Corticotropin-releasing Factor Receptor Subtype Research Articles

Stress-mediated heart rate dynamics after deletion of the gene encoding corticotropin-releasing factor receptor 2.

The objective of the present study was to investigate the potential role of corticotropin-releasing factor receptor subtype 2 (CRFR2) in autonomic regulation of heart rate and heart rate variability under physiological conditions in conscious mice. Heart rate dynamics during novelty exposure and auditory fear conditioning were assessed by radiotelemetry. Heart rate responses and heart rate variability values were not different in CRFR2+/+ and CRFR2-/- mice during novelty exposure, which was associated with similar locomotor activity exhibited by both genotypes. The heart rate responses during retention of conditioned auditory fear were similar and the exponential relationship between heart rate and heart rate variability was independent of genotype. Pharmacological stimulation of the peripheral CRFR2beta by intraperitoneal injection of 200 ng human/rat corticotropin-releasing factor yielded a sustained tachycardia in wildtype control (CRFR2+/+) mice which was absent in CRFR2-deficient (CRFR2-/-) mice. Similarly, the tachycardia was effectively blocked by preinjection of the CRFR2 antagonist antisauvagine-30. In conclusion, the results indicate the involvement of CRFR2 in heart rate dynamics upon pharmacological stimulation but demonstrate that CRFR2 is not involved in baseline heart rate regulation and stress-mediated modulation of heart rate responses.

Dopamine-dependent responses to cocaine depend on corticotropin-releasing factor receptor subtypes.

The effects on locomotor response to cocaine challenge, acquisition of cocaine conditioned place preference and cocaine-induced dopamine (DA) release in nucleus accumbens and ventral tegmental area by the non-specific corticotropin-releasing factor (CRF) receptors antagonist alpha-helical CRF, the selective CRF receptor subtype 1 antagonist CP-154,526 and the selective CRF receptor subtype 2 antagonist anti-sauvagine-30 (AS-30) were investigated in rats. Both alpha-helical CRF (10 microg, i.c.v.) and CP-154,526 (3 microg, i.c.v.) decreased the cocaine-induced distance travelled, whereas AS-30 (3 microg, i.c.v.) did not show such an effect. The CRF receptor antagonists also have significant effects on stereotype counts induced by cocaine injection, in which the alpha-helical CRF or CP-154,526 but not AS-30 did significantly reduce the stereotype counts. alpha-Helical CRF (10 microg) prior to each injection of cocaine blocked cocaine conditioned place preference with no significant difference observed in the time spent in the drug-paired side between post- and pre-training and both 1 and 3 microg CP-154,526 also had significant inhibitory effects on cocaine-induced place preference. However, pre-treatment with an i.c.v. infusion of AS-30 (1 or 3 microg) prior to each injection of cocaine did not affect the acquisition of conditioned place preference. The alpha-helical CRF and CP-154,526 reduced extracellular DA levels of nucleus accumbens and ventral tegmental area in response to the injection of cocaine. However, both alpha-helical CRF and CP-154,526 did not modify extracellular DA levels under basal conditions. In contrast, the i.c.v. infusion of AS-30 had no effects on either the basal DA or the cocaine-induced increase in DA release in nucleus accumbens and ventral tegmental area. These findings demonstrate that activation of the CRF receptor is involved in behavioral and neurochemical effects of cocaine challenge and cocaine reward and that the role of CRF receptor subtypes 1 and 2 in cocaine-induced locomotion, reward and DA release is not identical. The CRF receptor subtype 1 is largely responsible for the action of the CRF system on cocaine locomotion and reward. These results suggest that the CRF receptor antagonist, particularly the CRF receptor subtype 1 antagonist, might be of some value in the treatment of cocaine addiction and cocaine-related behavioral disorders.

Corticotropin-releasing factor (CRF) rapidly suppresses apoptosis by acting upstream of the activation of caspases.

