Alexis C. Thompson

Delta-Opioid Receptor Antagonists Prevent Sensitization to the Conditioned Rewarding Effects of Morphine

BACKGROUND Functional interactions between mu- and delta-opioid receptors (MOPr and DOPr, respectively) are implicated in morphine tolerance and dependence. The contribution of DOPr to the conditioned rewarding effects of morphine and the enhanced conditioned response that occurs after repeated morphine administration is unknown. This issue was addressed with the conditioned place preference procedure (CPP). METHODS Rats received home cage injections of saline or morphine (5.0 mg/kg/day x 5 days) before conditioning. For sensitization studies, DOPr antagonists (DOPr1/2: naltrindole, DOPr2: naltriben, DOPr1: 7-benzylidenenaltrexone) were administered before morphine injections. Conditioning sessions (2 morphine; 2 saline) commenced 3 days later. To assess the influence of acute DOPr blockade on the conditioning of morphine reward in naïve animals, 3 morphine and 3 saline conditioning sessions were employed. Antagonists were administered before morphine conditioning sessions. RESULTS Morphine was ineffective as a conditioning stimulus after two conditioning sessions in naïve rats. However, doses > or = 3.0 mg/kg produced significant CPP in morphine pre-exposed rats, confirming that sensitization develops to the conditioned rewarding effects of morphine. In animals that received morphine pre-exposure with naltrindole or naltriben but not 7-benzylidenenaltrexone, sensitization was prevented. No attenuation of morphine CPP was observed in animals that received DOPr antagonists acutely, before conditioning sessions. CONCLUSIONS These data indicate a critical role of DOPr systems in mediating sensitization to the conditioned rewarding effects of morphine. The efficacy of naltrindole and naltriben in preventing the enhanced response to morphine suggest the specific involvement of DOPr2 in the sensitization process.

Placenta ingestion enhances opiate analgesia in rats.

Analgesia, produced by either a morphine injection or footshock, was monitored (using a tail-flick test) in nonpregnant female rats. Analgesia was induced within minutes of having the rats eat one of several substances. When the substance eaten was rat placenta, both the morphine- and shock-induced types of analgesia were significantly greater than in controls that ingested other substances (or nothing). When footshock (hind-paw) was administered in conjunction with the opiate antagonist naltrexone, the analgesia produced was attenuated but detectable; in this case, placenta ingestion did not enhance the analgesia, suggesting that the effect of placenta is specific to opiate-mediated analgesia. Placenta ingestion, in the absence of an analgesia-producing manipulation, did not elevate pain threshold. It is possible that this enhancement of analgesia is one of the principal benefits to mammalian mothers of ingesting the placenta and birth fluids (placentophagia) at delivery.

Long-Term Reduction in Ventral Tegmental Area Dopamine Neuron Population Activity Following Repeated Stimulant or Ethanol Treatment

BACKGROUND Drugs of abuse exert profound effects on the mesolimbic/mesocortical dopaminergic (DA) systems. Few studies have investigated the long-term adaptations in ventral tegmental area (VTA) DA neuron activity after repeated exposure to drugs of abuse. We investigated changes in the electrical activity of VTA DA neurons after cessation from treatment with several stimulants and ethanol. METHODS Adult rats were treated with stimulants (amphetamine: 2 mg/kg per day, 5 days/week, 2 weeks; cocaine: 15 mg/kg per day, 5 days/week, 2 weeks; nicotine: .5 mg/kg per day, 5 days; ethanol: 10 g/kg per day, 3 weeks) and the single-unit activity of VTA DA neurons was studied in vivo 3 to 6 weeks later. RESULTS Stimulant and ethanol treatment decreased basal VTA DA neuron population activity but not firing rate or firing pattern. This effect was reversed by acute apomorphine, suggesting that the underlying mechanism for reduced population activity was depolarization inactivation. Anesthesia did not confound this result, as similar effects were observed in amphetamine-treated rats recorded in a conscious preparation. CONCLUSIONS Reduced basal VTA DA neuron population activity presumably due to depolarization inactivation is a common and long-term neuroadaptation to repeated treatment with stimulants and ethanol. This change in VTA DA neuron activity could underlie the persistent nature of addiction-associated behaviors.

Ingestion of amniotic fluid enhances opiate analgesia in rats

Placenta ingestion has recently been shown to enhance opiate-mediated analgesia produced by morphine injection, footshock, or vaginal/cervical stimulation. The enhancement of the effect of endogenous opiates (especially analgesia) may be one of the principal benefits to mammalian mothers of placentophagia at delivery. During labor and delivery, however, mothers also ingest amniotic fluid (AF) which, unlike placenta, becomes available during, or even before expulsion of the infant. The present experiments were undertaken to determine whether AF ingestion, too, enhances analgesia; if so, whether the effect requires ingestion of, or merely exposure to, AF; whether the effect can be produced by AF delivered directly to the stomach by tube; and whether the enhancement, if it exists, can be blocked by administering an opiate antagonist. Nulliparous Long-Evans rats were tested for analgesia using tail-flick latency. We found that rats that ingested AF after receiving a morphine injection showed significantly more analgesia than did rats that ingested a control substance; AF ingestion, alone, did not produce analgesia; ingestion of AF, rather than just smelling and seeing it, was necessary to produce analgesia enhancement; AF produced enhancement when oropharyngeal factors were eliminated by delivering it through an orogastric tube; and treatment of the rats with naltrexone blocked the enhancement of morphine-induced analgesia that results from AF ingestion.

