Naslov
Delayed excitotoxicity evoked by kainic acid in spinal cord organotypic cultures as simple in vitro model of spinal injury.

Autori
Mazzone GL ; Mladinić Pejatović, Miranda ; Nistri A.

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Skup
FENS 2010

Mjesto i datum
Amsterdam, Nizozemska, 03.-07.07.10

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
excitotoxicity; metabolic dysfunction; spinal cord injury.

Sažetak
Spinal organotypic cultures represent a useful model to investigate certain molecular and cellular alterations, as well as elementary circuit characteristics following experimental conditions that mimic a spinal cord lesion (Sibilla and Ballerini, Prog Neurobiol 2009, 89:46). In our previous studies using the neonatal rat spinal cord in vitro, a potent excitotoxic stimulus with the glutamate analogue kainate significantly reduced the number of neurons and irreversibly suppressed fictive locomotion (Taccola et al., Neuroscience 2008, 155:538). For longer term studies and pharmacological applications, we investigated organotypic cultures to evaluate the consequences of kainate application on cell survival with particular focus on dose- dependence, timeframe and topographical distribution of kainate excitotoxicity. Rat organotypic spinal slices (22 days in vitro) were treated with kainate (0.1 or 1 mM) for 1 h, and then washed with standard medium for variable times prior to analysis. Both doses of kainate significantly increased the percent of pyknotic nuclei (observed with DAPI staining): this process was already detected at 1 h and gradually intensified 4 and 24 h later. However, the rate of cell loss was faster with the higher dose of kainate. To characterize the topography of kainate-mediated effects, cells were counted in three regions, namely dorsal, central, and ventral. The largest number of pyknotic nuclei was found in dorsal areas. We next examined if kainate toxicity preferentially affected neurons or motoneurons using selective immunocytochemical markers. Impaired metabolic activity of these cultures, evaluated by mitochondrial toxicity test with methyl-tiazol-tetrazolium, suggested bioenergetic collapse. Our data indicate delayed excitotoxicity, thus implying a time window for development of strong damage and for devising new neuroprotective strategies

Izvorni jezik
Engleski

POVEZANOST RADA