Don Shin

Depression after status epilepticus: behavioural and biochemical deficits and effects of fluoxetine

Depression represents one of the most common comorbidities in patients with epilepsy. However, the mechanisms of depression in epilepsy patients are poorly understood. Establishment of animal models of this comorbidity is critical for both understanding the mechanisms of the condition, and for preclinical development of effective therapies. The current study examined whether a commonly used animal model of temporal lobe epilepsy (TLE) is characterized by behavioural and biochemical alterations involved in depression. Male Wistar rats were subjected to LiCl and pilocarpine status epilepticus (SE). The development of chronic epileptic state was confirmed by the presence of spontaneous seizures and by enhanced brain excitability. Post-SE animals exhibited increase in immobility time under conditions of forced swim test (FST) which was indicative of despair-like state, and loss of taste preference in saccharin solution consumption test which pointed to the symptomatic equivalence of anhedonia. Biochemical studies revealed compromised serotonergic transmission in the raphe-hippocampal serotonergic pathway: decrease of serotonin (5-HT) concentration and turnover in the hippocampus, measured by high performance liquid chromatography, and decrease of 5-HT release from the hippocampus in response to raphe stimulation, measured by fast cyclic voltammetry. Administration of fluoxetine (FLX, 20 mg/kg/day for 10 days) to naive animals significantly shortened immobility time under conditions of FST, and inhibited 5-HT turnover in the hippocampus. In post-SE rats FLX treatment led to a further decrease of hippocampal 5-HT turnover; however, performance in FST was not improved. At the same time, FLX reversed SE-induced increase in brain excitability. In summary, our studies provide initial evidence that post-SE model of TLE might serve as a model of the comorbidity of epilepsy and depression. The finding that behavioural equivalents of depression were resistant to an antidepressant medication suggested that depression in epilepsy might have distinct underlying mechanisms beyond alterations in serotonergic pathways.

Epileptogenesis after status epilepticus reflects age- and model-dependent plasticity

Although epilepsy often begins in childhood, factors that contribute to the development of epilepsy as a consequence of status epilepticus (SE) during early development are poorly understood. We investigated animal models in which seizure‐induced epileptogenicity could be studied. Rats undergoing self‐sustaining SE induced by perforant path stimulation (PPS) at the ages of postnatal day 21 (P21) and P35 were compared with those subjected to SE by lithium and pilocarpine (LiPC). Although only one animal subjected to PPS at P21 developed chronic spontaneous seizures by several months of observation, all the animals subjected to PPS at P35 became epileptic. In the LiPC model, however, most of the rat pups subjected to SE at P21 became epileptic. Animals with spontaneous seizures showed increased inhibition in the dentate gyrus, a characteristic of the epileptic brain, with evidence of mossy fiber synaptic reorganization. Examination of circuit recruitment by c‐Jun immunohistochemistry showed activation restricted to the hippocampus in P21 animals subjected to PPS, although extensive activation of hippocampal and extrahippocampal structures was seen in pups subjected to PPS‐induced self‐sustaining SE at P35 or LiPC SE at P21. These results demonstrate that the appearance of epilepsy as a consequence of SE is influenced by the type of insult as well as by age‐dependent circuit recruitment. Ann Neurol 2000;48:580–589

Elevated plasma corticosterone level and depressive behavior in experimental temporal lobe epilepsy

Depression is frequently reported in epilepsy patients; however, mechanisms of co-morbidity between epilepsy and depression are poorly understood. An important mechanism of depression is disinhibition within the hypothalamo-pituitary-adrenocortical (HPA) axis. We examined the functional state of the HPA axis in a rat model of co-morbidity between temporal lobe epilepsy and depression. Epilepsy was accompanied by the interictal elevation of plasma corticosterone, and by the positively combined dexamethasone/corticotropin releasing hormone test. The extent of the HPA hyperactivity was independent of recurrent seizures, but positively correlated with the severity of depressive behavior. We suggest that the observed hyperactivity of the HPA axis may underlie co-morbidity between epilepsy and depression.