The physiological role of the corticotropin-releasing factor (CRF) family of peptides has recently been extended by emerging evidence of their cytoprotective effects. To determine whether CRF-mediated cytoprotection is linked to caspase-dependent apoptosis, the effect of CRF on the activation of caspases was investigated in detail in Y79 human retinoblastoma cells. The results presented here demonstrate that the cytoprotective effect of CRF against the actions of camptothecin (CT) was mediated by CRF receptor subtype 1, but not subtype 2. The observed CRF-mediated cytoprotection involved rapid and pronounced suppression of proteolytic processing and activation of procaspase-3, exerted even when CRF was added hours after the application of the cytotoxic agent. Surprisingly, activation of procaspase-3 preceded activation of the initiator procaspases 2, 8, 9 and 10 during CT-induced apoptosis of Y79 cells. The mechanism of the effect of CRF was examined using inhibitors of signalling pathways such as Wortmannin (Akt), cyclic AMP-dependent protein kinase (PKA), extracellular signal-regulated kinase (ERK), protein kinase c (PKC), p38 mitogen-activated protein kinase (p38 MAPK), phospholipase c (PLC), nuclear factor-kappaB (NF-kappaBeta) and c-jun N-terminal kinase (JNK). The involvement of PKA in the mediation of the anti-apoptotic effect of CRF has been established. Taken together, these results demonstrate for the first time that the cytoprotective effect of CRF involved suppression of pro-apoptotic pathways at a site upstream of activation of procaspase-3.

Effect of chronic administration of a CRF 1 receptor antagonist, CRA1000, on locomotor activity and endocrine responses to stress

Corticotropin-releasing factor (CRF) is involved in the regulation of stress responses. The actions of CRF in the brain are mediated through two distinct CRF receptor subtypes, CRF 1 and CRF 2 receptors. In the present study, we examined the effects in rat of chronic administration of a nonpeptidic CRF 1 receptor-selective antagonist, CRA1000, 2-[ N-(2-methylthio-4-isopropylphenyl)- N-ethylamino]-4-[4-(3-fluorophenyl)-1,2,3,6-tetrahydropyridin-1-yl]-6-methylpyrimidine), on locomotor activity, feeding behavior and the hypothalamic–pituitary–adrenal axis. Chronic CRA1000 treatment significantly decreased locomotor activity in the dark phase of the diurnal cycle. However, chronic CRA1000 treatment showed no effect on food and water intake, or on body weight. After a 10-day period of CRA1000 treatment, plasma concentrations of adrenocorticotropic hormone (ACTH) and corticosterone in basal conditions and under immobilization stress were no different from those in rats treated with vehicle. However, CRA1000 administered 2 h before immobilization stress significantly reduced ACTH and corticosterone responses to stress with no effect on basal ACTH and corticosterone concentrations. These results suggest that CRF 1 receptors are involved in the regulation of locomotor activity during the dark period, but are not involved in the regulation of feeding behavior under non-stressful conditions. Furthermore, the results suggest that a 10-day treatment with CRA1000 does not affect hypothalamic–pituitary–adrenal axis activity either under basal conditions or after acute stress.

Fractal dynamics of heart beat interval fluctuations in corticotropin-releasing factor receptor subtype 2 deficient mice.

Non-linear fractal analysis of cardiac interbeat time series was performed in corticotropin-releasing factor receptor subtype 2 (CRFR2) deficient mice. Heart rate dynamics in mice constitutes a self-similar, scale-invariant, random fractal process with persistent intrinsic long-range correlations and inverse power-law properties. We hypothesized that the sustained tachycardic response elicited by intraperitoneal (ip) injection of human/rat CRF (h/rCRF) is mediated by CRFR2. In wildtype control animals, heart rate was increased to about maximum levels (approximately 750 bpm) while in CRFR2-deficient animals baseline values were retained (approximately 580 bpm). The tachycardic response elicited by ip-application is mediated by CRFR2 and is interpreted to result from sympathetic stimulation. However, the functional integrity of CRFR2 would not present a prerequisite to maintaining the responsiveness and resiliency of cardiac control to external environmental perturbations experimentally induced by extrinsic ip-application of h/rCRF or under physiological conditions that may be associated with an increased peripheral release of CRF. Under stressful physiological conditions achieved by novelty exposure, CRFR2 is not involved in the cardiodynamic regulation to external short-term stress. While the hypothesis of involvement of CRFR2 in cardiac regulation upon pharmacological stimulation cannot be rejected, the present findings suggest that the mechanism of action is by sympathetic stimulation, but would not unambiguously allow to draw any conclusions as to the physiological role of CRFR2 in the control of cardiac dynamics.

