone without the BBB Transwell ® compartment and where for the most part the formulations were only at least as good as the free drug conditions at protecting against rotenone, when using the Transwell ® model the P68 + DQA nanoformulated treatments were found to be generally more protective than the free drugs. ...
As the blood-brain barrier (BBB) prevents most compounds from entering the brain, nanocarrier delivery systems are frequently being explored to potentially enhance the passage of drugs due to their nanometer sizes and functional characteristics. This study aims to investigate whether Pluronic® F68 (P68) and dequalinium (DQA) nanocarriers can improve the ability of curcumin, n-acetylcysteine (NAC) and/or deferoxamine (DFO), to access the brain, specifically target mitochondria and protect against rotenone by evaluating their effects in a combined Transwell® hCMEC/D3 BBB and SH-SY5Y based cellular Parkinson’s disease (PD) model. P68 + DQA nanoformulations enhanced the mean passage across the BBB model of curcumin, NAC and DFO by 49%, 28% and 49%, respectively (p < 0.01, n = 6). Live cell mitochondrial staining analysis showed consistent co-location of the nanocarriers within the mitochondria. P68 + DQA nanocarriers also increased the ability of curcumin and NAC, alone or combined with DFO, to protect against rotenone induced cytotoxicity and oxidative stress by up to 19% and 14% (p < 0.01, n = 6), as measured by the MTT and mitochondrial hydroxyl radical assays respectively. These results indicate that the P68 + DQA nanocarriers were successful at enhancing the protective effects of curcumin, NAC and/or DFO by increasing the brain penetrance and targeted delivery of the associated bioactives to the mitochondria in this model. This study thus emphasises the potential effectiveness of this nanocarrier strategy in fully utilising the therapeutic benefit of these antioxidants and lays the foundation for further studies in more advanced models of PD.
... Particularly, motor abnormalities in Parkinson's patients are thought to be due to the death of dopaminergic neurons in the SNpc (substantia nigra pars compacta) [36]. In this study, the dUCH knockdown of DA neuron-specific also led to DA neuron degeneration similar to previous studies [19,20] and other Parkinson's disease models [37][38][39]. The present results showed that this adverse condition significantly reduced with PLE treatment in both larval and adult stages. ...
Parkinson’s disease (PD) is characterized by progressive locomotive defects and loss of dopaminergic neurons. Polyscias fruticosa leaves are used by Vietnamese as herbal medicines to support the treatment of some diseases related to neurodegeneration such as Parkinson’s and Alzheimer’s diseases. However, recent scientific data have not provided sufficient evidence for the use of P. fruticosa leaves to treat PD or decelerate PD progression. In the present study, the capacity of P. fruticosa leaf extract for PD treatment on the dietary supplementation was investigated using dUCH-knockdown Drosophila model. The results indicated that P. fruticosa leaf extract decelerated dopaminergic neuron degeneration induced by dUCH knockdown in not only the larval stage but also the adult stage, which might result in the amelioration in locomotor ability of dUCH-knockdown larvae and flies. Furthermore, antioxidant activities and some key phytochemicals such as saponins, polyphenols, and flavonoids that might contribute to the effects of the P. fruticosa leaf extract were identified.
... ROS lead to the formation of 4-HNE aggresomes, which we demonstrated were rescued by using the MnSOD mimetic, MnTnBuOE-2-PyP 5+ . This impact of MnP-driven rescue supports the data that rotenone causes an increase in mitochondrial ROS [34,35]. In our study, 4-HNE protein aggresomes co-localized with ubiquitin, HDAC6, and γ-tubulin. ...
Reactive oxygen species (ROS) cause oxidative stress by generating reactive aldehydes known as 4-hydroxynonenal (4-HNE). 4-HNE modifies protein via covalent adduction; however, little is known about the degradation mechanism of 4-HNE-adducted proteins. Autophagy is a dynamic process that maintains cellular homeostasis by removing damaged organelles and proteins. In this study, we determined the role of a superoxide dismutase (SOD) mimetic MnTnBuOE-2-PyP5+ (MnP, BMX-001) on rotenone-induced 4-HNE aggresome degradation in HL-1 cardiomyocytes. A rotenone treatment (500 nM) given for 24 h demonstrated both increased ROS and 4-HNE aggresome accumulation in HL-1 cardiomyocytes. In addition, cardiomyocytes treated with rotenone displayed an increase in the autophagy marker LC3-II, as shown by immunoblotting and immunofluorescence. A pre-treatment with MnP (20 µM) for 24 h attenuated rotenone-induced ROS formation. An MnP pre-treatment showed decreased 4-HNE aggresomes and LC3-II formation. A rotenone-induced increase in autophagosomes was attenuated by a pre-treatment with MnP, as shown by fluorescent-tagged LC3 (tfLC3). Rotenone increased tubulin hyperacetylation through the ROS-mediated pathway, which was attenuated by MnP. The disruption of autophagy caused HL-1 cell death because a 3-methyladenine inhibitor of autophagosomes caused reduced cell death. Yet, rapamycin, an inducer of autophagy, increased cell death. These results indicated that a pre-treatment with MnP decreased rotenone-induced 4-HNE aggresomes by enhancing the degradation process.
... It was also reported that systemic administration of RT may reproduce other features of PD in experimental animal models such as the alterations of the retina, loss of testosterone and sleep disorders (26)(27)(28). Other studies performed in mice have demonstrated that the selective toxicity of RT can be related to inhibition of the NADH gene dehydrogenase ubiquinone proteins Fe-S4 (NDUFS4) which encode the mitochondrial complex I which is inhibited by RT. ...
