Kainic Acid-lesioned Rats Research Articles

In vitro and ex vivo autoradiography studies on peripheral-type benzodiazepine receptor binding using [11C]AC-5216 in normal and kainic acid-lesioned rats

AC-5216 was reported as a novel ligand for peripheral-type benzodiazepine receptor (PBR) with a different chemical structure from DAA1106 analogues. This ligand had potent affinity for PBR and selectivity for PBR over other neurotransmitters. We have previously labeled AC-5216 using positron-emitter (11)C. The aim of this study was to evaluate [(11)C]AC-5216 in a rat brain model with neuroinflammation using an autoradiography (ARG) technique. In vitro ARG of normal rat brain showed that [(11)C]AC-5216 accumulated highly in the olfactory bulb, choroid plexus and cerebellum. The distribution pattern agreed with the localization of PBR in the rodent brain. Infusion of kainic acid (KA: 1, 2.5 and 5 nmol) into the rat striatum resulted in neuroinflammation. In vitro and ex vivo ARG revealed that the radioactivity level of [(11)C]AC-5216 was increased significantly in the KA-lesioned striatum compared to the non-lesioned striatum. Increasing the amount of KA infused into the striatum augmented radioactivity in the striatum as well as the cerebral cortex and hippocampus of the lesioned side. Treatment with a large amount of non-radioactive AC-5216 or PK11195 inhibited the binding of [(11)C]AC-5216 and diminished the difference of radioactivity levels between the lesion and non-lesioned sides. These results demonstrated that [(11)C]AC-5216 had high specific binding to PBR in the KA-lesioned rat brain. Thus, [(11)C]AC-5216 is a promising PET ligand for imaging PBR in a brain with neuroinflammation.

Incorporation of embryonic CA3 cell grafts into the adult hippocampus at 4-months after injury: Effects of combined neurotrophic supplementation and caspase inhibition

As receptivity of the injured hippocampus to cell grafts decreases with time after injury, strategies that improve graft integration are necessary for graft-mediated treatment of chronic neurodegenerative conditions such as temporal lobe epilepsy. We ascertained the efficacy of two distinct graft-augmentation strategies for improving the survival of embryonic day 19 hippocampal CA3 cell grafts placed into the adult hippocampus at 4-months after kainic acid induced injury. The donor cells were labeled with 5′-bromodeoxyuridine, and pre-treated and grafted with either brain-derived neurotrophic factor, neurotrophin-3 and a caspase inhibitor or fibroblast growth factor and caspase inhibitor. The yield of surviving grafted cells and neurons were quantified at 2-months post-grafting. The yield of surviving cells was substantially greater in grafts treated with brain-derived neurotrophic factor, neurotrophin-3 and caspase inhibitor (84%) or fibroblast growth factor and caspase inhibitor (99% of injected cells) than standard cell grafts (26%). Because ∼85% of surviving grafted cells were neurons, increased yield in augmented groups reflects enhanced survival of grafted neurons. Evaluation of the mossy fiber synaptic re-organization in additional kainic acid-lesioned rats receiving grafts enriched with brain-derived neurotrophic factor, neurotrophin-3 and caspase inhibitor at 3-months post-grafting revealed reduced aberrant dentate mossy fiber sprouting in the dentate supragranular layer than “lesion-only” rats at 4 months post-kainic acid, suggesting that some of the aberrantly sprouted mossy fibers in the dentate supragranular layer withdraw when apt target cells (i.e. grafted neurons) become available in their vicinity. Thus, the yield of surviving neurons from CA3 cell grafts placed into the adult hippocampus at an extended time-point after injury could be enhanced through apt neurotrophic supplementation and caspase inhibition. Apt grafting is also efficacious for reversing some of the abnormal synaptic reorganization prevalent in the hippocampus at later time-points after injury.