Non-peptidic CRF1 receptor antagonists for the treatment of anxiety, depression and stress disorders.

Anxiety and depression are psychiatric disorders that constitute a major health concern worldwide, and new pharmacological approaches with the potential for improved efficacy and decreased side effect profiles relative to currently marketed drugs are desired. Since the identification of corticotropin releasing factor (CRF) by Vale and colleagues in 1981, an extensive research effort has solidified the importance of this 41 amino acid peptide in mediating the body's behavioral, endocrine, and autonomic responses to stress. The further identification of CRF receptor subtypes has provided compelling targets for novel pharmaceutical agents. The present review focuses on the potential of non-peptidic antagonists of the CRF(1) receptor subtype as a novel therapeutic approach for the treatment of anxiety and depression. The first section reviews preclinical and clinical evidence implicating CRF, in general, and CRF(1) receptors, in particular, in anxiety and depression. Clinical studies have demonstrated a dysfunctional hypothalamic-pituitary-adrenal (HPA) axis and/or elevated CRF levels in depression and in some anxiety disorders. Preclinical data utlilizing correlational methods, genetic models, and exogenous CRF administration techniques in rodents and non-human primates supports a link between hyperactive CRF pathways and anxiogenic and depressive-like symptoms. Studies employing the use of receptor knockouts and selective, non-peptidic antagonists of the CRF(1) receptor have demonstrated anxiolytic and antidepressant effects under certain types of laboratory conditions. A Phase II, open-label, clinical trial in major depressive disorder has reported that a CRF(1) receptor antagonist was safe and effective in reducing symptoms of anxiety and depression. In the second section, a topological approach is used to describe the design strategies employed to produce potent, non-peptidic CRF(1) receptor antagonists. Two main topologies, featuring a center core, a top side-chain, and a pending aromatic ring, can be used to characterize the vast majority of currently known CRF(1) receptor antagonists. By exploiting some of these structural elements, pharmacological, physicochemical, and pharmacokinetic properties can be modulated and optimized. However, as a result of a relatively conservative iteration process during the structural optimization, the chemical space presently defined by the existing CRF(1) receptor antagonists still remains fairly narrow. Expanding these structural and topological boundaries, while optimizing the "drug-like" properties of the CRF(1) receptor antagonists, seems to be a common objective across pharmaceutical companies to maximize the chances for a clinical success in the near future.

Urocortin shares the memory modulating effects of corticotropin-releasing factor (CRF): mediation by CRF 1 receptors

Intracerebroventricular (i.c.v.) administration of corticotropin-releasing factor (CRF) biphasically affects performance in tests of learning and memory. In the present study, we used CRF, urocortin (Ucn), a recently cloned CRF homologue, and CRF receptor antagonists, to determine which CRF receptor subtype(s) mediate the memory modulating effects of CRF receptor agonists in male Wistar rats. Under difficult learning conditions (massed trials), i.c.v. pretreatment with CRF or Ucn facilitated the acquisition of spatial navigation in the Morris water maze in a non-dose-dependent fashion (optimal doses of 0.1 and 0.03 μg, respectively). Under less difficult learning conditions (spaced trials), both peptides impaired water maze performance. In addition, with i.c.v. posttraining treatment, the peptides were equipotent (1.0 μg) in facilitating the consolidation of passive avoidance learning. The performance-enhancing effects of Ucn in both water maze and passive avoidance paradigms were reversed by i.c.v. pretreatment with d-Phe CRF 12–41 (2.5, 5 μg), a broad CRF 1/CRF 2 receptor antagonist, or antalarmin (10 μg), a potent, nonpeptide, CRF 1 selective receptor antagonist. Thus, Ucn shares CRF’s memory-modulating effects, and these effects appear to be mediated via the CRF 1 receptor. These findings are consistent with the hypothesis that CRF receptor agonists affect performance in tests of learning and memory by increasing arousal.

Pharmacological evidence supporting a role for central corticotropin-releasing factor(2) receptors in behavioral, but not endocrine, response to environmental stress.

