Brain neurotoxicity of Penitrem A: electrophysiological, behavioral and histopathological study.

The neurotoxicity of Penitrem A (PA) in rats was assessed against neurophysiological, behavioral and histopathological parameters. Animals were acutely given intracerebroventricular (22-45 mg) or intraperitoneal injections (0.5-1.5 mg/kg) of PA. A typical trembling syndrome associated with PA was always noted. Depending on the dose administered, animals may convulse and eventually die (1-1.5 mg/kg). PA-induced transient alterations of the EEG involving an increase in the frequency and voltage of electrical activity recorded from the cerebral cortex. Hippocampal activity was not modified and some pathologic activities may be recorded at the thalamus. Generally these EEG alterations disappeared at d 3 after the injection and the animals progressively recovered. However in the most severe cases, neuromotor disturbances were maintained at d 7 (rotarod test). Coronal sections of the brain at the striatal, thalamic, hippocampal and pons levels mainly revealed that PA was able to induce dose related injuries in the cerebellum with massive degeneration of Purkinje cells and a significant vacuolization within the molecular layer. The neurotoxic mechanism remains unclear. Action of the mycotoxin on the cerebello-thalamo-cortical tract is discussed.

Different localizations of Met-enkephalin-like immunoreactivity in rat forebrain and spinal cord using hydrogen peroxide and Triton X-100. Light microscopic study.

The histological distribution of met-enkephalin-like immunoreactivity was studied in the forebrain (particularly the striatum) and the spinal cord of the rat using the indirect peroxidase-labelled antibody method. In most experiments, vibratome sections of formaldehyde-fixed tissues and purified antibodies were used. The search for optimal conditions for the immunohistochemical reaction lead us to establish that met-enkephalin-containing perikarya of both untreated and colchicinized animals were better demonstrated when tissue were pre-treated with diluted hydrogen peroxide only. The additional treatment of these sections with Triton X-100 (or some other detergents) resulted in the near disappearance of the perikaryal immunoreactivity; on the contrary, numerous met-enkephalin containing nerve fibres and varicosities were then demonstrated in the same region. Using only the hydrogen peroxide treatment, we found numerous met-enkephalin-containing perikarya in the medial and ventral regions of the neostriatum. This distribution was prolonged caudally by the existence of a prominent group of stained somata in the ventral putamen-central nucleus of the amygdala. When intraventricular injections of colchicine were used, positive perikarya were more numerous within the striatum (the globus pallidus excepted) but their distribution was largely the same as in non injected animals. However, some new groups of somata were stained in this case in the forebrain (in the lateral septum, the olfactory tubercle and the hypothalamus particularly). In control animals only few met-enkephalin-containing perikarya were observed in the dorsal horn of the spinal cord when H2O2 pretreatment was used alone and they were numerous only when intraspinal injections of colchicine were performed. Met-enkephalin-containing fibres and varicosities, which were scattered in the whole neostriatum in the conditions used above, became very numerous when the tissue sections were incubated in the presence of Triton X-100. Their density increased markedly from the latero-dorsal to the medio-ventral regions but, in addition, an organization under the form of islands of stronger immunoreactivity was also evidenced. These islands were more numerous ventrally in the anterior neostriatum and in the central region of the "putamen." The dense plexus of immunoreactive nerve fibers forming "tube-like structures" which was always observed in the paleostriatum and in the cranial medial forebrain bundle (islands of Calleja) appeared more diffuse when detergents were used.(ABSTRACT TRUNCATED AT 400 WORDS)

Met-enkephalin-like immunoreactivity in rat forebrain and spinal cord using hydrogen peroxide and Triton X-100. Ultrastructural study.

Using two immunocytochemical methods, we have shown in light microscopy that the met-enkephalin-like immunoreactivity within striatum and spinal cord of the rat is differentially distributed in either perikarya or nerve terminals according to the technical conditions used [1]. The present electron microscopic study has been undertaken in order to elucidate the subcellular localization of immunoprecipitates according to the same technical conditions. In the neostriatum, numerous met-enkephalin-containing perikarya were stained (principally at the level of rough endoplasmic reticulum) when tissue sections were treated with hydrogen peroxide (H2O2) only, prior to the immunocytochemical procedure. However, injections of colchicine were required to demonstrate perikarya in the dorsal horn of the spinal cord. At variance with previous results, numerous dendritic profiles and nerve terminals were also reactive in this condition. Neurotubules, mitochondria, large granular vesicles (LGVs) and small synaptic vesicles were stained within these structures. The addition of a low concentration of Triton-X-100 (0.02%) in the first incubation medium often resulted in the disappearance of most perikarya and in the staining of only LGVs in nerve terminals. The addition of a higher concentration of Triton-X-100 (0.1%) produced diffusion of immunoprecipitates at the level of nerve terminals, which was probably responsible for the increased intensity of staining and, subsequently, for the better demonstration of fibre varicosities in light microscopy. On the contrary, the disappearance of reactive perikarya seemed to result from the diffusion of the non-protected peptide out of the cytoplasm. The diverse ultrastructural localizations of met-enkephalin-like immunoreactivity in striatum and spinal cord are finally discussed in light of intrinsic connections or afferents described in the literature.

