RESUMEN
The organotin compounds trimethyltin (TMT), triethyltin (TET) and tributyltin (TBT) show different organotoxicities in vivo. While TMT and TET induce a strong neurotoxicity accompanied by microglial and astroglial activation, TBT rather effects the immune system. Previously, we have shown in an in vitro co-culture model that microglial cells can be activated by TMT in the presence of astrocytes. In this study, we wanted to investigate (a) if the neurotoxic organotin compound TET can also activate microglial cells in vitro similar to TMT and (b) if differences between the neurotoxicants TMT and TET on the one side and TBT on the other exist concerning microglial activation. Therefore, purified microglial and astroglial cell cultures from neonatal rat brains were treated either alone or in co-cultures for 24h with different concentrations of TMT, TET or TBT and the basal cytotoxicity and nitric oxide formation was determined. Furthermore, morphological changes of astrocytes were examined. Our results show that microglial activation can be increased in subcytolethal concentrations, but only in the presence of astrocytes and not in microglial cell cultures alone. This increase was induced by the neurotoxicants TMT and TET but not by TBT. Taken together, the differing microglia activating effect of the organotin compounds may contribute to the differing neurotoxic potential of this group of chemicals in vivo. In addition, our results emphasize the need for co-culture systems when studying interactions between different cell types for toxicity assessment.
Asunto(s)
Astrocitos/efectos de los fármacos , Microglía/efectos de los fármacos , Compuestos de Trialquiltina/toxicidad , Compuestos de Trietilestaño/toxicidad , Compuestos de Trimetilestaño/toxicidad , Animales , Células Cultivadas , Técnicas de Cocultivo , Relación Dosis-Respuesta a Droga , Óxido Nítrico/biosíntesis , Ratas , Ratas WistarRESUMEN
The potent antidepressant effect of moclobemide, a selective and reversible type A monoamine oxidase (MAO) inhibitor, is clinically established. In view of the ongoing debate on the neuroprotective properties of MAO inhibitors, the present study was undertaken to further define the protective effect of moclobemide in a rat model of neurotoxicant-induced edema. In this model, daily oral triethyltin (TET) administration for 5 consecutive days strongly perturbed the rat behaviour and induced a cerebral edema at the 5th day. Oral coadministration of moclobemide (2 x 100 mg.kg-1.day-1) with TET blocked the development of brain edema and the increase in the cerebral chloride content induced by TET. Moreover, moclobemide reduced the increase in the cerebral sodium content and attenuated the neurological deficit. In conclusion, moclobemide possesses potent protective properties in this rat model of cerebral edema, suggesting potential clinical utility as a neuroprotectant.
Asunto(s)
Edema Encefálico/prevención & control , Moclobemida/farmacología , Compuestos de Trietilestaño/toxicidad , Administración Oral , Animales , Índice de Masa Corporal , Edema Encefálico/inducido químicamente , Edema Encefálico/fisiopatología , Calcio/metabolismo , Corteza Cerebral/química , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/patología , Cloruros/metabolismo , Relación Dosis-Respuesta a Droga , Intubación Gastrointestinal , Masculino , Moclobemida/administración & dosificación , Moclobemida/uso terapéutico , Monoaminooxidasa/efectos de los fármacos , Inhibidores de la Monoaminooxidasa/administración & dosificación , Inhibidores de la Monoaminooxidasa/farmacocinética , Inhibidores de la Monoaminooxidasa/uso terapéutico , Actividad Motora/efectos de los fármacos , Síndromes de Neurotoxicidad/etiología , Síndromes de Neurotoxicidad/fisiopatología , Síndromes de Neurotoxicidad/prevención & control , Potasio/metabolismo , Ratas , Ratas Wistar , Sodio/metabolismo , Factores de Tiempo , Resultado del Tratamiento , Compuestos de Trietilestaño/administración & dosificación , Agua/metabolismoRESUMEN
The neurotoxic effects of chemical agents are often investigated in controlled studies on rodents, with multiple binary and continuous endpoints routinely collected. One goal is to conduct quantitative risk assessment to determine safe dose levels. Such studies face two major challenges for continuous outcomes. First, characterizing risk and defining a benchmark dose are difficult. Usually associated with an adverse binary event, risk is clearly definable in quantal settings as presence or absence of an event; finding a similar probability scale for continuous outcomes is less clear. Often, an adverse event is defined for continuous outcomes as any value below a specified cutoff level in a distribution assumed normal or log normal. Second, while continuous outcomes are traditionally analyzed separately for such studies, recent literature advocates also using multiple outcomes to assess risk. We propose a method for modeling and quantitative risk assessment for bivariate continuous outcomes that address both difficulties by extending existing percentile regression methods. The model is likelihood based; it allows separate dose-response models for each outcome while accounting for the bivariate correlation and overall characterization of risk. The approach to estimation of a benchmark dose is analogous to that for quantal data without the need to specify arbitrary cutoff values. We illustrate our methods with data from a neurotoxicity study of triethyl tin exposure in rats.