Corticotropin-releasing factor (CRF) is one of the principle components of the stress response. The physiological effects of CRF are mediated by two receptor subtypes, CRF(1) and CRF(2). Recent data obtained with the selective CRF(2) antagonist antisauvagine-30 (ASV-30) has begun to suggest that both CRF receptor subtypes may play a role in stress-related behaviors. Exactly how these two receptor subtypes interact to modulate the behavioral and endocrine responses to stress is not clear, however. We have attempted to understand the role of the CRF(2) receptor in the behavioral and endocrine responses to stress by comparing the effects of ASV-30 with the mixed CRF(1)/CRF(2) receptor antagonist astressin. Centrally administered ASV-30 reduced anxiety-like behavior in BALB/c mice in three models of anxiety: marble burying [minimal effective dose (MED) = 3 nmol], open field (MED = 3 nmol), and elevated plus maze (MED = 0.1 nmol). ASV-30 did not change locomotor activity or the adrenocorticotropic hormone (ACTH) response to restraint stress. The potent mixed CRF(1)/CRF(2) antagonist astressin not only reduced anxiety-like behavior in all three models with equivalent potency but also blunted the ACTH response to restraint stress. Finally, the new selective CRF(2) receptor agonist urocortin-II produced a dose-dependent increase in anxiety-like behavior in the plus maze test. Therefore, our data suggest that the CRF(2) receptor plays a role in the behavioral, but not the hypothalamic-pituitary-adrenal axis, response to stress.

Differential actions of peripheral corticotropin-releasing factor (CRF), urocortin II, and urocortin III on gastric emptying and colonic transit in mice: role of CRF receptor subtypes 1 and 2.

Peripheral CRF inhibits gastric emptying and stimulates colonic motor function in rats. We investigated the role of CRF(1) and CRF(2) receptors in i.p. CRF-induced alterations of gut transit in conscious mice using selective CRF(1) and CRF(2) ligands injected i.p. Gastric emptying 2 h after ingestion of a solid chow meal and colonic transit (time to expel a bead inserted into the distal colon) were determined simultaneously. Rat/human (r/h)CRF, which has CRF(1) > CRF(2) binding affinity, decreased distal colonic transit time at lower doses (6-12 microg/kg) than those inhibiting gastric emptying (20-60 microg/kg). Ovine CRF, a preferential CRF(1) receptor agonist (6-60 microg/kg), reduced significantly the colonic transit time without altering gastric emptying, whereas the selective CRF(2) receptor agonists mouse urocortin II (20-60 microg/kg) and urocortin III (120 microg/kg) inhibited significantly gastric emptying without modifying colonic transit. The CRF(1)/CRF(2) receptor antagonist, astressin (30-120 microg/kg), dose dependently prevented r/hCRF (20 microg/kg)-induced inhibition of gastric emptying and reduction of colonic transit time. The selective CRF(1) receptor antagonists, NBI-27914 (C(18)H(20)Cl(4)N(4)C(7)H(8)O(3)S) and CP-154,526 (butyl-[2,5-dimethyl-7-(2,4,6-trimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]ethylamine) (5-30 mg/kg), dose dependently blocked r/hCRF action on the colon without influencing the gastric response, whereas the CRF(2) receptor antagonist, antisauvagine-30 (30-100 microg/kg), dose dependently abolished r/hCRF-induced delayed gastric emptying and had no effect on colonic response. These data show that i.p. r/hCRF-induced opposite actions on upper and lower gut transit in conscious mice are mediated by different CRF receptor subtypes: the activation of CRF(1) receptors stimulates colonic propulsive activity, whereas activation of CRF(2) receptors inhibits gastric emptying.

Differential responsiveness of CRF receptor subtypes to N-terminal truncation of peptidic ligands

The role of the N-terminal domains of corticotropin-releasing factor (CRF) and CRF-like peptides in receptor subtype selectivity, ligand affinity and biological potency was investigated. Therefore, human CRF 12–41, human URP 12–38 and antisauvagine-30 (aSvg) were N-terminally prolonged by consecutive addition of one or two amino acids. The peptides obtained were tested for their binding affinities to rat CRF 1 and murine CRF 2β receptor, and their capability to stimulate cAMP-release by HEK cells producing either receptor. It was observed that human CRF N-terminally truncated by eight residues was bound with high affinity to CRF 2 receptor ( K i =5.4 nM), whereas affinity for CRF 1 receptor was decreased ( K i =250 nM). A similar shift of affinity was found with sauvagine (Svg) analogs. Truncation of human URP analogs did not affect their preference for CRF 2β receptor, but reduced their affinity. Changes in affinity were positively correlated with changes in potency. These results indicated that CRF 1 receptor was more stringent in its structural requirements for ligands to exhibit high affinity binding than CRF 2β receptor.

