Ultrastructural features of aberrant glial cells isolated from the spinal cord of paralytic rats expressing the amyotrophic lateral sclerosis-linked SOD1G93A mutation

Jiménez-Riani, Marcie; Díaz-Amarilla, Pablo; Isasi, Eugenia; Casanova, Gabriela; Barbeito, Luis; Olivera-Bravo, Silvia

doi:10.1007/s00441-017-2681-1

Cited by 26 publications

(66 citation statements)

References 51 publications

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“…Previous morphological studies on AbGC with MitoTracker Green evidenced that mitochondria appear clustered in the perinuclear area and that their main axis was shorter than that observed in neonatal nTg astrocytes as assessed in transmission electron microscopy (TEM) images [21]. Our measurements of mitochondrial length in MitoTracker Green-stained AbGC and nTg neonatal astrocytes were in the range of these previous TEM quantitative data.…”

Section: Discussionsupporting

confidence: 76%

“…AbGC simultaneously express astrocytic glial fibrillary acidic protein (GFAP), S100β, and Connexin-43 (Cx-43) [19] and microglial Iba-1, CD11b, and CD163 markers [20], at least at initial stages of culture. Furthermore, they exhibit features of cellular stress and inflammatory activation [21], a highly proliferative phenotype [19], and induce extensive gliosis after focal transplantation into the spinal cord of nTg rats [22]. Thus, detailed characterization of their toxic phenotype may contribute to understanding pathogenic mechanisms and therapeutic development in ALS.…”

Section: Introductionmentioning

confidence: 99%

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Mitochondrial Modulation by Dichloroacetate Reduces Toxicity of Aberrant Glial Cells and Gliosis in the SOD1G93A Rat Model of Amyotrophic Lateral Sclerosis

et al. 2019

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Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by motor neuron (MN) degeneration and gliosis. Neonatal astrocytes obtained from the SOD1G93A rat model of ALS exhibit mitochondrial dysfunction and neurotoxicity that can be reduced by dichloroacetate (DCA), a metabolic modulator that has been used in humans, and shows beneficial effects on disease outcome in SOD1G93A mice. Aberrant glial cells (AbGC) isolated from the spinal cords of adult paralytic SOD1G93A rats exhibit highly proliferative and neurotoxic properties and may contribute to disease progression. Here we analyze the mitochondrial activity of AbGC and whether metabolic modulation would modify their phenotypic profile. Our studies revealed fragmented mitochondria and lower respiratory control ratio in AbGC compared to neonatal SOD1G93A and nontransgenic rat astrocytes. DCA (5 mM) exposure improved AbGC mitochondrial function, reduced their proliferative rate, and importantly, decreased their toxicity to MNs. Furthermore, oral DCA administration (100 mg/kg, 10 days) to symptomatic SOD1G93A rats reduced MN degeneration, gliosis, and the number of GFAP/S100β double-labeled hypertrophic glial cells in the spinal cord. DCA treatment of AbGC reduced extracellular lactate levels indicating that the main recognized DCA action, targeting the pyruvate dehydrogenase kinase/pyruvate dehydrogenase complex, may underlie our findings. Our results show that AbGC metabolic phenotype is related to their toxicity to MNs and indicate that its modulation can reduce glial mediated pathology in the spinal cord. Together with previous findings, these results further support glial metabolic modulation as a valid therapeutic strategy in ALS.

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Section: Discussionsupporting

confidence: 76%

Section: Introductionmentioning

confidence: 99%

Mitochondrial Modulation by Dichloroacetate Reduces Toxicity of Aberrant Glial Cells and Gliosis in the SOD1G93A Rat Model of Amyotrophic Lateral Sclerosis

et al. 2019

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“…Further evidence for the aberrant nature of cultured aberrant glial cells isolated from SOD1 G93A symptomatic rats has been obtained from ultrastructural analysis . Cells show an absence of intermediate filaments, an abundance of microtubules together with an important production of extracellular matrix components, suggesting a pro‐fibrotic activity.…”

Section: From Activated Glial Cells To the Emergence Of Aberrant Phenmentioning

confidence: 99%

“…Further evidence for the aberrant nature of cultured aberrant glial cells isolated from SOD1 G93A symptomatic rats has been obtained from ultrastructural analysis. 67 Cells show an absence of intermediate filaments, an abundance of microtubules together with an important production of extracellular matrix components, suggesting a pro-fibrotic activity. In addition, cells showed exacerbated endoplasmic reticulum stress together with a significant abundance of lipid droplets, autophagy images and many heterogeneous formations including vesicles, suggesting a role in secretion.…”

