The regulation of hippocampal dynorphin by neural/neuroendocrine pathways: models for effects of aging on an opioid peptide system

Previous research has demonstrated increased messenger RNA expression and peptide content in an opioid system localized to hippocampal dentate granule cells in aged rats. This altered regulation of dynorphin was correlated with the emergence of an age-related impairment in spatial learning. Considerable evidence exists for additional effects of aging on systems that provide input to the dynorphin-containing dentate granule cells. Such changes have been well documented for loss of perforant path innervation from entorhinal cortex, deterioration in septohippocampal cholinergic neurons, and high amounts of glucocorticoids that have, among their targets, receptors located in the dentate gyrus. Similar to the effects of aging on hippocampal dynorphin, age-related changes in each of these systems correlate with the severity of spatial learning impairment in aged rats. This raises the possibility that dysregulation of dynorphin in the aged brain is a reactive response to antecedant change(s) in this circuitry, a hypothesis that was examined by separately manipulating in young rats the three neural/neuroendocrine systems identified above. Of the three models examined only removal of the perforant path reproduced the effect of aging on dynorphin in the hippocampal formation. An immunotoxin was used in Experiment 1 to selectively remove septo-hippocampal cholinergic neurons in young rats. No alteration in hippocampal opioid peptides was produced by this treatment. Experiment 2 examined effects of exposure to excess corticosterone. Adrenalectomized rats exhibited a significant decrease in hippocampal dynorphin-A (1-8) content, which was reversed by corticosterone replacement at a concentration approximating normal basal levels. Dynorphin-A (1-8) content, however, was not reliably increased by exposure to excess corticosterone. In contrast, perforant path removal was found to reproduce the effect of aging on dynorphin content; either aspiration of the entorhinal cortex or knife-cut transections of the perforant path reliably increased hippocampal dynorphin content. These results support the conclusion that age-related deterioration in the septohippocampal cholinergic system and evaluated exposure to corticosterone are not sufficient to induce an elevation in hippocampal dynorphin content. Only removal of the perforant path innervation was found to reproduce the elevation in hippocampal dynorphin content observed in aged rats with hippocampal-dependent learning impairment.     

Cocaine, dopamine and the endogenous opioid system

Cocaine addiction and opiate addiction are both major health problems in the United States today. Prospective studies from our Laboratory, which were able to detect the advent of the HIV-1 epidemic in parenteral drug abusers in New York City beginning around 1978, also showed that, from the beginning of the AIDS epidemic, cocaine abuse was a very important co-factor significantly increasing the risk for developing cocaine dependency. Fundamental studies from many laboratories including our own have shown that cocaine has profound effects on dopaminergic function, primarily from its well-established primary action of blocking the reuptake of dopamine from the synaptic cleft, an action of cocaine directed at the specific dopamine transporter. It has also been well-established by others that cocaine similarly blocks the reuptake of serotonin and norepinephrine. However, recent studies from our laboratory have shown that chronic cocaine administration profoundly disrupts the endogenous opioid system. Extensive studies have been conducted using an animal model which we have developed in our laboratory, the "binge" pattern cocaine administration model. Findings from these studies have led us to recognize the profound disruption of both dynorphin gene expression and kappa opioid receptor gene expression in a setting of chronic cocaine administration and, in turn, have led us to question a possible role of disruption of this system in the acquisition and persistence of cocaine addiction. These findings may have significance for the development of new pharmacotherapeutic agents which may be directed to specific components of the endogenous opioid system and, in particular, possibly the kappa opioid receptor system. Therefore, we have initiated studies to examine further the role of the dynorphin peptide-kappa opioid receptor system in normal physiologic function in humans.     