Asunto(s)
Modelos Estadísticos , Enfermedades del Sistema Nervioso Periférico/inducido químicamente , Medición de Riesgo/métodos , Pruebas de Toxicidad/métodos , Animales , Relación Dosis-Respuesta a Droga , Humanos , Funciones de Verosimilitud , Análisis Multivariante , Sistema Nervioso Periférico/efectos de los fármacos , Ratas , Compuestos de Trietilestaño/toxicidadRESUMEN
Zhu et al. (Zhu, Y., Wessel, M., Liu, T., Moser, V.C., 2005. Analyses of neurobehavioral screening data: dose-time-response modeling of continuous outcomes. Regul. Toxicol. Pharmacol. 41, 240-255) have recently applied dose-time-response models to longitudinal or time-course neurotoxicity data, and have illustrated the modeling process using continuous data from a functional observational battery (FOB). Following the work of these authors, the purpose of this paper is to show that the benchmark dose (BMD) method for single time point dose-response data can be generalized and applied to longitudinal data such as those generated in neurotoxicity studies. We propose a statistical procedure called bootstrap method for computing the lower confidence limits for the BMD. We demonstrate the method using three previously published FOB datasets of triethyltin (Moser, V.C., Becking, G.C., Cuomo, V., Frantik, E., Kulig, B., MacPhail, R.C., Tilson, H.A., Winneke, G., Brightwell, W.S., DeSalvia, M.A., Gill, M.W., Haggerty, G.C., Hornychova, M., Lammers, J., Larsson, J., McDaniel, K.L., Nelson, B.K., Ostergaard, G., 1997a. The IPCS study on neurobehavioral screening methods: results of chemical testing. Neurotoxicology 18, 969-1056.) and the models of Zhu et al. (Zhu, Y., Wessel, M., Liu, T., Moser, V.C., 2005. Analyses of neurobehavioral screening data: dose-time-response modeling of continuous outcomes. Regul. Toxicol. Pharmacol. 41, 240-255).
Asunto(s)
Algoritmos , Conducta Animal/efectos de los fármacos , Intoxicación del Sistema Nervioso por Metales Pesados/fisiopatología , Compuestos de Trietilestaño/toxicidad , Animales , Benchmarking/métodos , Benchmarking/estadística & datos numéricos , Relación Dosis-Respuesta a Droga , Miembro Anterior/efectos de los fármacos , Miembro Anterior/fisiopatología , Intoxicación del Sistema Nervioso por Metales Pesados/etiología , Miembro Posterior/efectos de los fármacos , Miembro Posterior/fisiopatología , Modelos Biológicos , Ratas , Factores de TiempoRESUMEN
Trimethyltin (TMT) and triethyltin (TET) caused cell death in cultures of primary human neurons and astrocytes, rat neurons and human neuroblastoma cell lines. Human neurons and astrocytes showed a delayed response to TMT cytotoxicity. After 24h of TMT exposure, LC50 values were 148.1, 335.5 and 609.7 microM for SK-N-MC neuroblastoma cell line, neurons and astrocytes, respectively. Over 5 days of exposure, the cytotoxic potency of TMT increased about 70-fold in human cortical neurons. Rat hippocampal neurons were the most vulnerable cells to TMT cytotoxicity, exhibiting an LC50 value 30-fold lower (1.4 microM) than that of rat cerebellar granule cells (44.28 microM). With the exception of rat hippocampal neurons, TET was more potent than TMT in inducing cell death (LC50 values of 3.5-16.9 microM). Moreover, TET was more effective than TMT in increasing intracellular free Ca2+ concentration in human and rat neurons. This work shows that human fetal neuron and astrocyte cultures are a useful model for studying the neurotoxic effects of these environmental contaminants and, thus, predicting their impact on human health.