Neuronal electrical high frequency stimulation modulates presynaptic GABAergic physiology

Electrical high frequency deep brain stimulation (DBS) of the globus pallidus internus (GPi) or the subthalamic nucleus (STN) has dramatic beneficial motor effects in advanced Parkinson's disease (PD). However, the mechanisms underlying these clinical results remain unclear. It is proposed that the γ-aminobutyric acid (GABA) system is involved in the effectiveness of DBS. To prove this hypothesis, rat striatal slices were stimulated electrically (130 Hz) in vitro; GABA and glutamate (GLU) outflow from striatal slices of normal or kainic acid-lesioned rats were measured after o-phthaldialdehyde sulphite derivatization using HPLC with electrochemical detection. Our results could demonstrate that high frequency stimulation (HFS) did not modulate basal GABA outflow in the perfusate. In the presence of submaximal concentrations of the voltage-gated sodium channel opener veratridine, HFS significantly enhanced GABA outflow. When the GABA transporter inhibitor, nipecotic acid, was added to the incubation medium, the HFS effects decreased to nearly control values. Destruction of striatal GABAergic neurons by kainic acid completely reversed the effects of HFS on GABA outflow. In the present study no effect of HFS on glutamate outflow was observed under any condition. These results suggest that HFS has a specific effect on GABAergic neuronal terminals resulting in an enhancement of extracellular GABA in the caudate nucleus. This effect is probably due to an inhibitory effect of HFS on the GABA uptake system rather than to stimulation of vesicular GABA release from GABAergic neurons, which are both associated with the presynaptic GABAergic physiology.

Effect of γ-hydroxybutyrate in two rat models of focal cerebral damage

γ-Hydroxybutyrate (GHB) and its lactone, γ-butyrolactone (GBL) have been previously shown to produce a protective effect in animal models of cerebral ischaemia/hypoxia, as well as in human conditions of head injury-induced coma. The aim of the present research was to study the effect of GHB in experimental conditions of focal cerebral damage, either induced by ischaemia or excitotoxicity. Under general anaesthesia, rats were injected into the right striatum with either endothelin-1 (ET-1, 0.43 nmol) or kainic acid (7.5 nmol) in a volume of 1 μl. Sham-lesioned rats received 1 μl of the solvent. Both ET-1- and kainic acid-lesioned rats were randomly assigned to one of the following intraperitoneal (i.p.) treatments: (i) and (ii) GHB, 100 or 300 mg kg −1 2 h after the lesion, followed by 50 or 100 mg kg −1, respectively, every 12 h; (iii) saline, 2 ml kg −1, same schedule. Sham animals were treated with saline, 2 ml kg −1, same schedule. Treatments lasted for 10 days. The higher dose of GHB produced a significant protection against the ET-1-induced impairments in sensory-motor orientation and coordinated limb use (evaluated 24 and 42 days after the lesion) and in place learning and memory (Morris test, performed 19 and 39 days after the lesion). The same dose regimen reduced the circling behaviour induced by apomorphine in kainate-lesioned rats (10 days after the lesion), and limited or prevented at all the histological damage produced either by ET-1 or by kainic acid (evaluated 43 or 10 days after the lesion, respectively). These results show that GHB limits both histological and functional consequences of a focal ischaemic or excitotoxic insult of the brain, in rats, even if the treatment is started 2 h after the lesion.

Reduced Mg 2+ blockade of synaptically activated N-methyl- d-aspartate receptor-channels in CA1 pyramidal neurons in kainic acid-lesioned rat hippocampus

