Central-peripheral Distal Axonopathy Research Articles

In vitro study of the neuropathic potential of the organophosphorus compounds fenamiphos and profenofos: Comparison with mipafox and paraoxon

Organophosphorus-induced delayed neuropathy (OPIDN) is a central-peripheral distal axonopathy that develops 8–14days after poisoning by a neuropathic organophosphorus compound (OP). Several OPs that caused OPIDN were withdrawn from the agricultural market due to induction of serious delayed effects. Therefore, the development of in vitro screenings able to differentiate neuropathic from non-neuropathic OPs is of crucial importance. Thus, the aim of this study was to evaluate the differences in the neurotoxic effects of mipafox (neuropathic OP) and paraoxon (non-neuropathic OP) in SH-SY5Y human neuroblastoma cells, using the inhibition and aging of neuropathy target esterase (NTE), inhibition of acetylcholinesterase (AChE), activation of calpain, neurite outgrowth, cytotoxicity and intracellular calcium as indicators. Additionally, the potential of fenamiphos and profenofos to cause acute and/or delayed effects was also evaluated. Mipafox had the lowest IC50 and induced the highest percentage of aging of NTE among the OPs evaluated. Only mipafox was able to cause calpain activation after 24h of incubation. Concentrations of mipafox and fenamiphos which inhibited at least 70% of NTE were also able to reduce neurite outgrowth. Cytotoxicity was higher in non-neuropathic than in neuropathic OPs while the intracellular calcium levels were higher in neuropathic than in non-neuropathic OPs. In conclusion, the SH-SY5Y cellular model was selective to differentiate neuropathic from non-neuropathic OPs; fenamiphos, but not profenofos presented results compatible with the induction of OPIDN.

Chlorpyrifos-induced Delayed Myelopathy and Pure Motor Neuropathy

Organophosphate (OP) poisoning is known to cause delayed neurological manifestations. Chlorpyrifos, an OP, causes a delayed syndrome that is characterized by a motor sensory polyneuropathy. Pure motor neuropathy with intact sensory conduction is rarely documented. Rapidly evolving delayed myelopathy is extremely uncommon. A healthy 15-year-old female was admitted to hospital with cholinergic crisis due to ingestion of a large dose of chlorpyrifos (OP). She was treated with atropine and recovered completely without any neurological symptoms or signs. She came to hospital 6 weeks later with upper and lower motor neuron signs involving the lower limbs without sensory loss. By the end of 7 weeks, there was urinary incontinence. At 2-month follow-up, she had progressive spasticity. Electrophysiological studies revealed a pure motor neuropathy. Spine magnetic resonance imaging showed early signs of thoracic cord atrophy. Other causes of myelopathy were excluded. Chronic neurotoxicity due to OP poisoning is dependent on several factors: chemical composition of the OP, dose systematized, and the administration of anitcholinergics for cholinergic crisis. The pathology of OP-induced delayed neuropathy involves a central-peripheral distal axonopathy. Peripheral distal axonopathy results in a predominantly motor polyneuropathy. Axonopathy of the central nervous system results in myelopathic features that makes for a poorer prognosis.

Recovery of axonal myelination sheath and axonal caliber in the mouse corpus callosum following damage induced by N,N‐diethyldithiocarbamate