Asunto(s)
Astrocitos/efectos de los fármacos , Astrocitos/patología , Neuronas/efectos de los fármacos , Neuronas/patología , Compuestos de Trietilestaño/toxicidad , Compuestos de Trimetilestaño/toxicidad , Animales , Calcio/metabolismo , Línea Celular , Feto/citología , Humanos , Masculino , Neuroblastoma/patología , Ratas , Ratas WistarRESUMEN
Triethyltin (TET)-induced neurotoxicity in the brain causes the formation of myelin edema and loss. Myelin deficits produced by early postnatal exposure to TET are permanent and cannot be repaired as the brain matures. The underlying causes have not been resolved. To investigate whether TET directly affects oligodendrocytes, the myelin-forming cells of the central nervous system, cultured rat brain oligodendrocytes were prepared and treated with TET. The data show that TET was cytotoxic for oligodendrocytes and led to the onset of programmed cell death, as indicated by DNA fragmentation. Cellular membranous extensions were severely damaged, and the nuclei appeared to be condensed and fragmented. Concomitantly, the small heat shock protein HSP32, also known as heme oxygenase-1 (HO-1), and an indicator of oxidative stress, as well as the activation of extracellular signal-regulated kinases 1 and 2 (ERK1,2), were observed. ERK1,2 have been implicated to participate in the regulation of cell death and survival. Myelin-specific proteins MBP and CNP were not affected. In TET-treated cells mitochondria redistributed from the processes to the cell somata near the nucleus, possibly as a consequence of microtubule disorganization. A disturbance of the mitochondrial membrane potential and mitochondrial fragmentation occurred. Hence, it might be hypothesized that oligodendroglial PCD, rather than axonal degeneration, contributes to myelin damage and deficits observed in rats after treatment with TET in vivo.
Asunto(s)
Apoptosis/efectos de los fármacos , Apoptosis/fisiología , Oligodendroglía/efectos de los fármacos , Oligodendroglía/metabolismo , Compuestos de Trietilestaño/toxicidad , Animales , Muerte Celular/efectos de los fármacos , Muerte Celular/fisiología , Células Cultivadas , Oligodendroglía/patología , Ratas , Ratas WistarRESUMEN
The toxicity and antitumor activity of the novel organotin compound triethyltin(IV)lupinylsulfide hydrochloride (IST-FS 29), administered by the oral route, have been evaluated against three transplantable murine tumor models: P388 lymphocytic leukemia, B16F10 melanoma and 3LL Lewis lung carcinoma. Mild and reversible signs of acute toxicity such as behavioral symptoms, weight loss and histological alterations were mainly reported at the highest single dose of 28 mg/kg. Conversely, lower concentrations of compound ranging from 7 to 21 mg/kg did not result in major toxic effects, even after repeated dosing. The antitumor activity studies showed that fractionation dosing, rather than single bolus administration, over 1 week, might prove more active and better tolerated by allowing the achievement of the highest therapeutic total dose of IST-FS 29 (42 mg/kg). Indeed, repeated administrations of IST-FS 29 resulted in marked significant improvement of antitumor activity against B16F10 (50% of tumor volume inhibition, p = 0.0003) and, to a greater extent, 3LL (90% of tumor volume inhibition, p = 0.0001) tumors. These results indicate that IST-FS 29 might be a suitable candidate as an orally administrable anticancer drug and support its further development in human tumor xenografts.
Asunto(s)
Antineoplásicos/farmacología , Neoplasias Experimentales/tratamiento farmacológico , Compuestos Orgánicos de Estaño/farmacología , Compuestos de Trietilestaño/farmacología , Animales , Antineoplásicos/toxicidad , Carcinoma Pulmonar de Lewis/tratamiento farmacológico , Femenino , Leucemia P388/tratamiento farmacológico , Melanoma Experimental/tratamiento farmacológico , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Trasplante de Neoplasias , Neoplasias Experimentales/patología , Compuestos Orgánicos de Estaño/toxicidad , Compuestos de Trietilestaño/toxicidad , Células Tumorales CultivadasRESUMEN
The effects of triethyltin on Ca2+ mobilization in human PC3 prostate cancer cells have been explored. Triethyltin increased [Ca2+]i at concentrations larger than 3 microM with an EC50 of 30 microM. Within 5 min, the [Ca2+]i signal was composed of a gradual rise and a sustained phase. The [Ca2+]i signal was reduced by half by removing extracellular Ca2+. The triethyltin-induced [Ca2+]i increases were inhibited by 40% by 10 microM nifedipine, nimodipine and nicardipine, but were not affected by 10 microM of verapamil or diltiazem. In Ca2+-free medium, pretreatment with thapsigargin (1 microM), an endoplasmic reticulum Ca+ pump inhibitor, reduced 200 microM triethyltin-induced Ca+ increases by 50%. Pretreatment with U73122 (2 microM) to inhibit phospholipase C did not alter 200 microM triethyltin-induced [Ca2+]i increases. Incubation with triethyltin at a concentration that did not increase [Ca2+]i (1 microM) in Ca2+-containing medium for 3 min potentiated ATP (10 microM)- or bradykinin (1 microLM)-induced [Ca2+]i increases by 41 +/- 3% and 51 +/- 2%, respectively. Collectively, this study shows that the environmental toxicant triethyltin altered Ca2+ handling in PC3 prostate cancer cells in a concentration-dependent manner: at higher concentrations it increased basal [Ca2+]i; and at lower concentrations it potentiated agonists-induced [Ca2+]i increases.