Neural mechanisms of tolerance to the effects of cocaine

Chronic use of cocaine in high doses can produce tolerance as assessed by various behavioral, neurochemical, cellular and molecular measures in specific brain regions. Tolerance to cocaine is indicated by drug discrimination and intracranial self-stimulation models, which show the development of tolerance after approximately 1 week of frequent cocaine treatment, with recovery after a similar period of cocaine abstinence. Tolerance to the reinforcing properties of cocaine depends on dose, duration and frequency of cocaine self-administered by experimental animal or human subjects. The mechanism underlying this effect may involve an absolute or relative attenuation of dopamine response to cocaine challenge after frequent or repeated treatment in the nucleus accumbens (NAc). Similarly, afferent and efferent NAc circuits exhibit reduced metabolic activity, which lasts throughout the early period of withdrawal following repeated treatment. Attenuation of immediate early gene response also occurs, which might be related to a functional desensitization of dopamine D1-like receptors. Furthermore, intracellular adaptive responses to chronic cocaine exposure induce striatal dynorphin expression decreasing the behavioral potency of subsequent drug treatment. Thus, a combination of various pharmacodynamic mechanisms and the attenuation of dopamine response induced by sufficient dose, duration and frequency of cocaine exposure ultimately invoke the transient development of tolerance to the reinforcing effects of cocaine.     

Methamphetamine alters prodynorphin gene expression and dynorphin A levels in rat hypothalamus

Chronic administration of morphine or cocaine affects opioid gene expression. To better understand the possible existence of common neuronal pathways shared by different classes of drugs of abuse, we studied the effects of methamphetamine on the gene expression of the opioid precursor prodynorphin and on the levels of peptide dynorphin A in the rat brain. Acute (6 mg/kg, intraperitoneally, i.p.) and chronic (6 mg/kg, i.p. for 15 days) methamphetamine markedly raised prodynorphin mRNA levels in the hypothalamus, whereas no effect was observed in the hippocampus. Dynorphin A levels increased after chronic treatment in the hypothalamus and in the striatum, whereas no significant changes were detected after acute treatment. These results indicate that methamphetamine affects prodynorphin gene expression in the hypothalamus, which may be an important site (also for its relevant neuroendocrine correlates) for opioidergic mechanisms activated by addictive drugs.     

Specific reductions of striatal prodynorphin and D1 dopamine receptor messenger RNAs during cocaine abstinence

It is well established that the opioid neuropeptide and dopamine systems are altered following the use of cocaine. However very little information is available about their possible involvement during cocaine abstinence. In the present study, the mRNA expression of the dopamine receptors, D1 and D2, and the opioid peptides, prodynorphin and proenkephalin, were analyzed in the rat striatum using in situ hybridization histochemistry. Saline or cocaine (30 mg/kg, i.p.) were administered to rats once daily for 1 or 10 days. To examine cocaine abstinence, animals were treated for 10 days as described followed by a 10-day drug free period. Acute and intermittent cocaine administration elevated the prodynorphin mRNA expression in the dorsal striatum, consistent with previous reports, while the abstinent phase resulted in a significant reduction of prodynorphin mRNA levels in the ventrorostral striatum. The D1-receptor mRNA was decreased in the caudorostral striatum during cocaine withdrawal, a finding opposite to the increase observed following a single injection of the drug. Proenkephalin and the D2-receptor mRNAs were not altered during cocaine abstinence, though proenkephalin was elevated following acute but not repeated cocaine administration. These results show long-term suppression on prodynorphin and D1-receptor systems in specific striatal populations localized mainly in rostral areas during withdrawal from cocaine.     

Quantitative electroencephalographic characteristics of crack cocaine dependence

This study replicates preliminary findings reporting a quantitative electroencephalographic (QEEG) profile of crack cocaine dependence in abstinence. All subjects (n = 52) met criteria for DMS-III-R cocaine dependence (in the form of crack), and were residing in a drug-free therapeutic community. Baseline QEEG evaluations were conducted at intake (5-10 days after last use of crack, and at follow-up (1 month after last reported use). Previous findings of significant excess of relative alpha power and deficit of absolute and relative delta and theta power were replicated in this expanded group. Abnormalities were greater in anterior than posterior regions, and disturbances in interhemispheric relationships were also observed. Further, QEEG showed little change in the interval between the first and second evaluations. This QEEG profile may reflect persistent alterations in neurotransmission as a possible consequence of chronic cocaine exposure.     

