David K. Pitts

Chronic cocaine reduces α2-adrenoceptor elicited mydriasis and inhibition of locus coeruleus neurons

The effects of chronic cocaine (50 mg/kg per day for two weeks) administration on two alpha 2-adrenoceptor-mediated responses were studied in rats. Chronic administration of cocaine significantly (compared to sham controls) attenuated the alpha 2-adrenoceptor-mediated inhibition of noradrenergic locus coeruleus (LC) neurons as well as alpha 2-adrenoceptor elicited mydriasis. Noradrenergic LC neurons from the cocaine treated and sham sham groups differed significantly in their responsiveness to the inhibitory effects of clonidine (ED50 values micrograms/kg: sham 7.35 +/- 1.13 and cocaine-treated 17.17 +/- 4.40, P less than 0.05). The ED50 values for the mydriatic response were sham 5.71 +/- 0.49 and cocaine-treated 16.42 +/- 0.69 micrograms/kg, respectively, P less than 0.001. No differences in cardiovascular responses to systemically injected clonidine between the chronic cocaine- and sham-treated groups were observed. Chronic cocaine treatment attenuates the two alpha 2-adrenoceptor-mediated responses most likely via an interaction with central catecholaminergic neurotransmission.

Optical bioassay for measuring sublethal toxicity of insecticides in Daphnia pulex

Many emerging contaminants tend to be biologically active at very low concentrations, occur in water as part of complex mixtures, and impact biota in ways that are not detected using traditional toxicity tests (e.g., median lethal concentration). To evaluate emerging contaminants, the authors developed a method for detecting sublethal behavioral effects by quantifying the swimming behavior of Daphnia pulex, a model organism for studying aquatic toxicity. This optical tracking technique is capable of measuring many swimming parameters, 2 of which-cumulative distance and angular change-are presented. To validate this technique, 2 prototypical compounds that exhibit different modes of action as well as corresponding insecticides that are commonly found in surface waters were investigated. The acetylcholinesterase (AChE) inhibitor physostigmine was used as the prototypical compound for the large number of AChE inhibitor insecticides (e.g., chlorpyrifos). Nicotine was used as the prototypical compound for neonicotinoid insecticides (e.g., imidacloprid). Results demonstrate that this assay is capable of detecting sublethal behavioral effects that are concentration-dependent and that insecticides with the same mode of action yield similar results. The method can easily be scaled up to serve as a high-throughput screening tool to detect sublethal toxic effects of a variety of chemicals. This method is likely to aid in enhancing the current understanding of emerging contaminants and to serve as a novel water-quality screening tool.

Repeated amphetamine: reduced dopamine neuronal responsiveness to apomorphine but not quinpirole

Extracellular recordings from single nigrostriatal dopamine (DA) neurons in rats revealed significantly reduced neuronal sensitivity to the inhibitory effects of i.v. apomorphine following repeated amphetamine (4 mg/kg per day i.p., 14 days). This effect was reversed by acute SCH 23390. Quinpirole sensitivity was reduced in amphetamine-treated rats only following acute SKF 38393 pretreatment. These results suggest that, in amphetamine-treated animals, D-1 receptor activation is important for the expression of reduced nigrostriatal DA neuron sensitivity to apomorphine.

Prenatal haloperidol reduces the number of active midbrain dopamine neurons in rat offspring

The dopamine (DA) receptor antagonist, haloperidol (HAL, 1.25 or 5 mg/kg), or vehicle, dimethyl sulfoxide (DMSO), was administered (SC) daily to pregnant Sprague-Dawley dams from gestational day (GD) 8 to GD 20. The average body weight of 2-week-old male offspring was significantly lower in all of the HAL-treated groups relative to controls. In extracellular electrophysiological studies, the male 2-week-old offspring from all HAL treatment groups were found to have significantly reduced average numbers of spontaneously active midbrain dopamine (DA)-containing neurons in both the substantia nigra (A9) and the ventral tegmental area (A10) relative to controls. In DA neurons classified as bursting neurons, HAL exposure (5 mg/kg) caused a significantly increased level of burst activity in A10 but not A9 DA neurons relative to controls. For both the A9 and A10 regions, the proportion of DA neurons classified as bursting or nonbursting was unaffected by HAL treatment. These results suggest that prenatal HAL exposure influences the development of midbrain DA neurons.

