Bruce J. Albala

Kainic-acid-induced seizures: A developmental study

Developmental dose-response curves for kainic-acid-induced seizures were generated in rats. Rats at 15-18 days (pups), 33-37 days ( pubescents ) and over 90 days (adults) were administered kainic acid intraperitoneally. Seizures were elicited in all 3 age groups, but some of the behavioral manifestations differed in the pups. This group also had the lowest convulsive threshold, the most severe seizures and the highest mortality. Forelimb convulsions and status epilepticus were associated with the occurrence of necrotic lesions in the adults and pubescents but not in the pups. Deoxyglucose (DG) autoradiographic studies of the convulsing rats disclosed differences in the DG uptake pattern of the substantia nigra across the 3 age groups. Increases in the DG uptake were present in the two older age groups but not in the pups. Since recent data have implicated the substantia nigra as a crucial site in a seizure modifying circuitry in adult animals, our results suggest that the lack of substantia nigra involvement in the pups may account for the early generalization and the increased severity of seizures in this age group.

Increased seizure susceptibility of the immature brain

The ability of the CNS to generate seizures as a function of age was investigated utilizing the kindling model of epilepsy. Hourly electrical, low intensity stimulations of the amygdala induced kindling in adult rats, but stimulations delivered at 15 min intervals failed to or markedly retarded the development of kindled convulsions. In contrast, both types of stimulation induced consistent prolongation of the afterdischarges and repeated generalized seizures in suckling rat pups. The rate of development of the kindled convulsions in the pups was similar, irrespective of whether the stimulations were delivered at 15 or 60 min intervals, indicating that short (less than 15 min) seizure refractory periods exist in the immature brain. The data suggest that seizure susceptibility changes with age and is greater early in life.

Nigral muscimol infusions facilitate the development of seizures in immature rats.

Since recent data utilizing GABAergic stimulation of the substantia nigra (SN) suggest that the SN is a crucial site in a circuitry involved in the modification of seizures in adult rats, the role of the SN was investigated in seizures of rat pups. Bilateral nigral infusions of the GABA agonist muscimol partially protected adult rats against flurothyl-induced seizures, while similar infusions actually facilitated the development of flurothyl seizures in 15-day-old rat pups. These results suggest that age-related differences in the nigral GABA sensitive system may account for the increased susceptibility to generalized seizures of the developing brain.

Kindling in developing rats: Persistence of seizures into adulthood

The effects of kindling induced in developing rat pups were re-examined when the rats reached maturity. Eighteen-day-old rats, implanted with bipolar electrodes, were separated into the following 4 groups: the fully kindled group consisted of pups that developed generalized convulsions; the partially kindled group of pups that consistently displayed afterdischarges and occasionally behavioral focal seizures; the afterdischarge group of pups in which the afterdischarge threshold was measured and the non-stimulated group of pups implanted but not stimulated. Subsequently, the electrodes were removed and the rats were allowed to grow. At 65-70 days, the surviving rats and age matched controls were rekindled. The vulnerability to kindled convulsions at maturity, as expressed by the kindling rates, correlated directly with the severity of previous kindled seizures, but did not correlate with the presence of long standing gliotic lesions. These findings suggest that the neural alterations evoked by kindling persist and are not modified by the dynamic changes that occur with growth.

Infantile status epilepticus and future seizure susceptibility in the rat.

The long-term effects of infantile seizures on the development of seizures in adulthood were studied in rats. Infantile seizures of varying severity were induced with intraperitoneal injections of kainic acid in 15-day-old rats. In adulthood the seizure susceptibility of the rats was determined by kindling the left amygdala and by measuring their ability to resist recurrent seizures. The results suggest that infantile status epilepticus is associated with a very high mortality; however in the surviving rats, infantile seizures even as severe as status epilepticus do not cause neuronal brain damage and do not predispose to the development of convulsions later in life.

