Effects of repeated nerve injuries at different time intervals on functional recovery and nerve innervation.

Effects of repeated nerve injuries on functional recovery and nerve innervation were examined in rodents. Crush injuries of the sciatic nerve were inflicted on adult rats and repeated twice or thrice at different time intervals of 1, 2, 3, and 4 weeks. Motor function was assessed by the static sciatic index at 1, 7, 14, 21, 28, 35, 42, 49, and 56 days after the final crush. The rates of nerve innervation of the tibialis anterior muscle, a main muscle innervated by the common peroneal nerve, were evaluated by the quantification of ßIII-tubulin-positive nerve terminals and α-bungarotoxin-positive acetylcholine receptors 21 and 56 days after the final crush of triple nerve injuries at 1-, 2-, 3-, and 4-week intervals. Compared with single nerve crush injury, delayed recovery of motor function was observed in repeated crush injuries. In addition, recoveries in the triple crush groups were slower than those in the double crush groups. The rates of reinnervation were lower in the triple crush groups than in the single crush groups, both at 21 days (single: 59.7%; triple: 54.1%-56.1%) and 56 days (single: 88.8%; triple: 72.5%-83.0%) after the final crush, except in the groups with 1-week (triple: 73.8%) and 2-week (triple: 70.5%) intervals at 21 days after the final crush. We concluded that the recovery of motor function was delayed according to the number of repetitions of crush injuries, and that the rates of nerve innervation were still low in the triple crush groups 8 weeks after the final crush.

Crush injury induces NMDA-receptor-dependent delayed nerve cell death in rat entorhinal-hippocampal slice cultures.

The aim of the present study was to develop an in vitro model that produces an injury-induced delayed nerve cell death. We used entorhinal-hippocampal slices from 12-day-old rats, which were cultured for 14 days before experiments prior to a crush injury in the middle layers of the entorhinal cortex. Crush injury increased specific propidium iodide (PI) fluorescence at the site of primary injury at day 1. Specific PI fluorescence decreased over the following days, with a slower decrease taking place at days 4-7. At days 4-7, PI fluorescence increased in CA1 and CA3. Treatment with 30 microM MK-801 for 2 h at the time of crush injury prevented the increase in PI fluorescence at days 4-7 at the site of injury, in CA1, and in CA3. Crush injury induced TUNEL-positive cells at the site of injury at days 1 and 5. MK-801 markedly reduced the number of TUNEL-positive cells observed at day 5, but had no effect on the number of TUNEL-positive cells at day 1. The present data indicate that crush injury may induce two types of secondary cell death. The first, early, type of cell death is induced quickly and is maximal at day 1, after which the dead cells are gradually removed. The second, delayed, type of cell death appears at day 4. NMDA receptor antagonism prevents the induction of the delayed cell death but has no effect on the early cell death.