Role of dynorphin and enkephalin in the regulation of striatal output pathways and behavior

Projection neurons in the striatum give rise to two output systems, the "direct" and "indirect" pathways, which antagonistically regulate basal ganglia output. While all striatal projection neurons utilize GABA as their principal neurotransmitter, they express different opioid peptide co-transmitters and also different dopamine receptor subtypes. Neurons of the direct pathway express the peptide dynorphin and the D1 dopamine receptor, whereas indirect pathway neurons express the peptide enkephalin and the D2 receptor. In the present review, we summarize our findings on the function of dynorphin and enkephalin in these striatal output pathways. In these studies, we used D1- or D2-receptor-mediated induction of immediate-early genes as a cellular response in direct or indirect projection neurons, respectively, to investigate the role of these opioid peptides. Our results suggest that the specific function of dynorphin and enkephalin is to dampen excessive activation of these neurons by dopamine and other neurotransmitters. Levels of these opioid peptides are elevated by repeated, excessive activation of these pathways, which appears to be an adaptive or compensatory response. Behavioral consequences of increased opioid peptide function in striatal output pathways may include behavioral sensitization (dynorphin) and recovery of motor function (enkephalin).     

Event-related potentials in hypertriglyceridemia

Chronic morphine-treated dorsal-root ganglion (DRG) neurons in DRG/spinal cord explant cultures were previously shown to become supersensitive to the excitatory effects of remarkably low concentrations of the opioid agonists, morphine and dynorphin, and the opioid antagonist, naloxone. The present study demonstrates that this opioid excitatory supersensitivity of chronic morphine-treated DRG neurons (1 microM for > 1 week) is retained for periods > 3 months after return to control culture medium. Acute application of femtomolar dynorphin, as well as nanomolar naloxone, to the treated neurons after months in control medium evoked characteristic prolongation of the action potential duration (APD), as occurs in cells tested during or shortly after chronic opioid exposure. The threshold concentrations for eliciting these excitatory effects in naive DRG neurons are > 1000-fold higher. Furthermore, treatment of micromolar morphine-sensitized neurons with 1 nM etorphine (which is a potent excitatory opioid receptor antagonist) for I week prior to return to control medium blocked further expression of opioid excitatory supersensitivity when tested after an additional 1-7 weeks in culture. These results provide a unique in vitro model system for analyses of some of the cellular mechanisms underlying protracted opioid dependence in vivo.     

Immunocytochemical localization of enkephalins in the brain of the African lungfish, Protopterus annectens, provides evidence for differential distribution of Met-enkephalin and Leu-enkephalin

The distribution of various opioid peptides derived from proenkephalin A and B was studied in the brain of the African lungfish Protopterus annectens by using a series of antibodies directed against mammalian opioid peptides. The results show that both Metenkephalin- and Leu-enkephalin-immunoreactive peptides are present in the lungfish brain. In contrast, enkephalin forms similar to Met-enkephalin-Arg-Phe, or Met-enkephalin-Arg-Gly-Leu, as well as mammalian alpha-neoendrophin, dynorphin A (1-8), dynorphin A (1-13), or dynorphin A (1-17) were not detected. In all major subdivisions of the brain, the overwhelming majority of Met-enkephalin- and Leu-enkephalin-immunoreactive cells were distinct. In particular, cell bodies reacting only with Leu-enkephalin antibodies were detected in the medial subpallium of the telencephalon, the griseum centrale, the reticular formation, the nucleus of the solitary tract, and the visceral sensory area of the rhombencephalon. Cell bodies reacting only with Met-enkephalin antibodies were found in the lateral subpallium of the telencephalon, the caudal hypothalamus, and the tegmentum of the mesencephalon. The preoptic periventricular nucleus of the hypothalamus exhibited a high density of Metenkephalin-immunoreactive neurons and only a few Leu-enkephalin-immunoreactive neurons. The distribution of Met-enkephalin- and Leu-enkephalin-immunoreactive cell bodies and fibers in the lungfish brain showed similarities to the distribution of proenkephalin A-derived peptides described previously in the brain of land vertebrates. The presence of Met-enkephalin- and Leu-enkephalin-like peptides in distinct regions, together with the absence of dynorphin-related peptides, suggests that, in the lungfish, Met-enkephalin and Leu-enkephalin may originate from distinct precursors.     

