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Concurrent use of cocaine and ethanol is a common mode of abuse. Cocaine and ethanol have distinctive pharmacologies but both have been shown to cause uterine vasoconstriction and fetal hypoxia. We developed a paradigm of chronic ethanol exposure via liquid diet coupled with binge cocaine exposure on the last day of gestation. Lipton et al. demonstrated unequal segregation of cocaine in rat fetuses as a function of proximal-distal location in the uterus, indicating a differential vasoconstriction of the two main arteries supplying the uterus in rats receiving cocaine. By performing C-sections after exposure to cocaine, we were able to measure the cocaine content and immediate-early gene (IEG) induction in the brains of fetuses according to their intrauterine position and assess the potentially vasoconstrictive effect of ethanol. HPLC analysis of fetal brains exposed to cocaine supported the study of Lipton et al.: fetuses from the proximal (lower) end of the uterus had more cocaine than fetuses from the distal (upper) end. Concurrent ethanol decreased the amount of cocaine reaching the fetuses and diminished the proximal-distal gradient. There were increased numbers of Fos-immunoreactive cells in fetuses exposed to both ethanol and cocaine compared to cocaine binge only. Additionally, the gradient of c-fos induction observed as a function of intrauterine position in cocaine-treated rats was in the opposite direction: most distal fetuses generally had the most Fos-immunoreactive cells. These results indicate that IEG induction in fetal brains exposed to cocaine and ethanol may be more related to hypoxic consequences of prenatal drug exposure.

Background and Purpose: Increased activation of excitatory amino acid (EAA) receptors is considered a major cause of neuronal damage. Possible sources and mechanisms of ischemia-induced EAA release were investigated pharmacologically with microdialysis probes placed bilaterally in rat striatum., Methods: Forebrain ischemia was induced by bilateral carotid artery occlusion and controlled hypotension in halothane-anesthetized rats. During 30 minutes of ischemia, microdialysate concentrations of glutamate and aspartate were measured in the presence of a nontransportable blocker of the astrocytic glutamate transporter GLT-1, dihydrokinate (DHK), or an anion channel blocker, 4,4'-dinitrostilben-2,2'-disulfonic acid (DNDS), administered separately or together through the dialysis probe., Results: In control striata during ischemia, glutamate and aspartate concentrations increased 44+/-13 (mean+/-SEM) times and 19+/-5 times baseline, respectively, and returned to baseline values on reperfusion. DHK (1 mmol/L in perfusate; n=8) significantly attenuated EAA increases compared with control (glutamate peak, 9. 6+/-1.7 versus control, 15.4+/-2.6 pmol/ microL). EAA levels were similarly decreased by 10 mmol/L DHK. DNDS (1 mmol/L; n=5) also suppressed EAA peak increases (glutamate peak, 5.8+/-1.1 versus control, 10.1+/-0.7 pmol/ microL). At a higher concentration, DNDS (10 mmol/L; n=7) further reduced glutamate and aspartate release and also inhibited ischemia-induced taurine release. Together, 1 mmol/L DHK and 10 mmol/L DNDS (n=5) inhibited 83% of EAA release (glutamate peak, 2.7+/-0.7 versus control, 10.9+/-1.2 pmol/ microL)., Conclusions: These findings support the hypothesis that both cell swelling-induced release of EAAs and reversal of the astrocytic glutamate transporter are contributors to the ischemia-induced increases of extracellular EAAs in the striatum as measured by microdialysis.