Unilateral peri-substantia nigra catecholaminergic lesion and amygdala kindling

Recent evidence suggests that the substantia nigra (SN) may be involved in the modulation of kindled seizures in adult rats. In this report we investigated the role of the dopaminergic nigrostriatal pathway in mediating the SN effect on seizures by lesioning this pathway with unilateral infusions of 6-hydroxydopamine (6-OHDA) in the vicinity of the right SN with or without desmethylimipramine pretreatment. Our data suggest that the facilitation of amygdala kindling observed following 6-OHDA lesion in the vicinity of the ipsilateral SN is due to norepinephrine depletion of the ipsilateral forebrain. Selective destruction of the nigrostriatal dopaminergic neurons neither facilitates nor inhibits the development of amygdala-kindled convulsions in adult rats.

Perinatal hypoxia and subsequent development of seizures

Perinatal asphyxia has been implicated in the pathogenesis of persistent convulsive disorders later in life. Whether epilepsy is the result of oxygen deficiency alone or is due to the combined effect of hypoxia and ischemia is not known. In this report we studied the role of perinatal hypoxia alone on the development of epilepsy. One day and ten days old rat pups were exposed to prolonged hypoxia (6% O2). The subsequent susceptibility to focally elicited (kindled) or generalized (flurothyl) seizures was determined in the fourth week of age. Rats exposed to hypoxia were not more susceptible to the development of either type of seizures when compared to controls. Since the equivalent degree of hypoxia used for the 1 day old rat has previously been shown to result in lasting neurochemical and behavioral alterations and the degree of hypoxia used for the 10 day rat was lethal in over 35% of the animals, it is suggested that oxygen deficiency in and of itself may not be sufficient to lead to the development of epilepsy.

Restriction of enhanced [2-14C]deoxyglucose utilization to rhinencephalic structures in immature amygdala-kindled rats

Sixteen-day-old albino rat pups were kindled to varying degrees of seizure severity with amygdala stimulations spaced 15 to 20 min apart. Subsequently, each rat pup was injected (ip) with 10 microCi of [2-14C]-deoxyglucose, and received several additional kindled seizures at regular intervals throughout the following 80 min, at which time it was killed and processed for deoxyglucose autoradiography. Increased seizure severity was associated with correspondingly increased deoxyglucose utilization in many rhinencephalic limbic structures. However, unlike adults, rat pups did not show discernibly increased neocortical, thalamic, or substantia nigra utilization. We postulate that the apparent confinement of seizure activity to limbic structures in pups is related to their relative lack of postictal seizure refractoriness, as well as to other indices of increased seizure susceptibility in immature animals.

Ventromedial thalamic lesions and seizure susceptibility

Recent data indicate that the substantia nigra is an important site in a circuitry involved in the modification of various experimental seizures with neocortical and limbic involvement. Since there are no direct nigral projections to either area, we assumed that the nigral effects on seizures are relayed by other sites such as the thalamus. To evaluate this hypothesis we produced bilateral high-radiofrequency thermocoagulative lesions of the ventromedial (VM) thalamic nuclei which receive the nigral efferents in the rat. We determined the susceptibility of lesioned and control adult rats to the development of flurothyl seizures 2 and 4 weeks later. The latency to the onset of a generalized seizure was considered as the convulsive threshold. There were no differences in the mean latencies between the groups. The results suggest that bilateral destruction of the VM thalamic nuclei does not modify the susceptibility to the development of flurothyl seizures in the rat.

Unidirectional interaction between flurothyl seizures and amygdala kindling

In this report, the interaction between flurothyl convulsions and electrical kindling of the amygdala was investigated. Three flurothyl convulsions decreased the afterdischarge threshold of the amygdala and enhanced the rate of development of electrical kindling without affecting the intensity of postictal refractoriness. On the other hand, 3 generalized kindled convulsions did not alter the flurothyl convulsive threshold. The data suggest that the influence of generalized convulsions on future seizure susceptibility may depend on the agents used to induce the convulsions.