Primary Cultures from Rat Dorsal Root Ganglia: Responses of Neurons and Glial Cells to Somatosensory or Inflammatory Stimulation

Leisengang, Stephan; Ott, Daniela; Murgott, Jolanta; Gerstberger, Rüdiger; Rummel, Christoph; Roth, Joachim

doi:10.1016/j.neuroscience.2018.10.018

Cited by 25 publications

(57 citation statements)

References 57 publications

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“…At these time points, we also observed both elevated TNF gene expression in lumbar ganglions (L3-L5) and increased amounts of TNF in L4 DRG. Our observations are consistent with a previous study showing that TNF expression in the DRG environment is produced by macrophages (47,59). As a result, we suggested that articular hyperalgesia remaining after the resolving phase in AIA is triggered by overexpression of TNF, possibly from non-neuronal cells activated in the DRG.…”

Section: Discussionsupporting

confidence: 93%

“…Our results show an increase in Iba1 + cells in DRG associated with elevated TNF, IL-1β, IL-6, COX2, and iNOS gene expression in situ. Our observations are consistent with the role of inflammatory molecules for the sensitization of neurons in DRG and the development of hyperalgesia (47)(48)(49)(50)(51)(52)(53). Interestingly, this neuroinflammation within the DRG was corroborated with the activation of leukocyte-endothelium interactions in the lumbar intumescence.…”

Section: Discussionsupporting

confidence: 89%

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Sensory Ganglia-Specific TNF Expression Is Associated With Persistent Nociception After Resolution of Inflammation

et al. 2020

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systemic injection of etanercept, an agent clinically utilized for TNF neutralization, at day 7 post arthritis induction, alleviated the persistent joint hyperalgesia by specific action in DRG. Our data suggest that neuroinflammation in DRG after the resolution of acute joint inflammation drives continuous neural sensitization resulting in persistent joint nociception in a TNF-dependent mechanism.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 89%

Sensory Ganglia-Specific TNF Expression Is Associated With Persistent Nociception After Resolution of Inflammation

et al. 2020

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“…Primary rat cultures were prepared as previously described (Soares et al, 2013). To stimulate the cells, they were incubated with LPS (Sigma-Aldrich, St. Louis, MO, USA; 10 µg/ml) or recombinant rat Cygb (Cusabio Technology, Houston, TX, USA; 10 and 20 µg/ml) for 4 h. The concentrations of LPS and Cygb used were based on previous studies (Wen et al, 2017;Leisengang et al, 2018). PBS was used as a negative control.…”

Section: Preoptic Area Primary Cell Culturesmentioning

confidence: 99%

“…previously published protocols (Leisengang et al, 2018). Primary antibodies were used in the following concentrations: anti-NF-IL6 (1:4,000), anti-NF-κB (1:2,000), anti-STAT3 (1:4,000), anti-GFAP (1:1,000), anti-ED-1 (1:1,000).…”

Section: Immunocytochemistrymentioning

confidence: 99%

Cytoglobin Attenuates Neuroinflammation in Lipopolysaccharide-Activated Primary Preoptic Area Cells via NF-κB Pathway Inhibition

Gomes

Sousa

Ott

et al. 2019

Front. Mol. Neurosci.

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Cytoglobin (Cygb) is a hexacoordinate protein, associated with the transport of oxygen, nitric oxide scavenging, tumor suppression and protection against oxidative stress and inflammation. This protein is expressed in brain areas including the preoptic area (POA) of the anterior hypothalamus, the region responsible for the regulation of body temperature. In this study, we show that Cygb is upregulated in the rat hypothalamus 2.5 h and 5 h after intravenous administration of lipopolysaccharide (LPS). We investigated the effect of treatment with Cygb in POA primary cultures stimulated with LPS for 4 h. The levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were measured and the results showed that Cygb reduced the concentrations of both cytokines. We further observed a decrease in immunoreactivity of the inflammatory transcription factor nuclear factor-κB (NF-κB), but not NF-IL6 and STAT3, in the nucleus of Cygb-treated POA cells. These findings suggest that Cygb attenuates the secretion of IL-6 and TNF-α in LPS-stimulated POA primary cultures via inhibition of the NF-κB signaling pathway, indicating that this protein might play an important role in the control of neuroinflammation and fever.

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“…After incubation, the supernatants were removed from the cells and stored at − 20 °C for later measurement of IL-6. The LPS doses to stimulate neuro-glial primary cultures of chicken were chosen according to own previous studies (Ott et al 2010;Simm et al 2016;Leisengang et al 2018b) and to pilot experiments.…”

Section: Lps Stimulation Of Chicken Hypothalamus Primary Cultures Andmentioning

confidence: 99%

Neurons and astrocytes of the chicken hypothalamus directly respond to lipopolysaccharide and chicken interleukin-6

et al. 2020

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In 4-5-month-old chicken, intravenous injections of bacterial lipopolysaccharide (LPS) induced a dose-dependent fever response and a pronounced increase of circulating interleukin-6 (IL-6). To assess a possible role for IL-6 in the brain of birds, a hypothalamic neuro-glial primary culture from 1-day-old chicken was established. Each well of cultured hypothalamic cells contained some 615 neurons, 1350 astrocytes, and 580 microglial cells on average. Incubation of chicken hypothalamic primary cultures with 10 or 100 µg/ml LPS induced a dose-dependent release of bioactive IL-6 into the supernatant. Populations of hypothalamic neurons (4%) and astrocytes (12%) directly responded to superfusion with buffer containing 10 µg/ ml LPS with a transient increase of intracellular calcium, a sign of direct cellular activation. Stimulation of hypothalamic cultures with buffer containing 50 ng/ml chicken IL-6 induced calcium signaling in 11% of neurons and 22% of astrocytes investigated. These results demonstrate that IL-6 is produced in the periphery and in the hypothalamus in response to LPS in chicken. The observed cellular responses of hypothalamic cells to chicken IL-6 indicate that this cytokine may readily be involved in the manifestation of fever in the avian hypothalamus.

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Primary Cultures from Rat Dorsal Root Ganglia: Responses of Neurons and Glial Cells to Somatosensory or Inflammatory Stimulation

Cited by 25 publications

References 57 publications

Sensory Ganglia-Specific TNF Expression Is Associated With Persistent Nociception After Resolution of Inflammation

Sensory Ganglia-Specific TNF Expression Is Associated With Persistent Nociception After Resolution of Inflammation

Cytoglobin Attenuates Neuroinflammation in Lipopolysaccharide-Activated Primary Preoptic Area Cells via NF-κB Pathway Inhibition

Neurons and astrocytes of the chicken hypothalamus directly respond to lipopolysaccharide and chicken interleukin-6

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