myelin-basic-protein and trimethyltin

In our previous investigations, we demonstrated that CD4(+) antimyelin basic protein (MBP) T cells protect hippocampal neurons against trimethyltin-induced damage. We hypothesized involvement of T cells, interacting with the various glial populations activated during the neurodegeneration process. In this study, we employ immunocytochemical methods to investigate the influence of administration of T cells on the response of microglia and of NG2(+) cells to trimethyltin (TMT)-induced damage. Female Lewis rats were treated with anti-MBP CD4(+) T cells (4 million per animal, i.v) 24 hr after TMT (8 mg/kg, i.p) intoxication. TMT caused degeneration of CA4 hipppocampal neurons and evoked an abundant reaction of microglial and NG2(+) cells in the injured region. The cells changed morphology into the activated state, and the number of OX42(+) and NG2(+) cells increased about 4.5-fold and 3-fold, respectively, relative to controls as assessed on day 21 after TMT treatment. Additionally, the cells of ameboid morphology, which expressed NG2 or microglial antigens, appeared in the zone of neurodegeneration. Furthermore, certain cells of ameboid phenotype shared both antigens. In rats treated with T cells, down-regulation of the activation of both glial classes and reduction of formation of their ameboid forms was observed. The number of the total OX42(+) and NG2(+) cells decreased by 21% and 54%, respectively, and the number of their ameboid forms decreased by 46% and 73%, respectively. Our data suggest that the diminished activation of microglia and NG2(+) cells, particularly the reduced number of their ameboid forms, may contribute to the neuroprotective effect of T cells.

Topics: Animals; Antigens; CD4-Positive T-Lymphocytes; Cell Count; Cytoprotection; Down-Regulation; Female; Fluorescent Antibody Technique; Hippocampus; Microglia; Microscopy, Confocal; Myelin Basic Protein; Nerve Degeneration; Neurons; Neuroprotective Agents; Proteoglycans; Rats; Statistics, Nonparametric; Trimethyltin Compounds

Developing accessible biomarkers of neurotoxic effects which are readily applicable to human populations poses a challenge for neurotoxicology. In the past, the neurotoxic organometal trimethyltin (TMT) has been used as a denervation tool to validate the enhanced expression of GFAP as a biomarker of astrogliosis and neurotoxicity resulting from chemical exposures. In the present study, TMT was used to assess the detection of serum autoantibodies as biomarkers of neurotoxicity. Previous studies in both human and animals have demonstrated the presence of serum autoantibodies to neurotypic [e.g., neurofilament triplet (NF)] and gliotypic proteins [myelin basic protein (MBP) and glial fibrillary acidic protein (GFAP)] as a peripheral marker of neurodegeneration that may be applicable to humans and experimental studies. Male Long-Evans rats (45 days of age) were administered either TMT (8 mg/kg; s) or an equal volume of sterile 0.9% saline. At 1, 2, and 3 weeks post-administration, serum was collected, and rats were sacrificed for the collection of brains. Serum autoantibodies (both IgM and IgG isotypes) to NF68, NF160, NF200, MBP, and GFAP were assayed using an ELISA. Saline only rats did not have detectable levels of autoantibodies. Only sera from TMT-exposed rats had detectable titers of autoantibodies to NFs with IgG predominating starting week 2. Anti-NF68 titers were highest compared to NF160, or NF200. Autoantibodies to MBP and GFAP also were detected; however, there was no significant increase in their titers until week 3. Hippocampal GFAP, detected at these time points, was significantly (p<0.05) higher than in control brains, indicating the induction of astrogliosis as confirmed by immunostaining of brain sections. The detection of anti-NFs, as indicative of neuronal insult, was consistent with loss of hippocampal neurons in CA3 and CA1. Our results suggest that the detection of autoantibodies to neurotypic and gliotypic proteins may be used as peripheral biomarkers to reveal evidence of nervous system neurotoxicity.

Topics: Animals; Autoantibodies; Biomarkers; Brain; Enzyme-Linked Immunosorbent Assay; Glial Fibrillary Acidic Protein; Immunoglobulin G; Immunoglobulin M; Male; Myelin Basic Protein; Nerve Tissue Proteins; Neurofilament Proteins; Rats; Rats, Long-Evans; Time Factors; Trimethyltin Compounds

We investigated the influence of administration of autoimmune T cells on trimethyltin-induced degeneration of hippocampal neurons. Female Lewis rats received 8 mg/kg trimethyltin intraperitoneally alone, or followed 24 h later by a second intravenous injection of anti-myelin basic protein T cells (green fluorescent protein-tagged). Neurodegeneration was assessed by NeuN and Nissl cell counts 21 days after trimethyltin injection. We found that neurodegeneration in the CA4 region of the hippocampus was significantly reduced in the group receiving T cells. T cells also caused an augmentation of trimethyltin-induced hippocampal astrocytic activation and astrocytic TrkA expression, which was particularly intense in the CA4 region. Our study provides the first evidence of neuroprotection evoked by transferred T cells following a neurotoxic brain insult. The data suggest that mediation of the neuroprotective effects of T-cell-released nerve growth factor occurs mainly via hippocampal astroglial TrkA receptors.

Topics: Animals; Cell Count; Female; Glial Fibrillary Acidic Protein; Hippocampus; Myelin Basic Protein; Nerve Degeneration; Neurons; Phosphopyruvate Hydratase; Rats; Rats, Inbred Lew; Receptor, trkA; T-Lymphocytes; Trimethyltin Compounds

Trimethyltin (TMT) is an alkyltin that targets neurons of the limbic system. A gene probe (i.e., mRNA) for myelin basic protein (MBP), a major component of central nervous system myelin, was used to monitor this toxic neuropathy in Sprague-Dawley rats. Animals were administered a single intraperitoneal injection of TMT-hydroxide at a neuropathic (8.0 mg/kg/body wt) or nonneuropathic (0.8 mg/kg/body wt) dose and sampled at 1, 3, or 7 days postexposure to correlate the progression of hippocampal neuropathology with probe (i.e., MBP-mRNA) levels. Microscopic examination of the brain showed only moderate but progressive damage over the 7-day postexposure period in animals treated with the neuropathic dose. Neuronal loss was first observed in the dendate gyrus and CA4 at 1 day postexposure, and progressed to the CA3c sector at 3 and 7 days postexposure. Elsewhere in the brain, minimal involvement of the entorhinal cortex neurons occurred 3 days postexposure and intensified by 7 days. No histological damage was seen at the nonneuropathic (0.8 mg/kg) dose. For gene probe analysis, the brain was divided into anterior and posterior halves. In rats treated with the neuropathic dose of TMT, the anterior brain showed progressive depressions of MBP-mRNA levels over the 1-, 3-, and 7-day postexposure period that correlated with increasing hippocampal neuropathology. The posterior brain showed no significant changes in MBP-mRNA levels with respect to that of controls over the same time period. At the nonneuropathic dose (0.8 mg/kg) a significant depression of MBP-mRNA levels occurred in the anterior brain at 7 days postexposure in the absence of overt histological damage.

Topics: Animals; Blotting, Northern; DNA Probes; Hippocampus; Male; Myelin Basic Protein; Nervous System; Nucleic Acid Hybridization; Rats; Rats, Inbred Strains; RNA, Messenger; Trimethyltin Compounds