Wynn H.T. Pan

Regulatory Effects of D2 Receptors in the Ventral Tegmental Area on the Mesocorticolimbic Dopaminergic Pathway

Abstract: To investigate the regulatory effects of somatodendritic D2 receptors on the terminal’s extracellular dopamine (DA) concentration, a D2 antagonist (eticlopride) was infused directly into the ventral tegmental area via a microdialysis probe in chloral hydrate‐anesthetized rats. Extracellular DA changes in both the nucleus accumbens (N ACC) and the medial prefrontal cortex (mPFC) were monitored. Infusion of 10.0 fM eticlopride had no effect on DA in the mPFC (110.2 ± 10.0% of baseline) but significantly increased DA in the N ACC (150.1 ± 11.7%). Infusion of a higher dose of eticlopride (100.0 or 1,000.0 fM) significantly augmented the DA in the mPFC (121.1 ± 7.6 and 180.7 ± 25.8%, respectively) but surprisingly had no effect on DA in the N ACC (111.5 ± 7.3 and 104.1 ± 8.7%, respectively). To further investigate whether the bluntness of DA increase in the N ACC was due to DA receptor activation in the mPFC, eticlopride or SCH23390 was infused into the mPFC prior to and during intrategmental eticlopride infusion, and the change of DA in the N ACC was simultaneously monitored. During intra‐mPFC 1.0 nM eticlopride infusion but not during 10.0 nM SCH23390 administration (95.5 ± 6.1%), intrategmental 1,000.0 fM eticlopride infusion could further elevate DA in the N ACC (130.0 ± 4.6%). Our results indicated that (1) the mesolimbic and the mesocortical pathways were under tonic inhibition by somatodendritic D2 receptors; (2) the DA concentration in the N ACC first increased and then returned to baseline while the intrategmental infusion dose of eticlopride increased; and (3) the bluntness of DA increase in the N ACC resulted from the D2 receptor activation in the mPFC.

Striatal glutamate release is important for development of ischemic damage to striatal neurons during rat heatstroke

This study attempted to ascertain whether heatstroke-induced ischemia is associated with augmented striatal glutamate release and can be attenuated by NMDA receptor antagonists. Mean arterial pressure (MAP), striatal cerebral blood flow (CBF), striatal glutamate release and striatal neuronal damage score were assessed in saline-treated rats and in rats treated with NMDA receptor antagonists. Heatstroke was induced by exposing the animals to a high ambient temperature; the moment at which MAP and CBF began to decrease from their peak levels was taken as the onset of heatstroke. During onset of heatstroke, rats displayed higher values of colonic temperature, striatal glutamate release and striatal neuronal damage score, and lower values of MAP and striatal blood flow compared with normothermic control rats. The decreased MAP, the diminished striatal blood flow, the augmented striatal glutamate release and the increased striatal neuronal damage score during onset of heatstroke were significantly attenuated by pretreatment with an NMDA receptor antagonist such as MK-801 or ketamine. In addition, the survival time (interval between onset of heatstroke and death) of the rats was extended by pretreatment with one of these two NMDA receptor antagonists. These results suggest that marked accumulation of glutamate in the striatum is important for the development of ischemic damage to striatal neurons during the onset of heatstroke.

NF-kappaB involvement in the induction of high affinity CAT-2 in lipopolysaccharide-stimulated rat lungs

Background:  Endotoxemia stimulates nitric oxide (NO) biosynthesis through induction of inducible NO synthase (iNOS). Cellular uptake of l‐arginine, the sole substrate for iNOS, is an important mechanism regulating NO biosynthesis by iNOS. The isozymes of type‐2 cationic amino acid transporters, including CAT‐2, CAT‐2A, and CAT‐2B, constitute the most important pathways responsible for trans‐membrane l‐arginine transportation. Therefore, regulation of CAT‐2 isozymes expression may constitute one of the downstream regulatory pathways that control iNOS activity. We investigated the time course of enzyme induction and the role of nuclear factor‐κB (NF‐κB) in CAT‐2 isozymes expression in lipopolysaccharide‐(LPS) treated rat lungs.

Differential Effects of Chloral Hydrate and Pentobarbital Sodium on a Cocaine Level and Its Catecholamine Response in the Medial Prefrontal Cortex: A Comparison with Conscious Rats

Abstract: Adult male Sprague‐Dawley rats anesthetized with chloral hydrate and pentobarbital sodium were used as two different treatment groups. Conscious rats were used as a control group. By using baseline (precocaine) concentration as 100%, after cocaine administration (3.0 mg/kg i.v.), the maximal dopamine (DA) increase occurring at the first microdialysis collection period (20 min) in the medial prefrontal cortex was 299 ± 46% for the chloral hydrate group, 168 ± 12% for the pentobarbital sodium group, and 325 ± 23% for the conscious group. At the same time, norepinephrine (NA) increases reached a maximum and were 162 ± 20%, 100 ± 5%, and 141 ± 17%, respectively. The maximal changes of DA and NA in the chloral hydrate group and in the control group were both significantly higher than that in the pentobarbital sodium group. Meanwhile, the cocaine concentration was higher over a 100‐min period of time in the chloral hydrate group when compared with the pentobarbital group and the control group. The peak cocaine concentration in dialysate occurred in the same time slot of maximal DA and NA responses, which were 0.65 ± 0.08, 0.30 ± 0.02, and 0.41 ± 0.05 µM, respectively. Anesthetics suppress the pharmacologic response of neurons, which may explain the difference in catecholamine response between the pentobarbital sodium and the conscious groups. Conversely, because there was no significant difference in DA and NA response between the chloral hydrate group and the conscious group, it may possibly be due to the balancing effect between the higher existing cocaine concentration and the anesthetic suppression on pharmacological response of neurons in the chloral hydrate group. The effect of guide cannula implantation on the cocaine‐induced catecholamine response was also evaluated.