Postnatal development of mesoaccumbens dopamine neurons in the rat: electrophysiological studies

The postnatal development of antidromically identified mesoaccumbens dopamine (MADA) neurons were examined with single-unit electrophysiological techniques. Rats were anesthetized with chloral hydrate. The physiological characteristics of 1-, 2-, 4- and 5-week-old rat pups were compared to adults (7-9-weeks-old). The basal discharge rate, conduction velocity, antidromic latency and discharge patterns of MADA neurons were not significantly different among the 4- and 5-week-old and adult MADA neurons. MADA neurons from 1- and 2-week-old pups, however, had significantly lower mean basal discharge rates and significantly lower mean conduction velocities than MADA neurons from the older animals (i.e., 4-weeks old, 5-weeks old and adults). 1- and 2-week-old MADA neurons were also found to have significantly longer mean antidromic latencies than MADA neurons from older animals. Significantly fewer 1- and 2-week-old MADA neurons were found to discharge in a bursting pattern when compared to MADA neurons from older animals. These results indicate that during early postnatal development MADA neurons are spontaneously active, but still physiologically immature. The results of the present study are discussed in the context of previous developmental electrophysiological studies of nigrostriatal dopamine neurons.

Cardiovascular effects of centrally perfused clonidine

Earlier work has indicated that the systemic cardiovascular actions of clonidine might be mediated by caudal brainstem centers, especially the nucleus of the solitary tract (NTS). This study sought to define the mode of clonidine action on the NTS more explicitly using the technique of push-pull perfusion on urethane-anesthetized rats. The NTS of stereotaxically mounted subjects was unilaterally perfused with an artificial cerebrospinal fluid at 25 microliter/min. Clonidine was added to the medium at concentrations of 5 to 500 microM, without interruption of flow, for test periods of 10 min. Systemic drug actions were expressed in terms of mean arterial pressure (MAP) and heart rate (HR), both of which were recorded continuously throughout the experiment. Decreases occurred in both MAP and HR following clonidine perfusion at all concentrations. However, the dose-effect relationship for the blood pressure response was dependent to some extent on control pressure. When this was considered as a variable, the drug-induced pressure effects were significantly dose-dependent. Control HR values were more stable than pressure and dose-related decreases following clonidine administration were highly significant. The clonidine concentrations investigated here were considerably lower than those previously studied by microinjection. The observed dose-related depression of MAP and HR under basal conditions may be related to specific alpha 2-adrenergic receptor activation of the NTS.

Effects of chronic low-level oral lead exposure on prepulse inhibition of acoustic startle in the rat

Previous work has suggested that the behavioral effects of chronic low-level lead exposure on fixed interval (FI) operant behavior result from enhanced dopaminergic neurotransmission in the nucleus accumbens (Cory-Slechta et al., J Pharmacol Exp Ther 286: 794-805, 1998). The present studies were designed to further characterize the effects of chronic low-level oral lead exposure on another behavior that is modulated by dopaminergic neurotransmission in the nucleus accumbens. In these studies acoustic startle and the prepulse inhibition (PPI) of startle were studied in rats following chronic low-level oral lead exposure. Weanling male rats were treated for 5-6 weeks with lead via drinking water (250 ppm lead acetate; controls drank 250 ppm sodium acetate). Acoustic startle reactivity (95, 105, and 115 dB noise bursts) and PPI (prepulses of 1-8 dB over the 70-dB background) of startle were tested following lead exposure. Lead exposure did not affect body weight. Lead exposure also did not significantly affect baseline [i.e., no prepulse inhibition (NO-PPI)] acoustic startle as measured by 1) startle amplitude on the first startle trial (105 dB), 2) the average startle amplitude for the first ten trials (105 dB), or 3) the average startle amplitude for the NO-PPI trials during PPI testing (95, 105, and 115 dB). Lead exposure also did not affect the latency to onset for the startle response. In contrast, for both the 105 dB and 115 dB acoustic startle stimuli, chronic low-level oral lead exposure significantly attenuated the capacity of an acoustic prepulse to reduce the startle response. This effect was present whether the data were presented and analyzed as raw change from baseline or as the percentage of baseline startle. Given the strong link between the modulation of PPI and dopaminergic neurotransmission in the nucleus accumbens, the present data support the hypothesis that chronic low-level oral lead exposure facilitates dopamine neurotransmission in the nucleus accumbens.