Induction of the immediate-early gene c-fos by the stimulants cocaine and amphetamine (AMPH) was analyzed by Fos immunocytochemistry at different ages in the brains of prenatally cocaine-treated and control rats. Cocaine and AMPH induced c-fos in patches of striatal neurons during the first postnatal week, and thereafter produced a progressively more homogeneous pattern that was more dense medially. Quantification of Fos-immunoreactive cells in older rats revealed differences related to sex and prenatal cocaine treatment. Both cocaine and AMPH produced dose-dependent increases in the number of Fos-immunoreactive cells in striatum. Prenatal cocaine exposure resulted in increased Fos in males in response to AMPH (2 mg/kg) at P18 and cocaine (10 mg/kg) at 1-2 months. In females, prenatal cocaine treatment resulted in a reduced response to cocaine at 1-2 months. Increased c-fos induction was observed in control females compared to control males in response to low doses of stimulants; no such sex difference was observed in prenatally cocaine-treated rats. The dopamine D1 antagonist SCH23390 blocked cocaine-mediated c-fos induction in all groups. The NMDA antagonist MK-801 blocked cocaine-mediated c-fos induction in the medial striatum. In females only, MK-801 pretreatment resulted in a dramatic increase in the number of Fos-immunoreactive cells in lateral striatum. These findings indicate differences in the neural basis of c-fos induction in males and females, and changes in stimulant-mediated c-fos induction resulting from prenatal cocaine exposure., (Copyright 1998 Elsevier Science B.V. All rights reserved.)

It is common practice in microdialysis studies for probes to be "calibrated" in artificial CSF and in vitro recoveries determined for all substances to be measured in vivo. Dialysate concentrations of such substances are then "corrected" for in vitro recoveries to provide "estimates" of extracellular concentrations. At least for dopamine, in vitro and in vivo recoveries are significantly different and, therefore, an estimate of extracellular dopamine based on correction for in vitro recovery is likely to be erroneous. Generally, however, the relative relationships of such estimates among animals are of interest rather than the "true" extracellular values. Such relationships would be valid to the extent that estimated values are correlated with or predictive of true values. Using the "no net flux" procedure, the present study sought to determine, for both dopamine and its metabolite 3,4-dihydroxyphenylacetic acid (DOPAC), whether in vitro and in vivo recoveries would correlate with each other as well as whether respective estimated and true (no net flux) values of these substances would correlate with each other. Probes (3 mm; BAS/CMed MF-5393), previously calibrated, were lowered into both the nucleus accumbens and striatum of freely moving rats the day before sample collection was begun. In vitro and in vivo recoveries were not significantly correlated (r = 0.1-0.3), for either dopamine or DOPAC. For both dopamine and DOPAC, however, there were significant correlations (r = 0.7-0.8) between estimated and true values. Surprisingly, when using these commercial probes, absolute dialysate levels for both substances were even better correlated (r = 0.9-0.95) with true values.(ABSTRACT TRUNCATED AT 250 WORDS)

Pregnant rats were given injections of saline (0.5 ml/kg) or cocaine (10 mg/kg, 20 mg/ml, s.c.) twice daily between gestational days 7-21. Offspring were examined by microdialysis between postnatal days 10-125 to study the effects of prenatal cocaine exposure on the nigrostriatal dopamine (DA) system. Twenty-min dialysis samples were collected and assayed for DA, DOPAC and HVA. After four baseline samples, the rat was exposed to 20 min of intermittent tail pinch and monitored for four samples; then each rat received an acute injection of cocaine (20 mg/kg, i.p.) and six additional samples were collected. Basal dialysate concentrations of all DA markers, estimated from pre-implantation calibration of the probes, were markedly reduced in young rats ('pups', 10-30 days old) as compared with adult rats (40-125 days old). Compared to control pups, basal DA, as well as DOPAC and HVA, were elevated in the prenatal-cocaine pups. Tail pinch (a mild stressor) produced a significant increase in DA only in the pups prenatally exposed to cocaine. The increase in basal DA induced by an acute cocaine injection (20 mg/kg) was also greater and more prolonged in the prenatal-cocaine pups. In older rats (40-125 days) there were no group differences in any of the DA parameters. Thus prenatal exposure to cocaine produces an activation of the DA system which persists after birth but returns to normal in older rats.