Experimental ultrastructural modifications of the content of serotoninergic supra-ependymal nerve fibres in the rat.

1. In this study, evidence is presented that the electron-dense dot observed in the synaptic vesicles of axonal varicosities belonging to the supra-ependymal nerve fibres (SEF) of rats is related to the serotonin content of these vesicles. 2. The size of the dense dot and the relative number of small granular vesicles (SGV) were markedly increased by intraventricular administration of serotonin as well as by acute or chronic treatment with inhibitors of monoamine oxidases. 3. Conversely, the administration of reserpine diminished the percentage of SGV, as compared to untreated animals, and prevented the formation of granules in synaptic vesicles of chronic nialamide-treated rats. 4. With respect to large granular vesicles (LGV), it was found that chronic inhibition of MAO increased their number within the varicosities, whereas reserpine administration in untreated rats resulted in their disappearance. 5. Our results strongly imply that serotonin is present in both SGV and LGV of the SEF varicosities. This conclusion is supported by some radioautographic pictures obtained after intraventricular administration of 3H-5-HT which show silver grains over both populations of vesicles. Further support was obtained by the use of the Tranzer's cytochemical method in either untreated or nialamide treated rats. This method demonstrates that the reaction specific of monoamines occurs in both types of vesicles with either treatment but that larger stores of 5 HT (larger granules) are demonstrated after inhibition of MAO. 6. It is concluded that both SGV and LGV may represent storage organelles for exogenous and endogenous 5-HT.

Effects of intraventricular injection of 6-hydroxydopamine in the developing kitten. III. Histochemical fluorescence and radioautographic studies of the noradrenaline hyperinnervation in the pons.

In the present study, using neonatal intraventricular injections of 6-hydroxydopamine (6-OHDA) and the fluorescence histochemical method for monoamines, it is observed that an extensive plexus of noradrenaline (NA) fibres develops in the pontine region of the cat brain subsequently to the neonatal destruction of the ascending NA bundles and of the NA innervation in the cerebral cortex by the neurotoxin. This plexus is only partly conserved 10 months after the 6-OHDA treatment. Generally, only a limited number of NA perikarya degenerate in the region of the locus coeruleus, the others (nucleus subcoeruleus senso lato) exhibiting the same strong fluorescence as the new NA fibres. Using the radioautographic method after intraventricular injections of [3H]NA, our work demonstrates also the transient disappearance (at least one month) of the uptake of [3H]NA in the pons, whose NA cell bodies and nerve terminals are no longer labeled in the same number as in control animals. The possibility of again labeling significantly NA perikarya and numerous nerve terminals occurred between 3 and 5 months of age, probably indicating both a re-establishment of normal uptake properties in the preserved NA perikarya and nerve terminals and some maturation of the uptake mechanisms in the abnormal NA fibres of the pons. This last observation is at variance with data from newborn animals showing that the uptake of NA develops in parallel with the accumulation of endogenous NA in catecholamine nerve terminals. The present results, however, do corroborate and complement previous biochemical data obtained in the cat after neonatal injection of 6-OHDA.

[Electrocorticograms and the histochemical effects in the rat accompanying the reactivation of cholinesterases after the effects of an organophosphorus inhibitor]. / Electrocorticogrammes et effets histochimiques accompagnant chez le rat la réactivation des cholinestérases après action d'un inhibiteur organophosphoré.

The administration of paraoxon, an organophosphate inhibitor of cholinesterases, elicits theta rhythms, fast rhythms and paroxystical alterations on the electrocorticogram of the Rat. The disappearance of these abnormalities of the cortical activity after administration of aromatic oximes seems to be in good correlation with the restoration of cerebral cholinesterase activity revealed by histochemical method.

Penetration of oximes across the blood-brain barrier. A histochemical study of the cerebral cholinesterases reactivation.

The comparison of the effects of 4 oximes upon the cerebral cholinesterases reactivation after intoxication with paraoxon shows that the best results are obtained with toxogonine and 1574 [(carbaldoxime-4 pyridinium)-1(methyl-1 imidazolium-3)-3 propane]. The reactivation power of this latter compound seems due to the ease with which it can pass through the blood-brain barrier.