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    Consequences of Neurite Transection In Vitro
    (MARY ANN LIEBERT INC, 2012) Cengiz, Nurettin; Ozturk, Gurkan; Erdogan, Ender; Him, Aydin; Oguz, Elif Kaval
    In order to quantify degenerative and regenerative changes and analyze the contribution of multiple factors to the outcome after neurite transection, we cultured adult mouse dorsal root ganglion neurons, and with a precise laser beam, we transected the nerve fibers they extended. Cell preparations were continuously visualized for 24 h with time-lapse microscopy. More distal cuts caused a more elongated field of degeneration, while thicker neurites degenerated faster than thinner ones. Transected neurites degenerated more if the uncut neurites of the same neuron simultaneously degenerated. If any of these uncut processes regenerated, the transected neurites underwent less degeneration. Regeneration of neurites was limited to distal cuts. Unipolar neurons had shorter regeneration than multipolar ones. Branching slowed the regenerative process, while simultaneous degeneration of uncut neurites increased it. Proximal lesions, small neuronal size, and extensive and rapid neurite degeneration were predictive of death of an injured neuron, which typically displayed necrotic rather than apoptotic form. In conclusion, this in vitro model proved useful in unmasking many new aspects and correlates of mechanically-induced neurite injury.
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    A new method in CNS (Central nervous system) in vitro cultures in the mouse: Study of effectiveness
    (KAFKAS UNIV, VETERINER FAKULTESI DERGISI, 2010) Erdogan, Ender; Ozturk, Gurkan; Ragbetli, Murat Cetin
    In this study: to evaluate the effectiveness of collagen coating method, using in the peripheral nervous system cultures. and its involving factors caused from manipulations in central nervous system (CNS) cultures was aimed. Via frontal approach, brains, transected from young Swiss albino mice, were taken into artificial cerebro-spinal fluid immediately and made blocks in agarose gel. With a vibration microtome, 200 pm thickness horizontally live slices were taken in to the dishes filled with culture medium. Tissue sections were analyzed as two groups. In the group 1 (control): fresh slices were evaluated directly. In the group 2: sections were covered with collagen gel (Type I) and left in the incubator (5% CO(2)) for 3 days. These sections were dyed with calcein and propidium iodide for viability and non-viability and then observed with confocal laser scanning microscope. Images were captured digitally and examined. Since negative effects of high melting temperature of standard agar on the livability, using low melting agar to tissue blocking and high frequency - low speed vibrotome setting to cut were more preferably. In the 3 days cultures, viability/nonviability rates were indicated better values. It is concluded that, in the CNS slicing cultures, collagen coating method was an easier, effective, useful and alternative method to present techniques.