Systemic administration of ibogaine (40 mg/kg, i.p.) has been reported to induce both acute (1-3 h) and persistent (19-20 h) changes in extracellular levels of dopamine and its metabolites in the nucleus accumbens and striatum. In the present study, local administration of ibogaine to the striatum and nucleus accumbens produced effects that mimicked both the acute and persistent effects of systemic administration: perfusion with high concentrations (200 and 400 microM) of ibogaine mimicked the acute effects (decreased extracellular dopamine levels and increased extracellular metabolite levels) whereas perfusion with a low concentration (10 microM) of ibogaine mimicked the persistent effects (decreased extracellular levels of DOPAC). These results indicate that ibogaine acts directly in brain regions containing dopaminergic nerve terminals and that long-lasting effects of systemically administered ibogaine might be mediated by persisting low levels of ibogaine. Locally administered ibogaine (10 microM) was also found to enhance the effects of systemically administered D-amphetamine (1.25 mg/kg, i.p.) on extracellular dopamine levels, and conversely, systemically administered ibogaine (40 mg/kg, i.p.; 19 h pretreatment) enhanced the effects of locally administered D-amphetamine (1-10 microM). These results indicate that, in addition to a metabolic mechanism implicated previously, a pharmacodynamic mechanism contributes to the interaction between ibogaine and D-amphetamine. The relevance of such mechanisms to claims regarding ibogaine's anti-addictive properties is unclear.

The effect of prenatal cocaine on the anatomical development of the striatum was examined. The distribution and density of dopaminergic innervation of the striatum of animals exposed to cocaine during the second and third week of gestation was not noticeably different from prenatally saline-injected or untreated controls at any age. The patch/matrix organization of the striatum also appeared unaltered: neurons exhibiting dense substance P staining were localized to patches that overlapped dopamine terminal patches early in development, and enkephalin- and calbindin-immunoreactive neurons were found segregated to the matrix. Histochemical staining for acetylcholinesterase and NADPH diaphorase also revealed no differences between prenatally cocaine-treated and control brains. Whereas prenatal cocaine treatment failed to modify the basic compartmental organization of the striatum, it did lead to a hyperinnervation of serotonin-immunoreactive fibers which developed slowly after birth. Thus prenatal exposure to cocaine is capable of altering the ingrowth of serotonergic projections to the striatum while producing no change in the organization of neurons intrinsic to the striatum.

Repeated microdialysis measurements, conducted 1 week apart at the same tissue site, were used to investigate the changes in basal and cocaine-stimulated extracellular dopamine (DA) levels after a single prior exposure to either saline or cocaine. Dialysis probes were placed into rats previously implanted with guide cannulas and basal levels of dopamine (DA), and its metabolites (DOPAC and HVA) were estimated in 20-min fractions of the dialysate. Basal levels in the extracellular fluid (ECF), estimated from pre-implantation calibration of the probes, were 7.9 +/- 0.7 nM DA, 4.9 +/- 0.8 microM DOPAC, and 3.6 +/- 0.6 microM HVA. After a stable baseline was obtained saline (1 ml/kg, i.p.) or cocaine (20 mg/kg, i.p.) was injected. Saline produced no significant changes in any of the neurochemical markers. A cocaine injection produced a sixfold increase in DA, while DOPAC and HVA were unchanged. One week later the same procedure was repeated except this time both groups received cocaine. In rats that had received cocaine 1 week earlier, basal DA levels in the ECF were doubled, whereas they were unchanged in rats that received saline a week earlier. Furthermore, the dopamine release in response to acute cocaine during the second week was elevated in animals which had been previously exposed to cocaine. Rotation was also measured during both weeks and, while a tendency toward behavioral sensitization was observed, it did not reach significance.(ABSTRACT TRUNCATED AT 250 WORDS)