Disulfiram is an aldehyde dehydrogenase inhibitor used for the treatment of alcohol dependence and of cocaine addiction. It has been demonstrated that subchronic administration of disulfiram or N,N-diethyldithiocarbamate (DEDTC), the main derivative of disulfiram, to rats can produce central-peripheral distal axonopathy. However, few data regarding the axonal effects of these compounds in the central nervous system exist. Our previous studies have revealed DEDTC-induced axonal damage in the mouse brain during the course of postnatal development, together with alterations in axonal pathfinding and in the myelination process, with partial recovery during the post-treatment period. In order to gather new data about how this axonal damage and recovery occurs in the central nervous system, we performed an ultrastructural analysis of the axons located in the corpus callosum from mice treated with DEDTC during postnatal development. The axonal caliber throughout the axonal area, the maximum axonal diameter, the maximum fiber diameter, and the axonal circularity, at different postnatal stages [from postnatal day (P)9 to P30], were analyzed. In addition, parameters related to the myelinization process (number of myelinated axons, sheath thickness, and the ratio of myelinated axons to total axons) were evaluated. A reduction in the average value of axonal caliber during treatment and a delay in the axonal myelination process were detected. Whereas early recovery of individual axons occurred after treatment (P22), complete recovery of myelinated axons occurred at late postnatal stages (P42). Therefore, chronic treatment with dithiocarbamates requires periods of rest to encourage the recovery of myelinated axons.

Involvement of cyclin-dependent kinase 5 in 2,5-hexanedione-induced neuropathy

Occupational exposure to n-hexane produces a neuropathy characterized as a central-peripheral distal axonopathy, which is mediated by 2,5-hexanedione (HD). To investigate the mechanisms of the neuropathy induced by HD, the contents and activities of cyclin-dependent kinase 5 (CDK5) and activators (p35 precursor, p35 and p25) in rats’ cerebrum cortex (CC), spinal cord (SC) and sciatic nerve (SN) were determined. The results showed that the levels and activities of CDK5 in CC of 200 or 400 mg/kg HD-treated rats were significantly decreased in both the cytosolic and membrane fractions and negatively correlated with gait abnormality in the cytosolic fraction. However, CDK5 contents and activities in SN of rats treated with 200 or 400 mg/kg HD were significantly increased and positively correlated with gait abnormality in both the cytosolic and membrane fractions. Although increases of CDK5 contents in both the cytosolic and membrane fractions of SC in 200 and 400 mg/kg HD-treated rats were also observed, CDK5 activities were significantly decreased in the cytosolic fraction and negatively correlated with gait abnormality. The changes of p35 precursor, p35 and p25 contents in CC, SC and SN showed the same pattern with that of CDK5 activities. Thus, HD intoxication was associated with deregulation of CDK5 and its activator p35 or p25 in nerve tissues. The inconsistent changes of CDK5 activities in CNS and PNS might delegate the different mechanisms of HD-induced peripheral neuropathy.

Malondialdehyde and catalase as the serum biomarkers of allyl chloride-induced toxic neuropathy

Chronic exposure to allyl chloride (AC) is known to produce a central-peripheral distal axonopathy. To access the biomarker of exposure and elucidate the mechanism of neuropathy induced by AC, we performed a longitudinal observational study of malondialdehyde (MDA), anti-reactive oxygen species (anti-ROS), glutathione (GSH), catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) in rats serum and sciatic nerve after 0, 3, 6, 9 and 12 weeks of AC administration. AC was administrated to Wistar rats by gavage at a single dosage of 200 mg/kg/per dose (three times per week). Rats were sacrificed after 0, 3, 6, 9 and 12 weeks of AC treatment, serum and sciatic nerves were quickly collected at 4 °C. The results showed that MDA levels in serum (115.4 and 126.2%) and sciatic nerve (130.5 and 145.3%) significantly increased ( p < 0.05) on 3rd week of AC treatment and at gait score of 2, and further changes of MDA levels were observed after 6, 9 and 12 weeks and at gait score of 3 and 4. While a decrease ( p < 0.05) in the activities of CAT on 6th week of AC intoxication and at gait score of 2 was observed in serum (81.2 and 72.8%) and sciatic nerve (71.7 and 70.7%). The other antioxidants also decreased in serum and sciatic nerve after 3, 6 and 9, 12 weeks’ intoxication and at gait score of 2, 3 and 4. Significant ( p < 0.05) positive correlations were observed between serum and sciatic nerve in MDA levels ( r = 0.9162 and 0.9551, respectively) and CAT ( r = 0.9410 and 0.9557, respectively) activities as time went on and symptoms developed. Thus, AC intoxication was associated with elevation of lipid peroxidation and reduction of antioxidative status, and the time dependent changes of these indexes in Wistar rats’ serum and sciatic nerve occurred. The misbalance of lipid peroxidation and antioxidation status might be one of mechanisms of toxic neuropathy induced by AC. MDA and CAT could be served as the biomarkers of AC exposure to afford the early diagnosis of AC-induced toxic neuropathy.