Self-sustaining status epilepticus after brief electrical stimulation of the perforant path

We examined the duration of intermittent perforant path stimulation (PPS) needed to induce self-sustaining status epilepticus (SSSE) in rats. Seven-minute PPS did not induce SSSE. Some rats receiving 15 min and all animals after 30 min PPS developed SSSE that continued for hours. The animals killed 3 days after SSSE showed extensive neuronal damage. Those which were allowed to survive for 6 weeks after SSSE displayed spontaneous seizures.

Serum neuron-specific enolase is a marker for neuronal damage following status epilepticus in the rat

We determined the serum concentrations of neuron-specific enolase (s-NSE) in rat pups of 1, 2, 3, and 4 weeks of age and in adult rats that were subjected to lithium-pilocarpine status epilepticus (SE). Damage to brain regions was rated on a scale of 0 (no damage) to 5 (> 50% cell loss). Rat pups of 1-2 weeks of age had a higher baseline s-NSE than the adults. Following SE, 1 week old rat pups had no elevation of s-NSE and no histologic evidence of damage. At older ages the increases in NSE ranged from 18.9 +/- 0.8 ng/ml in the 2 week old (vs. 11.5 +/- 0.5 control) to 35.8 +/- 2.1 ng/ml in the 3 week old (vs. 12.1 +/- 0.8 control). In the adult rats s-NSE increased from 5.4 +/- 0.4 in the control animals to 30.4 +/- 1.3 after SE. The different brain regions examined had distinctive ontogenic profiles for SE-induced damage. Elevation of s-NSE after SE correlated with overall histologic evidence for damage.

Inflammation induced by LPS enhances epileptogenesis in immature rat and may be partially reversed by IL1RA.

Inflammatory signaling in the central nervous system (CNS) has been shown to exacerbate both seizure activity and seizure‐induced neuronal injury. However, it has not been firmly established whether neurodegeneration is a prerequisite of proconvulsant effect of neuroinflammation, or whether the latter may facilitate seizures without involving neuronal injury. We examined effects of inflammation in the rapid kindling model, where seizure progression occurs in the absence of neurodegeneration. P14 male Wistar rats were subjected to a rapid kindling procedure: 60 electrical stimulations of the hippocampus delivered every 5 min at the current that had been established to induce afterdischarge. Lipopolysaccharide (LPS) was injected (50 μg/kg, i.p., 2 h prior to the rapid kindling protocol [RKP]); IL‐1Ra was injected (25 mg/kg, i.p., 2 h prior to the RKP). The effects of treatments were examined on baseline hippocampal excitability, on the progression of rapid kindling, and on the retention of rapid kindling. LPS increased baseline hippocampal excitability, evident as the decrease of hippocampal ADT. LPS also increased kindling progression. Twenty‐four hours after the completion of kindling procedure, LPS‐treated animals exhibited increased excitability as compared with saline‐treated kindling controls. The kindling progression was blocked by IL1RA when given in combination with LPS. IL1RA was able to reverse the effect of LPS on afterdischarge duration (ADD) while IL1RA alone decreased ADT. We showed that inflammation provoked by LPS enhanced rapid kindling epileptogenesis in immature rat brains. IL1RA was also able to mitigate this augmentation of epileptogenesis enhanced by LPS.

Granule Cell Neurogenesis After Status Epilepticus in the Immature Rat Brain

Summary: Purpose: Several experimental paradigms of seizure induction that produce epilepsy as a consequence have been shown to be associated with the proliferation of dentate granule cells. In developing animals, the acute sequela of hilar damage and the chronic sequelae of spontaneous seizures and mossy fiber synaptic reorganization, in response to status epilepticus, occur in an age‐dependent manner. We investigated seizure‐induced granule cell neurogenesis in developing rat pups to study the association between hilar injury, granule cell neurogenesis, and epilepsy.

