alpha-Sialylcholesterol enhances the depolarization-induced release of acetylcholine and glutamate in rat hippocampus: in vivo microdialysis study.

The effects of alpha-sialylcholesterol (alpha-SC), a synthetic ganglioside analogue, on synaptic neurotransmission were studied using in vivo microdialysis technique. Application of alpha-SC through a microdialysis probe enhanced high potassium-evoked release of acetylcholine and glutamate in the hippocampal CA3 region of Wistar rats. The experiments using synaptosomes and FM1-43, a fluorescent styryl dye used for studies of neurotransmitter release mechanisms, showed that alpha-SC increased depolarization-induced loss of dye but it did not evoke the dye loss at resting condition. These results indicate that alpha-SC promotes a depolarization-induced exocytotic neurotransmitter release in the brain under in vivo conditions. Application of alpha-SC increased the level of glutamate but not that of acetylcholine, suggesting that alpha-SC affects spontaneous glutamate release and/or transport system at the brain region.

Perfusate oxygen and carbon dioxide concentration influence basal microdialysate levels of striatal glucose and lactate in conscious rats.

O(2) concentration ([O(2)]) in air equilibrated solutions at room temperature is three fold higher than that in brain extracellular fluid (ECF), and CO(2) concentration ([CO(2)]) is 100 times lower. Using microdialysis the ECF is routinely dialyzed against glucose free isotonic perfusates containing 200 microM O(2) and 10 microM CO(2). In conscious rats, 2 days after probe implantation, decreasing perfusate [O(2)] from 200 to 68 microM (physiologic level) for 60 min, while maintaining a low [CO(2)] (10 microM), increased striatal dialysate glucose and lactate by 12% and 33%, respectively. The same protocol on the third day essentially had no effect on monoamine metabolites. Decreasing [O(2)] concurrent with increasing [CO(2)] to 1.3 mM (physiologic level) for 60 min increased glucose and lactate by 17% and 37%, respectively. This study demonstrates for dialysis studies of glucose and lactate, perfusates that mimic physiologic ECF [O(2)] and [CO(2)] are more appropriate.

Determination of carbohydrates as their 3-aminophthalhydrazide derivatives by capillary zone electrophoresis with on-line chemiluminescence detection.

A method based on pre-capillary derivatization with luminol (3-aminophthalhydrazide) for carbohydrate analysis using capillary electrophoresis with on-line chemiluminescence (CL) detection was developed. The derivatives of seven monosaccharides were separated and detected by using 200 mM borate buffer containing 100 mM hydrogen peroxide at pH 10.0 as separation electrolyte and 25 mM hexacyanoferrate in 3 M sodium hydroxide solution as post-capillary chemiluminescence reagent with separation efficiencies ranging from 160,000 to 231,000 plates per metre. The minimum amount of carbohydrate derivatized was 2 pmol (corresponding to the concentration of 2 microM). The method also provided a linear response for glucose in the concentration range of 0.1-250 microM with a mass detection limit of 420 amol or a concentration detection limit of 0.1 microM. Preliminary work using the CE-CL format to determine glucose in a rat brain microdialysis sample is presented as a typical case.

Analysis of glucose and lactate in hippocampal dialysates of rats during the operant conditioned reflex using microdialysis.

Changes of extracellular glucose and lactate in hippocampus for freely moving rats during the operant conditioned reflex were examined simultaneously. Samples of the dialysate were assayed for both glucose and lactate using in vivo microdialysis and a microbore flow injection analysis-immobilized enzyme reactor-electrochemical detection (FIA-IMER-ECD) system. Microdialysis samplings were conducted in a Skinner box where lights were delivered as conditioned stimuli (CS) paired with foot shocks as unconditioned stimuli (US). In the treatment group the concentration of glucose and lactate showed no fluctuations during the whole process. However, in the control group in which the rats were exposed to many foot shocks, lactate levels decreased by 19% below baseline during the behavioral session and glucose showed a delayed decrease (by 18%). Compared with glucose, lactate can immediately indicate the dynamic changes in brain.

Hyperoxia caused by microdialysis perfusion decreased striatal monoamines: involvement of oxidative stress.

