Anatomy of CNS opioid receptors.
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Animals↗
Biomedical subjects
H Akil
Publications and source records attributed to H Akil.
Paradoxical effect of corticosteroids on pituitary ACTH/beta-endorphin release in stressed animals.
We have previously demonstrated a number of changes in the anterior lobe proopiomelanocortin (POMC) system in chronically stressed rats. The purpose of the present experiments was to investigate whether chronically stressed rats demonstrate changes in pituitary sensitivity to glucocorticoid negative feedback. To study this question we compared the effects of glucocorticoids on ovine corticotropin releasing factor (oCRF)-stimulated ACTH and beta-endorphin release from cell suspensions prepared from naive unhandled rats versus chronically stressed rats. After dexamethasone, there was a 50% decrease in oCRF-stimulated hormone release in control rats but no inhibition of oCRF-stimulated hormone release in anterior lobe suspension from chronically stressed rats. Rather, the chronically stressed group exhibited a 50% hormone increase above the oCRF-stimulated baseline. The same pattern was observed after the addition of corticosterone to the medium. These findings suggest that there may be a positive feedback effect of glucocorticoids at the pituitary level under some conditions of chronic stress.
Adrenocorticotropic Hormone↗
Role of steric interactions in the delta opioid receptor selectivity of [D-Pen2, D-Pen5]enkephalin.
In order to assess the individual effects of each of the 3-methyl groups in residue 2 of [D-Pen2, D-Pen5]enkephalin on binding affinity to mu and delta opioid receptors, (2S,3S)methylcysteine ((3S)Me-D-Cys) and (2S,3R)methylcysteine ((3R)Me-D-Cys) were synthesized and incorporated into the analogs, [(3S)Me-D-Cys2, D-Pen5] enkephalin and [(3R)Me-D-Cys2, D-Pen5]enkephalin. Of these analogs, [(3S)Me-D-Cys2, D-Pen5]enkephalin appears from 1H n.m.r. spectra to assume a conformation similar to those of [D-Pen2, D-Pen5]enkephalin and the less delta receptor-selective, but more potent, [D-Cys2, D-Pen5]enkephalin. Assessment of binding affinity to mu and delta receptors revealed that [(3S)Me-D-Cys2, D-Pen5]enkephalin exhibits delta receptor affinity intermediate between [D-Pen2, D-Pen5]enkephalin and [D-Cys2, D-Pen5]enkephalin while its mu receptor affinity is similar to that of [D-Cys2, D-Pen5]enkephalin. These results suggest that, for [D-Pen2, D-Pen5]enkephalin, adverse steric interactions between the D-Pen2 pro-R methyl group and the mu receptor binding site lead to the low mu receptor binding affinity observed for this analog. By contrast, both the pro-R and pro-S D-Pen2 methyl groups lead to minor steric interactions which contribute to the somewhat lower delta receptor affinity of this compound.
Analgesics↗
Effects of low dose ovine corticotropin-releasing hormone in humans: endocrine relationships and beta-endorphin/beta-lipotropin responses.
The effects of low doses (0.03 and 0.1 microgram/kg) of ovine CRH (oCRH) on plasma beta-endorphin/beta-lipotropin (beta End/beta LPH), ACTH, and corticosteroid levels were studied in normal men. The 0.03 microgram/kg oCRH dose produced a reproducible response, with a rapid increase in plasma oCRH to peak levels between 45 and 95 fmol/mL and an appropriate doubling of plasma peptide and corticosteroid concentrations. The relationship between the corticosteroid rise and the rapid beta End/beta LPH and ACTH declines suggested negative feedback by corticosteroids on the release of these pituitary products. Plasma oCRH levels were proportionate to those reported in studies using much higher oCRH doses, and produced plasma oCRH levels in the reported range for the hypophyseal portal circulation. Molecular sieving of the beta End-immunoreactive materials in basal and post-oCRH (0.1 microgram/kg) plasma samples revealed an average basal beta End to beta LPH ratio of 1:1.5; 15 min after oCRH stimulation the average ratio was 4:1. We conclude that a low (threshold) dose of oCRH can reliably stimulate POMC peptide secretion and may preferentially release beta End from the anterior pituitary.
