Reduced kainic acid binding in rat hippocampal formation after limbic kindling

The specific binding of [3H]kainic acid to hippocampal membranes was examined autoradiographically in rats kindled by tetanic stimulation of the amygdala or angular bundle. One day after the last of 3 class 4-5 kindled seizures, the specific binding of [3H]kainic acid in stratum lucidum of area CA3 was 47-61% less than in electrode-implanted unstimulated controls. Specific binding in the inner third of the dentate molecular layer was reduced to a lesser degree. These observations demonstrate that kainic acid receptors are down-regulated by kindling stimulation.     

Loss and reacquisition of hippocampal synapses after selective destruction of CA3–CA4 afferents with kainic acid

Intraventricular injections of kainic acid were used to destroy the hippocampal CA3–CA4 cells bilaterally in rats, thus denervating the inner third of the molecular layer of the fascia dentata and stratum radiatum of area CA1. Electron microscopic studies showed that this lesion reduced the synaptic density of the CA1 stratum radiatum by an average of 86%. The synaptic density of the inner third of the dorsal dentate molecular layer declined by two-thirds and the corresponding zone of the ventral dentate molecular layer by about half. Within 6–8 weeks the synaptic density of these laminae had been restored to the control value or nearly so. In the CA1 stratum radiatum about 72% of the synaptic contacts destroyed by the lesion were replaced, the inner third of the ventral dentate molecular layer recovered 75% of its lost synapses and the inner third of the dorsal dentate molecular layer apparently recovered virtually all of them. The newly formed synapses did not differ noticeably from those normally present.A kainic acid lesion reduced the synaptic density of the outer two-thirds of the dentate molecular layer by 30% within 3–5 days, despite a virtual absence of presynaptic degeneration in that zone. This result implies a substantial disconnection of perforant path synapses. It did not appear to depend on the extent of denervation of the inner zone. The loss of perforant path synapses was completely reversible. We suggest that the dentate granule cells shed a portion of their synapses in response to a substantial loss of neurons to which they project and regained them when their axons had formed new synaptic connections.     

Seizures down-regulate muscarinic cholinergic receptors in hippocampal formation

Muscarinic cholinergic receptors (MCR) have been previously shown to decline in the hippocampal formation (HPF) of amygdala-kindled rats. Seizures have been proposed as the process responsible for this down-regulation. We now demonstrate similar down-regulation of MCR within HPF in 3 additional methods of inducing seizures: electroconvulsive shock, entorhinal kindling and entorhinal lesion. Two key parameters which causally link the MCR declines with seizures are their time course and reversal with anticonvulsants. The transient decline of MCR induced by entorhinal lesion-induced seizures parallels the time course established in amygdala kindling. Further, phenobarbital could block both these seizures and the MCR declines. Together, this supports the relationship of seizures causing the declines. We postulate that the MCR down-regulation represents an endogenous inhibitory response of neurons that are intensely and repeatedly depolarized during the seizures. neurons that are intensely and repeatedly depolarized during the seizures.     

Spatial memory following damage to hippocampal CA3 pyramidal cells with kainic acid: impairment and recovery with preoperative training

The behavioral function of an intrinsic component of the hippocampus was investigated. Neurotoxic lesions of the CA3 region of the hippocampus impaired performance of a spatial memory task, and produced both hyperactivity and hyperreactivity to sensory stimulation. Both the magnitude and duration of these behavioral alterations depended on the amount of preoperative training received, and on the extent and locus of the lesion within the CA3 subfield. This study indicates that the CA3 subfield contributes to the performance of at least three behavioral functions known to be mediated by the hippocampus. In addition, the lack of enduring deficits in the behavioral tests in which preoperative training was received suggests that, after loss of the CA3 subfield, some mechanism for recovery of function occurs within the hippocampus.     

Changes in neurofilament protein-immunoreactivity after kainic acid treatment of organotypic hippocampal slice cultures.