Regulation of Kindling Epileptogenesis by Hippocampal Galanin Type 1 and Type 2 Receptors: The Effects of Subtype-Selective Agonists and the Role of G-Protein-Mediated Signaling

The search for antiepileptic drugs that are capable of blocking the progression of epilepsy (epileptogenesis) is an important problem of translational epilepsy research. The neuropeptide galanin effectively suppresses acute seizures. We examined the ability of hippocampal galanin receptor type 1 (GalR1) and type 2 (GalR2) to inhibit kindling epileptogenesis and studied signaling cascades that mediate their effects. Wistar rats received 24-h-long intrahippocampal infusion of a GalR1/2 agonist galanin(1-29), GalR1 agonist M617 [galanin(1-13)-Gln14-bradykinin(2-9)-amide], or GalR2 agonist galanin(2-11). The peptides were administered alone or combined with an inhibitor of Gi protein pertussis toxin (PTX), Gi-protein activated K+ channels (GIRK) inhibitor tertiapin Q (TPQ), Gq/11 protein inhibitor [d-Arg1,d-Trp5,7,9,Leu11]-substance P (dSP), or an inhibitor of intracellular Ca2+ release dantrolene. Sixteen hours into drug delivery, the animals were subjected to rapid kindling—60 electrical trains administered to ventral hippocampus every 5 min. M617 delayed epileptogenesis, whereas galanin(1-29) and galanin(2-11) completely prevented the occurrence of full kindled seizures. TPQ abolished anticonvulsant effect of M617 but not of galanin(2-11). PTX blocked anticonvulsant effects of M617 and inversed the action of galanin(1-29) and galanin(2-11) to proconvulsant. dSP and dantrolene did not modify seizure suppression through GalR1 and GalR2, but eliminated the proconvulsant effect of PTX + galanin(1-29) and PTX + galanin(2-11) combinations. We conclude that hippocampal GalR1 exert their disease-modifying effect through the Gi-GIRK pathway. GalR2 is antiepileptogenic through the Gi mechanism independent of GIRK. A secondary proconvulsant pathway coupled to GalR2 involves Gq/11 and intracellular Ca2+. The data are important for understanding endogenous mechanisms regulating epileptogenesis and for the development of novel antiepileptogenic drugs.

Inflammation exacerbates seizure-induced injury in the immature Brain

Summary:  We examined the hypothesis that the introduction of an inflammatory agent would augment status epilepticus (SE)‐induced neuronal injury in the developing rat brain in the absence of an increase in body temperature. Postnatal day 7 (P7) and P14 rat pups were injected with an exogenous provocative agent of inflammation, lipopolysaccharide (LPS), 2 h prior to limbic SE induced by either lithium‐pilocarpine (LiPC) or kainic acid. Core temperature was recorded during the SE and neuronal injury was assessed 24 h later using profile cell counts in defined areas of the hippocampus. While LPS by itself did not produce any discernible cell injury at either age, it exacerbated hippocampal damage induced by seizures. In the LiPC model, this effect was highly selective for the CA1 subfield, and there was no concomitant rise in body temperature. Our findings show that inflammation increases the vulnerability of immature hippocampus to seizure‐induced neuronal injury and suggest that inflammation might be an important factor aggravating the long‐term outcomes of seizures occurring early in life.

Kindling epileptogenesis in immature rats leads to persistent depressive behavior.

Depression is a frequent comorbidity in epilepsy patients. A variety of biological factors may underlie epilepsy-associated depression. We examined whether kindling-induced chronic increase in seizure susceptibility is accompanied by behavioral symptoms of depression. Three-week-old Wistar rats underwent rapid kindling: 84 initially subconvulsant electrical stimulations of ventral hippocampus delivered every 5 minutes, followed by depression-specific behavioral tests performed 2 and 4 weeks later. Kindled animals exhibited a sustained increase in immobility time in the forced swim test and the loss of taste preference toward calorie-free saccharin, as compared with controls. Initial loss of preference toward the intake of calorie-containing sucrose was followed by the increased consumption at 4 weeks. At both time points, animals exhibited enhanced seizure susceptibility on test stimulations of the hippocampus. We conclude that neuronal plastic changes associated with the kindling state are accompanied by the development of depressive behavior.