Axotomy induces a transient and localized elevation of the free intracellular calcium concentration to the milimolar range. Ziv, Noam E., Micha E. Spira. From.
APStracts 2:0197N, 1995.
SUMMARY AND CONCLUSIONS
1. Axonal transection triggers a cascade of pathological processes that frequently lead to the degeneration of the injured neuron. It is generally believed that the degenerative process is triggered by an overwhelming influx of calcium through the cut end of the axon. 2. Theoretical considerations and indirect observations suggest that axotomy is followed by an increase in the free intracellular calcium concentration ([Ca 2+ ] i ) to the milimolar level. In contrast, only relatively modest and transient elevation in [Ca 2+ ] i to the micromolar level was revealed by recent fura-2 studies. 3. In the current study we have used the low-affinity Ca 2+ indicator mag-fura-2 (furaptra) to reexamine the spatiotemporal distribution pattern of Ca 2+ following axotomy and to map the free intracellular Mg 2+ concentration gradients. 4. We report that axotomy elevates [Ca 2+ ] i well beyond the "physiological" range of calcium concentrations, to levels exceeding 1 milimolar near the tip of the cut axon and to hundreds of micromolars along the axon further away from the cut end. Nevertheless, [Ca 2+ ] i recovers to the control levels within 2-3 minutes following the resealing of the cut end. 5. A comparison of the behavior of fura-2 and mag-fura-2 in the cytosol of the axotomized neurons reveals that the determination of [Ca 2+ ] i by fura-2 largely underestimates the actual intracellular Ca 2+ concentrations. 6. Experiments in which one branch of a bifurcated axon was transected revealed that the elevation in [Ca 2+ ] i is confined to the transected axonal branch, and does not spread beyond the bifurcation point. 7. Following axotomy, the intracellular Mg 2+ concentration equilibrates rapidly with the external concentration, and then recovers at a rate somewhat slower than that of [Ca 2+ ] i . 8. To the best of our knowledge, this study is the first direct demonstration that axotomy elevates [Ca 2+ ] i to the milimolar range and that neurons are able to recover from these extreme calcium concentrations.

Received 10 January 1995; accepted in final form 20 June 1995.
APS Manuscript Number J24-5.
Article publication pending J. Neurophysiol.
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on 30 July 1995.