Asunto(s)
Calcio/metabolismo , Neoplasias de la Próstata/metabolismo , Compuestos de Trietilestaño/toxicidad , Calcio/deficiencia , ATPasas Transportadoras de Calcio/antagonistas & inhibidores , Relación Dosis-Respuesta a Droga , Antagonismo de Drogas , Fura-2/metabolismo , Humanos , Masculino , Nicardipino/farmacología , Nifedipino/farmacología , Nimodipina/farmacología , Tapsigargina/farmacología , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/metabolismoRESUMEN
The neurotoxic organotin compounds trimethyl (TMT) and triethyltin (TET) are known to induce astrogliosis in vivo, which is indicated by an increased synthesis of glial fibrillary acidic protein (GFAP) in astrocytes. In contrast, tributyltin (TBT) does not induce astrogliosis. The aim of this study was to investigate whether trialkyltin derivatives can induce an increased GFAP synthesis in astrocyte cultures in the absence of neurons and whether differences between the action of TMT, TET, and TBT can be detected. Primary cultures of rat cortical astrocytes from 2-day-old rats were grown in 96-well plates until confluency and then exposed to various concentrations of TMT, TET, and TBT for 40 h. Effects on basal cell functions were measured by colorimetric determination of cell protein contents and by assessment of viability by means of the MTT assay. An indirect sandwich ELISA for 96-well plates was used for quantitative measurements of the GFAP content of the cells. All three compounds induced a concentration-dependent cytotoxicity indicated by parallel decreases of protein contents and MTT reduction. Half-maximum cytotoxic concentrations were 3 micromol/L (TBT), 30 micromol/L (TET), and 800 micromol/L (TMT). Cellular GFAP contents were reduced in parallel to cytotoxic action but no increase in GFAP expression at subcytotoxic concentrations could be observed. Thus, the astrocytes were not able to respond to TMT or TET exposure by an increased synthesis of GFAP in the absence of neuronal signals.
Asunto(s)
Astrocitos/efectos de los fármacos , Compuestos de Trietilestaño/toxicidad , Compuestos de Trimetilestaño/toxicidad , Animales , Astrocitos/citología , Astrocitos/metabolismo , Recuento de Células , Células Cultivadas , Corteza Cerebral/citología , Proteína Ácida Fibrilar de la Glía/metabolismo , Ratas , Ratas Endogámicas F344 , Compuestos de Trialquiltina/toxicidadRESUMEN
The cytotoxic effect and antitumor activity induced by the novel organotin compound triethyltin(IV)lupinyisulfide hydrochloride, have been investigated. Different patterns of antiproliferative effects have been observed in a panel of human tumor cell lines in vitro. Toxicity studies in mice reported acute toxicity at the doses of 21 and 17.5 mg/kg which progressively disappeared at lower concentrations. On this basis, the doses of 3.5, 7 and 14 mg/kg were selected to assess the antitumor activity in vivo against the P388 leukemic cells xenografted in mice. This compound was able to induce a dose-dependent significant reduction of tumor volume, up to 46%, at the highest concentration (p = 0.0062) without important toxicity, as also confirmed by histological analysis of the main organ tissues. This preliminary study seems to hold interest for further investigations in different tumor models as well as for the evaluation of optimal drug route and schedule.