Acrylamide-Induced Changes in the Neurofilament Protein of Rat Cerebrum Fractions

Acrylamide (ACR) is known to produce central-peripheral distal axonopathy, which is characterized by distal swellings and secondary degeneration both in experimental animals and human. Ultrastructurally, excessive accumulation of neurofilaments (NFs) in the distal swollen axon is a major pathological hallmark. However, the mechanisms of ACR axonopathy remain unknown. Twenty seven male Wistar rats were randomly divided into three groups. Lower and higher ACR groups were received 20 and 40 mg/kg ACR by i.p. injection respectively. The control group received physiological saline. All rats were sacrificed after 8 weeks of treatment and their cerebrums were dissected, homogenized and used for the determination of the NF proteins. In general, the levels of light NF (NF-L) and medium NF (NF-M) subunits increased consistently in the supernatant, whereas they decreased consistently in the pellet from rats treated with ACR. Compared to that of the control group, the levels of NF-L increased respectively by 104% and 45% (P<0.01) in the supernatant and decreased by 16% and 11% (P<0.01) in the pellet of rat cerebrums in lower and higher groups. The enhancement of NF-M was 76% and 147% (P<0.05, P<0.01) in supernatant, and the reduction was 26% and 36% (P<0.01) in pellet in lower and higher group respectively. The heavy NF (NF-H) level changed slightly. The present results suggested that the change of NF-L and NF-M levels in cerebrum might be relevant to the mechanisms of the neurofilamentous axonopathies induced by ACR.

MR-Pathologic Comparison of the Upper Spinal Cord in Different Motor Neuron Diseases

This MRI study was performed to evaluate in vivo alterations of the spinal cord in defined subgroups of motor neuron diseases. Standard MRI examinations of the cervical and thoracic spinal cord in sporadic amyotrophic lateral sclerosis (ALS; n = 39), sporadic lower motor neuron disease (LMND; n = 19), Kennedy’s disease (KD; n = 19) and a control group (n = 96) were analyzed with respect to spinal cord signal changes and the thickness of the spinal cord. No significant changes in thickness or signal alterations were observed when comparing ALS, LMND and control groups with one another. However, in KD patients significant upper spinal cord atrophy was detected at the cervical level as compared with all other groups. At the thoracic level, KD patients had significant upper cord atrophy as compared with controls and LMND. Marked atrophy of the upper spinal cord seems to be a feature of the KD-associated central-peripheral distal axonopathy.

Acrylamide axonopathy revisited

Distal swelling and eventual degeneration of axons in the CNS and PNS have been considered to be the characteristic neuropathological features of acrylamide (ACR) neuropathy. These axonopathic changes have been the basis for classifying ACR neuropathy as a central–peripheral distal axonopathy and, accordingly, research over the past 30 years has focused on the primacy of axon damage and on deciphering underlying mechanisms. However, based on accumulating evidence, we have hypothesized that nerve terminals, and not axons, are the primary site of ACR action and that compromise of corresponding function is responsible for the autonomic, sensory, and motor defects that accompany ACR intoxication (NeuroToxicology 23 (2002) 43). In this paper, we provide a review of data from a recently completed comprehensive, longitudinal silver stain study of brain and spinal cord from rats intoxicated with ACR at two different daily dosing rates, i.e., 50 mg/kg/day, ip or 21 mg/kg/day, po. Results show that, regardless of dose-rate, ACR intoxication was associated with early, progressive nerve terminal degeneration in all CNS regions and with Purkinje cell injury in cerebellum. At the lower dose-rate, initial nerve terminal argyrophilia was followed by abundant retrograde axon degeneration in white matter tracts of spinal cord, brain stem, and cerebellum. The results support and extend our nerve terminal hypothesis and suggest that Purkinje cell damage also plays a role in ACR neurotoxicity. Substantial evidence now indicates that axon degeneration is a secondary effect and is, therefore, not pathophysiologically significant. These findings have important implications for future mechanistic research, classification schemes, and assessment of neurotoxicity risk.