Self-administration of cocaine, alfentanil, and nalbuphine under progressive-ratio schedules: Consumer demand and labor supply analyses of relative reinforcing effectiveness.

Progressive-ratio (PR) schedules of intravenous (IV) drug self-administration are useful for establishing the relationships between reinforcing effectiveness and pharmacological actions of abused drugs. The authors compared the reinforcing effects of the high-efficacy opioid alfentanil, the low-efficacy opioid nalbuphine, and cocaine using a PR schedule of IV drug injection in rhesus monkeys in which the response requirement increased during the experimental session and the initial response requirement (IRR) was varied. Analyses based on either consumer demand or labor supply models of behavioral economics revealed that the relative reinforcing effectiveness of cocaine and alfentanil was greater than that of nalbuphine. These results suggest that PR schedules with varying IRRs can provide meaningful estimates of the relative reinforcing effectiveness of abused drugs. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Classical conditioning in the fetal rat: Reinforcing properties of dynorphin A (1–13).

This study examined the reinforcing properties of dynorphin A (1-13) in a single-trial classical conditioning paradigm in the E20 rat fetus. Injection of dynorphin into the cisterna magna increased fetal motor activity and reduced facial wiping in a test of perioral cutaneous responsiveness. Dynorphin was effective as an unconditioned stimulus (US) in a classical conditioning paradigm using an artificial nipple conditioned stimulus (CS) and dynorphin A (1-13) US. The association between CS and US was dependent on activity in the kappa opioid system. Re-exposure to the artificial nipple CS after a single pairing of the nipple with dynorphin resulted in conditioned activation of the kappa opioid system. Dynorphin A (1-13) functions as a reinforcer for classical conditioning in the rat fetus after intracisternal or intrahemispheric injection with the conditioned response depending on route of administration and site of injection. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Distinct mechanisms for activation of the opioid receptor-like 1 and kappa-opioid receptors by nociceptin and dynorphin A

To understand how two structurally analogous ligand-receptor systems, the nociceptin/opioid receptor-like 1 (ORL1) and dynorphin A/kappa-opioid receptor 1 (KOR1) systems, achieve selectivity, receptor chimeras were generated and analyzed. Replacing discrete domains located between the N-terminus and top of the third transmembrane helix of the KOR1 by the homologous domains of the ORL1 receptor yields hybrid receptors, which, in comparison with the parent KOR1, display up to 300-fold increased affinity but low sensitivity toward nociceptin, and unaltered (high) affinity and sensitivity toward dynorphin A. These substitutions contribute elements for binding of nociceptin but do not suppress determinants necessary for binding and potency of dynorphin A. More importantly, further replacement in these chimeras of the second extracellular loop with that of the ORL1 receptor fully restores responsiveness to nociceptin without impairing responsiveness to dynorphin A. A bifunctional hybrid receptor has thus been identified that binds and responds to both nociceptin and dynorphin A as efficiently as the ORL1 receptor does to nociceptin and the KOR1 to dynorphin A. Together, these results suggest that distinct peptide activation mechanisms operate in the two receptor systems. In particular, the second extracellular receptor loop appears to be an absolute requirement for activation of the ORL1 receptor by nociceptin, but not for activation of the KOR1 by dynorphin A.     

Stimulus contingencies that permit classical conditioning of opioid activity in the rat fetus.