Power spectral analysis of the electroencephalographic and hemodynamic correlates of propofol anesthesia in the rat: Intravenous bolus administration

Based on the tail-flick response to noxious thermal stimuli, we determined in the present study that the minimal effective antinociceptive dose of propofol in adult, male Sprague-Dawley rats, given in an intravenous bolus manner, was 10 mg/kg. Simultaneous power spectral analysis of the electroencephalographic (EEG) and systemic arterial pressure signals further revealed a concomitant transient suppression of the EEG activity, primarily in the theta and sigma bands, alongside minor hypotensive and negative inotropic and chronotropic actions, but with maintained vasomotor tone. These alterations followed a time course that paralleled the plasma concentration of propofol in the arterial blood, as detected by high-performance liquid chromatography.

Power spectral analysis of the electroencephalographic and hemodynamic correlates of propofol anesthesia in the rat: Intravenous infusion

Based on the tail-flick response to noxious thermal stimuli, we determined in the present study that effective antinociception could be achieved in adult male Sprague-Dawley rats 15 min after intravenous infusion of propofol at 60 mg/kg/h. Simultaneous power spectral analysis of the electroencephalographic (EEG) and systemic arterial pressure signals further revealed a concomitant depression of the activity of all EEG frequency bands (delta, theta, alpha, beta), alongside hypotension, negative inotropic and chronotropic actions, and attenuated baroreceptor reflex and vasomotor activity. These effects were congruent with a plasma concentration of propofol in the arterial blood of 1.70 +/- 0.13 micrograms/ml, as determined by high-performance liquid chromatography.

Preparative supercritical fluid extraction of pyrethrin I and II from pyrethrum flower

A preparative supercritical fluid extraction system is described and was used with supercritical carbon dioxide to extract the active insecticide components pyrethrin I (PI) and pyrethrin II (PII) successfully from pyrethrum flower. A high-performance liquid chromatography method was developed and was used to separate and analyze the supercritical carbon dioxide extracts. Extraction efficiencies under several different extraction conditions were examined. Under the conditions examined, the most effective extractions of PI and PII (140 +/- 18 mg and 55 +/- 9 mg per 100 g of dry pyrethrum flower powder) were performed at 40 degrees C and 1200 psi. The results showed that extraction efficiencies of supercritical carbon dioxide are much better than those of n-hexane for pyrethrins I and II. During the extraction process, the most efficient extraction period was the first 3 h of the experiment.

Prefrontal dopamine efflux during exposure to drug-associated contextual cues in rats with prior repeated methamphetamine

Conditioned stimulus-reward response and prefrontal dopamine efflux under context previously paired with methamphetamine administration were assessed in rats with or without prior sensitizing regimen. Sensitizing pretreatment was administered with methamphetamine (1mg/kg, every other day for six sessions) for behavioral sensitization. The animals received methamphetamine (1mg/kg) or saline injection (each for six sessions) to pair with distinct contexts on alternate days to induce conditioned place preference. Then, dopamine outflows in the medial prefrontal cortex were analyzed on the next day via microdialysis study as animals exposed to the methamphetamine or saline-paired context, respectively. Prefrontal DA efflux increased in those rats without sensitizing pretreatment, while they occupied the methamphetamine-paired chamber. The rats with prior sensitizing regimen demonstrated more robust conditioned place preference than those without pretreatment, however, their dopamine efflux was attenuated, while remaining in methamphetamine-paired context. It is suggested that the attenuated responsiveness of mesocortical dopamine transmission in prior sensitized rats may, at least in part, be responsible for their augmented conditioned place preference, which resulted from activation of related brain areas that together strengthen the associative learning to drug-related stimuli. This paradigm may reflect a dysregulated prefrontal function in the methamphetamine abusers.

Determination of free-form of cocaine in rat brain by liquid chromatography–electrospray mass spectrometry with in vivo microdialysis

A rapid liquid chromatography-electrospray mass spectrometry (LC-ES-MS) method with in vivo microdialysis for the determination of free-form of cocaine (COC) in rat brain has been developed. A C18 column and a gradient elution were employed for the separation. The [M+H]+ (m/z=304) and a fragmented ion (m/z=182) were detected using positive ion mode detection. Selective ion monitoring was utilized for quantitative measurement. The linearity of this assay was good ranging from 0.01 to 1.0 microM (r2=0.999). The inter- and intra-day precisions showed relative standard deviations ranging from 1.0% to 3.3% and 1.0% to 3.6%, respectively. In addition, the detection of one COC metabolite, benzoylecgonine (BE), by this assay was also investigated. The linearity, precision, and detection limit associated with this method for BE were determined. The application of this newly developed method was demonstrated by examining the pharmacokinetics of COC in rat brain.

Effects of ascorbate in microdialysis perfusion medium on the extracellular basal concentration of glutamate in rat's striatum

There are many evidences suggest that ascorbate in the extracellular space can affect glutamate concentration in the rats brain. In this report, we studied how ascorbate in microdialysis perfusion medium affected glutamate level at the striatum in freely-moving rats. Three perfusion mediums were used: 0, 250, and 400 microM of ascorbate in perfusion medium. The extracellular basal concentrations of glutamate were determined to be 1.29+/-0.52 microM for the no ascorbate group, 0.86+/-0.35 microM for the low ascorbate group and 4.76+/-1.48 microM for the high ascorbate group. By using 400 microM of ascorbate in a perfusion medium, we found that the extracellular basal concentration of glutamate significantly increased and its in vivo recovery significantly decreased. This indicated that ascorbate concentration in a perfusion medium was important and must be carefully considered while using microdialysis technique to monitor glutamate concentration in vivo.