Due to complex influence, such as utilization and permeability of arterial vessels to oxygen, there is a considerable difference of oxygen tension between extracellular fluid and perfusate usually used in microdialysis (30-60 Torr versus 145 Torr). Dialysate dopamine and monoamine metabolites-3,4-dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindoleacetic acid were measured under different kinds of oxygen tension solutions (145, 72, 48 Torr). In the acute and anesthetized group, dopamine, 3,4-dihydroxyphenylacetic acid, 5-hydroxyindoleacetic acid and homovanillic acid increased 72, 93, 86 and 65%, respectively when changing the perfusate from 145 Torr to near physiological 48 Torr, while in chronic and conscious group, carried out 72 h after surgery, these compounds showed obscure increases (only homovanillic acid produced a significant change of 14%). The different effect of perfusate oxygen tension on dialysate levels of monoamines in anesthetized and conscious rats might be caused by oxidative stress triggered by hyperoxia combined with anesthesia and surgical trauma.

Effects of salicylate on serotoninergic activities in rat inferior colliculus and auditory cortex.

In vivo microdialysis offers a unique approach to monitor biochemical events related to brain function and metabolism, and has been used extensively in many systems to measure the release of endogenous transmitters and other neuroactive substances during normal and pathological conditions. The characterization of neurotransmitters' changes induced by salicylate in the inferior colliculus (IC) and the auditory cortex (AC) may provide insight into the action of salicylate on the auditory system and, through this, provide a better understanding of neurological mechanism of salicylate-induced tinnitus. In the present study, the effect of salicylate on 5-HT system in IC and AC has been monitored by microdialysis in salicylate-induced tinnitus animal models. Glucose and lactate levels in IC and AC were significantly increased after application of salicylate (350 mg/kg, i.p.), indicating a salicylate-related increase in regional neuronal activity. The 5-HT level increased to a maximum of 268+/-27% basal level in IC 2 h after application and of 277+/-24% basal level in AC around 3 h after application. These data suggest that the increases of 5-HT levels in IC and AC may be involved in the tinnitus generation.

[Study of neurological mechanism of lidocaine's suppression to tinnitus via microdialysis].

OBJECTIVE: To elucidate the neurological mechanism of lidocaine's suppression to tinnitus. METHODS: Thirty-four Wistar rats weighing 300-350 grams were randomly divided into IC group (n = 17) and AC group (n = 17), according to microdialysis region. Each group was randomly subdivided into saline treatment group (n = 4), salicylate treatment group (n = 6), and salicylate + lidocaine treatment group (n = 7). Using in vivo microdialysis technique coupled with microbore HPLC-electrochemical detection, the present study first monitored the 5-HT release in IC and AC in salicylate-induced tinnitus animal models, and then, examined the effects of lidocaine on salicylate-induced 5-HT changes in IC and AC. The statistical analysis was performed using two-way ANOVA for repeated measures of raw data with time and treatment condition as main effects. Individual time-point values between no more than two groups were compared with the unpaired Student's t-test. The accepted level of significance was 0.05, two-tailed. RESULTS: The 5-HT level increased to a maximum of 268% +/- 27% (mean +/- s) basal level in IC 2 h after salicylate application and of 277% +/- 24% basal level in AC around 3 h after application. And then, the 5-HT level gradually decreased to 157% +/- 16% of baseline in IC and 180% +/- 18% of baseline in AC by the end of the sixth hour. Saline did not alter the IC and AC dialysate 5-HT level in control rats. Two-way ANOVA with repeated measures indicated a significant effect of the condition factor [F (1, 8) = 413.949, P < 0.000001 in IC group; F(1,8) = 192.184, P < 0.000001 in AC group]. The increases of 5-HT levels in salicylate treatment groups were significantly reduced to 85% +/- 8% basal level in IC and 92% +/- 26% basal level in AC after local infusion of 1% lidocaine (P < 0.05). Compared with corresponding control value at that time (unpaired student t-test). Two-way ANOVA with repeated measures showed a significant difference between the salicylate group and salicylate + lidocaine group [P < 0.000001 with F(1, 11) = 329.267 for the condition factor in IC subgroup; P < 0.000001 with F(1, 11) = 133.844 for the condition factor in AC subgroup]. CONCLUSION: The suppression of lidocaine to tinnitus may be associated with the decrease of 5-HT level in IC and AC.