Altered expression of transcripts for alpha-tubulin and an unidentified gene in the spinal cord of phenyl saligenin phosphate treated hens (Gallus gallus).

Phenyl saligenin phosphate (PSP) induces a central-peripheral distal axonopathy in domestic fowl that develops 7-21 days after a single exposure. Neurotoxic esterase (NTE) is the initial molecular target for this neurotoxicity. PSP has to covalently bind to NTE and chemically "age" for induction of axonopathy. It was hypothesized that exposure to PSP results in early changes in spinal cord gene expression that do not occur with phenylmethylsulfonyl fluoride, a non-neuropathic compound that also inhibits NTE, or DMSO controls. Targeted display was used to screen approximately 15,000 gel bands. Three candidate genes were identified, but only the transcript designated P1 showed decreased expression following PSP exposure (2 mg/kg i.m.) in subsequent Northern blot and in situ hybridization experiments in samples taken <48 h after exposure. Additional experiments revealed that a approximately 2.5 kb alpha-tubulin transcript had decreased expression at 12-48 h after PSP exposure, with maximum change at 48 h (33%, p = 0.0479). A approximately 4.5 kb alpha-tubulin transcript had increased expression at 12 h (38%, p = 0.0125) and decreased expression at 48 h (28%, p = 0.0576). In situ hybridization on spinal cord revealed neuronal expression of P1 and alpha-tubulin transcripts. Decreased expression of transcripts for P1 and alpha-tubulin was present at 12 and 48 h, respectively. This decrease occurred in all neurons, not just those whose axons degenerate. Results suggest that (1) in PSP-induced OPIDN (organophosphorus-induced delayed neurotoxicity) some gene transcript expression changes are associated with initiation of axonopathy, and (2) PSP modulates spinal cord gene expression in neuronal types that do not undergo axonal degeneration.

Nerve Terminals as the Primary Site of Acrylamide Action: A Hypothesis

Acrylamide (ACR) is considered to be prototypical among chemicals that cause a central–peripheral distal axonopathy. Multifocal neurofilamentous swellings and eventual degeneration of distal axon regions in the CNS and PNS have been traditionally considered the hallmark morphological features of this axonopathy. However, ACR has also been shown to produce early nerve terminal degeneration of somatosensory, somatomotor and autonomic nerve fibers under a variety of dosing conditions. Recent research from our laboratory has demonstrated that terminal degeneration precedes axonopathy during low-dose subchronic induction of neurotoxicity and occurs in the absence of axonopathy during higher-dose subacute intoxication. This relationship suggests that nerve terminal degeneration, and not axonopathy, is the primary or most important pathophysiologic lesion produced by ACR. In this hypothesis paper, we review evidence suggesting that nerve terminal degeneration is the hallmark lesion of ACR neurotoxicity, and we propose that this effect is mediated by the direct actions of ACR at nerve terminal sites. ACR is an electrophile and, therefore, sulfhydryl groups on presynaptic proteins represent rational molecular targets. Several presynaptic thiol-containing proteins (e.g. SNAP-25, NSF) are critically involved in formation of SNARE (soluble N-ethylmaleimide (NEM)-sensitive fusion protein receptor) complexes that mediate membrane fusion processes such as exocytosis and turnover of plasmalemmal proteins and other constituents. We hypothesize that ACR adduction of SNARE proteins disrupts assembly of fusion core complexes and thereby interferes with neurotransmission and presynaptic membrane turnover. General retardation of membrane turnover and accumulation of unincorporated materials could result in nerve terminal swelling and degeneration. A similar mechanism involving the long-term consequences of defective SNARE-based turnover of Na +/K +-ATPase and other axolemmal constituents might explain subchronic induction of axon degeneration. The ACR literature occupies a prominent position in neurotoxicology and has significantly influenced development of mechanistic hypotheses and classification schemes for neurotoxicants. Our proposal suggests a reevaluation of current classification schemes and mechanistic hypotheses that regard ACR axonopathy as a primary lesion.