Asunto(s)
Antineoplásicos/toxicidad , Supervivencia Celular/efectos de los fármacos , Riñón/patología , Leucemia P388/tratamiento farmacológico , Compuestos Orgánicos de Estaño/toxicidad , Compuestos de Trietilestaño/toxicidad , Animales , Antineoplásicos/uso terapéutico , Femenino , Humanos , Células Jurkat , Riñón/efectos de los fármacos , Leucemia P388/patología , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Compuestos Orgánicos de Estaño/uso terapéutico , Trasplante Heterólogo , Compuestos de Trietilestaño/uso terapéutico , Células Tumorales CultivadasRESUMEN
The environmental contaminants trimethyltin (TMT) and triethyltin (TET) stimulated the spontaneous release of [(3)H]noradrenaline ([(3)H]NA) from hippocampal slices in a time- and concentration-dependent manner. TMT was the most potent compound, exhibiting an EC50 value 10-fold lower (3.8 microM) than that of TET (39.5 microM). Metal-evoked [(3)H]NA release did not increase in the absence of desipramine and was completely blocked by reserpine preincubation, indicating a vesicular origin of [(3)H]NA release but not a mechanism involving reversal of the transmitter transporter. The voltage-gated Na(+) channel blocker tetrodotoxin (TTX) did not affect metal-evoked [(3)H]NA release. [(3)H]NA release elicited by TMT was partially extracellular Ca(2+)-dependent, since it was significantly decreased in a Ca(2+)-free EGTA-containing medium, whereas TET induced an extracellular Ca(2+)-independent release of [(3)H]NA. Neither inhibitors of Ca(2+)-entry through Na(+)/Ca(2+)exchanger and voltage-gated calcium channels, nor agents that interfere with Ca(2+)-mobilization from intracellular stores affected [(3)H]NA release induced by TMT. TET-evoked [(3)H]NA release was reduced by ruthenium red, which depletes mitochondrial Ca(2+)stores, but was not modified by caffeine and thapsigargin, which interfere with Ca(2+)mobilization from endoplasmic reticulum. The fact that TET effect was also attenuated by DIDS, an inhibitor of anion exchange, indicates that the effect of TET on spontaneous [(3)H]NA release may be mediated by intracellular mobilization of Ca(2+) from mitochondrial stores through a Cl(-) dependent mechanism.
Asunto(s)
Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Norepinefrina/metabolismo , Compuestos de Trietilestaño/toxicidad , Compuestos de Trimetilestaño/toxicidad , Ácido 4,4'-Diisotiocianostilbeno-2,2'-Disulfónico/farmacología , Inhibidores de Captación Adrenérgica/farmacología , Animales , Transporte Biológico/efectos de los fármacos , Calcio/metabolismo , Bloqueadores de los Canales de Calcio/farmacología , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Quelantes/farmacología , Cloruros/metabolismo , Desipramina/farmacología , Ácido Egtácico/farmacología , Contaminantes Ambientales/toxicidad , Iones , Masculino , Norepinefrina/antagonistas & inhibidores , Ratas , Ratas Wistar , Sodio/metabolismo , Bloqueadores de los Canales de Sodio , Canales de Sodio/metabolismo , Intercambiador de Sodio-Calcio/antagonistas & inhibidores , Intercambiador de Sodio-Calcio/metabolismo , Tetrodotoxina/farmacología , Tritio , omega-Conotoxina GVIA/farmacologíaRESUMEN
It has been demonstrated that exposure to mercury or cadmium compounds causes alterations in the glutathione system in a model glial cell line, C6. Here we report that two organic tin compounds, triethyltin (TET) and trimethyltin (TMT), are also toxic to these cells with EC50 values for cell death of c. 0.02 microM and 0.8 microM respectively. Exposure for 24 h to either of these compounds at sub-toxic concentrations caused increases in the amount of reduced glutathione (GSH) per cell. Increases in glutathione-S-transferase enzyme activity were also demonstrated after TET or TMT exposure. This suggests that glutathione increases occur in glial cells after toxic insults below that required to cause cell death, possibly acting as a protective mechanism. To test whether GSH plays a role in organotin-induced cell death we manipulated GSH in the culture media or via intracellular GSH and looked at the effects on sensitivity to TET or TMT toxicity. Adding GSH to the culture media did not protect the cells. Depletion of intracellular GSH with buthionine-[S,R] sulphoximine did not alter cytotoxicity of TET or TMT. However, pre-treatment with (-)-2-oxo-4-thiazolidine carboxylic acid (OTC), which increases intracellular GSH levels, protected the cells against both compounds. The EC50 for TMT was increased from 0.77 to 1.8 microM, a 2.3-fold shift, whereas the EC50 for TET was increased > 20-fold, from 0.022 to 0.47 microM. One interpretation of these results is that GSH protects cells against the toxicity of organic tin compounds without reacting directly with them to any significant extent. Under conditions where GSH is depleted, additional protective mechanisms may be active.
Asunto(s)
Astrocitos/efectos de los fármacos , Glutatión/metabolismo , Compuestos de Trietilestaño/toxicidad , Compuestos de Trimetilestaño/toxicidad , Animales , Astrocitos/metabolismo , Glioma , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
The effect of triethyltin-chloride (TET), a highly neurotoxic compound, on the cellular metabolism of rat brain astrocytes in vitro was examined by nuclear magnetic resonance (NMR) spectroscopy. 5-week-old cultures were exposed to TET (0.2-40 microM) either for (1) acute (3h), (2) 24 h, or (3) chronic treatment (8 d). Cells were labeled with 1-(13)C-glucose, cell extracts were prepared and 31P, 1H, and 13C spectra were analyzed. Cytotoxic effects of TET were assessed by vital dye uptake assay using neutral red (NR) and by exclusion of trypan blue (TB). Cells were examined ultrastructurally by electron microscopy. The data show that the major target of TET at concentrations already causing morphological effects on cultured astrocytes is not the energy metabolism, but that TET rather alters the intracellular concentrations of organic osmolytes, such as myo-inositol, taurine and hypotaurine, which are part of the control of ion and volume regulation and osmotic balance in astrocytes.
