Tissue sparing, behavioral recovery, supraspinal axonal sparing/regeneration following sub-acute glial transplantation in a model of spinal cord contusion.

BACKGROUND: It has been shown that olfactory ensheathing glia (OEG) and Schwann cell (SCs) transplantation are beneficial as cellular treatments for spinal cord injury (SCI), especially acute and sub-acute time points. In this study, we transplanted DsRED transduced adult OEG and SCs sub-acutely (14 days) following a T10 moderate spinal cord contusion injury in the rat. Behaviour was measured by open field (BBB) and horizontal ladder walking tests to ascertain improvements in locomotor function. Fluorogold staining was injected into the distal spinal cord to determine the extent of supraspinal and propriospinal axonal sparing/regeneration at 4 months post injection time point. The purpose of this study was to investigate if OEG and SCs cells injected sub acutely (14 days after injury) could: (i) improve behavioral outcomes, (ii) induce sparing/regeneration of propriospinal and supraspinal projections, and (iii) reduce tissue loss. RESULTS: OEG and SCs transplanted rats showed significant increased locomotion when compared to control injury only in the open field tests (BBB). However, the ladder walk test did not show statistically significant differences between treatment and control groups. Fluorogold retrograde tracing showed a statistically significant increase in the number of supraspinal nuclei projecting into the distal spinal cord in both OEG and SCs transplanted rats. These included the raphe, reticular and vestibular systems. Further pairwise multiple comparison tests also showed a statistically significant increase in raphe projecting neurons in OEG transplanted rats when compared to SCs transplanted animals. Immunohistochemistry of spinal cord sections short term (2 weeks) and long term (4 months) showed differences in host glial activity, migration and proteoglycan deposits between the two cell types. Histochemical staining revealed that the volume of tissue remaining at the lesion site had increased in all OEG and SCs treated groups. Significant tissue sparing was observed at both time points following glial SCs transplantation. In addition, OEG transplants showed significantly decreased chondroitin proteoglycan synthesis in the lesion site, suggesting a more CNS tolerant graft. CONCLUSIONS: These results show that transplantation of OEG and SCs in a sub-acute phase can improve anatomical outcomes after a contusion injury to the spinal cord, by increasing the number of spared/regenerated supraspinal fibers, reducing cavitation and enhancing tissue integrity. This provides important information on the time window of glial transplantation for the repair of the spinal cord.

Phospholipase B activity and organophosphorus compound toxicity in cultured neural cells.

Organophosphorus compounds (OP) such as phenyl saligenin phosphate (PSP) and mipafox (MPX) which cause delayed neuropathy, inhibit neuropathy target esterase (NTE), while OPs such as paraoxon (PXN) react more readily with acetylcholinesterase. In yeast and mammalian cell lines, NTE has been shown to have phospholipase B (PLB) activity which deacylates intracellular phosphatidylcholine to glycerophosphocholine (GroPCho) and can be detected by metabolic labeling with [(14)C]choline. Here we investigated PLB activity in primary cultures of mouse neural cells. In cortical and cerebellar granule neurons and astrocytes, [(14)C]GroPCho labeling was inhibited by PSP and MPX: phenyl dipentylphosphinate (PDPP), a non-neuropathic NTE inhibitor, was more potent, while PXN, was substantially less so. In all three cell types, conversion of [(14)C]phosphatidylcholine to [(14)C]GroPCho over 24 h was relatively small (2.3-14%). Consequently, even with >80% inhibition of [(14)C]GroPCho production, increased [(14)C]phosphatidylcholine was not detected. At concentrations of 1-10 microM, only PSP was cytotoxic to cortical and cerebellar granule neurons after 24-h exposure. Moreover, dramatic changes in glial cell morphology were induced by PSP, but not PDPP or MPX, with rapid (2-3 h) rounding up of astrocytes and of Schwann cells in cultures of dissociated mouse dorsal root ganglia. We conclude that PLB activity is present in a variety of cultured mouse neural cell types but that acute loss of this activity is not cytotoxic. Conversely, the rapid toxic effects of PSP in vitro suggest that a serine hydrolase distinct from NTE is required continuously by neurons and glia.

Retinal characteristics of the ornate dragon lizard, Ctenophorus ornatus.

The retina of a diurnal insectivorous lizard, Ctenophorus ornatus (Agamidae) was investigated using microspectrophotometry and light and electron microscopy. A prominent broad yellow band was observed that extended across the mid-retina. The yellow coloration was found to originate from both oil droplets and diffuse pigmentation within cone inner segments. Microspectrophotometric analysis revealed yellow oil droplets with variable absorption of wavelengths below 520 nm and transparent oil droplets with no detectable absorptance between 350 and 750 nm. Cones with transparent oil droplets lacked the diffuse yellow pigmentation. The mean wavelengths of maximum absorbance of visual pigments in the isolated cone outer segments were at 440, 493, and 571 nm. The retina was found to possess a deep convexiclivate fovea located within the yellow band, slightly dorsotemporal of the retinal midpoint. The topography of the retinal ganglion cells revealed that the fovea was contained within an area centralis. Photoreceptors were either single (80%) or unequal double (20%) cones. Within the region of the fovea, the cones were approximately 20% the diameter of those in the peripheral retina. Colored oil droplets and yellow pigment may increase visual acuity by absorbing short wavelength light scattered either by the atmosphere or the optical structures of the eye. The presence of a fovea containing slender cone photoreceptors and three visual pigments suggests that the lizard has high acuity and the potential for color vision.