Adrenal Cortex Hormones↗
Regulation of hypothalamic magnocellular neuropeptides and their mRNAs in the Brattleboro rat: coordinate responses to further osmotic challenge.
A paradigm was developed for the chronic osmotic stimulation of homozygous diabetes insipidus rats of the Brattleboro strain, a strain that fails to synthesize vasopressin. This study examines the adaptation of 2 sets of coexisting peptide hormone magnocellular neurons in the hypothalamoneurohypophyseal system (HNS) of Long Evans (LE), Brattleboro heterozygote (HZ), and Brattleboro homozygote (DI) rats: (1) the arginine8-vasopressin (AVP)/dynorphin (DYN) neurons, and (2) the oxytocin (OT)/cholecystokinin (CCK8) neurons of the paraventricular and supraoptic nuclei, which project to the posterior pituitary. The regimen of chronic intermittent salt-loading (CISL) involved the replacement of 2% saline for normal drinking water for 18 hr/d. This protocol effectively increased plasma levels of AVP and OT in LE and HZ rats, oxytocin in DI rats, and maintained the posterior pituitary in a state depleted of AVP, OT, CCK, and peptides derived from pro-dynorphin: DYN A 1-17, DYN A 1-8, and DYN B 1-13. The ratio of pituitary DYN A 1-17 to DYN A 1-8 content in DI rats or in LE, HZ, and DI rats following 6 d of CISL suggests a preferential release of DYN A 1-17 during periods of chronic secretory activity. In response to chronic secretory activity, mRNAs for AVP, OT, DYN, and CCK increased 1.5-2-fold in all 3 AVP rat strains, with mRNAs for coexisting peptide hormones displaying parallel increases. Mutant AVP mRNA in the DI rat was expressed at very low levels and DYN mRNA in very high levels, with each of these mRNAs continuing to be regulated by CISL in a normal manner. These results suggest a regulatory relationship between AVP and OT neurons, in which vasopressin neurons are feedback-regulated by AVP, most likely via plasma osmolarity, and that oxytocin neurons are modulated by peptides derived from pro-dynorphin.
Animals↗
Effects of morphine treatment on endogenous opioid biosynthesis.
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Animals↗
A possible interface between autonomic function and pain control: opioid analgesia and the nucleus tractus solitarius.
Opioid peptides appear to be important neurochemical mediators in central nervous system mechanisms of analgesia, cardiovascular control, and many endocrinological responses to stress. The nucleus tractus solitarius (NTS), a brain region expressing all 3 opioid peptide families, is also associated with regulation of autonomic and endocrine functions. We now report that electrical stimulation of the NTS causes pronounced analgesia in rats. This analgesia appears to involve opioids and is pharmacologically dissociable from the hemodynamic changes elicited by NTS stimulation. These results suggest the NTS as a neural substrate for inter-relationships between stress, cardiovascular function, alterations in respiration, and pain sensitivity.
Analgesia↗
Postdexamethasone plasma cortisol and beta-endorphin levels in depression: relationship to severity of illness.
The hypothalamic-pituitary-adrenal (HPA) axis is dysregulated in many patients with depression, probably at all levels of the axis. To determine if HPA dysregulation is associated with severity of depression, we studied a group of 66 patients with major depressive disorder. Each patient underwent a pretreatment Dexamethasone Suppression Test, with plasma postdexamethasone cortisol determination at 8:00 AM, 4:00 PM, and 11:00 PM. All three postdexamethasone cortisol levels were significantly correlated with the Hamilton Rating Scale for Depression (HRSD) scores. We also examined the "profile" measures of mean, maximum, and minimum of the three cortisol values; again, all three were significantly correlated with HRSD scores. To evaluate associations between clinical severity and HPA dysregulation at the pituitary level, we studied a second group of 44 patients with major depressive disorder. Each had postdexamethasone cortisol determinations at 4:00 PM and 11:00 PM as well as pre- and postdexamethasone beta-endorphin determinations at 4:00 PM. The cortisol data from this group followed the same pattern as in the first sample, and there was a significant relationship between HRSD score and degree of beta-endorphin nonsuppression as well. These results suggest that severity of depression is one of the determinants of dysregulation at both adrenal and pituitary levels of the HPA axis, accounting for 10%-20% of the observed variance.