Asunto(s)
Astrocitos/metabolismo , Metabolismo Energético/efectos de los fármacos , Compuestos de Trietilestaño/farmacología , Animales , Animales Recién Nacidos , Astrocitos/efectos de los fármacos , Muerte Celular , Células Cultivadas , Espectroscopía de Resonancia Magnética , Microscopía Electrónica , Ratas , Factores de Tiempo , Compuestos de Trietilestaño/administración & dosificación , Compuestos de Trietilestaño/toxicidadRESUMEN
Neonatal Long-Evans rats dosed with TET (5 mg/kg; IP) or saline on postnatal day (PND) 10 were examined across the life span for neural damage and performance on spatial learning tasks. A subset of rats were sacrificed to assess early damage with Nissl-staining, Timm's histochemistry, and glial fibrillary acidic protein (GFAP) immunohistochemistry 2, 7, or 14 days after dosing. Littermates were tested behaviorally in a T-maze spatial delayed alternation task on PND 23 or PND 90, and in a Morris water maze place learning task at 3, 12, or 24 months postdosing and then sacrificed for histological analysis. In neonatal rats, histological analysis indicated gliosis in discrete cortical regions, loss of Nissl-stained neurons in the hippocampal formation, entorhinal cortex and piriform cortex, and loss of Timm's staining in the entorhinal cortex. The behavioral assessment at PND 23 indicated a significant impairment in the T-maze. However, no significant impairments were observed in the T-maze at 3 months or the water maze at 3 or 12 months postdosing. At 24 months, TET-treated rats showed significant deficits in acquisition and retention of the water maze task compared with age-matched controls. Both groups of 24 months old rats were significantly impaired compared with young controls. At 24 months, there was a general age-related decrease in the optical density of Timm's staining in cortical regions (9%), compounded by a further decrease in the entorhinal cortex and outer molecular layer of the dentate gyrus of the hippocampus in TET treated rats (30%). These data indicate that early developmental exposure to an organometal resulted in morphological damage that was apparent behaviorally only during early postnatal development and with advanced aging.
Asunto(s)
Envejecimiento/fisiología , Sistema Nervioso/efectos de los fármacos , Compuestos de Trietilestaño/toxicidad , Envejecimiento/patología , Análisis de Varianza , Animales , Animales Recién Nacidos , Conducta Animal/efectos de los fármacos , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/patología , Giro Dentado/efectos de los fármacos , Giro Dentado/patología , Femenino , Proteína Ácida Fibrilar de la Glía/metabolismo , Gliosis , Inmunohistoquímica , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Sistema Nervioso/metabolismo , Sistema Nervioso/patología , Ratas , Ratas EndogámicasRESUMEN
RATIONALE AND OBJECTIVES: We investigated the multicompartmental nature of T2 decay in a specific white matter edema model. METHODS: Triethyltin (TET) intoxication was produced in six male New Zealand White rabbits. Images were obtained over the 23-day study duration using a 64-echo Carr-Purcell-Meiboom-Gill (CPMG) sequence (repetition time = 3000 msec, echo time = 20 msec). T2 decay curves were extracted from 0.7 x 0.7 x 3.0 mm3 voxels in the corpus callosum and contiguous white matter tracts, cortex, thalamic nuclei, hypothalamic nuclei, and the masseter muscles. The curves were fit with biexponential functions. RESULTS: Increased signal intensity in the corpus callosum was evident 2-3 days after the first TET injection. At this time, a substantial slowly relaxing component appeared in the decay curves of the corpus callosum and, to a lesser extent, in the thalamus and hypothalamus. Changes in the rabbits' body weight, general physical condition, and neurologic state paralleled the growth and regression of the second, slowly relaxing component. CONCLUSION: The appearance and regression of a slowly decaying second component in the T2 decay curve is consistent with the formation and shrink-age of intracellular vesicles in the intramyelin sheaths of central white matter.
