Little Helpers or Mean Rogue—Role of Microglia in Animal Models of Amyotrophic Lateral Sclerosis

Cihankaya, Hilal; Theiß, Carsten; Matschke, Veronika

doi:10.3390/ijms22030993

Cited by 9 publications

(4 citation statements)

References 109 publications

(209 reference statements)

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“…Various mouse models with different ALS-associated mutations are currently available to study the pathomechanisms of ALS (Stephenson and Amor, 2017 ; Cihankaya et al, 2021 ), including the Wobbler mouse, which was first described by Falconer in 1956 (Andrews et al, 1974 ; Giorgio et al, 2019 ). The Wobbler mouse displays almost the same symptoms as human ALS patients at the clinical-symptomatic, cellular and metabolic levels (Boillée et al, 2003 ; Moser et al, 2013 ; Ott et al, 2015 ).…”

Section: Introductionmentioning

confidence: 99%

ROS scavengers decrease γH2ax spots in motor neuronal nuclei of ALS model mice in vitro

Junghans

John

Cihankaya

et al. 2022

Front. Cell. Neurosci.

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Background: Amyotrophic lateral sclerosis (ALS) is an incurable neurodegenerative disease characterized by the loss of motor neurons in cerebral cortex, brainstem and spinal cord. Numerous studies have demonstrated signs of oxidative stress in postmortem neuronal tissue, cerebrospinal fluid, plasma and urine of ALS patients, without focusing on the specific processes within motor neurons. Thus, we aimed to investigate the relevance of reactive oxygen species (ROS) detoxification mechanisms and its consequences on the formation of toxic/lethal DNA double strand breaks (DSBs) in the ALS model of the Wobbler mouse.Methods: Live cell imaging in dissociated motor neuronal cultures was used to investigate the production of ROS using Dihydroethidium (DHE). The expression levels of ROS detoxifying molecules were investigated by qPCR as well as Western blots. Furthermore, the expression levels of DNA damage response proteins p53bp1 and H2ax were investigated using qPCR and immunofluorescence staining. Proof-of-principle experiments using ROS scavengers were performed in vitro to decipher the influence of ROS on the formation of DNA double strand breaks quantifying the γH2ax spots formation.Results: Here, we verified an elevated ROS-level in spinal motor neurons of symptomatic Wobbler mice in vitro. As a result, an increased number of DNA damage response proteins p53bp1 and γH2ax in dissociated motor neurons of the spinal cord of Wobbler mice was observed. Furthermore, we found a significantly altered expression of several antioxidant molecules in the spinal cord of Wobbler mice, suggesting a deficit in ROS detoxification mechanisms. This hypothesis could be verified by using ROS scavenger molecules in vitro to reduce the number of γH2ax foci in dissociated motor neurons and thus counteract the harmful effects of ROS.Conclusion: Our data indicate that maintenance of redox homeostasis may play a key role in the therapy of the neurodegenerative disease ALS. Our results underline a necessity for multimodal treatment approaches to prolong the average lifespan of motor neurons and thus slow down the progression of the disease, since a focused intervention in one pathomechanism seems to be insufficient in ALS therapy.

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Section: Introductionmentioning

confidence: 99%

ROS scavengers decrease γH2ax spots in motor neuronal nuclei of ALS model mice in vitro

Junghans

John

Cihankaya

et al. 2022

Front. Cell. Neurosci.

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“…One of the primary objectives of this study was to determine whether disruption of scat expression in adult flies caused phenotypes like those observed in the wobbler mouse. While mutations in Vps54 have not been directly linked to human disease, the wobbler mouse has been used by many researchers as a model for the sporadic form of ALS because of striking similarities to ALS pathology ( Ikeda et al, 1998 ; Dennis and Citron, 2009 ; Nieto-Gonzalez et al, 2011 ; Moser et al, 2013 ; Dahlke et al, 2015 ; Schmitt-John, 2015 ; Klatt et al, 2019 ; Rohm et al, 2019 ; Cihankaya et al, 2021 ; De Nicola et al, 2021 ). It has previously been shown that scat mutants share spermatogenesis defects caused by Golgi dysfunction with the wobbler mouse ( Castrillon et al, 1993 ; Fari et al, 2016 ).…”

