title: Cellular changes in motor neuron cell culture produced by cytotoxic cerebrospinal fluid from patients with amyotrophic lateral sclerosis.
abstract: BACKGROUND
  The neurotoxic effects of cerebrospinal fluid (CSF) from patients with amyotrophic lateral sclerosis (ALS) have been reported by various authors who have attributed this neurotoxicity to the glutamate in CSF-ALS.
  
  
  METHODS
  Cultures of rat embryonic cortical neurons were exposed to CSF from ALS patients during an incubation period of 24 hours. Optical microscopy was used to compare cellular changes to those elicited by exposure to 100μm glutamate, and confocal microscopy was used to evaluate immunohistochemistry for caspase-3, TNFα, and peripherin.
  
  
  RESULTS
  In the culture exposed to CSF-ALS, we observed cells with nuclear fragmentation and scarce or null structural modifications to the cytoplasmic organelles or to plasma membrane maintenance. This did not occur in the culture exposed to glutamate. The culture exposed to CSF-ALS also demonstrated increases in caspase-3, TNFα, and in peripherin co-locating with caspase-3, but not with TNFα, suggesting that TNFα may play an early role in the process of apoptosis.
  
  
  CONCLUSIONS
  CFS-ALS cytotoxicity is not related to glutamate. It initially affects the nucleus without altering the cytoplasmic membrane. It causes cytoplasmic apoptosis that involves an increase in caspase-3 co-located with peripherin, which is also overexpressed.
authors.affiliation: Instituto de Neurociencias, IdISSC, Hospital Clínico San Carlos, Universidad Complutense, Madrid, España. Electronic address: inc.hcsc@salud.madrid.org.
authors.forename: U
authors.initials: U
authors.lastname: Gomez-Pinedo
authors.affiliation: Instituto Teófilo Hernando, Departamento de Farmacología, Universidad Autónoma de Madrid, Madrid, España.
authors.forename: M
authors.initials: M
authors.lastname: Yáñez
authors.affiliation: Instituto de Neurociencias, IdISSC, Hospital Clínico San Carlos, Universidad Complutense, Madrid, España.
authors.forename: J
authors.initials: J
authors.lastname: Matías-Guiu
authors.affiliation: Instituto de Neurociencias, IdISSC, Hospital Clínico San Carlos, Universidad Complutense, Madrid, España.
authors.forename: L
authors.initials: L
authors.lastname: Galán
authors.affiliation: Instituto de Neurociencias, IdISSC, Hospital Clínico San Carlos, Universidad Complutense, Madrid, España.
authors.forename: A
authors.initials: A
authors.lastname: Guerrero-Sola
authors.affiliation: Instituto de Neurociencias, IdISSC, Hospital Clínico San Carlos, Universidad Complutense, Madrid, España.
authors.forename: M S
authors.initials: MS
authors.lastname: Benito-Martin
authors.affiliation: Instituto de Neurociencias, IdISSC, Hospital Clínico San Carlos, Universidad Complutense, Madrid, España.
authors.forename: A
authors.initials: A
authors.lastname: Vela
authors.affiliation: Instituto Teófilo Hernando, Departamento de Farmacología, Universidad Autónoma de Madrid, Madrid, España.
authors.forename: J A
authors.initials: JA
authors.lastname: Arranz-Tagarro
authors.affiliation: Instituto Teófilo Hernando, Departamento de Farmacología, Universidad Autónoma de Madrid, Madrid, España.
authors.forename: A G
authors.initials: AG
authors.lastname: García
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000134:cerebrospinal fluid*; D000818:Animals; D002478:Cells, Cultured; D002555:Cerebrospinal Fluid / Q000737:chemistry / Q000378:metabolism*; D003603:Cytotoxins / Q000494:pharmacology; D018698:Glutamic Acid / Q000494:pharmacology*; D006801:Humans; D009046:Motor Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051381:Rats
chemical_list: D003603:Cytotoxins; D018698:Glutamic Acid
keywords: Amyotrophic lateral sclerosis; Caspasa-3; Caspase-3; Cerebrospinal fluid; Esclerosis lateral amiotrófica; Glutamate; Glutamato; Líquido cefalorraquídeo; Neurotoxicidad; Neurotoxicity; Periferina; Peripherin; TNFα