Expression of GAP‐43 mRNA in Mouse Spinal Cord Following Unilateral Peripheral Nerve Damage: is There a Contralateral Effect?

Abstract

Two new oligonucleotide anti-sense probes and their corresponding sense probes specific for mouse GAP-43 mRNA were synthesized and end-labelled with digoxygenin. They were used to localize GAP-43 mRNA in the spinal cords of normal mice and in mice 3 and 7 days following unilateral sciatic nerve cut. GAP-43 mRNA was found to be expressed at low levels in motor and other neurons of the normal spinal cords. As expected from other studies, up-regulation occurred in the cell bodies of axotomized motor neurons but, in addition, up-regulation was also observed in the cell bodies of intact motor neurons contralateral to the lesion. Densitometer measurements showed that the up-regulation of GAP-43 mRNA was less in the intact, contralateral motor neuron cell bodies than in the axotomized motor neuron cell bodies and furthermore was transient, being higher at 3 days than at 7 days following axotomy. Both anti-sense probes gave the same result, although differences in cellular localization was observed, and the two sense probes were negative. Probe binding was abolished by pretreatment of the sections with ribonuclease and hybridization was carried out under different conditions of stringency in order to ascertain whether the contralateral expression of GAP-43 mRNA was a true reflection of its distribution in vivo. There is conflicting evidence on the presence or absence of contralateral effects following unilateral peripheral nerve injury in the literature, and it is suggested that these differences can be accounted for by the methodology and type of probe used.