Neurofilament (NF) proteins are expressed in the majority of neurons in the central nervous system, and play a crucial role in the organization of neuronal shape and function. In the present study, we have used immunoblotting and immunocytochemical methods to study the light (NF-L), medium (NF-M ), and heavy (NF-H) molecular weight NF proteins in cultured organotypic hippocampal slices during the in vitro maturation and the changes after kainic acid (KA) treatment. In control cultures at 11 DIV throughout 25 DIV, CA3 pyramidal neurons and their proximal dendrites were heavily labeled with the antibodies against all three NF proteins. In CA1 pyramidal neurons, no staining was detected in any age group. A few weakly NF-L positive granule cells with fibers were detected in each age group, whereas NF-M and NF-H positive granule cells first appeared in the older cultures. The application of KA (5 microM) to the cultures for 48 hr, induced a pronounced cell death in the CA3 cell layers, and also moderately damaged granule cells. After the treatment, the immunoblot signal of NF-L and NF-M markedly decreased, whereas that of NF-H almost completely disappeared. The amount of NF-L positive fibers, however, dramatically increased in the molecular and hilar regions of the dentate gyrus in both age groups. Our results show the cellular heterogeneity in the distribution of NF protein triplet in cultured organotypic hippocampal slices. Kainic acid treatment induced changes, which mimicked those observed in the hippocampal region of epileptic animals.     

Modifications in recognition sites for neurotransmitters in rat hippocampus by kainic acid lesion

The specific binding of the tritiated radioligands of dexetimide, serotonin, clonidine, prazosin, WB-4101 and dihydroalprenolol to hippocampal membranes was determined two weeks after producing a virtual complete degeneration of perikarya by the local application of 0.5 μg of kainic acid in the dorsal and ventral parts of the hippocampus. Afferent terminals were unaffected by the neurotoxin since the contents of noradrenaline, serotonin and acetylcholine, as well as the activity of choline acetyltransferase, were not modified.Scatchard analysis is revealed that the kainic acid lesion produced a 60% decrease in the density of both cholinergic muscarinic binding sites and serotonin binding sites. A significant portion ofα₁-andα₂-adrenoceptor binding sites are also associated with intrinsic neurons of the hippocampus, as shown by the approximately 30% reduction in the densities of tritiated WB-4101, prazosin and clonidine produced by the action of kainic acid. By contrast, the affinity and density of β-adrenoceptor binding sites were unaffected by the lesion. It is suggested that the recognition sites of the different receptor populations surviving the lesion most likely reside on homologous and/or heterologous nerve terminals.     

Evidence for an agonist independent down regulation of hippocampal muscarinic receptors in kindling

Kindling induces a decline of hippocampal muscarinic cholinergic receptors. To test the hypothesis that the decline was mediated by the agonist, acetylcholine, adult male Sprague-Dawley rats were lesioned in the medial septum prior to kindling. Despite the marked destruction of presynaptic cholinergic terminals in the hippocampus, amygdala kindling proceeded normally and the hippocampal muscarinic receptor decline was not blocked. A small but significant decline in choline acetyltransferase activity was demonstrated in non-lesioned kindled rats. It is proposed that the kindling induced decline of hippocampal muscarinic receptors is mediated by repeated neuronal depolarization.     

Cellular and synaptic basis of kainic acid-induced hippocampal epileptiform activity

The effects of kainic acid (KA) were studied using extracellular and intracellular recordings in the hippocampal slice preparation. In sufficient concentrations, KA led to a loss of all evoked responses. However, the amount of drug needed for this varied according to anatomic region. CA3 was more sensitive (1 microM) than CA1 or the dentate gyrus (10 microM). These results can be understood in terms of a profound and long-lasting depolarization of neurons. Lower concentrations of KA (0.05-0.1 microM) did not change the resting membrane potential or input resistance of hippocampal pyramidal cells but produced spontaneous epileptiform activity which originated in CA3 and propagated to CA1. Epileptiform discharges were not present in the dentate gyrus. Coincident with the induction of paroxysms, the following changes were observed: (1) an increase in the excitability of CA3 and CA1 pyramidal cells as measured by a left shift in the input-output curves of evoked responses and a lowered threshold stimulus intensity necessary for activation of action potentials in single neurons; (2) augmentation and synchronization of bursting in pyramidal cells; and (3) prolonged EPSPs without an increase in their amplitude. These findings indicate that multiple changes, involving both the properties of single neurons and synaptic connections, are involved in the development of hippocampal paroxysms and that CA3 and CA1 have different roles in the generation of these discharges.     