Unilateral kainic acid lesion in the hippocampus caused a long-term change in the balance between excitatory and inhibitory drive onto CA1 pyramidal cells, making these cells hyperexcitable several weeks post-lesion. In this study, we have shown an enhanced N-methyl- d-aspartate receptor-mediated component in the excitatory synaptic transmission together with a reduced GABA A receptor-mediated inhibition in CA1 pyramidal cells one-week post kainic acid lesion. In these cells, pharmacologically isolated N-methyl- d-aspartate receptor-mediated whole-cell excitatory postsynaptic currents were significantly larger at negative holding potentials, and the voltage-dependence of N-methyl- d-aspartate receptor channels was shifted in the hyperpolarizing direction. The plot of relative conductance (g/g Max) shifted significantly ( P<0.01) to more negative holding potentials by 19 mV (−28±4 mV in control slices and −47±4 mV in kainic acid slices) at the half maximal conductance point (g/g Max=0.5). This shift gives a larger N-methyl- d-aspartate receptor-mediated component in the excitatory synaptic transmission at resting membrane potentials (around −60 mV). The shifted voltage dependence is highly sensitive to extracellular Mg 2+ ions. Moderate increases in [Mg 2+] o from 1 mM to 2.6 mM more than compensated for the negative shift and effectively suppressed the population epileptiform bursting activity. Fitting the voltage dependence to an ionic block model revealed a higher dissociation constant of N-methyl- d-aspartate receptor channels for Mg 2+ in kainic acid-lesioned slices (52 mM at 0 mV; 330 μM at −60 mV) than in control slices (7.7 mM at 0 mV; 93 μM at −60 mV). While a simple single site model adequately fitted the control data for [Mg 2+] o at 1 mM and 2.6 mM, no consistent model of this form was found for the kainic acid-lesioned slices. These results revealed changed properties of N-methyl- d-aspartate receptor channels in the kainic acid-lesioned model of epilepsy. The reduced Mg 2+ blockade of N-methyl- d-aspartate receptor channels contributed significantly to the epileptiform bursting activity.

Radial maze learning deficits in striatal kainic acid-lesioned rats and effects of GABA receptor agonists

Cholecystokinin (CCK) is deeply involved in the control of learning and emotional behaviors. The authors characterize the behavioral properties of Otsuka Long Evans Tokushima Fatty (OLETF) rats, which lack the CCK-A receptor because of a genetic abnormality. In the Morris water-maze task, the OLETF rats showed an impaired spatial memory. In the inhibitory avoidance test, they showed facilitating response 24 h after training. Hypoalgesia was observed in a hot-plate test. In the elevated plus-maze and food neophobia test, OLETF rats showed an anxiety-like response. In addition, OLETF rats were hypoactive in the Morris water-maze and the elevated plus-maze. The results suggest that the OLETF rats showed a spatial memory deficit, hypoactivity and anxiety due, at least in part, to the lack of CCK-A receptors.

Differential expression of apolipoprotein D and apolipoprotein E in the kainic acid-lesioned rat hippocampus

Expression of apolipoprotein D, a member of the lipocalin superfamily of transporter proteins, was investigated in the kainic acid-lesioned rat hippocampus. Using an anti-rat apolipoprotein D antibody and biotin–avidin-enhanced immunocytochemistry, in the normal rat hippocampus there was little apolipoprotein D expression, that was restricted mainly to scattered astrocytes. By contrast, kainic acid-injected rats showed apolipoprotein D immunoreactivity in the pyramidal neurons of the affected CA fields 24–48 h after injection of the excitotoxin, at a time when there was no histological evidence of cell death. Apolipoprotein D immunoreactivity peaked by day 3, coincident with neuronal cell death, and declined thereafter, reaching very low levels by day 7. Besides pyramidal neurons, apolipoprotein D immunoreactivity was also observed in a small number of reactive glial cells in the affected CA fields, but not in the vascular compartments at any time-point. In contrast to the neuronal expression of apolipoprotein D, apolipoprotein E immunoreactivity was observed predominantly in degenerating astrocytes. In conclusion, following excitotoxic injury with kainic acid, apolipoprotein D is expressed in hippocampal pyramidal neurons destined for subsequent cell death.

A role for synaptic and network plasticity in controlling epileptiform activity in CA1 in the kainic acid-lesioned rat hippocampus in vitro.

