Presence of Titanium and Toxic Effects Observed in Rat Lungs, Kidneys, and Central Nervous System in vivo and in Cultured Astrocytes in vitro on Exposure by Titanium Dioxide Nanorods.

BACKGROUND: Non-spherical titanium dioxide (TiO2) nanoparticles have been increasingly applied in various biomedical and technological fields. Their toxicological characterization is, however, less complete than that of roundish nanoparticles. MATERIALS AND METHODS: Anatase form TiO2 nanorods, ca. 15x65 nm in size, were applied to cultured astrocytes in vitro and to the airways of young adult Wistar rats in vivo in 5, 10, and 8 mg/kg BW dose for altogether 28 days. Presence of nanorods and cellular damage was investigated in the astrocytes and in rat lungs and kidneys. Functional damage of the nervous system was studied by electrophysiological methods. RESULTS: The treated astrocytes showed loss of viability without detectable apoptosis. In rats, TiO2 nanorods applied to the airways reached the blood and various organs including the lungs, kidneys, and the central nervous system. In lung and kidney samples, nanorods were observed within (partly damaged) phagolysosomes and attached to organelles, and apoptotic cell death was also detected. In cortical and peripheral electrophysiological activity, alterations corresponding to energy shortage (resulting possibly from mitochondrial damage) and astrocytic dysfunction were detected. Local titanium levels and relative weight of the investigated organs, apoptotic cell death in the lungs and kidneys, and changes in the central and peripheral nervous activity were mostly proportional to the applied doses, and viability loss of the cultured astrocytes was also dose-dependent, suggesting causal relationship of treatments and effects. CONCLUSION: Based on localization of the visualized nanorods, on neuro-functional changes, and on literature data, the toxic mechanism involved mitochondrial damage, oxidative stress, and apoptotic cell death. These indicate potential human toxicity and occupational risk in case of exposure to rod-shaped TiO2 nanoparticles.

[Investigation of the effect of titanium dioxide nanorods on the lungs in a subacute rat model]. / Titán-dioxid-nanopálcikák tüdore kifejtett hatásának állatkísérletes vizsgálata szubakut patkánymodellben.

INTRODUCTION: The development of nanotechnology increases the risk of occupational and population-level exposure to nanoparticles nowadays. However, scientifically based knowledge relating to the toxicity of heavy metal nanoparticles and potential health damage is insufficient. AIM: Investigation of lung tissue damage induced by titanium dioxide (TiO2) nanorods in subacute intratracheal instillation by morphological, chemical and biochemical methods in rat model. METHOD: General toxicity (changes of body and organ weights), local acute and chronic cellular toxicity (in alveolar spaces and epithelium, in hilar lymph nodes) and oxidative stress were examined using light and electron microscopy, and biochemical methods (reactive oxygen species, lipid peroxidation, expression of pro-inflammatory cytokines). RESULTS: No dose- and time-dependent alteration was found in the body weight of the treated groups; but the mass and Ti content of lungs increased with dose. Light and electron microscopy of the lung tissue verified the presence of nanoparticles, free in the alveolar space and within phagosomes of macrophages not attached to alveolar epithelium. Chronification of local acute alveolitis was supported by dose-dependent increase of macrophage count in the alveolar region, oedema and thickening of interstitium, and increased expression of certain pro-inflammatory cytokines (interleukin-1a, LIX, L-selectin, vascular endothelial growth factor). Oxidative stress and lipid peroxidation increased substantially in the treated rats' lungs, and correlation was found between Ti content and lipid peroxidation. Insufficiency of the alveolar epithelial and capillary endothelial barrier was indicated by nanoparticle-laden phagocytes in hilar lymph nodes, suggesting nanoparticles reaching systemic circulation and distant organs, inducing systemic acute inflammation. CONCLUSION: TiO2 nanoparticles, reaching lower airways, may be etiological factors in the causation or aggravation of pulmonary diseases with acute and chronic airways inflammation and/or progressive fibrosis and obstruction (e.g., chronic obstructive pulmonary disease or asthma). Autophagy and damaged immune response (lymphocytic activity) may have here a role. Orv Hetil. 2019; 160(2): 57-66.

Pulmonary impact of titanium dioxide nanorods: examination of nanorod-exposed rat lungs and human alveolar cells.

