Prolonged epileptiform bursting induced by 0-Mg(2+) in rat hippocampal slices depends on gap junctional coupling.

The transition from brief interictal to prolonged seizure, or 'ictal', activity is a crucial event in epilepsy. In vitro slice models can mimic many phenomena observed in the electroencephalogram of patients, including transition from interictal to ictaform or seizure-like activity. In field potential recordings, three discharge types can be distinguished: (1) primary discharges making up the typical interictal burst, (2) secondary bursts, lasting several hundred milliseconds, and (3) tertiary discharges lasting for seconds, constituting the ictal series of bursts. The roles of chemical synapses in these classes of burst have been explored in detail. Here we test the hypothesis that gap junctions are necessary for the generation of secondary bursts. In rat hippocampal slices, epileptiform activity was induced by exposure to 0-Mg(2+). Epileptiform discharges started in the CA3 subfield, and generally consisted of primary discharges followed by 4-13 secondary bursts. Three drugs that block gap junctions, halothane (5-10 mM), carbenoxolone (100 microM) and octanol (0.2-1.0 mM), abolished the secondary discharges, but left the primary bursts intact. The gap junction opener trimethylamine (10 mM) reversibly induced secondary and tertiary discharges. None of these agents altered intrinsic or synaptic properties of CA3 pyramidal cells at the doses used. Surgically isolating the CA3 subfield made secondary discharges disappear, and trimethylamine under these conditions was able to restore them.We conclude that gap junctions can contribute to the prolongation of epileptiform discharges.

Second messenger modulation of electrotonic coupling between region CA3 pyramidal cell axons in the rat hippocampus.

Gap junction coupling between hippocampal cell axons has been implicated in high frequency oscillations. We used antidromic activation of region CA3 from the fimbria to test the hypothesis that, if gap junctions exist between CA3 pyramidal cell axons, they should cause cross-talk between cells. Agents known to open gap junctions, including 8-Br-cAMP and forskolin (analogue and activator of the cAMP 2nd messenger system respectively) augmented the antidromic population spike and uncovered fast oscillations in the extracellular field. Increasing 2nd messenger concentration reduced the threshold stimulation for antidromic triggering of action potentials, suggesting an improved capability to conduct the electrical impulse retrogradely to the soma. Our studies support the existence of gap junction coupling between CA3 pyramidal cell axons in the fimbria that can be acutely modulated by 2nd messengers.

Functional phenotype in transgenic mice expressing mutant human presenilin-1.

Mutations in the presenilin-1 (PS1) gene cause approximately 50% of cases of early onset familial Alzheimer's disease. The function of this protein remains unknown. We have made an electrophysiological study of hippocampal slices from transgenic mice expressing either a normal human PS1 transgene (WT) or one of two human PS1 transgenes bearing pathogenic mutations at codon M146 (M146L and M146V). Medium and late afterhyperpolarizations in CA3 pyramidal cells were larger in mice expressing either mutant form compared with WT and nontransgenic controls. Calcium responses to depolarization were larger in M146L mice compared with nontransgenic littermates; synaptic potentiation of the CA3 to CA1 projection was also stronger. These results demonstrate disruption of the control of intracellular calcium and electrophysiological dysfunction in PS1 mutant mice.

D(1)-like dopamine receptors on retrogradely labelled sympathoadrenal neurones in the thoracic spinal cord of the rat.

Cellular localization of dopamine D(1)-like receptors was accomplished on target-specified sympathoadrenal preganglionic neurones using the radioligand [(3)H]SCH23390. Sympathoadrenal neurones were retrogradely labelled with cholera B subunit conjugated to horseradish peroxidase and were detected in segments T(1) to T(13) with a predominance at T(8)/T(9). Binding of the selective D(1)-like radioligand [(3)H]SCH23390 was associated with the retrogradely labelled sympathoadrenal neurones in longitudinal/horizontal sections of thoracic spinal cord. D(1)-like receptor localization on target-specific neurones was determined in more than half of the spinal cord sections and was associated predominantly with the cell soma and principal proximal dendrites in the intermediolateral cell column of the spinal grey matter. D(2)-like receptor localization was not associated with retrogradely labelled sympathoadrenal neurones but a higher degree of specific binding was noted in more medial aspects of the spinal grey matter. This is the first successful demonstration of receptor localization combining two quite different techniques and provides conclusive anatomical evidence for D(1)-like receptor localization on sympathetic preganglionic neurones that project to the adrenal medulla.

Fast excitatory post synaptic potentials and their response to catecholaminergic antagonists in rat sympathetic preganglionic neurones in vitro.

In an in vitro slice preparation from neonatal rats intracellular recordings were made from electrophysiologically identified sympathetic preganglionic neurones. Electrical stimulation in the lateral funiculus (>500 microm) from the recording site elicited a mono- or polysynaptic excitatory post synaptic potential. The latter potential was blocked with the dopamine D2 antagonist haloperidol but not with the dopamine D1 antagonist SCH 23390. We therefore report the first showing of a functional descending pathway in an in vitro slice preparation describing both the transmitter and the receptor subtype involved and physiologically show that dopamine may exert an indirect excitatory influence on sympathetic preganglionic neurones possibly via interneurones present in the spinal cord.

Inhibitory and indirect excitatory effects of dopamine on sympathetic preganglionic neurones in the neonatal rat spinal cord in vitro.

Regions of the thoraco-lumbar spinal cord containing sympathetic preganglionic neurones are rich in dopamine terminals. To determine the influence of this innervation intracellular recordings were made from antidromically identified sympathetic preganglionic neurones in (400 micrometers) transverse neonatal rat spinal cord slices. Dopamine applied by superfusion caused a slow monophasic hyperpolarisation in 46% of sympathetic preganglionic neurones, a slow monophasic depolarisation in 28% of sympathetic preganglionic neurones and a biphasic effect consisting of a slow depolarisation followed by a slow hyperpolarisation or vice-versa in 23% of sympathetic preganglionic neurones. Three percent of sympathetic preganglionic neurones did not respond to the application of dopamine. Low Ca2+/high Mg2+ Krebs solution or TTX did not change the resting membrane potential but abolished the slow depolarisation elicited by dopamine, indicating this was synaptic and did not prevent the dopamine induced hyperpolarisation. The dopamine induced slow hyperpolarisation was mimicked by the selective D1 agonists SKF 38393 or SKF 81297-C and blocked by superfusion with the D1 antagonist SCH 23390. It was not prevented by superfusion of the slices with alpha1 or alpha2 or beta-adrenoceptor antagonists, whereas the inhibitory or excitatory actions of adrenaline were prevented by alpha1 or alpha2 antagonists, respectively. The dopamine induced slow depolarisation occurring in a sub-population of sympathetic preganglionic neurones was mimicked by quinpirole, a D2 agonist, and blocked by haloperidol, a D2 antagonist. Haloperidol did not block the dopamine induced hyperpolarisations. Dopamine also induced fast synaptic activity which was mimicked by a D2 agonist and blocked by haloperidol. D1 agonists did not elicit fast synaptic activity.