Chronic failure of inhibition of the CA1 area of the hippocampus following kainic acid lesions of the CA3/4 area

The chronic effects of the lesioning agent, kainic acid, on paired pulse inhibition in the CA1 area were investigated in the hippocampus both in vivo and in vitro. Pretreatment of animals with a unilateral intracerebroventricular (i.c.v.) injection of kainic acid resulted in a lesion of the CA3/4 area of the hippocampus ipsilateral to the injection site. On activating the surviving Schaffer collateral afferents in the contralateral hippocampus, normal paired-pulse inhibition of the extracellularly recorded population spike in CA1 was observed. On activating the surviving commissural afferents to the CA1 area ipsilateral to the lesion, no such inhibition could be observed. However, paired-pulse inhibition was recorded in the dentate gyrus ipsilateral to the lesion in response to stimulation of the perforant path. The chronic failure of inhibition following the unilateral i.c.v. injection of kainic acid further supports the use of this method to provide a chronic model in the rat for the study of epileptogenesis in the hippocampus.     

Evidence from lesion studies for epileptogenic and non-epileptogenic neurotoxic interactions between kainic acid and excitatory innervation

The toxicity of kainic acid toward rat hippocampal neurons depends on the presence of specific excitatory afferents. Acute destruction of the critical pathway essentially abolishes the neurotoxicity of intraventricular kainic acid, but some or all hippocampal neurons continue to be destroyed by locally-injected kainic acid until the critical pathway(s) degenerates. These results support the view that kainic acid destroys hippocampal neurons in two ways: (1) by initiating a lethal status epilepticus; and (2) by interacting with certain pathways independently of on-going electrical activity within those pathways.     

Fast and slow depolarizing potentials induced by short pulses of kainic acid in hippocampal neurons

Responses of hippocampal neurons to short pulses of alpha-kainic acid (KA) were studied intracellularly in thin brain slices of the guinea pig. A KA pulse induced a slow depolarizing potential either with or without a preceding much faster depolarization. Large fast responses were induced only at the center of glutamate-sensitive spots, where short pulses of L-glutamate (Glu) induced large depolarizations in the impaled neuron. The fast responses resembled Glu-induced depolarizations in time-course, in sensitivity to movement of the tips of amino acid-pipettes, in sensitivity to Glu-antagonists and in reversal potential. The slow response was much more resistant to movement of the amino acid-pipettes and to Glu-antagonists. Mn2+ was without effect on the fast as well as the slow responses. Glu-induced depolarizations super-imposed on the slow response were simply depressed. These results indicate that two different types of receptors are activated by administration of KA, and suggest that the slow response results from a direct action of KA and the fast response is produced as a consequence of either the direct action of KA on the Glu receptors or a calcium-independent release of Glu by KA.     

The effect of kainic acid on the hippocampal content of putative transmitter amino acids

The time courses of the changes in the contents of GABA (γ-aminobutyric acid), glutamate, aspartate, taurine, glycine and alanine were measured in both hippocampi of rats which were injected in one hippocampus with kainic acid (KA). The contents of these amino acids were measured with high performance liquid chromatography. The hippocampal contents of GABA, glutamate, aspartate and taurine were reduced homolaterally to the KA injection; in contrast, the contents of glycine and alanine failed to change. The extent of the reductions of GABA, glutamate, aspartate and taurine was dependent on the size of the lesion caused by KA. The greatest decrease occurred after two simultaneous injections of KA, in dorsal and ventral hippocampus. An analysis of the amino acids at different levels of the hippocampus after dorsal injections of KA showed that in the hippocampus the amino acidergic axons do not travel longitudinally. This study suggests that GABA, glutamate, aspartate and taurine are preferentially located in intrinsic hippocampal neurons with short axons.     