Parkinson’s disease (PD) is the second most prevalent neuro-degenerative disease after Alzheimer´s disease. It is characterized by motor symptoms such as akinesia, bradykinesia, tremor, rigidity, and postural abnormalities, due to the loss of nigral dopaminergic neurons and a decrease in the dopa-mine contents of the caudate-putamen structures. To this date, there is no cure for the disease and available treatments are aimed at controlling the symptoms. Therefore, there is an unmet need for new treatments for PD. In the past decades, animal models of PD have been proven to be valuable tools in elucidating the nature of the pathogenic processes involved in the disease, and in designing new pharmacological approaches. Here, we review the use of neurotoxin-induced and pesticide-induced animal models of PD, specifically those induced by rotenone, paraquat, maneb, MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) and 6-OHDA (6-hydroxydopamine), and their application in the development of new drug delivery systems for PD.
... Finally, cellular viability was assayed by the MTT method. Curcumin has shown neuroprotective activity in several studies with rotenone, some of which were performed in the same SH-SY5Y cell line [81,82], and similar results were reported in PC12 cells [83]. Rotenone is employed in our case in combination with oligomycin A, but both toxics target the electron transport chain of the mitochondria, thus leading to oxidative stress and similar cellular effects. ...
Curcumin shows a broad spectrum of activities of relevance in the treatment of Alzheimer’s disease (AD); however, it is poorly absorbed and is also chemically and metabolically unstable, leading to a very low oral bioavailability. A small library of hybrid compounds designed as curcumin analogues and incorporating the key structural fragment of piperlongumine, a natural neuroinflammation inhibitor, were synthesized by a two-step route that combines a three-component reaction between primary amines, β-ketoesters and α-haloesters and a base-promoted acylation with cinnamoyl chlorides. These compounds were predicted to have good oral absorption and CNS permeation, had good scavenging properties in the in vitro DPPH experiment and in a cellular assay based on the oxidation of dichlorofluorescin to a fluorescent species. The compounds showed low toxicity in two cellular models, were potent inductors of the Nrf2-ARE phase II antioxidant response, inhibited PHF6 peptide aggregation, closely related to Tau protein aggregation and were active against the LPS-induced inflammatory response. They also afforded neuroprotection against an oxidative insult induced by inhibition of the mitochondrial respiratory chain with the rotenone-oligomycin A combination and against Tau hyperphosphorylation induced by the phosphatase inhibitor okadaic acid. This multitarget pharmacological profile is highly promising in the development of treatments for AD and provides a good hit structure for future optimization efforts.
... The prior studies have revealed CUR neuroprotective properties in both in-vitro and in-vivo PD models [42][43][44][45][46][47][48][49][50][51][52][53]; and its inhibition effectiveness in the aggregation of αS by binding to its monomers [54,55] has been shown, in addition to binding to its oligomers [56]. ...
Alpha-Synuclein (αS) is a protein involved in Parkinson’s disease (PD) and is probably the main cause of the pathology of the disease. During pathogenesis, αS monomers aggregate, leading to the formation of a variety of oligomeric species. Recent research studies suggest that the oligomeric toxic species may be one of the main processes for pathology and disease. Here, we studied influence of two natural polyphenolic compounds, Curcumin (CUR) and Rosmarinic acid (RA), on disrupting the general properties of αS oligomer by molecular dynamics (MD) simulation method. The hydrophobic central domain of αS (NAC), is the most essential district responsible for protein self-aggregation; so, in this study, our systems have been developed to form a quintuplet NAC region of αS called 5mer; they have 10 and 20 CUR and RA molecules and a 5mer with no ligand. The several important and efficient analyzes were performed to investigate the effect of ligands on the structural properties of αS oligomers. The results indicated that both ligands can be successful in disrupting the original structure of αS oligomers; therefore, they can be considered suitable candidates for designing Parkinson's drugs.
... [25] In a recent study conducted; chronic dietary supplementation with turmeric protected against MPTP-mediated neurotoxicity in vivo in a mice model of PD. [26] In another study to explored the protective effects of curcumin against rotenone-induced toxicity in SH-SY5Y cells, scientist also showed that curcumin administration (1 mM) protected against rotenen-induced cell death in a dose-dependent manner by reducing the intracellular ROS levels, mitochondrial depolarization, cytochrome c release, and caspase-9 and caspase-3 activation. [27] Figure 2A and B convolvulus pluricaulis, is among the frequently prescribed rasayanas for improvement of learning and memory and also treatment of mental health problem. CP-MEX, CP-EEX, and CP-WEX have been reported to protect the IMR32 neuroblastoma cell from induce toxic Figure 1], a naturally occurring flavonone major flavanone that exist in citrus and other plants and can isolated in large amount from peels of Citrus aurantium (bitter orange). ...
Inhibition of soluble epoxide hydrolase (sEH) is considered as a promising target to reduce blood pressure, improve insulin sensitivity, and decrease inflammation. Material and Method: In this study, a series of some novel quinazoline-4(3H)- one derivatives (3a-t) with varying steric and electronic properties was designed, synthesized, and evaluated as sEH Inhibitors (sEHI). Most of the synthesized compounds had similar inhibitory activity to the commercial reference inhibitor, 12-(3-adamantan-1-ylureido) dodecanoic acid, and among them, 4-chloro-N-(4-(4-oxo-3,4-dihy- droquinazoline-2-yl)phenyl)benzamide (3g) was identified as the most active sEHI (IC50 = 0.5 nM), about two-fold more potent compared to the reference inhibitor. Conclusion: The results of molecular modeling followed by biological studies indicate that a quinazolinone ring serves as a suitable scaffold to develop novel small molecule candidates to inhibits EH and the nature of substituent on the amide moiety has a moderate effect on the activity.
