Interaction between glutamate and GABA on H-noradrenaline release from rat hypothalamus

Glutamate has been shown to stimulate noradrenaline (NA) release from hypothalamic nerve terminals. In the present study, we evaluated the possible interaction between the excitatory amino acid glutamate and gamma-aminobutyric acid (GABA), an inhibitory transmitter, on noradrenaline (NA) release from mediobasal hypothalamus (MBH) of adult male rats. Hypothalamic slices loaded in vitro with ³H-NA were superfused and exposed to glutamate, N-methyl- -aspartic acid (NMDA), or kainate (KA). We found that ³H-NA release evoked by the excitatory amino acids glutamate and NMDA was dramatically decreased by GABA. The facilitatory effects of NMDA and KA were prevented concentration-dependently by the GABA_B receptor antagonist 2-hydroxy saclofen which restored the NMDA effect. In addition, beclofen blocked K⁺-induced ³H-NA release. Activation of GABA_A receptors by muscimol and THIP was ineffective. In conclusion, glutamate and GABA, through GABA_B receptors, may interact to modulate NA release from the rat mediobasal hypothalamus.     

Mechanism underlying potentiation by progesterone of the kainate-induced current in cultured neurons

The effect of the neurosteroid progesterone on the kainate receptor-mediated response was studied in cultured chick spinal cord neurons using the whole-cell voltage-clamp recording technique. Progesterone rapidly and reversibly potentiates the kainate-induced current in a dose-dependent manner, with an EC₅₀ of 35 μM and a maximal potentiation of 30%. Potentiation of the kainate response by extracellularly applied progesterone is not significantly affected by inclusion of a saturating concentration of progesterone in the electrode buffer, indicating that progesterone acts at the extracellular surface of the membrane. Furthermore, progesterone enhances the kainate maximal response with little effect on the kainate EC₅₀.     

Evidence for increased dorsal hippocampal adenosine release and metabolism during pharmacologically induced seizures in rats

There is growing pharmacological evidence from several animal models of seizure disorder that adenosine possesses endogenous anticonvulsant activity. In order to further evaluate the role of adenosine in seizure activity, we monitored adenosine and its major biochemical metabolites inosine, xanthine, and hypoxanthine in the dorsal hippocampus by in vivo microdialysis before and during the induction of generalized seizures. Seizures were induced pharmacologically in groups of urethane-anesthetized rats by the administration of bicuculline (0.5 mg/kg, i.v.), kainic acid (12.0 mg/kg, i.v.) or pentylenetetrazol (100–250 mg/kg, i.p). Seizure activity was monitored electrophysiologically from the dorsal hippocampus. Dialysate hippocampal purine levels increased during all three seizure types. The largest increases were for the adenosine metabolites hypoxanthine and inosine, with smaller increases observed for adenosine and xanthine. Intra-hippocampal perfusion with the adenosine deaminase inhibitor erythro-9-(2-hydroxy-3-nonyl-adenine, (EHNA, 300 μM), only slightly increased basal hippocampal adenosine. Guanosine levels in the hippocampus, a purine not directly related to adenosine metabolism, were unaffected by all treatments. These findings demonstrate that an increase in hippocampal adenosine release and metabolism is associated with seizure activity and support the hypothesis that the increased adenosine levels may attenuate hippocampal seizure activity, possibly by terminating ongoing seizures and altering the pattern of subsequent seizures.     

The locus coeruleus noradrenergic system in the rat brain studied by dual-probe microdialysis

