Paul A. Dijkhuizen

NT-3 delivered by an adenoviral vector induces injured dorsal root axons to regenerate into the spinal cord of adult rats

Sensory axons interrupted in the dorsal roots of adult mammals are normally unable to regenerate into the spinal cord. We have investigated whether the introduction of a neurotrophin gene into the spinal cord might offer an approach to otherwise intractable spinal root injuries. The dorsal roots of the 4th, 5th, and 6th lumbar spinal nerves of adult rats were severed and reanastomosed. Fourteen to nineteen days later, adenoviral vectors containing either the LacZ or NT‐3 genes were injected into the ventral horn of the lumbar spinal cord, resulting in strong expression of the transgenes in glial cells and motor neurons between 4 and 40 days after injection. When dorsal root axons were transganglionically labelled with HRP conjugated to cholera toxin subunit B, 16 to 37 days after dorsal root injury, large numbers of labelled axons could be seen to have regenerated into the cord, but only in those animals injected with vector carrying the NT‐3 gene. The regenerated axons were found at the injection site, mainly in the grey matter, and had penetrated as deep as lamina V. Gene therapy with adenoviral vectors encoding a neurotrophin has therefore been shown to be capable of enhancing and directing the regeneration of a subpopulation of dorsal root axons (probably myelinated A fibres), into and through the CNS environment. J. Neurosci. Res. 54:554–562, 1998.

Intercostal nerve implants transduced with an adenoviral vector encoding neurotrophin-3 promote regrowth of injured rat corticospinal tract fibers and improve hindlimb function

Following injury to central nervous tissues, damaged neurons are unable to regenerate their axons spontaneously. Implantation of peripheral nerves into the CNS, however, does result in axonal regeneration into these transplants and is one of the most powerful strategies to promote CNS regeneration. In the present study implantation of peripheral nerve bridges following dorsal hemisection is combined with ex vivo gene transfer with adenoviral vectors encoding neurotrophin-3 (Ad-NT-3) to examine whether this would stimulate regeneration of one of the long descending tracts of the spinal cord, the corticospinal tract (CST), into and beyond the peripheral nerve implant. We chose to use an adenoviral vector encoding NT-3 because CST axons are sensitive to this neurotrophin and Schwann cells in peripheral nerve implants do not express this neurotrophin. At 16 weeks postimplantation of Ad-NT-3-transduced intercostal nerves, approximately three- to fourfold more of the anterogradely traced corticospinal tract fibers had regrown their axons through gray matter below the lesion site when compared to control animals. Regrowth of CST fibers occurred over more than 8 mm distal to the lesion site. No regenerating CST fibers were, however, observed into the transduced peripheral implant. Animals with a peripheral nerve transduced with Ad-NT-3 also exhibited improved function of the hindlimbs when compared to control animals treated with an adenoviral vector encoding LacZ. Thus, transient overexpression of NT-3 in peripheral nerve tissue bridges is apparently sufficient to stimulate regrowth of CST fibers and to promote recovery of hindlimb function, but does not result in regeneration of CST fibers into such transplants. Taken together, combining an established neurotransplantation approach with viral vector-gene transfer promotes the regrowth of injured CST fibers through gray matter and improves the recovery of hindlimb function.

Regulation of dendritic growth by calcium and neurotrophin signaling

The development of cortical dendrites is regulated by both activity-dependent and activity-independent signaling. Activity-dependent dendritic growth involves calcium-dependent gene expression. Both CREB and CREST are transactivators that contribute to calcium-dependent dendritic growth. Dendritic development is also regulated by extracellular factors such as neurotrophins. Neurotrophin-dependent dendritic growth is mediated by the MAP kinase and PI 3-kinase pathways. Selective responsiveness to activity cues and neurotrophins may contribute to morphological diversity in the nervous system.

The use of neurotrophic factors to treat spinal cord injury: Advantages and disadvantages of different delivery methods

Neurotrophic factors are well known for their use in stimulating regeneration in the adult central nervous system. However, delivery of neurotrophic factors to e.g. the injured spinal cord is notoriously difficult, since these proteins do not cross the blood brain barrier. Several strategies have been developed that now allow for convenient delivery of neurotrophic factors to injured neurons. These include: 1. methods to deliver the protein itself, 2. the use of transplanted cells genetically engineered to secrete neurotrophic factors and 3. direct transfer of neurotrophic factor genes using viral vectors. In this review, the advantages and disadvantages of these delivery methods that have been applied in spinal cord injury models will be discussed.