In a prenatal model of classical conditioning, rat fetuses received presentations of an artificial nipple (conditioned stimulus; CS) paired with milk (unconditioned stimulus). Infusion of milk promotes activity in the kappa opioid system of the fetus, but after 2, 3, or 6 pairings with the artificial nipple, milk evoked both kappa and mu opioid activity. The nipple CS has no effect on opioid activity, but after pairing with milk evoked a mu opioid response. Conditioned mu opioid activity was evident in 60% of subjects tested after I paired conditioning trial. Significantly more fetal subjects (90%) exhibited conditioned opioid activity if preexposed to the nipple twice before conditioning. CS preexposure altered behavior during the conditioning trial, with preexposed fetuses showing more pronounced responses to milk infusion. Exposure to familiar stimuli facilitates classical conditioning of physiological responses, including opioid activity, during the first suckling episode. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Respiratory and thermoregulatory responses of rabbits breathing carbon dioxide during heat exposure

1. Rabbits were clipped and exposed in turn to three environmental conditions: control (C), cold exposure (CE) and water deprivation (WD). Following each type of treatment, the rabbits were exposed to an ambient temperature (Ta) of 35 degrees C for 1 hr. Throughout this period they breathed either normal atmospheric air or 6% CO2 in air. 2. During heat exposure, measurements were made of the respiratory responses and of the O2 consumption (Vo2) of the rabbits. Rectal temperature (Tre) was measured immediately before and again immediately after heat exposure. 3. When subjected to cold exposure or water deprivation the rabbits showed an initial decrease in respiratory frequency (RF) and an initial increase in VT when compared with controls. There was no difference in VE. Rabbits breathing 6% CO2 showed an increase in VT and VE and a decrease in RF when compared with rabbits breathing atmospheric air. In all cases a change in VT or RF was associated with a reciprocal change in the other parameter. 4. The respiratory responses to breathing 6% CO2 were essentially similar in treated and control rabbits, from which it is concluded that neither cold exposure nor water deprivation alter the sensitivity of the medullary respiratory centre to the respiratory drive from the central chemosensors. 5. The increase in Tre during heat exposure was significantly less in rabbits breathing 6% CO2 than in rabbits breathing atmospheric air. However, there was no significant over-all difference in VO2 between rabbits breathing CO2 and those breathing air. From this it is concluded that increased ventilation induced by CO2 causes a greater dissipation of heat than does thermally-induced panting. 6. It is concluded that VT is controlled by the level of blood PCO2 whereas RF is controlled by thermoregulatory requirements. It is further concluded that the reciprocal relationship between VT and RF is regulated in such a way as to maintain VE at the appropriate level for effecting gaseous exchange and evaporative heat loss.     

Repeated dosing with oral cocaine in humans: Assessment of direct effects, withdrawal, and pharmacokinetics.

Cocaine withdrawal symptoms are thought to play a role in relapse; studies characterizing the symptomatology have yielded mixed findings. This study sought to examine the pharmacodynamic/pharmacokinetic profile of repeated high dose exposure to oral cocaine and characterize acute and protracted withdrawal in cocaine abusers. This study employed a repeated-dosing, single-blind design in which subjects (n = 9), resided for 40 days on a closed ward. They were maintained for two 4-day cocaine exposure periods (Days 1–4 & Days 9–12, cocaine 175 mg, p.o.; 5 hourly doses; 875 mg/day) separated by a 4-day matched placebo exposure period (Days 5–8). After these 12 days, an additional period of 28 days of placebo maintenance followed (Days 13–40). Test sessions were conducted during each phase; measures of mood, drug effects, sleep, pharmacokinetics, and prolactin were collected throughout the study. The dosing regimen produced cocaine plasma concentrations (Cmax of 680 ng/mL) two to threefold higher than typically seen in acute dose studies. Prototypic psychostimulant effects, including subjective ratings of euphoric effects (liking, high, good effects) and significant cardiopressor effects, were sustained during the active dosing periods, corresponding to the rise and fall of plasma cocaine. Withdrawal-like symptoms (i.e., disruptions of sleep, increased ratings of anxiety, irritability, crashing) were observed within 24-hr after cessation of dosing. Cocaine reduced prolactin acutely, but no sustained alterations were observed for this measure or for other signs or symptoms during the 28-day abstinence period. These findings indicate that exposure to controlled high doses of cocaine produces modest symptoms consistent with cocaine withdrawal within hours of cessation of dosing but provide no evidence of symptoms persisting beyond 24 hours. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Aging and the Ranschburg effect: No evidence of reduced response suppression in old age.

