Axon regeneration is arrested in the injured central nervous system (CNS) by axon growth-inhibitory ligands expressed in oligodendrocytes/myelin, NG2-glia, and reactive astrocytes in the lesion and degenerating tracts, and by fibroblasts in scar tissue. Growth cone receptors (Rc) bind inhibitory ligands, activating a Rho-family GTPase intracellular signaling pathway that disrupts the actin cytoskeleton inducing growth cone collapse/repulsion. The known inhibitory ligands include the chondroitin sulfate proteoglycans (CSPG) Neurocan, Brevican, Phosphacan, Tenascin, and NG2, as either membrane-bound or secreted molecules; Ephrins expressed on astrocyte/fibroblast membranes; the myelin/oligodendrocyte-derived growth inhibitors Nogo, MAG, and OMgp; and membrane-bound semaphorins (Sema) produced by meningeal fibroblasts invading the scar. No definitive CSPG Rc have been identified, although intracellular signaling through the Rho family of G-proteins is probably common to all the inhibitory ligands. Ephrins bind to signalling Ephs. The ligand-binding Rc for all the myelin inhibitors is NgR and requires p75(NTR) for transmembrane signaling. The neuropilin (NP)/plexin (Plex) Rc complex binds Sema. Strategies for promoting axon growth after CNS injury are thwarted by the plethora of inhibitory ligands and the ligand promiscuity of some of their Rc. There is also paradoxical reciprocal expression of many of the inhibitory ligands/Rc in normal and damaged neurons, and NgR expression is restricted to a limited number of neuronal populations. All these factors, together with an incomplete understanding of the normal functions of many of these molecules in the intact CNS, presently confound interpretive acumen in regenerative studies.

Purpose: To evaluate manganese (Mn 2ϩ )-enhanced MRI in a longitudinal study of normal and injured rat visual projections. Materials and Methods:MRI was performed 24 hours after unilateral intravitreal injection of MnCl 2 (150 nmol) into adult Fischer rats that were divided into four groups: 1) controls (N ϭ 5), 2) dose-response (N ϭ 10, 0.2-200 nmol), 3) time-response with repeated MRI during 24 -168 hours post injection (N ϭ 4), and 4) optic nerve crush (ONC) immediately preceding the MnCl 2 injection (N ϭ 7). Control and ONC animals were reinjected with MnCl 2 20 days after the first injection, and MRI was performed 24 hours later. Results:In the control group, the optic projection was visualized from the retina to the superior colliculus, with indications of transsynaptic transport to the cortex. There was a semilogarithmic relationship between the Mn 2ϩ dose and Mn 2ϩ enhancement from 4 to 200 nmol, and the enhancement decayed gradually to 0 by 168 hours. No Mn 2ϩ -enhanced signal was detected distal to the ON crush site. In the control group, similar enhancement was obtained after the first and second MnCl 2 injections, while in the ONC group the enhancement proximal to the crush site was reduced 20 days post lesion (20dpl). Conclusion:Mn 2ϩ -enhanced MRI is a viable method for temporospatial visualization of normal and injured ON in the adult rat. The observed reduction in the Mn 2ϩ signal proximal to the ONC is probably a result of retrograde damage to the retinal ganglion cells, and not of Mn 2ϩ toxicity.

Restoration of lost neuronal function after spinal cord injury (SCI) still remains a big challenge for current medicine. One important repair strategy is bridging the SCI lesion with a supportive and stimulatory milieu that would enable axonal rewiring. Injectable extracellular matrix (ECM)-derived hydrogels have been recently reported to have neurotrophic potential in vitro. In this study, we evaluated the presumed neuroregenerative properties of ECM hydrogels in vivo in the acute model of SCI. ECM hydrogels were prepared by decellularization of porcine spinal cord (SC) or porcine urinary bladder (UB), and injected into a spinal cord hemisection cavity. Histological analysis and real-time qPCR were performed at 2, 4, and 8 weeks postinjection. Both types of hydrogels integrated into the lesion and stimulated neovascularization and axonal ingrowth into the lesion. On the other hand, massive infiltration of macrophages into the lesion and rapid hydrogel degradation did not prevent cyst formation, which progressively developed over 8 weeks. No significant differences were found between SC-ECM and UB-ECM. Gene expression analysis revealed significant downregulation of genes related to immune response and inflammation in both hydrogel types at 2 weeks post SCI. A combination of human mesenchymal stem cells with SC-ECM did not further promote ingrowth of axons and blood vessels into the lesion, when compared with the SC-ECM hydrogel alone. In conclusion, both ECM hydrogels bridged the lesion cavity, modulated the innate immune response, and provided the benefit of a stimulatory substrate for in vivo neural tissue regeneration. However, fast hydrogel degradation might be a limiting factor for the use of native ECM hydrogels in the treatment of acute SCI.

Purpose:To provide dose-response data for the safe and effective use of MnCl 2 for manganese (Mn 2ϩ ) -enhanced MRI (MEMRI) of the visual pathway. Materials and Methods:Retinal ganglion cell (RGC) toxicity, CNR in MEMRI, axon density resolution for MEMRI, mode of axonal transport and clearance of Mn 2ϩ from the vitreous after ivit were investigated. After 0, 30, 150, 300, 1500, and 3000 nmol ivit MnCl 2 , neural toxicity was measured by counting surviving RGC back-filled with FluroGold (FG), CNR of the vitreous body and visual pathway by three-dimensional (3D) MEMRI, resolution of ON axon density by correlating CNR with axon density, and axonal transport of Mn 2ϩ by studying CNR in 3D MEMRI of the ON after ion of 200 nmol MnCl 2 .Results: There were no changes in RGC density after ivit MnCl 2 Յ 150 nmol, and reductions of 12%, 57%, and 94% occurred after 300, 1500, and 3000 nmol MnCl 2 . CNR increased in the visual pathway with MnCl 2 Յ 300 nmol, and decreased when the dose was raised further. Minimum detectable ON axon densities were 125,000/mm 2 . After 200 nmol ion MnCl 2 , CNRϾ0 were recorded distally from the ion site, but there was no signal in the retina. At ivit doses Ͼ1500 nmol, clearance from the vitreous body was impaired. Conclusion:The optimal dose for MEMRI of the rat visual pathway was found to be 150 -300 nmol ivit MnCl 2 . Higher doses are toxic, causing RGC death, impair active clearance from the vitreous, and loss of Mn 2ϩ enhancement throughout the visual pathway. Mn 2ϩ traffic within RGC axons is mediated mainly by anterograde transport.