1. Stimulation of the surviving afferents in the stratum radiatum of the CA1 area in kainic acid-lesioned hippocampal slices produced graded epileptiform activity, part of which (> 20%) involved the activation of N-methyl-D-aspartate (NMDA) receptors. There was also a failure of synaptic inhibition in this region. In this preparation, we have tested the effects of low-frequency stimulation (LFS; 1 Hz for 15 min) on synaptic responses and epileptiform activity. 2. LFS resulted in long-term depression (LTD) of excitatory synaptic potentials (EPSPs), long-term decrease of population spike amplitudes (PSAs) and EPSP-spike (E-S) potentiation. Evoked epileptiform activity was reduced but neurons had a higher probability of discharge. LTD could be reversed by subsequent tetanic stimulation whereas E-S dissociation remained unchanged. Synaptic and network responses could be saturated towards either potentiation or depression. However, E-S potentiation was maximal following the first conditioning stimulus. 3. NMDA receptor-mediated responses were pharmacologically isolated. LFS resulted in LTD of synaptic responses, long-term decrease of PSAs and E-S depression. These depressions could not be reversed by subsequent tetanic stimulation. alpha-Amino-3-hydroxy-5-methylisoxazolepropionic acid (AMPA) and NMDA receptor-mediated responses were then measured in isolation before and following conditioning stimuli. LFS was shown to simultaneously produce LTD of AMPA and NMDA receptor-mediated responses. E-S potentiation of the AMPA component and E-S depression of the NMDA component occurred coincidentally. 4. LTD of AMPA and NMDA receptor-mediated responses were shown to be NMDA dependent. In contrast, E-S potentiation and depression occurred even when NMDA receptors were pharmacologically blocked. 5. These findings indicate that synaptic responses could be modified bidirectionally in the CA1 area of kainic acid-lesioned rat hippocampus in an NMDA receptor-dependent manner. However, E-S dissociations were independent of the activation of NMDA receptors, hinting at mechanisms different from those of synaptic LTD. We suggest that changes in E-S coupling were caused by a modification of the firing threshold of the CA1 pyramidal neurons. Furthermore, the firing mechanisms controlling NMDA and AMPA receptor-mediated network activity appeared to be different. The possible use of LFS applied to the hippocampus as a clinical intervention to suppress epileptiform activity is discussed.

Simultaneous expression of long-term depression of NMDA and long-term potentiation of AMPA receptor-mediated synaptic responses in the CA1 area of the kainic acid-lesioned hippocampus.

This study investigates the plasticity of the excitatory synapses in an experimental model of epilepsy, the kainic acid-lesioned rat hippocampus. Stimulation of afferents in the CA1 area of lesioned hippocampi produced an epileptiform burst of action potentials, with an underlying synaptic potential composed of mixed alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA; 80%) and N-methyl-D-aspartate (NMDA; 20%) receptor-mediated components. Tetanic stimulation yielded a long-term potentiation (LTP) of the mixed AMPA/NMDA receptor-mediated population excitatory postsynaptic potentials. However, the same type of tetanus resulted in a long-term depression (LTD) of pharmacologically isolated NMDA receptor-mediated responses. This LTD occurred independently of the antagonism of AMPA receptors. This suggests that tetanic stimulation produced LTP of AMPA and LTD of NMDA receptor-mediated responses simultaneously. Finally, both LTP and LTD were shown to be NMDA dependent. This property has profound functional implications for the control of excitatory networks in temporal lobe epilepsy.

Axons of passage may be responsible for fastigial nucleus pressor response.

Bilateral destruction of perikarya in the fastigial nucleus (FN) of the rat with the cytotoxic agent kainic acid (0.5 mg) did not alter the blood pressure (BP) increases observed during monopolar electrical stimulation (100 microA, 50 Hz, 0.5-ms pulse width) of this region. BP increases in control animals were 30 +/- 8 mm Hg, whereas BP increased 30 +/- 7 mmHg in kainic acid-lesioned rats. Furthermore, picrotoxin (100 ng) and muscimol (25 ng) microinjected unilaterally into the FN of conscious, unrestrained rats produced postural asymmetry but no change in BP or heart rate. These data suggest that the FN pressor response may be due, at least in part, to stimulation of axons of passage.