Two experiments tested 1 aspect of L. Hasher and R. T. Zacks's (1988) reduced inhibition hypothesis, namely, that old age impairs the ability to suppress information in working memory that is no longer relevant. In Experiment 1, young and older adults were asked to recall lists of letters in the correct order. Half of the lists contained repeated items while half were control lists. Recall of nonadjacent repeated items was worse than that of control items. This Ranschburg effect was larger (i.e., greater response suppression) in older than in young adults. In Experiment 2, young and older adults were required either to recall the list or to report if there was a repeated item. Repetition detection was high and similar in the 2 age groups. When age differences in overall performance were taken into account, there was evidence of increased repetition inhibition with age in both experiments. Thus, contrary to the general reduced inhibition hypothesis, the specific process of response suppression during serial recall is not reduced by aging. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Control of respiration in newborn babies. IV. Rib cage stability and respiratory regulation

The recordings from an earlier study regarding the respiratory depth and rate changes induced by exposure to 4% CO2 in air in 13 babies with PM age varying between 32 and 43 weeks were reexamined with regard to the pattern of thoracic abdominal breathing excursion in breathing immediately prior to the CO2 exposure and the type of response induced. The pattern was called "stable" when the thoracic breathing excursions were in phase and congruent with the abdominal ones. When the thoracic excursions in comparison with the abdominal excursions were totally inverted, or incongruous but in phase, or rapidly varying between those two, the pattern was called "unstable". "Unstable" pattern of the breathing prior to the CO2 exposures was followed in an incidence of 60% by the type of response to CO2 which is characterized by a prompt rate increase (the "Type B" response) and only in 16% by the type characterized by an increased breathing amplitude (the "Type A" response). When the excursion pattern of the breathing prior to the CO2 exposures was "stable" "Type A" responses were induced in 59% and "Type B" responses in only 14%. The excursion pattern present when a baby is exposed to 4% CO2 thus seems to affect the type of respiratory depth and rate changes achieved. With increasing postmenstrual age the excursion pattern of the spontaneous breathing is more often "stable" and respiratory depth and rate changes of the "Type B" induced by CO2 less common. The variabilities of the breathing seen preferably in the preterm baby regarding regularity, rate and tidal volumes (as they could be approximated by the registration methods used) were noted most when the excursion pattern was "unstable". The results can be hypothetically interpreted to indicate a dynamic interaction between the thoracic wall and pulmonary mechanoreceptor systems of respiratory regulation. The decreasing variability of the breathing seen with increasing maturation in the baby could be explained by an increasing maturation of the neuromuscular ability to provide stability to the rib cage which would act stabilizing on the pulmonary vagal afferent input to the respiratory center.     

Role of Fc gamma RIIa (CD32) in IgG anti-RhD-mediated red cell phagocytosis in vitro

1. The influence of central inspiratory drive on heart rate variability was investigated in young human subjects using power spectral analysis of R-R intervals. 2. The area of the high-frequency component occurring at the respiratory frequency (0.2-0.25 Hz) in the power spectral density curves was used as an index of respiratory sinus arrhythmia. 3. Central inspiratory drive was increased by breathing a CO2-enriched (5%) gas mixture and this condition was compared with a similar degree of ventilation produced voluntarily. 4. Tests were conducted on eight young subjects with and without low-dose scopolamine (scopoderm TTS) in a double-blind cross-over trial. 5. Scopolamine decreased heart rate and increased the high-frequency peak, suggesting that its main action on the cardiac vagal pathway was a peripheral one, possibly increasing the efficacy of vagal impulses on the cardiac pacemaker. 6. With scopolamine, CO2 breathing increased the area of the high-frequency component significantly more than a similar degree of ventilatory movements produced by voluntary hyperventilation. 7. It is concluded that respiratory sinus arrhythmia in humans is at least partly dependent on a central respiratory-cardiac coupling, most probably similar to that shown in animal studies.