In this study, we have examined the spontaneous efflux and stimulation-induced overflow of endogenous dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) from striatal slices prepared from adult rats. Partial destruction of striatal DA terminals with 6-hydroxydopamine reduced the electrically evoked overflow of DA and DOPAC, but by less than the DA content of the tissue slices. This resulted in an increase in the fractional overflow of DA and DOPAC, a measure of overflow from residual dopaminergic terminals. The increase in fractional DA overflow was related to the lesion size, reaching 7-fold with DA depletions of greater than 90%. Inhibition of DA re-uptake with nomifensine (10 microM) increased DA overflow and reduced DOPAC overflow by an equivalent amount, indicating that a portion of the DA released by electrical stimulation is recaptured and metabolized before appearing as DOPAC in the superfusate. However, 6-hydroxydopamine lesions further elevated fractional DA overflow above control even in the presence of nomifensine, suggesting that slices prepared from lesioned animals exhibit an increase in DA release from residual dopaminergic terminals. DA overflow was enhanced by the DA receptor antagonist sulpiride (1 microM) in control tissue but not in slices prepared from lesioned animals, suggesting that increased efflux per terminal was accompanied by reduced autoinhibition of release. Over a range of firing frequencies typical of nigrostriatal neurons in vivo (2-8 Hz), fractional DA overflow per pulse from lesioned slices was 3-fold higher than control overflow; however, fractional overflow per pulse was reduced from lesioned but not control slices when slices were exposed to a higher frequency (12 Hz). Thus, the lesion appeared to have increased DA release at moderate frequencies, but had reduced the effective range of frequencies over which the DA terminals could operate. Finally, 3-iodotyrosine (2 mM), an inhibitor of tyrosine hydroxylase, reduced DA overflow from intact slices, but completely abolished overflow from lesioned slices, suggesting that 6-hydroxydopamine had increased the dependence of DA efflux on a sustained rate of DA synthesis. Taken together, these data suggest that after lesioning with 6-hydroxydopamine, DA released per pulse from residual terminals is increased relative to control, so long as the stimulation frequency is within the physiological range. This increase in release may serve a compensatory function, maintaining dopaminergic control over striatal function despite extensive loss of DA neurons.

Using in vivo voltammetry in rats, we examined the relationship between the electrochemical signal measured in the striatum and the behavioral responses associated with various types of stimulation. Three patterns emerged. First, a series of homeostatic challenges, including abrupt decreases in glucose utilization, blood volume, or arterial blood pressure, were ineffective in altering the electrochemical signal despite the sympathoadrenal response produced by each. Second, intense exteroceptive stimuli, such as an electric shock applied to the tail or placing animals in a shallow ice-water bath, provoked large and abrupt rises in the signal which decayed rapidly. Third, rats eating after a 24-h fast, drinking after a period of dehydration, or presented with novel olfactory or visual stimuli, exhibited much smaller and more gradual rises in the electrochemical signal which were more long-lasting. In each case, the magnitude of the change in electrochemical signal was generally related to the level of behavioral activation, being most prominent when treatments produced a startle response. Those large increases in signal were markedly attenuated by pretreatment with alpha-methyl-p-tyrosine or gamma-butyrolactone, drugs known to decrease the release of dopamine, suggesting that the signal observed was associated with an increase in the activity of central dopaminergic neurons.

Dopaminergic control over striatal targets appears to be retained in rats sustaining lesions of the nigrostriatal dopamine (DA) system as long as 5-10% of that projection remains. Similarly, during postnatal development, dopaminergic control over striatal neurons matures well before the innervation of striatum by the nigrostriatal bundle is attained. These observations suggest that enhanced efficacy of dopaminergic transmission may compensate for hypoinnervation of striatum after lesions or during development. To examine this hypothesis, striatal slices were superfused with Krebs bicarbonate buffer and effluent was collected and analyzed for endogenous DA. Electrical field stimulation (2 Hz) continuously delivered to slices prepared from intact adult rats increased DA efflux to 3-5 times the prestimulation rate within 10 min. Efflux then fell to approximately twice the basal rate over the next 20 min. DA efflux was also examined using slices prepared from adult animals given 6-hydroxydopamine 2-3 weeks earlier, and from 7-10-d-old rat pups. In each group, striatal DA levels were 10-40% of adult control values. Nevertheless, stimulated DA efflux from these slices attained the same rate as that observed with intact, adult slices. Thus, fractional DA efflux from these slices was several times higher than the control rate by the end of the stimulation period. This increased DA efflux appeared to be a consequence of both increased release and decreased reuptake of DA, as the fractional DA efflux from control striatal slices could not be increased to the rate seen in hypoinnervated slices using nomifensine (10 microM), an inhibitor of DA efflux.(ABSTRACT TRUNCATED AT 250 WORDS)

We have examined the impact of L-dihydroxyphenylalanine (L-DOPA) on the in vivo release of dopamine (DA) in rat striatum using carbon fiber voltammetry. L-DOPA caused a large increase in the DA released by nigrostriatal bundle stimulation. This was reduced by 94% in animals pretreated with intraventricular 6-hydroxydopamine. We conclude that L-DOPA increases depolarization-induced DA release from DA neurons.