Additive effect of NOS inhibitor and neurotrophic factors on the survival of injured Clarke's neurons.

The present study examined the effect of treatment with the NOS inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) together with peripheral nerve (PN) graft or brain-derived neurotrophic factor (BDNF) on the survival of CN neurons at the L1 level of the spinal cord following hemisection at T11. In control animals 41% of CN neurons survived 15 days after the hemisection, and 48% of these expressed NOS. Treatment with either PN graft implantation or continuous infusion of BDNF increased the survival rate of CN neurons to 70%; 70% of these expressed NOS. Combined L-NAME and PN graft or L-NAME and BDNF improved the rescue rate up to 94%, but only approximately 33% expressed NOS. Our results suggest that the expression of NOS might adversely influence the neuroprotective function of neurotrophic factors on injured CN neurons in the spinal cord.

Chondroitinase ABC promotes axonal regeneration of Clarke's neurons after spinal cord injury.

We examined whether enzymatic digestion of chondroitin sulfate (CS) promoted the axonal regeneration of neurons in Clarke's nucleus (CN) into a peripheral nerve (PN) graft following injury of the spinal cord. After hemisection at T11, a segment of PN graft was implanted at the lesion site. Either vehicle, brain-derived neurotrophic factor (BDNF) or chondroitinase ABC was applied at the implantation site. The postoperative survival period was 4 weeks. Treatment with vehicle or BDNF did not promote the axonal regeneration of CN neurons into the PN graft. Application of 2.5 unit/ml chondroitinase ABC resulted in a significant increase (12.8%) in the number of regenerated CN neurons. Double labeling with Fluoro-Gold and NADPH-diaphorase histochemistry showed that the regenerated CN neurons did not express nitric oxide synthase (NOS). Our results suggest that CS is inhibitory to the regeneration of CN neurons following injury of the spinal cord.

Time course of NOS expression and neuronal death in Clarke's nucleus following traumatic injury in adult rat spinal cord.

The objective of this study was to investigate the influence of the length of the remaining axons on neuronal survival following axonal injury in the adult rat spinal cord. The relationship between the expression of nitric oxide synthase (NOS) and neuronal death was also studied. The time course of NOS expression and neuronal loss was observed in Clarke's nucleus at segment L1 following hemisection of the cord at either segment C3, T1, or T11. NOS-positive neurons were first observed 3 days after injury, rising significantly after 5 days, and reaching a maximum by 10-20 days post-injury. During the same period the number of neurons surviving in the nucleus decreased. The decrease began at 3 days, rapidly decreased after 5 days, and reached a minimum by 40 days post-injury. Among the three lesioned groups, the highest percentage of NOS expression and neuronal loss was observed in the group with lesions at T11, while the lowest percentage was in the group with lesions at C3. Although the time course of neuronal loss varied following lesion at different spinal levels, in all cases the majority of neurons in Clarke's nucleus had degenerated by 40 days after axotomy.

Peripheral nerve graft and neurotrophic factors enhance neuronal survival and expression of nitric oxide synthase in Clarke's nucleus after hemisection of the spinal cord in adult rat.

The present study examined the effects of peripheral nerve (PN) graft and neurotrophic factors on the expression of nitric oxide synthase (NOS) and the survival of Clarke's nucleus (CN) neurons at the first lumbar spinal segment (L1) 15 days after hemisection of the spinal cord at T11. Normal intact CN neurons did not express NOS. Forty-one percent of the ipsilateral CN neurons survived after hemisection at T11, and 48% of the surviving neurons expressed NOS. Transplantation of PN graft at the lesion site promoted the survival of CN neurons to 71% and increased the expression of NOS to 70%. Continuous infusion of brain-derived neurotrophic factor, ciliary neurotrophic factor, and neurotrophic-3, but not glial cell-derived neurotrophic factor, at the lesion site enhanced the survival of CN neurons to about 65%. Among the surviving neurons about 70% were NOS-positive. These results indicated that transplantation of autologous PN graft or continuous infusion of neurotrophic factors could enhance the survival of axotomized CN neurons. In addition, the survival-promoting function of the neurotrophic agents was coincided with the upregulation of the expression of NOS. However, whether the upregulation of NOS expression in injured CN neurons is related to the rescue function or is a side effect of the neurotrophic factors is not clear and needed further investigation.