Progressive myelopathy and neuropathy in New Zealand Huntaway dogs

Aim: To investigate the nature and cause of a progressive ataxia in three 20-month-old Huntaway dogs that were litter mates. Methods: Affected dogs were examined before they were humanely killed and submitted to necropsy. Selected formalin-fixed tissues were examined by light and electron microscopy. Results: The lesions were those of axon and myelin degeneration within sensory, proprioceptive and motor tracts of the spinal cord and to a lesser degree some peripheral nerves. Conclusion: A progressive myelopathy and neuropathy, tentatively described as a central-peripheral distal axonopathy, was present in all 3 dogs.The cause was not determined but was likely to be either genetic or nutritional. Clinical relevance: In the early stages of this disease, careful examination maybe necessary to distinguish the signs of ataxia from orthopaedic disease such as hip dysplasia. Affected animals are unlikely to be of use as working dogs.

Peripheral and central conduction abnormalities in diabetes mellitus.

To investigate peripheral and central somatosensory conduction in patients with diabetes. The authors recorded sensory nerve action potentials and 5-channel somatosensory evoked potentials (SEPs) with noncephalic reference after median nerve stimulation in 55 patients with diabetes and 41 age- and height-matched normal subjects. The authors determined onset or peak latencies of the Erb's potential (N9) and the spinal N13-P13 and the cortical N20-P20 components, and obtained the central conduction time (CCT) by onset-to-onset and peak-to-peak measurements. Both onset and peak latencies of all SEP components were prolonged in patients with diabetes. The mean onset CCT in the diabetic group was 6.3 +/- 0.5 msec (mean +/- SD)-significantly longer than that in the control group (6.1 +/- 0.2 msec)-whereas no significant difference was found in the peak CCT. The amplitudes of N9 and N13-P13 components (but not N20-P20) were significantly smaller in the diabetic group. The peripheral sensory conduction velocity was also decreased in the diabetic group, but there was no significant correlation between peripheral conduction slowing and the onset of CCT prolongation. Diabetes affects conductive function in the central as well as peripheral somatosensory pathways. The CCT abnormality does not coincide with lowering of the peripheral sensory conduction. The current results do not favor a hypothesis that a central-peripheral distal axonopathy plays an important role in development of diabetic polyneuropathy.

Redefining toxic distal axonopathies

Nerve damage classified as a central-peripheral distal axonopathy is produced by a variety of chemicals (e.g. acrylamide, n-hexane). Historically, axon swelling and secondary degeneration have been considered the morphologic hallmarks of toxic axonopathies and substantial research has been devoted toward deciphering corresponding molecular mechanisms. However, recent studies from the author’s laboratory investigating rate (mg toxicant/kg/day) and route (i.p. vs gavage) of intoxication have shown that swelling and degeneration were related to neurotoxicant dosing conditions (i.e. low-dose, subchronic exposure) and not to development of neurophysiological deficits or classic behavioral toxicity. This suggests the presumed hallmarks of distal axonopathy are epiphenomena of uncertain pathophysiologic significance. Therefore, the current definition of and chemical classification scheme for toxic distal axonopathies requires re-evaluation.

Adrenomyeloneuropathy: a neuropathologic review featuring its noninflammatory myelopathy.