Section: Discussionmentioning

confidence: 99%

Vps54 Regulates Lifespan and Locomotor Behavior in Adult Drosophila melanogaster

2021

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Vps54 is an integral subunit of the Golgi-associated retrograde protein (GARP) complex, which is involved in tethering endosome-derived vesicles to the trans-Golgi network (TGN). A destabilizing missense mutation in Vps54 causes the age-progressive motor neuron (MN) degeneration, muscle weakness, and muscle atrophy observed in the wobbler mouse, an established animal model for human MN disease. It is currently unclear how the disruption of Vps54, and thereby the GARP complex, leads to MN and muscle phenotypes. To develop a new tool to address this question, we have created an analogous model in Drosophila by generating novel loss-of-function alleles of the fly Vps54 ortholog (scattered/scat). We find that null scat mutant adults are viable but have a significantly shortened lifespan. Like phenotypes observed in the wobbler mouse, we show that scat mutant adults are male sterile and have significantly reduced body size and muscle area. Moreover, we demonstrate that scat mutant adults have significant age-progressive defects in locomotor function. Interestingly, we see sexually dimorphic effects, with scat mutant adult females exhibiting significantly stronger phenotypes. Finally, we show that scat interacts genetically with rab11 in MNs to control age-progressive muscle atrophy in adults. Together, these data suggest that scat mutant flies share mutant phenotypes with the wobbler mouse and may serve as a new genetic model system to study the cellular and molecular mechanisms underlying MN disease.

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“…Motor neuron degeneration is generally accompanied by neuroinflammation in both ALS patients [ 7 , 8 ] and in the animal models of ALS [ 9 ]. Neuroinflammation is the natural response of the innate immune system in the central nervous system (CNS), and when left uncontrolled, chronic inflammation starts to pose a danger to motor neurons, causing premature cell death.…”

Section: Introductionmentioning

confidence: 99%

Elevated NLRP3 Inflammasome Activation Is Associated with Motor Neuron Degeneration in ALS

Cihankaya,

Bader,

Winklhofer

et al. 2024

Cells

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Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by motor neuron degeneration in the central nervous system. Recent research has increasingly linked the activation of nucleotide oligomerization domain-like receptor protein 3 (NLRP3) inflammasome to ALS pathogenesis. NLRP3 activation triggers Caspase 1 (CASP 1) auto-activation, leading to the cleavage of Gasdermin D (GSDMD) and pore formation on the cellular membrane. This process facilitates cytokine secretion and ultimately results in pyroptotic cell death, highlighting the complex interplay of inflammation and neurodegeneration in ALS. This study aimed to characterize the NLRP3 inflammasome components and their colocalization with cellular markers using the wobbler mouse as an ALS animal model. Firstly, we checked the levels of miR-223-3p because of its association with NLRP3 inflammasome activity. The wobbler mice showed an increased expression of miR-223-3p in the ventral horn, spinal cord, and cerebellum tissues. Next, increased levels of NLRP3, pro-CASP 1, cleaved CASP 1 (c-CASP 1), full-length GSDMD, and cleaved GDSMD revealed NLRP3 inflammasome activation in wobbler spinal cords, but not in the cerebellum. Furthermore, we investigated the colocalization of the aforementioned proteins with neurons, microglia, and astrocyte markers in the spinal cord tissue. Evidently, the wobbler mice displayed microgliosis, astrogliosis, and motor neuron degeneration in this tissue. Additionally, we showed the upregulation of protein levels and the colocalization of NLRP3, c-CASP1, and GSDMD in neurons, as well as in microglia and astrocytes. Overall, this study demonstrated the involvement of NLRP3 inflammasome activation and pyroptotic cell death in the spinal cord tissue of wobbler mice, which could further exacerbate the motor neuron degeneration and neuroinflammation in this ALS mouse model.

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Little Helpers or Mean Rogue—Role of Microglia in Animal Models of Amyotrophic Lateral Sclerosis

Cited by 9 publications

References 109 publications

ROS scavengers decrease γH2ax spots in motor neuronal nuclei of ALS model mice in vitro

ROS scavengers decrease γH2ax spots in motor neuronal nuclei of ALS model mice in vitro

Vps54 Regulates Lifespan and Locomotor Behavior in Adult Drosophila melanogaster

Elevated NLRP3 Inflammasome Activation Is Associated with Motor Neuron Degeneration in ALS

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