BACKGROUND: Titanium dioxide nanoparticles have numerous applications, resulting in human exposure. Nonetheless, available toxicological and safety data are insufficient regarding aspherical particles, such as rod-shaped nanoparticles. METHODS: In a combined in vitro-in vivo approach, cultured A549 lung alveolar adenocarcinoma cells were treated with approximately 15×65 nm TiO2 nanorod-containing medium, while young adult rats received the same substance by intratracheal instillation for 28 days in 5 and 18 mg/kg body-weight doses. Nanoparticle accumulation in the lungs and consequent oxidative stress, cell damage, and inflammation were assessed by biochemical and histopathological methods. RESULTS: Titanium was detected in tissue samples by single-particle inductively coupled plasma mass spectrometry. Nanoparticles were visualized inside cultured A549 cells, within pulmonary macrophages, and in hilar lymph nodes of the rats. A549 cells showed dose-dependent oxidative stress and lethality, and the observed nanoparticle-laden endosomes suggested deranged lysosomal function and possible autophagy. Strongly elevated Ti levels were measured in the lungs of nanorod-treated rats and moderately elevated levels in the blood of the animals. Numerous cytokines, indicating acute and also chronic inflammation, were identified in the lung samples of TiO2-exposed rodents. CONCLUSION: Several signs of cell and tissue damage were detected in both the cultured alveolar cells and in treated rats' lungs. Rod-shaped nanoparticulate TiO2 may consequently be more harmful than has generally been supposed. The occupational health risk suggested by the results calls for improved safety measures.

Functional neurotoxicity and tissue metal levels in rats exposed subacutely to titanium dioxide nanoparticles via the airways.

BACKGROUND AND PURPOSE: Nanoparticles of titanium dioxide are suspected neurotoxic agents and have numerous applications possibly resulting in human exposure by several ways including inhalation. In the present work, rats were exposed to spherical TiO2 nanoparticles of two different sizes by the intratracheal route. It was investigated how the neuro-functional alterations, detected by electrophysiological and behavioral methods, were related to the concentration of Ti in the tissue samples and what the influence of the size of the NPs was. METHODS: Rats (young adult Wistar males, 10/group) were exposed to TiO2 nanoparticles of ca. 10 and 100 nm diameter (suspension medium: neutral PBS with 1% hydroxyethyl cellulose) by intratracheal instillation in 5 and 18 mg/kg b.w. dose; 5 days per week for 6 weeks. Controls were instilled with saline, and vehicle controls, with the suspension medium. To see general toxicity, body weight was checked daily, and organ weights were measured at the end of experiment. Grip strength test, to assess motor function damage, was done before and after the 6-week treatment. Finally, the rats were anesthetized with urethane, spontaneous cortical activity and sensory evoked potentials were recorded, then the rats were dissected and tissue samples were taken for Ti level measurement. RESULTS: Body weight gain indicated no general toxicity, and no significant change in the relative organ weights, except that of the lungs, was seen. However, change of time-to-fall in the grip strength test, and latency of cortical evoked po-tentials, were altered in the treated groups, indicating functional damage. Correlation of these alterations with the cortical Ti level was dissimilar for the two sizes of nanoparticles. CONCLUSION: The results provided further support to the functional neurotoxicity of TiO2 nanoparticles. The exact role of particle size, and the mechanisms involved, remain to be elucidated.

Electrophysiological alterations and general toxic signs obtained by subacute administration of titanium dioxide nanoparticles to the airways of rats.

BACKGROUND AND PURPOSE: Particles of titanium dioxide (TiO2) with typical size below 100 nm have gained a broad range of application by now, partly involving direct human exposure. Their known properties - high specific surface, mobility within the organism, induction of oxidative stress, release of inflammation mediators etc. - raise the possibility of nervous system damage but the available data regarding this are scarce and contradictory. Based on that, and the experiences with other metal oxide nanoparticles, the aim of the present study was to investigate certain general end nervous system toxic effects of TiO2 nanoparticles applied in the airways of rats. METHODS: Young adult Wistar rats (5 groups of 10 rats each) received, daily for 28 days, intratracheal instillations of titanium dioxide nanoparticles of ca. 10 nm diameter, suspended in 1% hydroxyethyl cellulose dissolved in phosphate-buffered saline, in the doses of 1, 3, and 10 mg/kg b. w. Vehicle controls received the suspension medium and there was also an untreated control group. During treatment, the rats' body weight was measured, and their clinical state observed, daily. After the 28 days, spontaneous cortical activity, sensory evoked potentials and tail nerve action potential was recorded in urethane anesthesia, then the rats were dissected and tissue samples were taken for Ti level determination and biochemical measurements of some oxidative stress indicators. RESULTS: The two higher doses reduced the rate of body weight gain significantly. Sensory evoked potentials and tail nerve action potential were significantly slowed, but the change in the spectrum of spontaneous cortical activity was not significant. Correlation of moderate strength was found between certain evoked potential parameters and brain Ti level and oxidative stress data. CONCLUSION: Our results underlined the possible neurotoxicity of TiO2 NPs but also the need for further investigations.