A dual-probe microdialysis technique was applied to the locus coeruleus (LC) and prefrontal cortex (PFC) of the brain of conscious rats. One probe was implanted close to the LC and was used to apply receptor-specific compounds by retrograde microdialysis. The effects of the LC infusions were recorded by a sampling noradrenaline by a second probe that was implanted in the ipsilateral prefrontal cortex. Infusion of sodium channel blocker tetrodotoxin (1 μM: 90 min) into the LC decreased extracellular noradrenaline in the PFC to ≈ 20% of control values. Infusion of α₂-adrenoceptor agonist clonidine (100 μM, infused during 15 or 45 min) near to the LC, decreased extracellular noradrenaline in the PFC to 35 and 20% of controls, respectively. These results indicate that > 80% of the extracellular levels of noradrenaline in the PFC is derived from LC intervation, and confirms the importance of α₂-autoreceptors on noradrenergic neurons in the LC. Infusion of the cholinergic receptor agonist, carbachol (100 μM, 45 min) near to the LC increased extracellular noradrenaline in the PFC to ≈ 150% of controls. Infusions of the excitatory amino-acid agonists NMDA and kainate into the LC caused marked increases in extracellular noradrenaline in the PFC to 240 and 200% of controls, respectively. The experiments with clonidine, carbachol, NMDA and kainate were repeated in anesthetized rats. Clonidine and carbachol were similarly effective as in conscious animals but the effects of NMDA and kainate on extracellular noradrenaline in the PFC were clearly suppressed: 145 and 130% of controls, respectively. These results suggest that increased arousal or behavioural activation might have contributed to the increases in extracellular noradrenaline that was seen after infusion of the glutamate agonists. These results also provide evidence for localization of cholinergic-, NMDA-, non-NMDA-receptor on noradrenergic neurons in the LC. Finally it is concluded that dual-probe microdialysis is a useful method to further investigate the pharmacology of LC-noradrenergic neurons. Carbachol and clonidine are suitable tools for a rapid and reversible stimulation or inhibition, respectively, of noradrenergic LC neurons.     

Immunohistochemical evidence for flupirtine acting as an antagonist on theN-methyl-d-aspartate and homocysteic acid-induced release of GABA in the rabbit retina

When rabbit retinas are exposed in vitro to specific excitatory amino acid receptor agonists certain GABAergic amacrine cells are activated to cause a release of GABA. The GABA that is not released can be detected by immunohistochemistry. Exposure of tissues to kainate or NMDA each caused a characteristic change in the GABA immunoreactivity. CNQX antagonised the kainate effect specifically while MK-801 counteracted the influence of NMDA The effect produced by kainate was mimicked by domoic acid while the influence of homocysteic acid was identical with NMDA. Flupirtine alone did not influence the nature of the GABA immunoreactivity and so did not act as a kainate or NMDA agonist. However, flupirtine counteracted the influence produced by NMDA and homocysteic acid but had no effect on the kainate and domoic acid responses. Thus in this system flupirtine acts as an NMDA antagonist.     

Sex-dependent body weight changes after iontophoretic application of kainic acid into the LH or VMH

Body weight changes and food and water intakes were studied in CFY male and female rats after kainic acid (KA)-induced destruction of the lateral hypothalamic area (LH) or the ventromedial hypothalamic nucleus (VMH). To minimize the extent of damages, KA was iontophoretically applied by means of glass micropipettes. KA was ejected in 50 or 80 mM concentrations with 5-15 microA current for 5 min. Tip diameter of pipettes varied between 10-20 microns. Lesions were restricted to the LH or VMH. Effects were sex-dependent. LH lesions resulted in hypophagia, hypodipsia and body weight loss only in male rats. On the other hand, only female animals exhibited hyperphagia and weight increase when the VMH was destroyed. The role of sex-dependence in hypothalamic body weight regulation is discussed.     

Dynamic alterations in hippocampal morphology following intra-ventricular kainic acid

Several primary and secondary reactions were observed in rat hippocampus at various times following low doses of intraventricular kainic acid. The well documented fulminating lesion of subfield CA3a was evident, however, with the parameters of the present study, the acute neuronal death was restricted to areas which have been shown by others to have an exceptionally dense mossy fiber input. Neurons in subfields CA3c and CA4 also die, however, this was apparent only after a survival time of 2 weeks; prior to this, these cells went through a period of chromatolysis and dendritic swelling. In remaining subfields, principally 'CA2' and CA1, a third type of change was observed at longer intervals of 1 month. This was characterized by neuronal shrinkage and chromophilia and eventual cell loss. The present study provides additional observations on the actions of kainic acid in the hippocampus and may be relevant to the dynamic pathology of temporal lobe epilepsy.