Expression of urocortin and corticotropin-releasing factor receptor subtypes in the human heart.

Urocortin (Ucn) is a new member of the corticotropin-releasing factor (CRF) neuropeptide family and has positive inotropic actions and protective effects against ischemia in the rat heart. Ucn binds with very high affinity to both CRF receptor type 1 (CRF-R1) and CRF receptor type 2 (CRF-R2). However, to date, endogenous ligand(s) for CRF receptors expressed in the human heart have yet to be elucidated. In this study, we therefore examined the expression of Ucn and CRF receptors in human heart obtained at autopsy by RT-PCR, immunohistochemistry, and RIA. RT-PCR analysis demonstrated that Ucn and CRF-R2alpha mRNAs were detected in all four chambers. CRF-R1 mRNA was weakly present in some left atria, left ventricles, and in one right ventricle. CRF-R2beta mRNA was detected predominantly in the left atrium. CRF mRNA was not detected in any of the four chambers. Immunostaining for both Ucn and CRF receptors was detected in cardiac myocytes in all four chambers. Ucn-like immunoreactivity was detected in all four chambers by RIA, with the highest concentrations in the left ventricle (1.90 +/- 0.5 pmol/g wet weight, mean +/- SEM; n = 4). On the other hand, CRF-like immunoreactivity was very low or undetectable in the human heart. Sephadex G-50 column chromatography demonstrated that most of the Ucn-like immunoreactivity in the human heart was eluting earlier than the standard Ucn, with one minor peak in the position for Ucn. Ucn immunoreactivity was not detected in skeletal muscle by immunohistochemistry or RIA. These results suggest that Ucn is produced in the human heart and stored there mainly in the larger molecular weight forms. Endogenously produced Ucn may therefore exert its effects mostly through CRF-R2 in an autocrine and/or paracrine manner in the human heart.

Expression of Urocortin and Corticotropin-Releasing Factor Receptor Subtypes in the Human Heart

Expression of Urocortin and Corticotropin-Releasing Factor Receptor Subtypes in the Human Heart

Peripheral urocortin inhibits gastric emptying and food intake in mice: differential role of CRF receptor 2.

Intraperitoneal urocortin inhibits gastric emptying and food intake in mice. We investigated corticotropin-releasing factor receptor (CRF-R) subtypes involved in intraperitoneal urocortin actions using selective CRF-R antagonists. Gastric emptying was measured 2 h after a chow meal, and food intake was measured hourly after an 18-h fast in mice. Urocortin (3 microg/kg ip) inhibited gastric emptying by 88%. The CRF-R1/CRF-R2 antagonist astressin B (30 microg/kg ip) and the selective CRF-R2 antagonist antisauvagine-30 (100 microg/kg ip) completely antagonized urocortin action, whereas the selective CRF-R1 antagonist CP-154,526 (10 mg/kg ip) had no effect. Urocortin (1-10 microg/kg ip) dose dependently decreased the 2-h cumulative food intake by 30-62%. Urocortin (3 microg/kg)-induced hypophagia was completely antagonized by astressin B (30 microg/kg ip) and partially (35 and 31%) by antisauvagine-30 (100 or 200 microg/kg ip). The CRF-R1 antagonists CP-154,526 or DMP904 (10 mg/kg ip) had no effect. Capsaicin did not alter urocortin-inhibitory actions while blocking the satiety effect of intraperitoneal CCK. These data indicate that intraperitoneal urocortin-induced decrease in feeding is only partly mediated by CRF-R2, whereas urocortin action to delay gastric emptying of a meal involves primarily CRF-R2.