Behavioral and general effects of subacute oral arsenic exposure in rats with and without fluoride.

Consequences of oral arsenic and fluoride exposure on motor behavior and general toxicity were modeled in young adult male rats which received sodium (meta)arsenite (10 mg/kg b.w.), sodium fluoride (5 mg/kg b.w.), and their combination by gavage, once daily, 5 days a week for 6 weeks. After 6 weeks, 6 animals per group were dissected, while the other 6 were kept for 6 more weeks untreated. Body weight, together with food and water consumption, was measured daily. Arsenic, alone or along with fluoride, caused significant decrease in rearing, and increase in immobility and local activity in the open field in the 3rd and 6th week. By the 12th week, these changes mostly diminished. Weight gain, and food and water consumption were significantly reduced by arsenic but normalized post treatment. Fluoride had no own effect and mostly no influence on effects of arsenic. Massive deposition of arsenic in the rats' blood, cerebral cortex, and liver by the 6th week, and partial elimination by the 12th week, was seen. The results underline the risk of neuro-functional damage by arsenic and call for further investigations.

Potential toxic effects of iron oxide nanoparticles in in vivo and in vitro experiments.

The aim of this study was to determine the potential toxic effects of iron(II,III)oxide nanoparticles (IONPs). In in vivo experiments, the toxic effects of IONPs were monitored in adult male Wistar rats by morphological methods after a single intratracheal instillation. For the control group 1 ml of physiological saline per animal was given, and the treatment group received the same volume of a suspension containing 1 and 5 mg kg⁻¹ body weight IONPs. Lungs and internal organs underwent histopathological examination after 1, 3, 7, 14 and 30 days. The mutagenic effect of these nanoparticles was evaluated by the bacterial reverse mutation assay on Salmonella typhimurium TA98, TA100, TA1535 and TA1537 strains, and on Escherichia coli WP2uvrA strain, in the presence and absence of the mammalian metabolic activation system S9. The in vitro cytotoxic effect of IONPs was also examined in Vero cells after short-term (4 h) and long-term (24 h) exposure. There were no pathological changes in examined internal organs, except a very weak pulmonary fibrosis developing by the end of the first month in the treated rats. While in vitro MTT assay showed a moderate cytotoxic effect, IONPs proved to be devoid of mutagenic effect in the bacterial systems tested. The results may be a useful extension of our knowledge on the safety of magnetite nanoparticles in view of their possible medical applications, such as in hyperthermia and magnetic resonance imaging.

Nervous system effects in rats on subacute exposure by lead-containing nanoparticles via the airways.

CONTEXT AND OBJECTIVE: Lead (Pb) is a heavy metal harmful for human health and environment. From leaded gasoline (still used in certain countries), and in Pb processing and reprocessing industries, airborne particles are emitted which can be inhaled. In such exposure, the size of particles entering the airways is crucial. The nervous system is a primary target for Pb, and consequences like occupational neuropathy and delayed mental development of children are well-known. The aim of this work was to investigate the neurotoxicity of Pb nanoparticles (NPs) applied into the airways of rats. METHODS: Nano-sized lead oxide particles (mean diameter ca. 20 nm) were suspended in distilled water and instilled into the trachea of adult male Wistar rats (in doses equivalent to 2 and 4 mg/kg Pb), 5 times a week for 3 and 6 weeks. At the end, open field motility was tested, then central and peripheral nervous activity was recorded in urethane anesthesia. RESULTS AND CONCLUSION: The treated rats' body weight gain was significantly lower than that of the controls from the 3rd week onwards, and the weight of their lungs was significantly increased. Horizontal motility increased while vertical motility decreased. Spontaneous cortical activity was shifted to higher frequencies. The somatosensory cortical evoked potential showed increased latency and decreased frequency-following ability, and similar alterations were seen in the tail nerve. Significant Pb deposition was measured in blood, brain, lung and liver samples of the treated rats. The experiments performed seem to constitute an adequate model of the human effects of inhaled Pb NPs.

Functional neurotoxicity of Mn-containing nanoparticles in rats.

