The Effects of Ionotropic GABA Receptor Blockage on Brain in Sepsis-induced Rats

Encephalopathy develops following sepsis, which is defined as life-threatening organ failure due to the irregular response of a host to infection. It has high mortality and morbidity rates. In this study, we aimed to investigate the effects of inflammation on brain tissue, and the effects of the GABAA receptor antagonist bicuculline in rats with sepsis. Sepsis was experimentally generated in rats using LPS. The rats were divided into four groups as control, LPS (10 mg/kg i.p.), bicuculline (1.5 mg/kg bicuculline methiodide s.c.), and LPS + bicuculline. Electrophysiologic recordings and body temperature measurements were completed at the 24th hour after injection, and blood samples were taken from the heart for measurements of biochemical parameters. TNF-α, IL-10, and GABA levels were measured using ELISA, and MDA levels were measured using the Bouge method from brain tissue. Tissue imaging was performed with S100-ß, NEUN, and synaptophysin antibody using immunofluorescence staining. One-way ANOVA and the Tukey test were used in statistical analysis. Inflammatory parameters increased in brain tissue in the LPS group compared with the control group. The immunofluorescence staining results in brain tissues were as follows: S-100ß involvement increased, and NeuN and synaptophysin involvement decreased in the LPS group. In electrophysiologic recordings, activity consistent with acute non-focal seizures was observed in the LPS group; however, it was consistent with the resting status in other groups. We suggest that the GABAA antagonist bicuculline methiodide may be a prophylactic agent in sepsis, which caused the impaired neurotransmitter balance, increased pro-inflammatory cytokine and lipid peroxidation, and decreased anti-inflammatory cytokine levels.

Ontogenetic Development of Sensitivity of the Cerebral Cortex to an Antagonist of GABAA Receptor Bicuculline

Local application of four concentrations of bicuculline methiodide (a specific antagonist of GABAA receptors) was used to study a sensitivity of somatosensory cortex in four age groups of immature rats with implanted electrodes. Presence and latencies of two epileptic phenomena (focal discharges and seizures) were evaluated. Focal discharges exhibited moderate tendency to a decrease of sensitivity to bicuculline methiodide with maturation. Concentration-effect relation of incidence of focal discharges was observed only in 7- and 12-day-old but not in older animals. Results with incidence and latencies of seizures did not show relations to age or concentration of bicuculline. Neither of the epileptic phenomena can be used as a reliable index of cortical maturation.

An Antagonist of GABA-B Receptors Potentiates Activity of Cortical Epileptic Foci

Cortical epileptic foci elicited by local application of bicuculline methiodide represent a model of interictal epileptic activity with a transition into ictal phases. We studied a role of GABA-B receptors in this model using GABA-B receptor antagonist CGP35348 in adult rats with implanted cortical electrodes and cannula. CGP35348 (100 or 200 mg/kg i.p.) did not affect interictal discharges but it augmented ictal activity. Latency to the first ictal episode was decreased by the lower dose of CGP35348, duration of episodes was increased by the higher dose. GABA-B receptor antagonist did not influence purely cortical epileptic phenomenon but it is proconvulsant in ictal activity generated with participation of subcortical structures.

Bilateral Inhibition of γ-Aminobutyric Acid Type A Receptor Function within the Basolateral Amygdala Blocked Propofol-induced Amnesia and Activity-regulated Cytoskeletal Protein Expression Inhibition in the Hippocampus