The neuropathologic features of adrenomyeloneuropathy (AMN) are reviewed by supplementing those few previously published cases with 5 additional cases collected over the years. The endocrine involvement in AMN is briefly presented to serve as a pathogenetic backdrop and to emphasize that most of the lesions in AMN, as in adreno-leukodystrophy (ALD), are noninflammatory in the traditional sense of the word. The myeloneuropathy is emphasized, but the dysmyelinative/inflammatory demyelinative lesions also are presented. The preponderance of available data indicates that the myeloneuropathy of AMN is a central-peripheral distal (dying-back) axonopathy, as was originally proposed. The severity of the myeloneuropathy does not appear to correlate with the duration or severity of endocrine dysfunction. Microglia are the dominant participating cells in the noninflammatory myelopathy. Abnormalities in the ALD gene, which encodes a peroxisomal ABC half-transporter, do not correlate with clinical phenotypes. The relationship of the gene product, ALDP, to the peroxisomal very long chain fatty acid (VLCFA) synthetase, the activity of which is deficient in ALD/AMN, is unclear. An ALD-knockout mouse model has developed axonal degeneration, particularly in spinal cord, and is therefore more reminiscent of AMN than ALD. We continue to postulate that the fundamental defect in the myeloneuropathy of AMN is an axonal or neuronal membrane abnormality perhaps due to the incorporation of VLCFA-gangliosides, which perturbs the membrane's microenvironment and leads to dysfunction and atrophy.

Studies of multimodal evoked potentials in treated phenylketonuria: the pattern of vulnerability.

We performed studies of multimodal evoked potentials and peripheral sensory and motor nerve conductions in 33 early and 6 late treated patients with phenylketonuria. The studies revealed the following picture: 1. In 27% of early treated patients latencies of visual evoked potentials were increased. The cause for these changes in unknown. 2. Nerve conduction studies showed the presence of a minor sensory neuropathy which in rare cases may also affect peripheral motor nerves. This neuropathy did not have features of a central-peripheral distal axonopathy which argues against a toxic/nutritional causation. 3. Deficits in the central sensory, motor, and auditory pathways were present, but rare in early treated patients. If the results of electrophysiological studies reported by different groups are compared, the emerging picture is very similar and the majority of the-minor-differences is likely to be explained by technical aspects.

Severe sensory-autonomic neuropathy and endocrinopathy in insulin-dependent diabetes

A patient developed insulin-dependent diabetes mellitus at the age of 9 years, complicated by a sensory/autonomic polyneuropathy which presented with a respiratory arrest at the age of 41 years. The neuropathy increased in severity over the subsequent two decades. At the age of 52 years she had hypopituitarism, hypothyroidism and low normal adrenal function. Autopsy at the age of 59 years revealed loss of pituitary tissue with evidence of hypophysitis, a lymphocytic thyroiditis and severe adrenal atrophy with lymphocytic infiltration of the medulla. The pancreas showed reduced numbers and size of the islets of Langerhans with total loss of immunoreactivity for insulin but intact glucagon-producing cells. These features are consistent with a type 2 autoimmune polyendocrine syndrome, in which lymphocytic hypophysitis has not previously been recorded. There was severe loss of myelinated nerve fibres in the sural nerve and rostrally accentuated fibre degeneration in the gracile fasciculi, but only mild cell loss in the dorsal root ganglia. This combination suggests the presence of a central-peripheral distal axonopathy. The cervical sympathetic ganglia were severely atrophic. Minor inflammatory infiltration was observed in the dorsal root and sympathetic ganglia. Significant vascular abnormalities were not present in the peripheral nerves. This, and the pattern of nerve fibre degeneration, suggest that in this case the neuropathy was likely to have been related to metabolic rather than vascular causes. The inflammatory infiltrates in sensory and sympathetic ganglia raise the possibility of an autoimmune inflammatory contribution to the neuropathy.

Modifications of neurofilament proteins by possible metabolites of allyl chloride in vitro.