Airborne metal-containing particles represent a known source of health risk but the role of nano-sized particles in the pathogenicity of dust has been recognized only recently. As a model of inhalational exposure to manganese, adult male Wistar rats were treated with a suspension of MnO(2) nanoparticles of ca. 23 nm diameter, instilled into the trachea for 3, 6, and 9 weeks in daily doses of 2.63 and 5.26 mg Mn/kg, and endpoints of functional neurotoxicity (open field behavior and electrophysiology) and general toxicity (body and organ weights) were investigated. Weekly body weighing showed that control rats had normal weight gain but the treated rats' body weight failed to increase from the 6th week on. Dissection and organ weighing after the corresponding treatment periods revealed dose- and time-dependently increased relative lung weights. In brain and blood samples, significantly elevated Mn level was detected after 9 weeks exposure. The treated rats' open field behavior showed decreased ambulation and rearing, and increased local activity and immobility. Electrophysiological investigations after 9 weeks exposure indicated a shift of the spontaneous cortical activity to higher frequencies, lengthened cortical evoked potential latency, and slowed nerve conduction. Several of these general and neuro-functional parameters were significantly correlated to the tissue Mn levels. Instilled Mn in nanoparticle form was indeed absorbed and exerted neurotoxic effects, so the model seems suitable for studying the effects of airborne nanoparticles, relevant to human health.

Metal deposition and functional neurotoxicity in rats after 3-6 weeks nasal exposure by two physicochemical forms of manganese.

Airborne manganese represents a major risk of nervous system damage first of all in industrial settings. The resulting effects may depend on the dose and physicochemical form of Mn. To compare the effect of soluble and nanoparticulate Mn, adult male rats received daily instillation of MnCl(2) solution or MnO(2) nanoparticle suspension (dose: 2.53mg Mn per rat) into the nasal cavity for 3 and 6 weeks. At the end of treatment, spontaneous open field motility was tested, electrophysiological recording was done in anesthesia, and brain tissue Mn level was determined. Metal level increase in the rats' brain, body weight gain reduction, and decrease of open field motility was significant in the MnCl(2), but not nano-Mn, treated rats. Most evoked cortical activity parameters were significantly altered in both groups, but spontaneous cortical activity spectrum only in the rats receiving MnCl(2). There was fair correlation between brain Mn levels and certain neuro-functional parameters, underlining the causal relationship. Electrophysiological tests might be more sensitive to the effects of Mn than general toxicological or neurobehavioral tests.

Altered open field behavior in rats induced by acute administration of 3-nitropropionic acid: possible glutamatergic and dopaminergic involvement.

3-nitropropionic acid (3-NP), a substance used for modelling Huntington's disease, was given to male Wistar rats in a single 20 mg/kg b.w. dose, and the resulting behavioral alterations in spontaneous locomotor activity were measured after 30 minutes. To detect the involvement of neurotransmitter systems in this immediate effect, the NMDA antagonist MK-801 (0.8 mg/kg); as well as an agonist, quinpirole (QP, 5 mg/kg) and an antagonist, sulpiride (SP, 80 mg/kg) of the dopamine D2 receptors, were given before 3-NP to separate groups of rats. Controls were given saline. All substances were injected ip. 3-NP decreased the rats' locomotor, especially vertical, activity, whereas local activity was increased. Based on the further changes of 3-NP effects in the combination groups it could be concluded that dopaminergic rather than glutamatergic mechanisms were possibly involved in the acute behavioral effect of 3-NP.

Subacute intratracheal exposure of rats to manganese nanoparticles: behavioral, electrophysiological, and general toxicological effects.

The toxicity of manganese-containing airborne particles is an important occupational and environmental problem. In this work, adult male Wistar rats were treated with a nanosuspension of MnO(2) of approximately 23 nm nominal particle diameter, instilled into the trachea for 3, 6, and 9 wk in doses of 2.63 and 5.26 mg Mn/kg. The animals' body weight was checked weekly. At the end of treatment, the rats' spontaneous motility was tested in an open field box. Then, spontaneous and stimulus-evoked cortical activity and action potential of the tail nerve were recorded in urethane anesthesia. The rats were finally dissected, organs weights were measured, and the presence of excess Mn in lung and brain samples was determined using scanning electron microscopy with energy-dispersive x-ray spectroscopy. While control rats had normal weight gain, the body weights of the treated rats ceased to grow from wk 6 on. The relative weight of the lungs increased in the treated rats, and that of the liver decreased, in a dose- and time-dependent manner; Mn was detected in their lung and brain samples. In the open field activity, the percentage of ambulation and rearing decreased while local activity and immobility increased. The latency of the evoked potentials was lengthened, and the conduction velocity of the tail nerve decreased. These results indicate that the Mn content of instilled nanoparticles had access from the airways to the brain, and the resulting damage could be investigated in animals using neuro-functional and general toxicological endpoints.