Similarities vs. differences in place learning and circadian activity in rats after fimbria-fornix section or ibotenate removal of hippocampal cells

Damage to either the fimbria-fornix or to the hippocampus can produce a deficit in spatial behavior and change in locomotor activity but the extent to which the two kinds of damage are comparable is not known. Here we contrasted the effects of cathodal sections of the fimbria-fornix with ibotenic acid lesions of the cells of the hippocampus (Ammon's horn and the dentate gyrus) on place learning in a swimming pool and on circadian activity. Rats in both ablation groups were impaired relative to control rats in learning a single place response but they did acquire the response as measured by swim latencies, errors, and by enhanced searching on probe trials. They were also more active than the control group on the test of activity. Nevertheless, the fimbria-fornix group was initially more impaired on learning and was more active than the hippocampal group. Analysis of the strategies used in learning indicated that the lesion groups were very similar to each other but different from the control group especially in that at asymptotic performance, rats in both lesion groups made rather tight loops as they swam toward the platform. This strategy likely contributed to the greater proportion of time they spent swimming in the correct quadrant on the subsequent probe trial. These findings confirm that rats with fimbria-fornix or hippocampal damage display impairments in place learning and are hyperactive but also show that there are lesion differences. The results are discussed with respect to the relative effectiveness of the lesions and the possibility that fibers in the fimbria-fornix may mediate some functions that are not attributable to the hippocampus. © 1995 Wiley-Liss, Inc.     

Alterations in retinal neurotransmitter receptors and neuropeptides of the chick by kainic acid and acrylamide

The effects of intraocular injection of kainic acid and acrylamide upon retinal neuropeptides and high affinity binding sites have been determined in the chick. Kainic acid causes a sharp reduction in Met-enkephalin and somatostatin while neurotensin levels are unchanged. This treatment also lowers the extent of cholinergic muscarinic but not of [3H]naloxone or [3H]spiroperidol binding. In contrast, acrylamide treatment causes major increases of retinal Met-enkephalin and neurotensin concentrations. The binding of [3H]naloxone is also increased, and no reductions of any peptide or binding intensity were observed. The results indicate the plasticity of retinal neuropeptide levels and the selectivity with which these can be modulated.     

Amygdaloid lesion increases the toxicity of intrahippocampal kainic acid injection and reduces the late occurrence of spontaneous recurrent seizures in rats

Spontaneous recurrent seizures (SRS) following intrahippocampal kainic acid (KA) injection have been described in a previous paper from our laboratory. The SRSs are clinically similar to the seizures induced by kindling the amygdala and we suggested that the amygdala plays a role in initiating the SRSs. Accordingly, the present paper examines the effect of amygdaloid lesions on intrahippocampal KA-treated rats. There were short- and long-term effects. (1) Short-term: the toxicity of KA was increased in lesioned animals. Status epilepticus followed by death of the animals was evoked with half of the dose required to cause the same effect in intact rats. Moreover, a gross haematuria was encountered 6–12 h after KA injection. This was not observed in non-lesioned rats even following the highest KA doses. (2) Long-term: amygdaloid lesions delayed the occurrence of the SRSs, reduced their incidence and modified their expression. In lesioned animals seizures has began with a period of tonic immobility with no sign of the masticatory movements seen in intact animals. Histological examination of the KA-induced lesions did not show any major differences between lesioned and intact animals. It is suggested that the short-term effects are due to an unspecific effect on homeostatic mechanisms, whereas the long-term ones reflect a specific involvement of the amygdala in the late appearing seizures.