Toxic effects of combined stressors on Daphnia pulex: Interactions between diazinon, 4‐nonylphenol, and wastewater effluent

Contaminant exposure in aqueous systems typically involves complex chemical mixtures. Given the large number of compounds present in the environment, it is critical to identify hazardous chemical interactions rapidly. The present study utilized a prototype for a novel high-throughput assay to quantify behavioral changes over time to identify chemical interactions that affect toxicity. The independent and combined effects of 2 chemicals, diazinon (an insecticide) and 4-nonylphenol (a detergent metabolite), on the swimming behavior of the freshwater crustacean Daphnia pulex were examined. Cumulative distance and change in direction were measured repeatedly via optical tracking over 90 min. Exposure to low concentrations of diazinon (0.125-2 µM) or 4-nonylphenol (0.25-4 µM) elicited significant concentration- and time-dependent effects on swimming behavior. Exposure to 0.5 µM 4-nonylphenol alone did not significantly alter mean cumulative distance but did elicit a small, significant increase in mean angle, the measure of change in direction. When 0.5 µM 4-nonylphenol was used in combination with diazinon (0.125-0.5 µM), it augmented the adverse impact of diazinon on the swimming behavior of Daphnia. Additionally, enhanced sensitivity to diazinon was observed in animals exposed to treated wastewater effluent for 24 h prior to a diazinon challenge. The present experiments demonstrate that exposure to 4-nonylphenol and complex chemical mixtures (e.g., treated wastewater) can enhance the toxicity of exposure to the insecticide diazinon.

Repeated SKF 38393 and nigrostriatal system neuronal responsiveness: functional down-regulation is followed by up-regulation after withdrawal

SummaryThe effects of chronic administration of the D1 dopamine (DA) agonist SKF 38393 on the basal activity and electrophysiological and pharmacological responsiveness of nigrostriatal DA neurons were examined by means of extracellular, single-unit recording techniques. Chronic D1 stimulation failed to alter either the basal activity of DA neurons or the potency of quinpirole to induce inhibition of these cells. However, 28-day SKF 38393 treatment (but not 14-day treatment) eliminated the ability of subsequent (24 h later) acute SKF 38393 to alter the rate-dependent nature of quinpirole-induced inhibition. In contrast, one week after a 28-day SKF 38393 treatment we found that quinpirole-induced inhibition by itself was no longer rate-dependent, an effect which was reversed by acute pretreatment with the D1 antagonist SCH 23390. This latter finding is suggestive of enhanced endogenous D1 tone. Similarly, 28-day SKF 38393 treatment eliminated the effect of subsequent acute SKF 38393 on sciatic nerve stimulation-induced inhibition of nigrostriatal DA neurons, whereas one week after the chronic D1 regimen these cells were highly sensitive to acute D1 enhancement of the response to sciatic nerve stimulation.In order to address the postsynaptic effects of chronic D1 stimulation, the influence of iontophoretically administered SKF 38393 was examined on type I caudate neurons. Again, 28-day SKF 38393 treatment resulted in reduced sensitivity of caudate neurons tested 24 h later, and an enhanced sensitivity was observed one week after the completion of chronic SKF 38393 administration. Thus, chronic SKF 38393 induced functional desensitization of D1 receptors, but one-week withdrawal was followed by sensitization.

An electrophysiological evaluation of serotonergic dorsal raphe neurons in maudsley rats

Extracellular electrophysiological recording techniques were used to study serotonergic dorsal raphe (DRN) neurons in Maudsley Reactive (MR), Maudsley Non-Reactive (MNRA) and Sprague Dawley (SD; reference control strain) rats. No significant differences were observed in the average discharge rates of DRN neurons from SD, MR AND MNRA rats. The sensitivity of DRN neuron somatodendritic 5-HT1A autoreceptors to the inhibitory effects of i.v. 8-OH-DPAT or alpha 1-adrenoceptors to the excitatory effects of iontophoretic phenylephrine did not differ significantly among strains. These findings are discussed in light of the previously reported strain-dependent differences in anxiety-like behavior and noradrenergic locus coeruleus neurons.