Asunto(s)
Edema Encefálico/diagnóstico , Encéfalo/patología , Membranas Intracelulares/patología , Imagen por Resonancia Magnética , Animales , Agua Corporal/metabolismo , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Edema Encefálico/inducido químicamente , Edema Encefálico/metabolismo , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/metabolismo , Corteza Cerebral/patología , Cuerpo Calloso/efectos de los fármacos , Cuerpo Calloso/metabolismo , Cuerpo Calloso/patología , Modelos Animales de Enfermedad , Estudios de Seguimiento , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Hipotálamo/patología , Membranas Intracelulares/efectos de los fármacos , Membranas Intracelulares/metabolismo , Masculino , Conejos , Sensibilidad y Especificidad , Tálamo/efectos de los fármacos , Tálamo/metabolismo , Tálamo/patología , Compuestos de Trietilestaño/toxicidadRESUMEN
The question of what happens to cholesterol in the adult central nervous system during its slow turnover has been addressed using rats with brain and spinal cord labeled with [4-14C]cholesterol upon intracerebral injection of labeled cholesterol into rats at 10-12 days of age. At six months after injection, 14C was found only in the brain and spinal cord and was slowly released via the rat's urine. When labeled rats were given demyelinating agents (triethyl tin chloride, hexachlorophene, sodium cyanide) and when experimental allergic encephalomyelitis was induced, a measurable increase in urinary 14C label above control levels was found. It was concluded that there is a direct relationship between the experimental demyelination induced and the increased release of cholesterol metabolites into urine. The study suggests that a clinical method could be developed to determine the rate of central nervous system demyelination by measuring the amount of urinary cholesterol metabolites.
Asunto(s)
Enfermedades del Sistema Nervioso Central/orina , Colesterol/orina , Enfermedades Desmielinizantes/orina , Animales , Radioisótopos de Carbono , Enfermedades del Sistema Nervioso Central/inducido químicamente , Colesterol/metabolismo , Enfermedades Desmielinizantes/inducido químicamente , Modelos Animales de Enfermedad , Encefalomielitis Autoinmune Experimental/inducido químicamente , Encefalomielitis Autoinmune Experimental/orina , Hexaclorofeno/toxicidad , Ratas , Ratas Endogámicas Lew , Ratas Sprague-Dawley , Cianuro de Sodio/toxicidad , Inanición/orina , Compuestos de Trietilestaño/toxicidadRESUMEN
Long-Evans rat pups were injected i.p. on postnatal day 5 (PND5) or 12 with 0, 3, or 5 mg/kg triethyltin sulfate (TET) and then tested on T-maze delayed alternation on PND21 or 28. Delayed alternation learning was impaired on PND21 and 28 in pups given 5 mg/kg TET. Pups given 5 mg/kg TET on PND5 were more impaired on delayed alternation than pups given 5 mg/kg TET on PND12. Pups given 3 mg/kg TET on PND5 or 12 were unimpaired at either age of testing. On the day following training, pups were sacrificed for histological assessment employing Nissl- or immunohistochemical staining for glial fibrillary acidic protein (GFAP), a putative marker of gliosis. Pups given 5 mg/kg TET on PND5 showed increases in GFAP immunoreactivity (IR) in subiculum, amygdala, hippocampus, piriform cortex, and entorhinal cortex with concomitant decreases in Nissl-stained cells in these regions. Pups given 5 mg/kg TET on PND12 showed increases in GFAP IR in piriform cortex, amygdala and dorsal hippocampus with concomitant decreases in Nissl-stained cells in these regions. Exposure to 3 mg/kg TET on PND5 and PND12 produced a mild increase in GFAP IR in piriform cortex and amygdala but no discernible loss of Nissl-staining in these respective regions. TET-induced behavioral deficits appear related to damage of structural correlates of the human temporal lobe and not piriform cortical pathology. These results demonstrate that the day of exposure greatly influences the magnitude of the cognitive deficits and neuropathology associated with exposure to TET. There appears to be a critical period during postnatal development for the developmental neurotoxicity of this compound.
Asunto(s)
Encéfalo/efectos de los fármacos , Cognición/efectos de los fármacos , Conducta Espacial/efectos de los fármacos , Compuestos de Trietilestaño/toxicidad , Animales , Encéfalo/crecimiento & desarrollo , Química Encefálica/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Femenino , Proteína Ácida Fibrilar de la Glía/análisis , Inmunohistoquímica , Masculino , RatasRESUMEN
Brain edema can be classified into three categories: vasogenic, cytotoxic, and interstitial. The mechanism of edema is thought to be different in each type. The authors studied the movement of water molecules in each type of white matter edema in a rat model by using diffusion-weighted magnetic resonance imaging. Conventional T2-weighted imaging did not allow distinction between the three types of white matter edema; the three types of edema were, however, distinguished by using diffusion-weighted imaging. The apparent diffusion coefficient (ADC) of water was different in each type of edema. Water molecules in cytotoxic edema induced by triethyl-tin intoxication showed a smaller and less anisotropic ADC than in normal white matter. In contrast, water in vasogenic edema induced by cold injury had a larger and more anisotropic ADC than in normal white matter. Water in interstitial edema due to kaolin-induced hydrocephalus had an anisotropic and very large ADC.
