Post-Acute Alterations in the Axonal Cytoskeleton after Traumatic Axonal Injury (original) (raw)
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Axonal cytoskeletal changes after non-disruptive axonal injury
Journal of Neurocytology, 1997
In animal models of human diffuse axonal injury, axonal swellings leading to secondary axotomy occur between 2 and 6 h after injury. But, analysis of cytoskeletal changes associated with secondary axotomy has not been undertaken. We have carried out a quantitative analysis of cytoskeletal changes in a model of diffuse axonal injury 4 h after stretch-injury to adult guinea-pig optic nerves. The major site of axonal damage was the middle portion of the nerve. There was a statistically significant increase in the proportion of small axons with a diameter of 0.5 μm and smaller in which there was compaction of neurofilaments. Axons with a diameter greater than 2.0 μm demonstrated an increased spacing between cytoskeletal elements throughout the length of the nerve. However, in the middle segment of the nerve these larger axons demonstrated two different types of response. Either, where periaxonal spaces occurred, there was a reduction in axonal calibre, compaction of neurofilaments but no change in their number, and a loss of microtubules. Or, where intramyelinic spaces occurred there was an increased spacing between neurofilaments and microtubules with a significant loss in the number of both. Longitudinal sections showed foci of compaction of neurofilaments interspersed between regions where axonal structure was apparently normal. Neurofilament compaction was correlated with disruption of the axolemma at these foci present some hours after injury. We suggest that the time course of these axonal cytoskeletal changes after stretch-injury to central axons is shorter than those changes documented to occur during Wallerian degeneration.
Axonal Cytoskeletal Changes After Nondisruptive Axonal Injury. II. Intermediate Sized Axons
Journal of Neurotrauma, 1998
In animal models of human diffuse axonal injury, axonal swellings leading to secondary axotomy occur between 2 and 6 h after injury. But, analysis of cytoskeletal changes associated with secondary axotomy has not been undertaken. We have carried out a quantitative analysis of cytoskeletal changes in a model of diffuse axonal injury 4 h after stretch-injury to adult guinea-pig optic nerves. The major site of axonal damage was the middle portion of the nerve. There was a statistically significant increase in the proportion of small axons with a diameter of 0.5 m and smaller in which there was compaction of neurofilaments. Axons with a diameter greater than 2.0 m demonstrated an increased spacing between cytoskeletal elements throughout the length of the nerve. However, in the middle segment of the nerve these larger axons demonstrated two different types of response. Either, where periaxonal spaces occurred, there was a reduction in axonal calibre, compaction of neurofilaments but no change in their number, and a loss of microtubules. Or, where intramyelinic spaces occurred there was an increased spacing between neurofilaments and microtubules with a significant loss in the number of both. Longitudinal sections showed foci of compaction of neurofilaments interspersed between regions where axonal structure was apparently normal. Neurofilament compaction was correlated with disruption of the axolemma at these foci present some hours after injury. We suggest that the time course of these axonal cytoskeletal changes after stretch-injury to central axons is shorter than those changes documented to occur during Wallerian degeneration.
Acta Neuropathologica, 1990
A new model of focal axonal injury was reproduced by rapid and controlled elongation (uniaxial stretch) of the guinea pig optic nerve. Light microscopy study of optic nerve specimens after horseradish peroxidase injection into the vitreous of the animal's eye showed that axonal lesions were identical to those seen in human and primate post-traumatic diffuse axonal injury (DAI). The lesions were characterized by the formation of terminal clubs in severed axons and focal axonal enlargements in those axons that were lesioned-in-continuity. Visual-evoked potentials upon flash stimulation were recorded before and after injury. Mean amplitude and mean latency of occipital peaks were significantly elongated in the acute post-traumatic phase. Electron microscopy examination showed that the main axonal changes observed in this model were cytoskeleton disorganization, accumulation of axoplasm membrane-bound bodies at the site of terminal balls and dilatations-in-zontinuity and detachment of the axolemma from the myelin sheath. Such axonal alterations were similar to those found in many other biological models of central and peripheral axonal injuries in which the lesion was produced by invasive methods. This model is unique since it reproduces the same mechanism of injury and the identical lesions that have been demonstrated in humans and primates with post-traumatic (DAI).