Chronic exposure to allyl chloride (ALL) is known to produce a central-peripheral distal axonopathy. In relation to the mechanism(s), the present study was conducted to examine the abilities of ALL and its putative metabolites, i.e., epichlorohydrin, glycerol alpha-monochlorohydrin, allyl alcohol and acrolein to cross-link proteins in vitro. Neurofilament-riched cytoskeletal proteins (1mg/ml) and ovalbumin (10mg/ml) were incubated with 160 mM tested chemicals except for acrolein at 0.5 mM and 1 mM. Time-dependent studies by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed that not only ALL, but also acrolein and epichlorohydrin exerted chemical modifications on axonal cytoskeletal proteins; while only acrolein-treated ovalbumin could manifest evidence of polymerization of the protein. Immunoblotting of PAGE-separated proteins confirmed that the high molecular weight proteins on the top of SDS-PAGEs were NF antigen-contained covalent cross-linked materials.

Sensory conduction study in chronic sensory ataxic neuropathy.

Sensory conduction was studied in six patients with chronic sensory ataxic neuropathy of an idiopathic type and associated with Sjögren's syndrome. Motor nerve conduction velocities were normal in most cases, but sensory nerve potentials could not be evoked in a routine peripheral nerve conduction study. Cortical and cervical somatosensory evoked potentials (SEPs) and evoked potentials from Erb's point were barely recorded by median nerve stimulation at the wrist. When the median nerve was stimulated at more proximal points, clear potentials were recorded from Erb's point, but cortical SEPs were still hardly elicited. Thus the sensory nerves are centrally and peripherally involved in this condition, and the involvement is more prominent in the distal portion in the peripheral nerve. These findings suggest that central-peripheral distal axonopathy is a process involved in this illness and that the dorsal root ganglia may be primarily involved, in accord with previous pathological studies.

Neurotoxicity of glycidamide, an acrylamide metabolite, following intraperitoneal injections in rats.

Acrylamide (2-propenamide) monomer produces central-peripheral distal axonopathy in humans and some animal species. Its neurotoxicity is characterized by abnormal sensation, decreased motor strength, and ataxia. Acrylamide forms adducts with glutathione, proteins, and DNA. Recent studies demonstrated that acrylamide is metabolized to its epoxide, glycidamide (2,3-epoxy-1-propanamide). We studied the neurotoxicity potential of glycidamide in male Sprague-Dawley rats. Animals (groups of 6) were injected ip daily with either aqueous acrylamide or glycidamide at an acrylamide-equivalent dose of 50 mg/kg (0.70 mmol/kg). Both treatments resulted initially in the rats circling, which was followed by the onset of ataxia at 7-9 d and hindlimb paralysis at 12-14 d. Treated animals showed muscle wasting. At termination, acrylamide- and glycidamide-treated rats weighed 105% and 86% of initial weight, respectively, compared to 145% for controls. Animals were anesthetized and perfused with 10% neutral phosphate-buffered formalin 12 or 14 d after beginning of treatment. Both treatment groups exhibited similar neuropathologic changes in the central and peripheral nervous systems. More severe lesions were produced by glycidamide. A marked increase in the number of affected Purkinje cells in the cerebellum, which exhibited changes ranging from pyknosis to cell death, were present. The brainstem exhibited axonal degeneration with chromatolytic necrosis in midbrain medial and lateral reticular nuclei. The spinal cord was characterized by spongy form changes with vacuoles of different sizes in various levels. These results suggest that glycidamide is an active neurotoxic metabolite of acrylamide.

Effects of Neurotoxins on Brain Creatine Kinase Activity

The effects of ethylene oxide (EO), acrylamide, N,N′-methylene-bis-acrylamide (bis-acrylamide), and methyl mercury chloride (MMC) on brain creatine kinase (CK) activity were examined in vivo. EO and acrylamide, both of which cause central–peripheral distal axonopathy, inhibited CK activity in the brain and spinal cord. On the other hand, neither bis-acrylamide, a nonneurotoxic analogue, nor MMC which causes neuronopathy affected brain CK activity significantly. The inhibition of CK may play a role in the pathogenesis of distal axonal degeneration in the central and peripheral nervous systems.