Dramatic increase in nigral t-[ 35S]butylbicyclophosphorothionate binding sites elicited by the degeneration of the striato-nigral GABAergic pathway: reversal by diazepam

In rats, the degeneration of the striato-nigral GABAergic pathway caused by the intrastriatal injection of kainic acid induced a marked decrease (65%) of GABA content and glutamic acid decar☐ylase (GAD) activity and a dramatic increase (225%) in the binding of t-[ 35S]butylbicyclophosphorothionate ([ 35S]TBPS) to a membrane preparation from the substantia nigra homolateral to the injected striatum. The increase in [ 35S]TBPS binding in the denervated substantia nigra was exclusively due to an increased density of binding sites ( B max) with no change in the dissociation constant ( k d). The enhancement in [ 35S]TBPS binding was almost completely reversed by the intraperitoneal administration of diazepam (3 mg/kg) to kainic acid-lesioned rats. Moreover, diazepam produced a significant decrease (30%) in the density of [ 35S]TBPS binding sites also in the sham-operated side. In contrast the ‘in vitro’ addition of the GABA A receptor antagonist bicuculline (1 μM) to the membrane preparation from the denervated substantia nigra further increased [ 35S]TBPS binding. These findings suggest the view that the increase of nigral [ 35S]TBPS binding is directly related to the inhibition in the function of nigral GABAergic synapses following the loss of the striato-nigral GABAergic pathway. Our results indicate that [ 35S]TBPS binding to brain structure is a potential tool to reveal alteration in the function of GABA A receptor complex elicited by physiological, pharmacological and pathological conditions.

Differentiation of glioblasts from adult brain

Rat glial (RG) cells have been established in continuous culture using sections of kainic acid-lesioned rat striata as the primary cell source. Such cultures consistently provided at least 2 morphologically distinguishable cell types. More than 95% of this population was composed of large, flat cells which possessed ill-defined junctions and lacked cellular processes. The second class of cells comprised less than 5% of the population, was smaller and had several processes per cell. Dibutyryl cyclic-AMP (dB-cAMP), which has been shown to promote a maturation-stimulating effect on embryonic glioblasts in vitro, appears to have a similar effect on RG cells. In the presence of 1 mM dB-cAMP virtually all RG cells undergo morphological transformation, becoming smaller and denser, in addition to forming several processes per cell. Furthermore, two cell-type-specific markers could be detected in dB-cAMP-treated cultures: the surface oligodendrocyte-specific galactocerebroside (GC) moiety was demonstrated to be present on less than 10% of the cells, whereas the intracellular astrocyte-specific glial fibrillary acidic (GFA) protein could be readily detected in 80% of the population. These data indicate that glia derived from the striata of injured adult rat brain exhibit fetal characteristics.

Kainic acid injections in the striatum alter the cataleptic and locomotor effects of drugs influencing dopaminerig and cholinergic systems

Rats with bilateral injections of kainic acid into the striatum were tested for their motor responsiveness to drugs influencing dopaminergic and cholinergic systems. The kainic acid-induced lesions potentiated the locomotor response to both the dopaminergic agonist, d-amphetamine, and the cholinergic antagonist, scopolamine, attenuated the cataleptic response to the dopaminergic antagonist, haloperidol, and potentiated the cataleptic and convulsive responses to the cholinergic agonist, pilocarpine. The analogy of these pharmacological effects with those induced by similar drugs in patients with Huntington's disease supports the view that this animal preparation is a useful model of Huntington's disease. The opposite effects of haloperidol and pilocarpine on catalepsy in kainic acid-lesioned rats suggest that more work should be done to elucidate the mechanism behind this conflict before full support can be given to its use as a model system for evaluating possible pharmacotherapy in Huntington's disease.