Background It has been reported that bilateral lesions of the basolateral amygdala complex (BLA) blocked propofol-induced amnesia of inhibitory avoidance (IA) training. Based on these results, the authors hypothesized that the amnesia effect of propofol was partly due to its impairment of memory formation in the hippocampus through activating the BLA gamma-aminobutyric acid type A receptor function. The authors determined the changes in activity-regulated cytoskeleton-associated protein (Arc) expression to be an indicator of IA memory formation. Methods Male Sprague-Dawley rats received bilateral injection of bicuculline methiodide (10, 50, or 100 pmol/0.5 microl) or saline (0.5 microl) into the BLA. Fifteen minutes later, the rats were intraperitoneally injected with either propofol (25 mg/kg) or saline. After 5 min, the one-trial IA training was conducted. Rats intraperitoneally infused with saline served as controls and only received saline injections into the BLA. Twenty-four hours later, the IA retention latency was tested. Separate groups of rats treated the same way were killed either 30 min after IA training for hippocampal Arc mRNA measurement or after 45 min for protein level quantification. Results The largest dose of bicuculline methiodide (100 pmol) not only blocked the propofol-induced amnesia but also reversed the inhibition effect of propofol on Arc protein expression in the hippocampus (P < 0.05). However, the mRNA level of Arc showed no significant changes after propofol and bicuculline methiodide administration. Conclusions The amnesic effect of propofol seems to involve the modulation of Arc protein expression in the hippocampus, occurring through a network interaction with the BLA.

Inhibition of Trigeminal Respiratory Activity by Suckling

The trigeminal motor system is involved in many rhythmic oral-motor behaviors, such as suckling, mastication, swallowing, and breathing. Despite the obvious importance of functional coordination among these rhythmic activities, the system is not well-understood. In the present study, we examined the hypothesis that an interaction between suckling and breathing exists in the brainstem, by studying the respiratory activity in trigeminal motoneurons (TMNs) during fictive suckling using a neonatal rat in vitro brainstem preparation. The results showed that fictive suckling, which was neurochemically induced by bath application of N-methyl-D,L-aspartate and bicuculline-methiodide, or by local micro-injection of the same drugs to the trigeminal motor nucleus, inhibited the inspiratory activities in both respiration TMNs and respiratory rhythm-generating neurons. Under patch-clamp recording, fictive suckling caused membrane potential hyperpolarization of respiration TMNs. We conclude that the brainstem preparation contains an inhibitory circuit for respiratory activity in the trigeminal motor system via the rhythm-generating network for suckling. Abbreviations: BIC, bicuculline methiodide; GABA, gamma aminobutyric acid; NMA, N-methyl-D,L-aspartate; NMDA, N-methyl-D-aspartate; and TMN, trigeminal motoneuron.

Effects of Bicuculline Methiodide on Fast (>200 Hz) Electrical Oscillations in Rat Somatosensory Cortex

Fast oscillatory activity (more than ∼200 Hz) has been attracting increasing attention regarding its possible role in both normal brain function and epileptogenesis. Yet, its underlying cellular mechanism remains poorly understood. Our prior investigation of the phenomenon in rat somatosensory cortex indicated that fast oscillations result from repetitive synaptic activation of cortical pyramidal cells originating from GABAergic interneurons ( Jones et al. 2000 ). To test this hypothesis, the effects of topical application of the γ-aminobutyric acid-A (GABAA) antagonist bicuculline methiodide (BMI) on fast oscillations were examined. At subconvulsive concentrations (∼10 μM), BMI application resulted in a pronounced enhancement of fast activity, in some trials doubling the number of oscillatory cycles evoked by whisker stimulation. The amplitude and frequency of fast activity were not affected by BMI in a statistically significant fashion. At higher concentrations, BMI application resulted in the emergence of recurring spontaneous slow-wave discharges resembling interictal spikes (IIS) and the eventual onset of seizure. High-pass filtering of the IIS revealed that a burst of fast oscillations accompanied the spontaneous discharge. This activity was present in both the pre- and the postictal regimes, in which its morphology and spatial distribution were largely indistinguishable. These data indicate that fast cortical oscillations do not reflect GABAergic postsynaptic currents. An alternate account consistent with results observed to date is that this activity may instead arise from population spiking in pyramidal cells, possibly mediated by electrotonic coupling in a manner analogous to that underlying 200-Hz ripple in the hippocampus. Additionally, fast oscillations occur within spontaneous epileptiform discharges. However, at least under the present experimental conditions, they do not appear to be a reliable predictor of seizure onset nor an indicator of the seizure focus.