Anais da Academia Brasileira de Ciências, 2001
In this paper we report a qualitative morphological analysis of Wallerian degeneration in a marsupial. Right optic nerves of opossums Didelphis marsupialis were crushed with a fine forceps and after 24, 48, 72, 96 and 168 hours the animals were anaesthetized and perfused with fixative. The optic nerves were immersed in fixative and processed for routine transmission electron microscopy. Among the early alterations typical of axonal degeneration, we observed nerve fibers with focal degeneration of the axoplasmic cytoskeleton, watery degeneration and dark degeneration, the latter being prevalent at 168 hours after crush. Our results point to a gradual disintegration of the axoplasmic cytoskeleton, opposed to the previous view of an "all-or-nothing'' process (Griffin et al 1995). We also report that, due to an unknown mechanism, fibers show either a dark or watery pattern of axonal degeneration, as observed in axon profiles. We also observed fibers undergoing early myelin ...
Temporal Profiles of Cytoskeletal Protein Loss following Traumatic Axonal Injury in Mice
Neurochemical Research, 2007
To examine the time course and relative extent of proteolysis of neurofilament and tubulin proteins after traumatic axonal injury (TAI), anesthetized mice were subjected to optic nerve stretch injury. Immunohistochemistry confirmed neurofilament accumulation within axonal swellings at 4, 24, and 72 h postinjury (n = 4 injured and 2 sham per time point). Immunoblotting of optic nerve homogenates (n = 5 injured and 1 sham at 0.5, 4, 24 or 72 h) revealed calpain-mediated spectrin proteolytic fragments after injury. Protein levels for NF68 progressively decreased from 0.5 h to 24 h postinjury, while NF200 and alpha-tubulin levels decreased acutely (0.5-4 h), with a secondary decline at 72 h postinjury. These data demonstrate that diffusely distributed TAI is associated not only with a localized accumulation of neurofilament proteins, but also significant decreases in total cytoskeletal protein levels which may be mediated, in part, by calpains. Protection of the axonal cytoskeleton represents a potential therapeutic target for axonal damage associated with injury or neurodegenerative diseases.
BMC Neuroscience, 2010
Background: Excitotoxicity is involved in the pathogenesis of a number neurodegenerative diseases, and axonopathy is an early feature in several of these disorders. In models of excitotoxicity-associated neurological disease, an excitotoxin delivered to the central nervous system (CNS), could trigger neuronal death not only in the somatodendritic region, but also in the axonal region, via oligodendrocyte N-methyl-D-aspartate (NMDA) receptors. The retina and optic nerve, as approachable regions of the brain, provide a unique anatomical substrate to investigate the "downstream" effect of isolated excitotoxic perikaryal injury on central nervous system (CNS) axons, potentially providing information about the pathogenesis of the axonopathy in clinical neurological disorders. Herein, we provide ultrastructural information about the retinal ganglion cell (RGC) somata and their axons, both unmyelinated and myelinated, after NMDA-induced retinal injury. Male Sprague-Dawley rats were killed at 0 h, 24 h, 72 h and 7 days after injecting 20 nM NMDA into the vitreous chamber of the left eye (n = 8 in each group). Saline-injected right eyes served as controls. After perfusion fixation, dissection, resin-embedding and staining, ultrathin sections of eyes and proximal (intraorbital) and distal (intracranial) optic nerve segments were evaluated by transmission electron tomography (TEM). Results: TEM demonstrated features of necrosis in RGCs: mitochondrial and endoplasmic reticulum swelling, disintegration of polyribosomes, rupture of membranous organelle and formation of myelin bodies. Ultrastructural damage in the optic nerve mimicked the changes of Wallerian degeneration; early nodal/paranodal disturbances were followed by the appearance of three major morphological variants: dark degeneration, watery degeneration and demyelination. Conclusion: NMDA-induced excitotoxic retinal injury causes mainly necrotic RGC somal death with Wallerian-like degeneration of the optic nerve. Since axonal degeneration associated with perikaryal excitotoxic injury is an active, regulated process, it may be amenable to therapeutic intervention.
