Correction of humoral derangements from mutant superoxide dismutase 1 spinal cord.
OBJECTIVE
We sought to define molecular and cellular participants that mediate motor neuron injury in amyotrophic lateral sclerosis using a coculture system.


METHODS
We cocultured embryonic stem cell-derived motor neurons with organotypic slice cultures from wild-type or SOD1G93A (MT) mice. We examined axon lengths and cell survival of embryonic stem cell-derived motor neurons. We defined and quantified the humoral factors that differed between wild-type and MT organotypic cultures, and then corrected these differences in cell culture.


RESULTS
MT spinal cord organotypic slices were selectively toxic to motor neurons as defined by axonal lengths and cell survival. MT spinal cord organotypic slices secreted higher levels of nitric oxide, interleukin (IL)-1beta, IL-6, and IL-12p70 and lower levels of vascular endothelial growth factor. The toxicity of MT spinal cord organotypic cultures was reduced and axonal lengths were restored to near normal by coculturing in the presence of reactive oxygen species scavenger, vascular endothelial growth factor, and neutralizing antibodies to IL-1beta, IL-6, and IL-12.


CONCLUSIONS
MT spinal cord organotypic cultures overexpress certain factors and underexpress others, creating a nonpermissive environment for cocultured motor neurons. Correction of these abnormalities as a group, but not individually, restores axonal length to near normal. Such a "cocktail" approach to the treatment of amyotrophic lateral sclerosis should be investigated further.
Kim
Yun-Sook
YS
Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Martinez
Tara
T
Deshpande
Deepa M
DM
Drummond
Jennifer
J
Provost-Javier
Katie
K
Williams
Ariel
A
McGurk
Julie
J
Maragakis
Nicholas
N
Song
Hongjun
H
Ming
Guo-Li
GL
Kerr
Douglas A
DA
D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D000818:Animals; D000906:Antibodies / Q000494:pharmacology* / Q000627:therapeutic use; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000633:toxicity; D004195:Disease Models, Animal; D005260:Female; D016166:Free Radical Scavengers / Q000494:pharmacology* / Q000627:therapeutic use; D006801:Humans; D007378:Interleukins / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000633:toxicity; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009046:Motor Neurons / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D009416:Nerve Regeneration / Q000187:drug effects / Q000502:physiology; D009569:Nitric Oxide / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000633:toxicity; D009924:Organ Culture Techniques; D013116:Spinal Cord / Q000187:drug effects / Q000201:enzymology / Q000503:physiopathology; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D016896:Treatment Outcome; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology / Q000627:therapeutic use*
D000906:Antibodies; D017077:Culture Media, Conditioned; D016166:Free Radical Scavengers; D007378:Interleukins; D042461:Vascular Endothelial Growth Factor A; D009569:Nitric Oxide; C109415:SOD1 G93A protein; D013482:Superoxide Dismutase