Trauma-induced neurotoxicity in rat hippocampal neurons.
OBJECTIVE
We have previously shown that traumatic injury of hippocampal cells triggers release of a soluble neurotoxin that can be transferred to an uninjured culture. The mechanism of this trauma-induced neurotoxicity is independent of glutamate receptor activation. We extended this observation to study the mechanism of this neurotoxicity.


METHODS
Dissociated rat hippocampal neurons were traumatized by disrupting the culture by scratching the plate. The toxicity expressed by the injured culture was studied by transferring the medium to an uninjured culture and assessing the death rate by trypan blue exclusion.


RESULTS
This neurotoxin is stable in the medium at room temperature for several hours and withstands boiling. The molecular weight is between 100 and 500. The release and the effect of this toxin seem to be independent of glutamate receptor activation. The toxicity is unaffected by removal of extracellular calcium. However, dantrolene dose-dependently blocked the toxicity in the recipient culture, suggesting that the release of intracellular stores of calcium is involved in the toxic effect. This release of calcium is likely to be followed by an activation of nitric oxide synthase because competitive nitric oxide synthase inhibitors attenuated this toxicity. Consistent with this result, cholecystokinin octapeptide significantly reduced cell death when combined with this toxic medium.


CONCLUSIONS
Traumatic injury of dissociated cells can propagate neurotoxicity in uninjured cells by a soluble toxin released into the extracellular space. This toxin causes a rise in cytosolic calcium that activates nitric oxide synthase that can be blocked by cholecystokinin.
Yoon
K W
KW
Department of Surgery, St Louis, Mo University Health Science Center, MO 63110-0250, USA.
Mitchell
H L
HL
Broder
L D
LD
Brooker
R W
RW
Delisle
R K
RK
D000818:Animals; D002118:Calcium / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D004396:Coloring Agents; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D003620:Dantrolene / Q000008:administration & dosage / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006624:Hippocampus / Q000187:drug effects / Q000201:enzymology / Q000293:injuries* / Q000378:metabolism*; D008970:Molecular Weight; D009474:Neurons / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D018696:Neuroprotective Agents / Q000494:pharmacology; D009498:Neurotoxins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017470:Receptors, Glutamate / Q000378:metabolism; D012844:Sincalide / Q000494:pharmacology; D014343:Trypan Blue
D004396:Coloring Agents; D018696:Neuroprotective Agents; D009498:Neurotoxins; D017470:Receptors, Glutamate; D019001:Nitric Oxide Synthase; D003620:Dantrolene; D014343:Trypan Blue; D012844:Sincalide; D002118:Calcium