Although occupational exposure to n-hexane induces neurotoxic effects in the central and peripheral nervous systems, the mechanisms of its neurotoxicity remain unclear. n-Hexane is metabolized to 2,5-hexanedione (2,5-HD), which is the neurotoxic agent and the indicator chosen for the biological monitoring of exposed workers. It has been previously reported that chronic exposure to 2,5-HD impairs the glutamate-nitric oxide-cyclic GMP pathway at the level of activation of soluble guanylate cyclase (sGC) enzyme by nitric oxide (NO), both in cultured neurons and in the cerebellum of rats in vivo. The aim of this study was to assess whether the activation of sGC by NO is also altered in lymphocytes from rats treated with 2,5-HD and/or workers chronically exposed to n-hexane. Lymphocytes were isolated from male Wistar rats treated with 2,5-HD in drinking water, and from blood samples from shoe-factory workers environmentally and chronically exposed to n-hexane. Urine samples were also collected from workers at the end of the shift in order to measure the urinary levels of 2,5-HD. Activation of sGC by NO was significantly higher (p<0.05) in lymphocytes from rats treated with 2,5-HD than in control rats. In isolated lymphocytes from exposed workers the activation of sGC by NO also increases (p<0.05) in contrast to the controls. The results presented here indicate that the activation of lymphocytes could be an indicator of the toxicity produced by being exposed to n-hexane, since the effects observed in workers chronically exposed to n-hexane are similar to those found in rats chronically treated with 2,5-HD in drinking water.
Toxicology 02/2007; 229(1-2):73-8. DOI:10.1016/j.tox.2006.10.002 · 3.75 Impact Factor
Hexane is used in many industrial processes and induces neurotoxic effects in the central and peripheral nervous system. Hexane is metabolized to 2,5-hexanedione, which is the neurotoxic agent. Continued exposure to hexane or 2,5-hexanedione results in loss of sensorial and motor function in arms and legs and to alterations in axonal neurofilament proteins. The effects of 2,5-hexanedione on different cytoskeletal proteins in different brain areas have not been studied in detail. The aim of this work was to study the effects of chronic exposure of rats to 2,5-hexanedione (1% in the drinking water) on tubulin, neurofilament NF-L, microtubule-associated protein MAP-2, and on glial fibrillary acidic protein (GFAP), in cerebellum, hippocampus and cerebral cortex. The amount of each protein was determined by immunoblotting and its distribution was analysed by immunohistochemistry. The results obtained show a regional selectivity in the 2,5-hexanedione effects on cytoskeletal proteins. NF-L content decreased in all brain areas. MAP-2 decreased in cerebellum and hippocampus and tubulin decreased only in cerebellum. GFAP decreased only in cerebral cortex, but its distribution was altered in cerebellum, with increased content in the granular layer and decreased content in the molecular layer. The area most affected was the cerebellum, where all the proteins analysed were altered. These cytoskeletal proteins alterations may impair the transfer of information involved in the regulation by the cerebellum of motor function and contribute to the altered motor performance in rats exposed to 2,5-hexanedione and humans exposed to hexane.
Toxicology and Industrial Health 09/2002; 18(7):333-41. DOI:10.1191/0748233702th154oa · 1.71 Impact Factor
Glutamate is the main excitatory neurotransmitter in mammals. However, excessive activation of glutamate receptors is neurotoxic,
leading to neuronal degeneration and death. In many systems, including primary cultures of cerebellar neurons, glutamate neurotoxicity
is mainly mediated by excessive activation of NMDA receptors, leading to increased intracellular calcium which binds to calmodulin
and activates neuronal nitric oxide synthase (NOS), increasing nitric oxide (NO) which in turn activates guanylate cyclase
and increases cGMP. Inhibition of NOS prevents glutamate neurotoxicity, indicating that NO mediates glutamate-induced neuronal
death in this system. NO generating agents such as SNAP also induce neuronal death. Compounds that can act as “scavengers”
of NO such as Croman 6 (CR-6) prevent glutamate neurotoxicity.
The role of cGMP in the mediation of glutamate neurotoxicity remain controversial. Some reports indicate that cGMP mediates
glutamate neurotoxicity while others indicate that cGMP is neuroprotective. We have studied the role of cGMP in the mediation
of glutamate and NO neurotoxicity in cerebellar neurons. Inhibition of soluble guanylate cyclase prevents glutamate and NO
neurotoxicity. There is a good correlation between inhibition of cGMP formation and neuroprotection. Moreover 8-Br-cGMP, a
cell permeable analog of cGMP, induced neuronal death. These results indicate that increased intracellular cGMP is involved
in the mechanism of neurotoxicity. Inhibitors of phosphodiesterase increased extracellular but not intracellular cGMP and
prevented glutamate neurotoxicity. Addition of cGMP to the medium also prevented glutamate neurotoxicity. These results are
compatible with a neurotoxic effect of increased intracellular cGMP and a neuroprotective effect of increased extracellular
Neurotoxicity Research 04/2001; 3(2):179-188. DOI:10.1007/BF03033190 · 3.15 Impact Factor