Involvement of corticotropin-releasing factor receptor subtype 1 in morphine withdrawal regulation of the brain noradrenergic system

Effects of pretreatment with the selective corticotropin-releasing factor (CRF) subtype 1 (CRF 1) receptor antagonist, 2-( N-(2-methylthio-4-isopropylphenyl)- N-ethyl-amino-4-(4-(3-fluorophenyl)-1,2,3,6-tetrahydropyridin-1-yl)-6-methylpyrimidine (CRA1000) on the behavioral and biochemical changes after naloxone-precipitated morphine withdrawal were examined in ICR mice. Mice were chronically treated with morphine (8–45 mg/kg) for 5 days. Naloxone (3 mg/kg, s.c.) precipitated jumping, diarrhea, and body weight loss in morphine-dependent mice. In addition, 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG) and noradrenaline turnover (MHPG/noradrenaline) levels in the cerebral cortex were increased following naloxone challenge in morphine-dependent mice. However, 5-hydroxytriptamine turnover did not alter the increase following naloxone challenge in morphine-dependent mice. Pretreatment with CRA1000 (20 mg/kg, i.p.) attenuated the incidence of withdrawal signs and naloxone-precipitated increases in noradrenaline turnover. These results suggest that the activation of CRF 1 receptor may play an important role in the elevation of noradrenaline transmission, but not in 5-hydroxytriptamine transmission, in the cerebral cortex, which projects from the locus coeruleus during morphine withdrawal.

Pharmacological and chemical properties of astressin, antisauvagine-30 and α-helCRF: significance for behavioral experiments

Corticotropin releasing factor (CRF) represents an early chemical signal in the stress response and modulates various brain functions through G protein-coupled receptors. Two CRF receptor subtypes, CRF 1 and CRF 2, have been identified. Since the physicochemical properties of CRF receptor antagonists might influence their biological potency, the peptidic antagonists astressin, α-helical CRF 9–41 (α-helCRF) and antisauvagine-30 (aSvg-30) have been analyzed. The rank order of solubility of these compounds in artificial cerebrospinal fluid (aCSF, pH 7.4) was aSvg-30>α-helCRF>>astressin, whereas the rank order of relative lipophilicity as determined with RP–HPLC was α-helCRF>astressin>aSvg-30. The calculated isoelectric points were 4.1 (α-helCRF), 7.4 (astressin) and 10.0 (aSvg-30). According to Schild analysis of the CRF receptor-dependent cAMP production of transfected HEK cells, aSvg-30 exhibited a competitive antagonism and displayed a 340 fold selectivity for mCRF 2β receptor. For astressin, however, the pharmacodynamic profile could not be explained by a simple competitive mechanism as indicated by Schild slopes >1 for rCRF 1 or mCRF 2β receptor. Behavioral experiments demonstrated that after i.c.v. injection, α-helCRF reduced oCRF-induced anxiety-like behavior in the elevated plus-maze, whereas astressin, despite its higher in vitro potency, did not. These findings could be explained by different physicochemical properties of the antagonists employed.

Corticotropin-releasing factor receptor type 1 mediates stress-induced relapse to cocaine-conditioned place preference in rats

Corticotropin-releasing factor (CRF) has been suggested to play an important role in the development of drug dependence and withdrawal. Based on the recent finding that CRF receptor antagonists inhibit the stress-induced relapse to opiate dependence and attenuate anxiety-like responses related to cocaine withdrawal, the present experiment was performed to examine the possible effect of different CRF receptor antagonists on reactivation of cocaine-conditioned place preference induced by cocaine and stress in rats. The results show that a single injection of cocaine (10 mg/kg, i.p.) could reactivate cocaine-conditioned place preference following a 28-day extinction, and pretreatment with i.c.v. 10 μg α-helical CRF, a nonspecific CRF receptor antagonist, significantly attenuated this reactivation of conditioned place preference. However, pretreatment with i.p. 1 or 10 mg/kg CP-154,526 (butyl-[2,5-dimethyl-7-(2,4,6-trimethylphenyl)-7 H-pyrrolo[2,3- d]pyrimidin-4-yl]-ethylamine), a specific CRF receptor subtype 1 antagonist, and i.c.v. 1 or 10 μg AS-30 ([ d-Phe 11,His 12]Svg-(11-40)), a specific CRF receptor subtype 2 antagonist, failed to show the same effects. In addition, a single footshock stress also elicited the reactivation of cocaine-conditioned place preference following a 28-day extinction and pretreatment with α-helical CRF (10 μg, i.c.v.) and CP-154,526 (1 or 10 mg/kg, i.p.) significantly blocked this effect. In contrast, pretreatment with AS-30 at a dose of 1 or 10 μg (i.c.v.) did not affect the stress-induced reactivation of cocaine-conditioned place preference. The present study demonstrated that CRF receptor type 1, but not CRF receptor type 2, mediates the stress-induced reactivation of cocaine-conditioned place preference. These findings suggest that CRF receptor subtype 1 antagonists might be of some value in the treatment and prevention of stress-induced relapse to drug dependence long after detoxification.