Effects of kainic acid lesions in the lateral hypothalamus on behavior and hippocampal and neocortical electroencephalographic (EEG) activity in the rat

Electrolytic lesions of the lateral hypothalamus (LH) are known to produce severe and chronic behavioral and electrographic abnormalities. In order to assess the effects of damage to LH cells vs damage to fibers of passage in the LH, a comparison was made of the effects of electrolytic lesions and micro-injections of the neurotoxin kainic acid (kainate) in the LH. Behavior and neocortical and hippocampal electroencephalographic (EEG) activity were studied before, immediately after, and for 25 days after the lesions were made. Electrolytic-lesioned rats were aphagic and adipsic, showed an absence of normal atropine-resistant EEG activity, and a release of atropine-sensitive EEG activity in the hippocampus and neocortex. Kainic acid-lesioned rats showed some similar behavioral impairments but the kainate lesions produced different EEG abnormalities, including chronic slow-wave and seizure activity in both the neocortex and hippocampus. Following extended recovery hippocampal EEG was normal despite extensive cellular loss in areas CA3 and CA4. Understanding of the differences in EEG between the electrolytic and kainate effects was compounded by widespread cellular damage in areas outside the hypothalamus in the rats with kainate lesions. Thus, the kainate-produced abnormalities precluded a simple analysis of the contribution that cell damage alone makes to LH lesion-induced behavioral and EEG changes.

Effects of kainic acid lesions in the lateral hypothalamus on behavior and hippocampal and neocortical electroencephalographic (eeg) activity in the rat

Electrolytic lesions of the lateral hypothalamus (LH) are known to produce severe and chronic behavioral and electrographic abnormalities. In order to assess the effects of damage to LH cells vs damage to fibers of passage in the LH, a comparison was made of the effects of electrolytic lesions and micro-injections of the neurotoxin kainic acid (kainate) in the LH. Behavior and neocortical and hippocampal electroencephalographic (EEG) activity were studied before, immediately after, and for 25 days after the lesions were made. Electrolytic-lesioned rats were aphagic and adipsic, showed an absence of normal atropine-resistant EEG activity, and a release of atropine-sensitive EEG activity in the hippocampus and neocortex. Kainic acid-lesioned rats showed some similar behavioral impairments but the kainate lesions produced different EEG abnormalities, including chronic slow-wave and seizure activity in both the neocortex and hippocampus. Following extended recovery hippocampal EEG was normal despite extensive cellular loss in areas CA3 and CA4. Understanding of the differences in EEG between the electrolytic and kainate effects was compounded by widespread cellular damage in areas outside the hypothalamus in the rats with kainate lesions. Thus, the kainate-produced abnormalities precluded a simple analysis of the contribution that cell damage alone makes to LH lesion-induced behavioral and EEG changes.

Characterisation of denervation supersensitivity in the striatonigral GABA pathway of the kainic acid-lesioned rat and in Huntington's disease

Destruction of the rat Striatonigral GABA pathway by striatal kainic acid (KA) lesion resulted in both increased behavioural response to intranigral muscimol and elevated nigral 3H-GABA binding. Behavioural responses were correlated with both depletions of GABA and elevations in 3H-GABA binding. Elevated 3H-GABA binding was characterised by an increase in Bmax with no change in K d for the high affinity component. Following the proposal of the striatal KA lesion as an animal model for Huntington's disease (HD), 33H-GABA binding was studied in the nigra of post-mortem brains from 5 control and 4 HD patients. Bmax for the high affinitiy component was elevated by 136% ( p<0.01) with no change in K d. Low affinity binding parameters were unaltered. These results are consistent with the development of denervation supersensitivity of nigral GABA receptors in HD and its animal model. They demonstrate the adaptive capacity of the human GABA system and have implications for the therapeutic potential of GABA agonists in HD.