Ultrastructural Studies of Inh-Induced Neuropathy in Rats. I. Early Axonal Changes
The American journal of pathology, 1964
The inadequacy of histologic techniques for the simultaneous recording of the interaction of all peripheral nerve components during regeneration and repair has contributed to our present ignorance of several fundamental factors. Extensive investigations by light microscopy have resulted in disagreements over several fundamental aspects of these processes.' 2 The electron microscope offers the opportunity for a more accurate demonstration of the evolution of the reactions. Several observations of regeneration following wallerian degeneration have been made with this instrument.8-'0 The Isoniazid (isonicotinic acid hydrazide, INH) induced peripheral neuropathy of the rat is characterized by a primary axonal alteration and a subsequent breakdown of the myelin sheath." 12 These degenerative processes are accompanied and succeeded by reparative and regenerative phenomena of both parenchymal and mesenchymal elements. The specific activity of fibroblasts, axons, and Schwann cells in the degenerated nerve constitute the basis for this report. MATERIAL AND METHODS Twenty female Sprague-Dawley rats were fed 350 mg. Isoniazid per kg. daily per os for periods varying from 6 to i6 days. Three rats were treated for 14 days and sacrificed on the 44th, I20th and I54th days. The sciatic nerves of experimental and 4 normal control rats were removed and examined by light, phase and electron microscopy. The fixative for electron microscopy was i per cent buffered osmium tetroxide; methacrylate and Epon resin were used as embedding media. Part of the material was stained with lead hydroxide. RESULTS Light Microscopy An examination of the 6to i6-day old nerve lesion demonstrated a variable focal disruption of axons and myelin sheaths. Increased cel-This work was partially supported by United States Public Health Service Grant 5482 and Special Fellowship in Neuropathology (BT-794), National Institute of Neurological Diseases and Blindness.
Acta Neuropathologica, 2006
By means of a new head-injury apparatus, a 0.75-mm-deep depression was produced momentarily at a predetermined site of the rat calvaria. This immediately evoked ultrastructural (neurofilament) compaction in many myelinated axon segments in layers IV and V of the neocortex under the impact site. The affected axon segments run quasi-parallel to the brain surface in a diffuse distribution among normal axons. Other kinds of damage to the brain tissue were insignificant; the conditions were therefore favorable for investigation of the fate of the compacted axons. Quantitative analysis of the findings on groups of ten rats that were sacrificed either immediately after the head injury or following a 1 day or a 1 week survival period showed that around 50% of the compacted axons recovered in 1 day, and a further less than 10% did so in 1 week. Electron microscopy revealed that the non-recovering compacted axons underwent a sequence of degenerative morphological changes including homogenization, fragmentation and resorption of the fragments. However, the myelin sheaths around these degenerating axons remained apparently unchanged even in the long-surviving rats, and hardly any phagocytotic cells were encountered. On the other hand, many such myelin sheaths contained axolemma-bound, normal-looking axoplasm besides the above morphological signs of axon-degeneration. It is concluded that the non-recovering compacted axons undergo an uncommon (non-Wallerian) kind of degeneration, which is mostly reversible.
Brain Research, 1996
It has recently been demo~astrated [Pettus et al., J. Neurotraurna, 11 (1994) [507][508][509][510][511][512][513][514][515][516][517][518][519][520][521][522]] that moderate traumatic brain injury evokes alterations in axolemmal permeability associated with rapid local compaction of axonal neurofilaments (NF). The current communication fully characterizes these local[ NF changes, while also exploring the possibility of other related cytoskeletal abnormalities. A tracer normally excluded by the intact axolemma (horseradish peroxidase) was administered intrathecally in cats, which were then subjected to moderate/severe fluid percussion brain injury (FPI). After survival times ranging from 5 min to 6 h post-traumatic brain injury (TBI), the animals were perfused and processed for light microscopic (LM) and electron microscopic (EM) visualization of horseradish peroxidase (HRP). HRP-containing axons identified by LM, were investigated by EM in both the sagittal and coronal planes. Electron micrographs were videographically captured, digitized, and analyzed for cytoskeletal distribution. Local alterations in axolemmal permeability to HRP were detected, and consistently linked with distinct cytoskeletal changes. Within 5 min of injury, the injured HRP-containing axons displayed a significant decrease in inter-NF spacing associated with a lack of NF side arm projections. Density analysis proved a significant increase in NF packing in the HRP-containing axons, and further revealed an associated significant decrease in microtubule (MT) density. All ultrastructural changes were seen within 5 min of injury, and persisted unchanged for up to 6 h post-TBI. Collectively, these abnormalities suggest SLat altered axolemmal permeability triggers a rapid, yet persisting general cytoskeletal change most likely linked to local ionic disregulation. We posit that this local cytoskeletal collapse/alteration marks a site of impaired axonal transport, associated with upstream axoplasmic swelling and eventual axonal detachment.