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The mechanisms and brain circuitry that render genetically epilepsy-prone rats (GEPRs) susceptible to acoustically induced seizures are not completely known. The present study explores the neuroanatomy of acoustically induced seizures by immunohistochemical analysis of the proto-oncoprotein fos after intense acoustic stimulation (AS) with and without seizures. Acoustic stimulation induced tonic convulsions in GEPR-9s, but not in control rats. Locations of brain nuclei showing fos-like immunoreactive (FLI) neurons following AS with and without seizures were mapped. Semiquantitative methods were used to compare FLI neuron numerical densities in AS control rats and GEPRs. Many brain areas exhibited profound FLI in AS control rats and GEPRs. Unexpectedly, the cochlear nuclei and the central nucleus of the inferior colliculi (ICc), both of which are requisite for AGS initiation, exhibited a diminished fos expression in animals having seizures compared to AS controls. In contrast, GEPRs displayed a significant increase in FLI neurons within the dorsal cortex of the IC (ICd) compared to AS controls. This finding may suggest a seizure-related amplification of the auditory signal between the ICc and the ICd. Other nuclei, known to be involved in auditory transmission (i.e., superior olivary complex; trapezoid nucleus; dorsal nucleus of the lateral lemniscus, DNLL), did not show differential FLI densities between seizure and AS control animals. In contrast, seizure-induced FLI was observed in many nonauditory brain nuclei. Of particular interest was the identification of an intensely labeled nucleus in the GEPR. This nucleus resides in the most posterior and dorsal-lateral part of the pedunculopontine tegmental nucleus-pars compacta (PPTn-pc) immediately adjacent to the DNLL and extends posteriorly into the superior lateral subnucleus of the lateral parabrachial area (SLPBn). Therefore, we have tentatively termed this nucleus the PPSLPBn. The PPSLPBn lies in a region previously described as a mesencephalic locomotor region and a suspected functional involvement of this nucleus in display of seizure activity is under investigation. Other brain stem nuclei showing differential fos expression between GEPRs and AS control rats are also described.

The effects of pyrethroid insecticides and other sodium channel-specific neurotoxins on synaptosomal membrane potential were investigated in rat and trout brain synaptosomes using the membrane-permeant lipophilic cation [3H]tetraphenylphosphonium (TPP+). Concentration-dependent and tetrodotoxin-sensitive decreases in TPP+ accumulation, indicative of membrane depolarization, were produced by veratridine, aconitine, scorpion (Leiurus quinquestriatus) venom, and type I and type II pyrethroids, in both species. Veratridine, aconitine, and Leiurus venom were more potent and efficacious membrane-depolarizing agents in rat synaptosomes than in trout synaptosomes. Type II (deltamethrin, cypermethrin) pyrethroids produced similar depolarizing responses in rat and trout synaptosomes; however, the 1R-cis-alpha R isomer of deltamethrin, which had no effect on membrane potential in rat synaptosomes, depolarized trout synaptosomes. This isomer of deltamethrin was also shown to produce toxicity in trout, but not in rats. The type I pyrethroids, permethrin and NRDC 157, exhibited significantly greater intrinsic activity in trout brain synaptosomes, producing maximal membrane depolarizations that were three times greater than those observed in rat brain synaptosomes. These results provide evidence of species-specific differences in the membrane-depolarizing properties of pyrethroid insecticides and sodium channel-specific neurotoxins. They also suggest that some of the neurotoxin binding domains of the voltage-sensitive sodium channel in trout brain differ from those in mammalian brain. The hypersensitivity of fish to the neurotoxic actions of pyrethroid insecticides may be related to these differences.

The neuroexcitatory actions of two toxicologically distinct classes of pyrethroid insecticides were characterized in rat brain synaptosomes using [3H]tetraphenylphosphonium to measure changes in synaptosomal membrane potential and by measuring the release of [3H]acetylcholine. Both type I (permethrin) and type II (deltamethrin, cypermethrin and fenvalerate) pyrethroids produced a concentration-dependent tetrodotoxin-sensitive membrane depolarization which was stereospecific for the neurotoxic isomer of each pyrethroid. Deltamethrin was the most potent and efficacious pyrethroid in these studies, with an EC50 of 30 nM and a maximal estimated membrane depolarization of 27 mV, followed by cypermethrin, fenvalerate and permethrin. The phenoxybenzyl pyrethroids also increased the spontaneous release of [3H]acetylcholine from rat brain synaptosomes, further supporting a depolarizing action of these insecticides on nerve terminal membranes. Pyrethroid-induced release of [3H]acetylcholine was tetrodotoxin-sensitive and occurred over the same concentration range as membrane depolarization. These data indicate that type I and type II phenoxybenzyl pyrethroids act potently and stereoselectively on the voltage-sensitive sodium channel to increase sodium influx into synaptic terminals producing membrane depolarization and neurotransmitter release. Furthermore, they show that pyrethroid-induced alterations in synaptosomal membrane potential is a sensitive measure of pyrethroid action on the sodium channel and of pyrethroid toxicity.

The development and regional distribution of ribonucleotide reductase (EC 1.17.4.1) were determined in rat brain. Ribonucleotide reductase was partially purified by ammonium sulfate fractionation (20-40% saturation). Enzyme activity was measured by a specific radiochemical assay. This method involved the reduction of [14C]cytidine diphosphate (CDP) to [14C]deoxycytidine diphosphate with subsequent hydrolysis and separation of the product ([14C]deoxycytidine) from substrate ([14C]cytidine) by Dowex-1-borate ion-exchange chromatography. The specific activity of ribonucleotide reductase in whole brain of newborn rats was 3.78 +/- 0.55 units (pmol/h)/mg protein (SEM; n = 6) and declined to 0.17 +/- 0.01 units/mg protein (n = 7) at 10-12 weeks of age, with a further decline to 0.11 +/- 0.01 units/mg protein (n = 3) at 1 year. Ribonucleotide reductase activity in rat liver decreased from 4.58 +/- 0.62 units/mg protein (n = 3) in newborn animals to 0.06 +/- 0.01 units/mg protein (n = 7) at 10-12 weeks and was present at trace levels at 6 months of age. The decline in specific activity with age was not due to a change in the Km for CDP. The Km for CDP in brain of newborn and adult rats was 80-90 microM. In 10- to 12-week-old rats, the specific activity of ribonucleotide reductase was similar in the various regions of the brain tested except for the brainstem, which had 50% lower specific activity than the whole brain. These results indicate that ribonucleotide reductase activity is present and widely distributed in adult rat brain.