Adult↗
The effects of footshock stress on regional brain dopamine metabolism and pituitary beta-endorphin release in rats previously sensitized to amphetamine.
The repeated intermittent administration of amphetamine (AMP) produces an enduring enhancement in the response of dopamine (DA) systems in the brain to a subsequent "challenge" with amphetamine. However, former amphetamine addicts are not only hypersensitive to amphetamine, but also to "physical or psychological stress". This suggests that sensitization to amphetamine may change the response of DA neurons in brain to subsequent stress. To explore this idea, the effects of footshock stress on regional metabolism of DA in brain, and on the concentrations of plasma beta-endorphin and N-acetylated beta-endorphin, were studied in rats previously exposed to amphetamine or saline. It was found that: Prior treatment with amphetamine produced enduring (at least 7 days) changes in the dopaminergic response to footshock in the medial frontal cortex, hypothalamus, dorsolateral striatum and nucleus accumbens. Generally, rats pretreated with amphetamine showed a greater initial reduction in concentrations of DA in response to footshock, and a greater elevation in concentrations of metabolites of DA and/or metabolite/transmitter ratios, compared to nonhandled control rats. In some regions of the brain repeated injections of saline produced changes in the response to subsequent footshock that were comparable to those produced by amphetamine. Prior treatment with amphetamine enhanced the release of beta-endorphin and N-acetylated beta-endorphin from the pituitary elicited by footshock stress. It is concluded that repeated intermittent treatment with amphetamine or stress (injections of saline) produce enduring changes in the response of DA neurons and the pituitary to subsequent stress. These changes may be responsible for the hypersensitivity to stress reported in former amphetamine addicts, and in rats previously sensitized to amphetamine.
Amphetamine↗
Species-specific processing of prodynorphin in the posterior pituitary of mammals.
The steady state levels of the prodynorphin-derived opioid peptides, dynorphin A, dynorphin B, and alpha-neo-endorphin, have been extensively studied in the magnocellular/posterior pituitary system of the rat. To determine whether the rat system serves as a general model for prodynorphin processing in the mammalian posterior pituitary, we examined the steady state levels of prodynorphin-derived opioid peptides in the posterior pituitary systems of representatives of three diverse orders of mammals: guinea pig (order Rodentia), pig (order Artiodactyla), and rhesus monkey (order Primates). In each species studied there was evidence for species-specific deviations from the rat model. The most pronounced differences were observed with respect to the conversion of dynorphin A-(1-17) to dynorphin A-(1-8). In rodents, rats, and guinea pigs, under steady state conditions, the molar ratios of these forms are approximately 1:2 and 2.5:1, respectively. However, in the pig and rhesus monkey, the molar ratios of these forms are 10:1 and 14:1, respectively. Thus, under steady state conditions, the conversion of dynorphin A-(1-17) to dynorphin A-(1-8) appears to be a minor event in porcine and rhesus monkey posterior pituitary. Species-specific variations were also observed with respect to the steady state levels of alpha-neo-endorphin and dynorphin B-(1-13). In addition, the results of these studies suggest that the conversion of prodynorphin-derived opioids to leu-enkephalin probably represents a minor event in the species studied.
Animals↗
Functional aspects of multitransmitter neurons. Studies of interactions among pro-opiomelanocortin products.
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Adrenocorticotropic Hormone↗
Autoradiographic differentiation of mu, delta, and kappa opioid receptors in the rat forebrain and midbrain.
