title: Neurotoxic injury pathways in differentiated mouse motor neuron-neuroblastoma hybrid (NSC-34D) cells in vitro--limited effect of riluzole on thapsigargin, but not staurosporine, hydrogen peroxide and homocysteine neurotoxicity.
abstract: The neuroblastoma-spinal motor neuron fusion cell line, NSC-34, in its differentiated form, NSC-34D, permits examining the effects of riluzole, a proven treatment for amyotrophic lateral sclerosis (ALS) on cell death induction by staurosporine (STS), thapsigargin (Thaps), hydrogen peroxide (H(2)O(2)) and homocysteine (HCy). These neurotoxins, applied exogenously, have mechanisms of action related to the various proposed molecular pathogenetic pathways in ALS and are differentiated from endogenous cell death that is associated with cytoplasmic aggregate formation in motor neurons. Nuclear morphology, caspase-3/7 activation and high content imaging were used to assess toxicity of these neurotoxins with and without co-treatment with riluzole, a benzothiazole compound with multiple pharmacological actions. STS was the most potent neurotoxin at killing NSC-34D cells with a toxic concentration at which 50% of maximal cell death is achieved (TC(50)=0.01μM), followed by Thaps (TC(50)=0.9μM) and H(2)O(2) (TC(50)=15μM) with HCy requiring higher concentrations to kill at the same level (TC(50)=2200μM). Riluzole provided neurorescue with a 20% absolute reduction (47.6% relative reduction) in apoptotic cell death against Thaps-induced NSC-34D cell (p≤0.05), but had no effect on STS-, H(2)O(2)- and HCy-induced NSC-34D cell death. This effect of riluzole on Thaps induction of cell death was independent of caspase-3/7 activation. Riluzole mitigated a toxin that can cause intracellular calcium dysregulation associated with endoplasmic reticulum (ER) stress but not toxins associated with other cell death mechanisms.
authors: ALS Translational Neuroscience Laboratory, Carolinas Medical Center, Charlotte, NC 28203, USA. richelle.hemendinger@carolinashealthcare.org
authors: Richelle A
authors: RA
authors: Hemendinger
authors: Edward J
authors: EJ
authors: Armstrong
authors: Nick
authors: N
authors: Radio
authors: Benjamin Rix
authors: BR
authors: Brooks
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000188:drug therapy* / Q000503:physiopathology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002118:Calcium / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006710:Homocysteine / Q000008:administration & dosage / Q000633:toxicity; D006822:Hybrid Cells; D006861:Hydrogen Peroxide / Q000008:administration & dosage / Q000633:toxicity; D051379:Mice; D009046:Motor Neurons / Q000187:drug effects / Q000378:metabolism; D009447:Neuroblastoma / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009498:Neurotoxins / Q000008:administration & dosage / Q000633:toxicity*; D019782:Riluzole / Q000494:pharmacology*; D019311:Staurosporine / Q000008:administration & dosage / Q000633:toxicity; D019284:Thapsigargin / Q000008:administration & dosage / Q000633:toxicity
chemical_list: D018696:Neuroprotective Agents; D009498:Neurotoxins; D006710:Homocysteine; D019284:Thapsigargin; D019782:Riluzole; D006861:Hydrogen Peroxide; D053148:Caspase 3; D053179:Caspase 7; D019311:Staurosporine; D002118:Calcium