title: Astroglia acquires a toxic neuroinflammatory role in response to the cerebrospinal fluid from amyotrophic lateral sclerosis patients.
abstract: Non-cell autonomous toxicity is one of the potential mechanisms implicated in the etiopathogenesis of amyotrophic lateral sclerosis (ALS). However, the exact role of glial cells in ALS pathology is yet to be fully understood. In a cellular model recapitulating the pathology of sporadic ALS, we have studied the inflammatory response of astroglia following exposure to the cerebrospinal fluid from ALS patients (ALS-CSF).
  
  
  
  Various inflammatory markers including pro-inflammatory and anti-inflammatory cytokines, COX-2, PGE-2, trophic factors, glutamate, nitric oxide (NO), and reactive oxygen species (ROS) were analyzed in the rat astroglial cultures exposed to ALS-CSF and compared with the disease control or normal controls. We used immunofluorescence, ELISA, and immunoblotting techniques to investigate the protein expression and real-time PCR to study the messenger RNA (mRNA) expression. Glutamate, NO, and ROS were estimated using appropriate biochemical assays. Further, the effect of conditioned medium from the astroglial cultures exposed to ALS-CSF on NSC-34 motor neuronal cell line was detected using the MTT assay. Statistical analysis was carried out using one-way ANOVA followed by Tukey's post hoc test, or Student's t test, as applicable.
  
  
  
  Here, we report that the ALS-CSF enhanced the production and release of inflammatory cytokines IL-6 and TNF-α, as well as COX-2 and PGE-2. Concomitantly, anti-inflammatory cytokine IL-10 and the beneficial trophic factors, namely VEGF and GDNF, were down-regulated. We also found impaired regulation of glutamate, NO, and ROS in the astroglial cultures treated with ALS-CSF. The conditioned medium from the ALS-CSF exposed astroglial cultures induced degeneration in NSC-34 cells.
  
  
  
  Our study demonstrates that the astroglial cells contribute to the neuroinflammation-mediated neurodegeneration in the in vitro model of sporadic ALS.
authors: Department of Neurophysiology, National Institute of Mental Health and Neurosciences (NIMHANS), Bangalore, 560029, India.|Present address: Centre de Recherche de l'Institut Universitaire en Santé Mentale de Québec (CRIUSMQ), Québec, QC, G1J 2G3, Canada.
authors: Pooja-Shree
authors: PS
authors: Mishra
authors: Department of Neurophysiology, National Institute of Mental Health and Neurosciences (NIMHANS), Bangalore, 560029, India.|Present address: Institute of Pharmaceutical Sciences, UGC-Center of Advanced Study (UGC-CAS), Panjab University, Chandigarh, 160014, India.
authors: Dinesh K
authors: DK
authors: Dhull
authors: Department of Neurology, National Institute of Mental Health and Neurosciences (NIMHANS), Bangalore, 560029, India.
authors: A
authors: A
authors: Nalini
authors: Department of Neurophysiology, National Institute of Mental Health and Neurosciences (NIMHANS), Bangalore, 560029, India.
authors: K
authors: K
authors: Vijayalakshmi
authors: Department of Neurophysiology, National Institute of Mental Health and Neurosciences (NIMHANS), Bangalore, 560029, India.
authors: T N
authors: TN
authors: Sathyaprabha
authors: Department of Neurophysiology, National Institute of Mental Health and Neurosciences (NIMHANS), Bangalore, 560029, India.
authors: Phalguni Anand
authors: PA
authors: Alladi
authors: Department of Neurophysiology, National Institute of Mental Health and Neurosciences (NIMHANS), Bangalore, 560029, India. trraju.nimhans.@gmail.com.
authors: Trichur R
authors: 0000-0002-7742-4050
authors: TR
authors: Raju
mesh_terms: D000328:Adult; D000368:Aged; D000690:Amyotrophic Lateral Sclerosis / Q000134:cerebrospinal fluid* / Q000378:metabolism / Q000473:pathology; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000378:metabolism* / Q000473:pathology; D002478:Cells, Cultured; D002555:Cerebrospinal Fluid / Q000378:metabolism*; D005260:Female; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D051381:Rats; D017208:Rats, Wistar
chemical_list: D018836:Inflammation Mediators
keywords: ALS; Astrocytes; COX-2; Cytokines; Neuroinflammation; ROS; Trophic factors