Asunto(s)
Edema Encefálico/diagnóstico , Imagen por Resonancia Magnética , Animales , Encéfalo/patología , Edema Encefálico/etiología , Edema Encefálico/patología , Frío , Diagnóstico Diferencial , Difusión , Hidrocefalia/complicaciones , Caolín , Masculino , Modelos Estructurales , Ratas , Ratas Wistar , Compuestos de Trietilestaño/toxicidadRESUMEN
Trimethyltin (TMT) and triethyltin (TET) disrupt auditory function at doses far below those shown to be neurotoxic. In vivo studies suggest that the initial effect of TMT on hearing occurs at the inner hair cell/spiral ganglion cell synapse, while later, the outer hair cell (OHC) undergoes structural and functional damage. TET produces acute effects upon afferent neurotransmission similar to those observed following TMT, but TET's effects on OHC structure and function have not been examined. OHCs are motile elements within the cochlea, believed to modulate the sensitivity and tuning within the inner ear. Changes in OHC length may alter hearing function, and length changes have been reported following exposure to various ototoxic agents in vitro. In the present study, 77 OHCs from 45 pigmented male guinea pigs were isolated in primary culture and exposed for 90 min to concentrations between 30 microM and 1.0 mM of TMT or TET and then to bathing medium for 30 min to remove the toxicant. Significant shortening of the OHC cell body occurred at all doses to both organotins, with a mean reduction in length of 15.1 and 20.2% for 1.0 mM TMT and TET, respectively, at the end of testing; control cells were only 3.4% shorter at the end of 90 min of perfusion with bathing medium. The effect of organotin exposure on OHC volume was not consistently related to either TMT or TET concentration or altered cell length. In addition, disruption of the plasma membrane characterized by bleb formation, the forceful ejection of cytoplasm, or bursting was seen in 80% of cells exposed to 1.0 mM TET, although not TMT; lower concentrations of both organotins disrupted the cell membrane in 10-30% of cells. Membrane rupture was not reliably associated with either increased cell volume or decreased length, implicating a weakening of the plasma membrane or cortical lattice as the basis for this effect. Consistent with the irreversible structural weakening of the lateral wall, resorption of organotin-induced cytoplasmic blebs was never evidenced. Qualitatively, subcellular elements in the central core of many organotin-treated OHCs appeared pathological. These changes are similar to histopathological changes observed following in vivo organotin administration and may represent one target of acute alkyltin ototoxicity.
Asunto(s)
Células Ciliadas Auditivas/efectos de los fármacos , Compuestos de Trietilestaño/toxicidad , Compuestos de Trimetilestaño/toxicidad , Animales , Tamaño de la Célula/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Cobayas , Células Ciliadas Auditivas/citología , Masculino , Concentración OsmolarRESUMEN
We have extended our studies on the content of white matter derived coated vesicles (WMCVs) to show that they are enriched in membrane-bound carbonic anhydrase. Within the myelin complex membrane-bound carbonic anhydrase is concentrated in the periaxolemmal domain; however, this protein is enriched almost sevenfold in the bilayer of coated vesicles even relative to this myelin membrane region. These data suggest that some vesicles are derived from a site at which this enzyme is highly localized. The enrichment observed for membrane-bound carbonic anhydrase is unique since other periaxolemmal proteins such as CNPase and plasmolipin are only present in equal amounts in periaxolemmal-myelin fractions and WMCVs. Based on their known localization, the presence of CNPase coupled with the absence of MAG in WMCVs suggest that these vesicles are derived from the paranodal region. The identification in WMCVs of periaxolemmal-myelin proteins associated with ion and fluid movement, such as carbonic anhydrase, Na+,K+ ATPase, and the putative K+ channel protein plasmolipin, prompted us to examine the status of these vesicles in triethyl tin (TET)-induced myelin edema. Coated vesicles and other membrane fractions were isolated from whole brains of control and TET-treated rats. Whole brains were used so we could compare the effects of TET on WMCV proteins with the effect on proteins enriched in gray matter coated vesicles. The results indicated that TET had no detectable effect on compact or periaxolemmal-myelin, however, Western blot analysis showed that WMCV proteins, such as carbonic anhydrase, CNPase, and plasmolipin, were virtually absent or greatly diminished from the whole brain coated vesicle fraction.(ABSTRACT TRUNCATED AT 250 WORDS)