Amniotic-fluid ingestion by parturient rats enhances pregnancy-mediated analgesia

Amniotic fluid and placenta contain a substance (POEF, for Placental Opioid-Enhancing Factor) that, when ingested, enhances opioid-mediated analgesia in nonpregnant rats; ingestion of the substance by rats not experiencing opioid-mediated analgesia, however, does not produce analgesia. It is highly likely that periparturitional analgesia-enhancement is a significant benefit of ingestion of the afterbirth (placentophagia) during delivery. Here we report that prepartum ingestion of amniotic fluid (via orogastric infusion) does indeed enhance the endogenous-opioid-mediated analgesia evident at the end of pregnancy and during delivery; that the degree of enhancement is greater with 0.75 ml than with 0.25 ml; and that the prepartum enhancement of analgesia can be blocked with the opioid antagonist naloxone.

Placenta ingestion enhances analgesia produced by vaginal/cervical stimulation in rats

Ingestion of placenta has previously been shown to enhance opiate-mediated analgesia (measured as tail-flick latency) induced either by morphine injection or by footshock. The present study was designed to test whether placenta ingestion would enhance the partly opiate-mediated analgesia produced by vaginal/cervical stimulation. Nulliparous Sprague-Dawley rats were tested for analgesia, using tail-flick latency, during and after vaginal/cervical stimulation; the tests for vaginal/cervical stimulation-induced analgesia were administered both before and after the rats ate placenta or ground beef. Placenta ingestion, but not beef ingestion, significantly heightened vaginal/cervical stimulation-induced analgesia. A subsequent morphine injection provided evidence that, as in a previous report, placenta ingestion, but not beef ingestion, enhanced morphine-induced analgesia.

Placental opioid-enhancing factor (POEF): Generalizability of effects

A substance in amniotic fluid and placenta (POEF for Placental Opioid-Enhancing Factor) has been shown to enhance opiate- or opioid-mediated analgesia in rats. Recent studies have only touched on the generalizability of the phenomenon. The present studies further tested the generalizability of the POEF effect: they examined sex specificity of the mechanism; whether POEF activity exists in afterbirth material of species other than the rat; whether POEF activity exists in tissue other than afterbirth material; whether POEF activity could be demonstrated after injection rather than ingestion of afterbirth material; and whether POEF enhances all opioid-mediated phenomena. We found that (a) POEF is effective in male rats as well as in female rats; (b) POEF activity exists in human and dolphin afterbirth material; (c) ingestion of pregnant-rat liver does not produce enhancement of opioid-mediated analgesia; (d) POEF does not seem to be effective when amniotic fluid is injected either IP or SC; and (e) POEF does not modify morphine-induced hyperthermia.

Dose-dependent enhancement of morphine-induced analgesia by ingestion of amniotic fluid and placenta

Ingestion of amniotic fluid and placenta by rats has been shown to enhance opioid-mediated analgesia. The present studies were designed to examine the effect of several doses and volumes of placenta and amniotic fluid on tail-flick latency in rats treated with 3 mg/kg morphine. The optimal dose of amniotic fluid was found to be 0.25 ml, although 0.50 and 1.0 ml also produced significant enhancement. Doses of 0.125 and 2 ml of amniotic fluid were ineffective, as was a dose of 0.25 ml diluted to 2 ml with saline. The optimal dose of placenta was found to be 1 placenta, although the resulting enhancement was not significantly greater than that produced by 025, 0.50, 2.0 or 4.0 placentas. Doses smaller than 0.25 placenta or larger than 4.0 placentas were ineffective. The most effective doses of amniotic fluid and placenta correspond to the amounts delivered with each pup during parturition.

Quantitative microdialysis of neuropeptide Y.

The feasibility of using the difference method of quantitative microdialysis to measure neuropeptide Y (NPY) was evaluated in vitro and in vivo. The accuracy of this method was tested in vitro under steady-state conditions for 3 test solutions containing known concentrations of NPY. The estimated concentrations of NPY were 1.2 +/- 0.6, 3.7 +/- 0.9, and 15.1 +/- 0.7 pg/microliter (mean +/- SEM) in agreement with the actual concentrations of NPY in the test solutions which were 1.1 +/- 0.8, 4.6 +/- 0.6, and 14.6 +/- 0.5 pg/microliter (mean +/- SEM of solution samples), respectively. The responsiveness of the estimated NPYext measure to changes in the external concentration of NPY was also evaluated in vitro. An accurate estimate of NPYext was obtained within the first sampling period (within 15 min) after a 2-3-fold increase in the test solution concentration of NPY and within 2-3 sampling periods (15-45 min) in response to a 2-3-fold decrease in the test solution concentration of NPY. In vivo, the estimated basal concentration of NPY in dialysis samples from probes in the medial basal hypothalamus of anesthetized female rats (n = 4) was 4.0 +/- 1.6 pg/microliters and increased to 9.5 +/- 0.3 pg/microliter during K+ stimulation. Relative recovery was 22% in vivo under steady-state conditions and ranged from 14% to 30% during dynamic conditions. These results demonstrate that the difference method of quantitative microdialysis accurately estimates picomolar concentrations of NPY in vitro, and is sufficiently sensitive to detect basal and increasing concentrations of NPY in vivo.

Amniotic-Fluid Ingestion Enhances the Central Analgesic Effect of Morphine

Amniotic fluid and placenta contain a substance (POEF) that when ingested enhances opioid-mediated analgesia produced by several agents (morphine injection, vaginal/cervical stimulation, late pregnancy, footshock), but not that produced by aspirin injection. The present series of experiments employed quaternary naltrexone, an opioid antagonist that does not readily cross the blood-brain barrier, in conjunction with either peripheral or central administration of morphine, to determine whether amniotic-fluid ingestion (and therefore POEF ingestion) enhances opioid-mediated analgesia by affecting the central and/or peripheral actions of morphine. The results suggest that POEF affects only the central analgesic effects of morphine.