Stress and the gastrointestinal tract III. Stress-related alterations of gut motor function: role of brain corticotropin-releasing factor receptors.

Alterations of gastrointestinal (GI) motor function are part of the visceral responses to stress. Inhibition of gastric emptying and stimulation of colonic motor function are the commonly encountered patterns induced by various stressors. Activation of brain corticotropin-releasing factor (CRF) receptors mediates stress-related inhibition of upper GI and stimulation of lower GI motor function through interaction with different CRF receptor subtypes. CRF subtype 1 receptors are involved in the colonic and anxiogenic responses to stress and may have clinical relevance in the comorbidity of anxiety/depression and irritable bowel syndrome.

Localization and Characterization of a Short Isoform of the Corticotropin-Releasing Factor Receptor Type 2α (CRF2α-tr) in the Rat Brain

We have recently isolated a cDNA encoding a short isoform of the corticotropin-releasing factor (CRF) receptor subtype, referred to as CRF2α-tr, from the rat amygdala. The present study determined the localization of the truncated receptor mRNA in the rat brain by in situ hybridization histochemistry. The results showed significant levels of hybridization in the lateral septum, central nucleus of the amygdala, cortico-amygdaloid nucleus, ventromedial nucleus of the hypothalamus (VMH), and frontal cortex. In the physiological study, antidepressive drugs increased the expression of CRF2α-tr mRNA and the total binding activity to CRF in the rat amygdala. These findings suggest that CRF2α-tr may regulate endogeneous CRF release in the amygdala.

Differential roles of corticotropin‐releasing factor receptor subtypes 1 and 2 in opiate withdrawal and in relapse to opiate dependence

The possible effects on the morphine withdrawal signs of the nonspecific corticotropin-releasing factor (CRF) receptor antagonist alpha-helical CRF, the selective CRF receptor subtype 1 antagonist CP-154,526 and the selective CRF receptor subtype 2 antagonist antisauvagine-30 (AS-30) were investigated in rats. The most withdrawal signs, including jumping, teeth chatter, writhing, shakes, lacrimation, piloerection, irritability and diarrhoea, were attenuated by pretreatment with alpha-helical CRF (10 microg i.c.v.) and CP-154,526 (30 mg/kg i.p.). However, no morphine withdrawal signs except for diarrhea were significantly affected by pretreatment with AS-30 (10 microg, i.c.v.). To investigate the possible role of different CRFR antagonists (alpha-helical CRF, CP-154,526 and AS-30) in relapse to opiate dependence, the 28-day extinction of morphine-conditioned place preference (CPP) was used. The morphine-CPP disappeared following a 28-day extinction and then was reactivated by a single injection of 10 mg/kg morphine. Pretreatment with alpha-helical CRF (10 microg, i.c.v.) and CP-154, 526 (30 mg/kg, i.p.) could significantly block this reactivation of morphine-CPP. In contrast, pretreatment with AS-30 (1 or 10 microg i. c.v.) did not affect this reactivation of morphine-CPP. The present study demonstrated that activation of the CRF receptor is involved in morphine withdrawal signs and relapse to morphine dependence, and that the role of CRF receptor subtypes 1 and 2 in withdrawal and reactivation of morphine dependence is not identical. CRF receptor subtype 1, but not subtype 2, is largely responsible for the action of the CRF system on opiate dependence. These results suggest that the CRF receptor antagonists, particularly the CRF receptor subtype 1 antagonist, might be of some value in the treatment and prevention of drug dependence.

Differential roles of corticotropin-releasing factor receptor subtypes 1 and 2 in opiate withdrawal and in relapse to opiate dependence

Differential roles of corticotropin-releasing factor receptor subtypes 1 and 2 in opiate withdrawal and in relapse to opiate dependence