Activation of voltage-dependent sodium channels in cultured cerebellar poffule cells induces neurotoxicity that is not mediated by glutamate release.

Exposure of rat cerebellar granule cell cultures to neurotoxins that specifically enhance the open state probability of voltage-dependent Na+ channels, resulted in neuronal death as estimated by a cell viability assay based on fluorescent staining and 51Cr-uptake. Toxicity was detected within 1 h after addition of 100 microM veratridine and was complete within 10-18 h; it was dose-dependent and was found to be completely abolished by tetrodotoxin, an Na+ channel blocker. When veratridine was replaced by an alpha-scorpion toxin, similar observations were done. In contrast, when cultured neurons prepared ffom the cerebral hemisphere of fetal rat brain were exposed to either veratridine or alpha-scorpion toxin for 18 h or even for a longer time of incubation, no neuronal death was observed. DNA fragmentation analysis showed that the toxicity was not mediated by apoptosis. Neuronal death was neither prevented by glutamate receptor antagonists, nor by depletion of endogenous glutamate, nor by voltage sensitive calcium channel antagonists such as omega-Conotoxin-GVIA (N-type channels), omega-Agatoxin-IVA (P-type channels), nimodipine and nitrendipine (L-type channels). Our study indicates that prolonged opening of Na+ channels induced neuronal death of cerebellar granule cells which is not mediated by glutamate and reveals novel neurotoxic mechanism in addition to the well-established excitatory amino acid receptor pathway.

Increases in the effects of albendazole on Echinococcus multilocularis metacestodes by the dipeptide methyl ester (Phe-Phe-OMe).

Three months after infection with Echinococcus multilocularis, Mongolian gerbils were given either the dipeptide methyl ester (Phe-Phe-OMe) or a combination of Phe-Phe-OMe plus albendazole to treat alveolar echinococcosis. Each drug was given orally at the daily dose of 50 mg/kg of body weight following various administration regimens. Histologic and ultrastructural studies of parasites recovered from infected gerbil tissues showed that the dipeptide methyl ester increases the effect of albendazole.

Echinococcus granulosus:isoprinosine treatment of the metacestode stage.

The efficacy of short- and long-term treatments with Isoprinosine, an immunomodulatory compound, was studied in Echinococcus granulosus cysts developed in NMRI mice intraperitoneally infected with sheep pulmonary cysts. After treatment, a reduction in the size and number of cysts with macroscopic modifications was observed. The structural alterations included damage or destruction of the protoscoleces and partial destruction of the cyst wall, which predominated at the inner germinal layer level. The efficacy of this drug was evaluated after long-term and short-term treatment. Short-term treatment with a dose of 1 g/kg/day gave better results, with a loss of infectivity of the larval tissue. The well-tolerated long-term treatment with a dose of 2 g/kg/day showed the absence of toxicity of this compound. The survival time of treated animals was greater than that of untreated controls.

Histochemical observations of alkaline phosphatase activity of Echinococcus multilocularis during in vivo development in golden hamsters, an alternative definitive host.

Histochemical observations of alkaline phosphatase activity of Echinococcus multilocularis during the in vivo development in golden hamster, an alternative definitive host. The present work reports on the ability of protoscoleces from a European fox strain of E. multilocularis to differentiate and develop into the adult form in the small intestine of male golden hamsters treated with prednisolone. Detection of alkaline phosphatase activity on various stages of the developing worm was performed by histochemical methods. The enzyme activity was not demonstrable in the early stages of infection but occurred with strobilization. Age-related changes in the distribution of the enzyme activity took place during strobilization. Alkaline phosphatase activity was evident in the excretory ducts of 8 to 11 day old strobila and in the tegument of mature proglottis of 16 day old worms. This in vivo procedure with rodents as definitive hosts provides interesting preliminary results on the biology of the E. multilocularis adult. Further investigations on membrane-bound enzymes involved in physiological and nutritional processes are in progress.

Specific distribution of sodium channels in axons of rat embryo spinal motoneurones.

1. The distribution of Na+ channels and development of excitability were investigated in vitro in purified spinal motoneurones obtained from rat embryos at E14, using electrophysiological, immunocytochemical and autoradiographical methods. 2. One hour after plating the motoneurones (DIV0), only somas were present. They expressed a robust delayed rectifier K+ current (IDR) and a fast-inactivating A-type K+ current (IA). The rapid neuritic outgrowth was paralleled by the emergence of a fast-activating TTX-sensitive sodium current (INa), and by an increase in both K+ currents. 3. The change in the three currents was measured daily, up to DIV8. The large increase in INa observed after DIV2 was accompanied by the onset of excitability. Spontaneous activity was observed as from DIV6. 4. The occurrence of axonal differentiation was confirmed by the fact that (i) only one neurite per motoneurone generated antidromic action potentials; and (ii) 125I-alpha-scorpion toxin binding, a specific marker of Na+ channels, labelled only one neurite and the greatest density was observed in the initial segment. Na+ channels therefore selectively targeted the axon and were absent from the dendrites and somas. 5. The specific distribution of Na+ channels was detectable as soon as the neurites began to grow. When the neuritic outgrowth was blocked by nocodazole, no INa developed. 6. It was concluded that, in spinal embryonic motoneurone in cell culture, Na+ channels, the expression of which starts with neuritic differentiation, are selectively addressed to the axonal process, whereas K+ channels are present in the soma prior to the neuritic outgrowth.

Immunoassays fail to detect antibodies against neuronal calcium channels in amyotrophic lateral sclerosis serum.

Recent studies suggested that autoantibodies that bind to voltage-dependent calcium channels and activate calcium entry may play a role in the progressive degeneration of motoneurons in sporadic amyotrophic lateral sclerosis. Immunoassays were performed to assess autoantibody titer in patients with amyotrophic lateral sclerosis or Lambert-Eaton myasthenic syndrome, a disease in which the presence of anti-calcium channel antibodies is well documented. Based on immunoprecipitation assays for antibodies against N-type calcium channels, only 8% (2/25) of amyotrophic lateral sclerosis patients had marginally positive titers, whereas 58% (18/31) of patients with Lambert-Eaton myasthenic syndrome had positive titers. Enzyme-linked immunosorbent assays with purified neuronal N-type calcium channels revealed immunoreactivity in 2 of 25 amyotrophic lateral sclerosis sera and 12 of 31 Lambert-Eaton myasthenic syndrome sera, which is not compatible with suggestions that enzyme-linked immunosorbent assay is a more sensitive technique for the detection of autoantibodies in amyotrophic lateral sclerosis. Furthermore, based on immunoprecipitation assays, amyotrophic lateral sclerosis sera were totally negative for antibodies against L-type calcium channels from skeletal muscle or brain. These data do not support the hypothesis that an autoimmune response against calcium channels plays a primary role in amyotrophic lateral sclerosis.