While there is an abundance of pharmacological and biochemical evidence to suggest the existence of multiple opioid receptors, their precise localization within the brain is unclear. To help clarify this issue, the present study examined the distributions of the mu, delta, and kappa opioid receptor subtypes in the rat forebrain and midbrain using in vitro autoradiography. Mu and delta receptors were labeled with the selective ligands 3H-DAGO (Tyr- D-Ala-Gly-MePhe-Gly-ol), and 3H-DPDPE (D-Pen2, D-Pen5-enkephalin), respectively, while the kappa receptors were labeled with 3H-(-)bremazocine in the presence of unlabeled DAGO and DPDPE. Based on previous findings in our laboratory, the labeling conditions were such that each ligand selectively occupied approximately 75% of each of the opioid sites. The results demonstrated that all 3 opioid receptor subtypes were differentially distributed in the rat brain. Mu binding was dense in anterior cingulate cortex, neocortex, amygdala, hippocampus, ventral dentate gyrus, presubiculum, nucleus accumbens, caudate putamen, thalamus, habenula, interpeduncular nucleus, pars compacta of the substantia nigra, superior and inferior colliculi, and raphe nuclei. In contrast, delta binding was restricted to only a few brain areas, including anterior cingulate cortex, neocortex, amygdala, olfactory tubercle, nucleus accumbens, and caudate putamen. Kappa binding, while not as widespread as observed with mu binding, was densely distributed in the amygdala, olfactory tubercle, nucleus accumbens, caudate putamen, medial preoptic area, hypothalamus, median eminence, periventricular thalamus, and interpeduncular nucleus. While all 3 opioid receptor subtypes could sometimes be localized within the same brain area, their precise distribution within the region often varied widely. For example, in the caudate putamen, mu binding had a patchy distribution, while delta and kappa sites were diffusely distributed, with delta sites being particularly dense ventrolaterally and kappa sites being concentrated ventromedially. These results support the existence of at least 3 distinct opioid receptors with possibly separate functional roles.
Animals↗
Pharmacological and anatomical evidence of selective mu, delta, and kappa opioid receptor binding in rat brain.
While the distribution of opioid receptors can be differentiated in the rat central nervous system, their precise localization has remained controversial, due, in part, to the previous lack of selective ligands and insensitive assaying conditions. The present study analyzed this issue further by examining the receptor selectivity of [3H]DAGO (Tyr-D-Ala-Gly-MePhe-Gly-ol), [3H]DPDPE (2-D-penicillamine-5-D-penicillamine-enkephalin), [3H]DSLET (Tyr-D-Ser-Gly-Phe-Leu-Thr) and [3H](-)bremazocine, and their suitability in autoradiographically labelling selective subpopulations of opioid receptors in rat brain. The results from saturation, competition, and autoradiographic experiments indicated that the three opioid receptor subtypes can be differentiated in the rat brain and that [3H]DAGO and [3H]DPDPE selectively labelled mu and delta binding sites, respectively. In contrast, [3H]DSLET was found to be relatively non-selective, and labelled both mu and delta sites. [3H]Bremazocine was similarly non-selective in the absence of mu and delta ligands and labelled all three opioid receptor subtypes. However, in the presence of 100 nM DAGO and DPDPE, concentrations sufficient to saturate the mu and delta sites, [3H]bremazocine did label kappa sites selectively. The high affinity [3H]bremazocine binding sites showed a unique distribution with relatively dense kappa labelling in the hypothalamus and median eminence, areas with extremely low mu and delta binding. These results point to the selectivity, under appropriate conditions, of [3H]DAGO, [3H]DPDPE and [3H]bremazocine and provide evidence for the differential distribution of mu, delta, and kappa opioid receptors in rat brain.
Animals↗
Immunocytochemical localization of pro-opiomelanocortin-derived peptides in the adult rat spinal cord.
A dispersed descending pro-opiomelanocortin (POMC) fiber system has been demonstrated by peroxidase-antiperoxidase (PAP) immunocytochemistry in the adult rat spinal cord. beta-endorphin, adrenocorticotrophic hormone (ACTH), alpha-melanocyte-stimulating hormone (alpha-MSH) and 16K immunoreactive fibers exist in the spinal cord from cervical down to sacral level. Descending fibers running parallel in the dorsolateral and lateral funiculus send collaterals ventromedially or medially to terminate in the gray matter surrounding the central canal, where nociceptive neurons have recently been located, in addition to those nociceptive cells in the dorsal horn. After spinal transection at lower thoracic level, POMC peptide immunoreactivities disappeared below the lesion. Moreover, no POMC cell bodies were found in the spinal cord. Therefore, the descending fibers are most likely of supraspinal origin.
Adrenocorticotropic Hormone↗
Characterization of the forms of beta-endorphin and alpha-MSH in the caudal medulla of the rat and guinea pig.