The acute effects of 3-nitropropionic acid on the behavior and spontaneous cortical electrical activity of rats.

In this study, the acute effect of 3-nitropropionic acid was investigated on open field and startle behavior of rats, and on their cortical electrical activity. Spontaneous locomotor activity, acoustic startle response, and pre-pulse inhibition of acoustic startle were measured in male Wistar rats (10 weeks old, 180-200 g body weight) after a single dose of 10 or 20 mg/kg i.p. 3-nitropropionic acid. After the behavioral tests, the rats were anaesthetized, and spontaneous cortical electrical activity was recorded. The vertical, horizontal and local open field performance showed dose-dependent deterioration in the rats treated with 3-nitropropionic acid. The number of "noise-positive" startle responses showed non-significant changes, but the inhibition by pre-pulse was significantly reduced in the high dose animals. High dose also increased the proportion of low-frequencies in the cortical activity. 3-nitropropionic acid, known primarily to act in repeated doses (e.g., in animal models of Huntington's disease) had also some clear-cut acute effects on behavioral and electrophysiological parameters of the treated rats.

Effects on the central and peripheral nervous activity in rats elicited by acute administration of lead, mercury and manganese, and their combinations.

Adult male Wistar rats were treated with inorganic lead, mercury and manganese, and their double combinations, in acute application. The aim was to study the effects on spontaneous and stimulus-evoked cortical, and evoked peripheral, nervous activity, to detect any interaction of the metals and any correlation between the changes caused in the spontaneous and stimulus-evoked electrical activity of the primary somatosensory cortical area, and the compound action potential of the tail nerve. In the frequency distribution of the spontaneous cortical activity, a shift to lower frequencies was seen. The cortical responses evoked by whisker or tail stimulation showed an increase of the peak-to-peak amplitude and peak latency on administration of the metals and metal combinations. With the metal combinations, synergism was observed. Correlations found between alterations of the spontaneous and evoked, or between cortical and peripheral, activity were evaluated in terms of mechanism. According to the results, combined exposure to the three heavy metals studied might lead to synergistic action, indicating an increased health risk in settings with exposure to several heavy metals.

Alterations in the cortical and peripheral somatosensory evoked activity of rats treated with 3-nitropropionic acid.

In this study, the action of 3-nitropropionic acid (3-NP) on the parameters of the cortical and peripheral evoked potentials was investigated in rats in different administration schemes (20 mg/kg i.p. during recording or 24 h before, and 5x 15 mg/kg daily 28 days before recording) to elucidate some neurophysiological effects of the substance. Responses in the somatosensory cortex and in the tail nerve, evoked by peripheral electric stimulation, were recorded in acute preparation under urethane anaesthesia. Amplitude, latency, and duration of the responses were measured. In rats treated 28 days before recording, latency of the cortical response was significantly (and the duration slightly) increased by 3-NP. The frequency dependence of the tail nerve response was more pronounced than that of the cortical response. After acute administration of 3-NP, the amplitude of the somatosensory evoked potential decreased. With double stimuli, the ratio of the amplitudes of the two responses (relative fatigue) was treatment-dependent. The relative refractory period of the tail nerve was altered both by acute and subacute 3-NP treatment. These results may be relevant in 3-NP based disease models but it needs further studies to find possible connections between the known biochemical effects of 3-NP and the functional neurotoxical changes described. The mode of evoked response analysis used is, theoretically, applicable for other neurotoxic effects and can be the base of development of functional biomarkers.

Acute effects of lead, mercury and manganese on the central and peripheral nervous system in rats in combination with alcohol exposure.

Heavy metals, due to their numerous applications in industrial processes, agrochemicals and household articles, have caused a widespread pollution and can be found in different foods. One of their target organs is the central nervous system. The toxic effects of heavy metals can be modified by lifestyle-originated factors such as consumption of alcohol. The aim of this study was to investigate the changes in spontaneous cortical activity (ECoG), cortical sensory evoked potentials (EPs) and peripheral nerve action potentials, recorded in rats pre-treated with alcohol and acutely treated with lead, mercury and manganese by intraperitoneal injection. In the ECoG, Hg2+ caused a massive shift to lower frequencies while the effect of Mn2+ and Pb2+ was slight, and alcohol pre-treatment altered the effect of the metals minimally. The amplitude of EPs increased upon the application of heavy metals, and the peak latency lengthened. The effect of Hg2+ was the strongest and that of Pb2+ the weakest, and these effects were potentiated by alcohol. Exposure to heavy metals, together with alcohol consumption, can aggravate the known neurotoxic effects.