Section: Ultrastructural Features Of Aberrant Glial Cellsmentioning

confidence: 99%

Phenotypic heterogeneity of astrocytes in motor neuron disease

Trías

Barbeito

Yamanaka

2018

Clinical & Exp Neuroim

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Accumulating evidence has shown that astrocytes do not just support the function of neurons, but play key roles in maintaining the brain environment in health and disease. Contrary to the traditional understanding of astrocytes as static cells, reactive astrocytes possess more diverse functions and phenotypes than previously predicted. In the present focused review, we summarize the evidence showing that astrocytes are playing profound roles in the disease process of amyotrophic lateral sclerosis. Aberrantly activated astrocytes in amyotrophic lateral sclerosis rodents express microglial molecular markers and provoke toxicities to accelerate disease progression. In addition, TIR domain–containing adapter protein–inducing interferon‐β‐dependent innate immune pathway in astrocytes also has a novel function in terminating glial activation and neuroinflammation. Furthermore, heterogeneity in phenotypes and functions of astrocytes are also observed in various disease conditions, such as other neurodegenerative diseases, ischemia, aging and acute lesions in the central nervous system. Through accumulating knowledge of the phenotypic and functional diversity of astrocytes, these cells will become more attractive therapeutic targets for neurological diseases.

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“…described previously(Jiménez-Riani et al, 2017), and ultrathin sections were observed with a Jeol JEM1010 transmission electron microscope.3 | RESULTS3.1 | Effect of a coverslip hypoxia variant on LNCaP cell morphology, hypoxic state, and extracellular pHWe modified previous versions of the coverslip hypoxia method by culturing cells in the wells of 35-mm glass bottom dishes and sealing the wells with coverslips larger than the diameter of the well. The coverslips…”

mentioning

confidence: 99%

Using a variant of coverslip hypoxia to visualize tumor cell alterations at increasing distances from an oxygen source

Arocena

Landeira

Paolo

et al. 2019

Journal Cellular Physiology

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Early stages in tumor development involve growth in confined spaces, where oxygen diffusion is limited and metabolic waste products accumulate. This hostile microenvironment imposes strong selective pressures on tumor cells, leading eventually to the survival and expansion of aggressive subclones that condition further tumor evolution. To model features of this microenvironment in vitro, a diffusional barrier can be introduced in the form of a coverslip placed on top of cells, a method termed coverslip hypoxia. Using a variant of this method, with larger volume between coverslip and cells and with oxygen diffusion occurring only through a small hole in the center of the coverslip, we have visualized alterations in LNCaP tumor cells as a function of their distance to the oxygen source at the center. We observed remarkable morphological changes in LNCaP cells as the distance from the center increases, with cells becoming highly spread, displaying dynamic membrane protrusions and occasionally adopting a migratory phenotype. Concomitantly, cells farther from the center displayed marked increases in the hypoxia marker hypoxyprobe, whereas extracellular pH decreased in the same direction. Cells with altered morphology displayed prominent increases in fibrillar actin, as well as swollen mitochondria with distorted cristae and accumulation of neutral lipid‐containing intracellular vesicles. These results show that an in vitro microenvironment that models diffusional barriers encountered by tumors in situ can have profound effects on tumor cells. The coverslip hypoxia variant we describe can be used to characterize in vitro the response of tumor cells to environmental conditions that play crucial roles in early tumor development.

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Ultrastructural features of aberrant glial cells isolated from the spinal cord of paralytic rats expressing the amyotrophic lateral sclerosis-linked SOD1G93A mutation

Cited by 26 publications

References 51 publications

Mitochondrial Modulation by Dichloroacetate Reduces Toxicity of Aberrant Glial Cells and Gliosis in the SOD1G93A Rat Model of Amyotrophic Lateral Sclerosis

Mitochondrial Modulation by Dichloroacetate Reduces Toxicity of Aberrant Glial Cells and Gliosis in the SOD1G93A Rat Model of Amyotrophic Lateral Sclerosis

Phenotypic heterogeneity of astrocytes in motor neuron disease

Using a variant of coverslip hypoxia to visualize tumor cell alterations at increasing distances from an oxygen source

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