The post-translational processing of pro-opiomelanocortin (POMC) in brain remains controversial. Classically, there was thought to be a single cell group in the arcuate nucleus with long projections through limbic structures. More recently, a second cell group was discovered in the caudal medulla. This study addresses the question of POMC processing in this region. Steady-state analysis of acid extracts of dorsal caudal medulla from rat and guinea pig CNS by gel filtration chromatography and radioimmunoassay indicated that in both species the major POMC-related end products are alpha-MSH-sized material and beta-endorphin-sized. In this tissue beta-LPH and ACTH represent minor end products. Analysis of the alpha-MSH-sized material from both species by reverse-phase HPLC indicated that in the rat caudal medulla approximately 79% of the alpha-MSH-related material is acetylated, and in the guinea pig caudal medulla approximately 85% of the alpha-MSH-related material is acetylated. Analysis of the forms of beta-endorphin isolated from the rat caudal medulla by cation exchange chromatography revealed that acetylated and non-acetylated forms of beta-endorphin are present in this region of the rat CNS. Approximately 65% of the beta-endorphin in the rat caudal medulla is N-acetylated. Analysis of the forms of beta-endorphin isolated from the guinea pig caudal medulla indicated that approximately 63% of the beta-endorphin is N-acetylated in this region of the guinea pig CNS. These data indicate that the post-translational processing of POMC in the dorsal caudal medulla, the site of the nucleus tractus solitarius POMC cell group, is distinct from the processing patterns that have been reported for POMC systems in the mammalian anterior pituitary, intermediate pituitary and arcuate nucleus.
Acetylation↗
[3H]dynorphin A binding and kappa selectivity of prodynorphin peptides in rat, guinea-pig and monkey brain.
We have previously demonstrated that [3H]dynorphin A selectively labels kappa opioid receptors in guinea-pig whole brain. In these current studies, using protection from inactivation by beta-chloronaltrexamine (beta-CNA), we are able to demonstrate that although dynorphin A prefers kappa receptors, it will label mu receptors when kappa receptors are not available, or present in only a small number. Thus, differences in numbers of mu and kappa receptors present in brain preparations are critical in determining the receptor binding profile of [3H]dynorphin A across species. Additionally, although all the prodynorphin derived peptides show kappa preference, the ability of the other prodynorphin derived peptides to compete with [3H]dynorphin A for its receptor varies across species. Consequently, in a highly enriched kappa preparation such as monkey cerebral cortex, [3H]dynorphin A appears to label kappa receptors with substantial selectivity, and the other prodynorphin-derived peptides show less ability to compete with dynorphin A for its receptor. In contrast, in a kappa-poor tissue such as rat brain, all of the prodynorphin-derived peptides, including dynorphin A-(1-8), show very similar potency. Thus, differences in mu and kappa receptor numbers across brain regions and species lead to differences in the receptor binding profile of dynorphin A.
Alkylating Agents↗
beta-Endorphin/beta-lipotropin immunoreactivity in endogenous depression. Effect of dexamethasone.
This study addresses the question of whether pituitary peptides (ie, beta-endorphin) show regulatory disruption in endogenous depression and, if so, does it co-occur in the same subjects who show cortisol dysregulation. Endogenously depressed patients and psychiatric controls from three centers were evaluated, when not taking medications, and studied for plasma cortisol and beta-endorphin levels. Plasma samples were taken at four time points over one hour, on the basal day, and 16 hours after 1 mg of dexamethasone. From 33% to 69% of the endogenous patients were abnormal in their postdexamethasone cortisol levels, and from 50% to 69% were abnormal on postdexamethasone beta-endorphin values (vs 0% and 8%, respectively, for controls). When endogenous subjects were evaluated for abnormality on both cortisol and beta-endorphin, after dexamethasone, it was found that the two measures of hypothalamic-pituitary-adrenal dysfunction did not necessarily co-vary. In fact when having either abnormal beta-endorphin or cortisol levels (or both) was used as a biological marker a larger number of the endogenous patients were detected than with either measure alone. Our conclusions are as follows: Plasma beta-endorphin shows a circadian rhythm similar to that seen with corticotropin (ACTH) and is suppressable by dexamethasone. In many endogenous patients plasma beta-endorphin levels escape from dexamethasone suppression. Many of these subjects are not cortisol escapers. When abnormality of either the beta-endorphin or cortisol is considered it is clear that both levels of the hypothalamic-pituitary-adrenal axis can be dysregulated in endogenous depression.
Adolescent↗