Behavioral and neurotoxic effects seen during and after subchronic exposure of rats to organic mercury.

Young adult male Wistar rats (24/group) were treated for 5 weeks with methyl mercury(II)chloride (corresponding to 0.5 and 2.0mgHg°/kg b.w., control: distilled water) by gavage, followed by a 19 weeks post-treatment period. Spontaneous motility, psychomotor performance and sensorimotor gating was repeatedly tested, electrophysiological recordings done, in the rats throughout the whole experiment. Decreased horizontal open field activity, reduced number of "noise positive" startle responses, as well as increase of startle response onset latency and peak time, and decrease of peak amplitude, was seen in the treated animals. Most changes disappeared in the post-treatment period. In the spontaneous cortical and hippocampal activity, altered distribution of the frequency bands was seen after 5 weeks of treatment but not at the end of the post-treatment period. Hippocampal population spikes in the treated animals were depressed and showed less potentiation, which effect was still present 19 weeks after finishing the treatment. The duration of the sensory cortical evoked potentials was shorter than in the controls. In the treated rats, tyrosine hydroxylase-immunoreactive boutons in the substantia nigra pars reticulata were shrunk; blood and brain Hg levels were significantly higher and decreased only slowly. Considering the continuous presence of low levels of mercurials in the human environment, effects of this kind may be supposed as the background of some human neurobehavioral abnormalities.

Behavioral and neurotoxicological effects of subchronic manganese exposure in rats.

In male Wistar rats, behavioral and electrophysiological investigations, and blood and brain manganese level determinations, were performed; during 10 weeks treatment with low-dose manganese chloride and a 12 weeks post-treatment period. Three groups of 16 animals each received daily doses of 14.84 and 59.36mg/kg b.w. MnCl(2) (control: distilled water) via gavage. During treatment period, Mn accumulation was seen first in the blood, then in the brain samples of the high-dose animals. Short- and long-term spatial memory performance of the treated animals decreased, spontaneous open field activity (OF) was reduced. The number of acoustic startle responses (ASR), and the pre-pulse inhibition (PPI) of these, diminished. In the cortical and hippocampal spontaneous activity, power spectrum was shifted to higher frequencies. The latency of the sensory evoked potentials increased, and their duration, decreased. By the end of the post-treatment period, Mn levels returned to the control in all samples. The impairment of long-term spatial memory remained, as did the number of acoustic startle responses. Pre-pulse inhibition, however, returned to the pre-treatment levels. The changes of the open field activity disappeared but a residual effect could be revealed by administration of d-amphetamine. The electrophysiological effects were partially reversed. By applying a complex set of methods, it was possible to obtain new data for a better-based relationship between the known effects of Mn at neuronal level and the behavioral and electrophysiological outcomes of Mn exposure.

Subchronic mercury treatment of rats in different phases of ontogenesis: functional effects on the central and peripheral nervous system.

Electrophysiological changes caused by inorganic mercury administration during the pre- and/or postnatal development were studied. Pregnant female Wistar rats were treated, by gavage, with 0.4, 0.8 or 1.6 mg/kg mercury (HgCl2 diluted in distilled water): 1/ from day 5 to 15 during pregnancy (P protocol); 2/ from day 5 to 15 of pregnancy+for 4 weeks of lactation (P+L protocol); 3/ from day 5 to 15 of pregnancy+for 4 weeks of lactation, and the offspring were further treated for 8 weeks post-weaning (P+L+P protocol). Electrophysiological parameters (electrocorticogram, cortical evoked potentials, conduction velocity and refractory periods of peripheral nerve) of the male offspring from dams in the groups treated according to the above protocols were investigated at the age of 12 weeks. The rats' spontaneous and evoked electrophysiological activity underwent dose- and treatment-dependent changes following the treatment (increased frequency of spontaneous activity, lengthened latencies and duration of evoked potentials, lower conduction velocity of the peripheral nerve, etc.). In the same rats, however, the treatment failed to cause major signs of general intoxication. The results emphasize the functional neurotoxic risk arising from the continuous presence of inorganic mercury in the human environment, and point to possible use of early functional changes for monitoring the effects of mercury.