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1.
Endocrinology ; 161(5)2020 05 01.
Article in English | MEDLINE | ID: mdl-32181477

ABSTRACT

Progesterone can block estrogen-induced luteinising hormone (LH) surge secretion and can be used clinically to prevent premature LH surges. The blocking effect of progesterone on the LH surge is mediated through its receptor in the anteroventral periventricular nucleus (AVPV) of the hypothalamus. However, the underlying mechanisms are unclear. The preovulatory LH surge induced by estrogen is preceded by a significant reduction in hypothalamic dynorphin and gamma-aminobutyric acid (GABA) release. To test the detailed roles of dynorphin and GABA in an LH surge blockade by progesterone, ovariectomized and 17ß-estradiol capsule-implanted (OVX/E2) mice received simultaneous injections of estradiol benzoate (EB) and progesterone (P) or vehicle for 2 consecutive days. The LH level was monitored from 2:30 pm to 8:30 pm at 30-minute intervals. Progesterone coadministration resulted in the LH surge blockade. A continuous microinfusion of the dynorphin receptor antagonist nor-BNI or GABAA receptor antagonist bicuculline into the AVPV from 3:00 pm to 7:00 pm reversed the progesterone-mediated blockade of the LH surge in 7 of 9 and 6 of 10 mice, respectively. In addition, these LH surges started much earlier than the surge induced by estrogen alone. However, 5 of 7 progesterone-treated mice did not show LH surge secretion after microinfusion with the GABAB receptor antagonist CGP-35348. Additionally, peripheral administration of kisspeptin-54 promotes LH surge-like release in progesterone treated mice. These results demonstrated that the progesterone-mediated suppression of the LH surge is mediated by an increase in dynorphin and GABAA receptor signaling acting though kisspeptin neurons in the AVPV of the hypothalamus in female mice.


Subject(s)
Dynorphins/metabolism , Hypothalamus/drug effects , Luteinizing Hormone/metabolism , Progesterone/pharmacology , Receptors, GABA-A/metabolism , Signal Transduction/drug effects , Animals , Bicuculline/pharmacology , Dynorphins/antagonists & inhibitors , Estradiol/pharmacology , Female , GABA Antagonists/pharmacology , Hypothalamus/cytology , Hypothalamus/metabolism , Hypothalamus, Anterior/cytology , Hypothalamus, Anterior/drug effects , Hypothalamus, Anterior/metabolism , Kisspeptins/metabolism , Mice, Inbred C57BL , Naltrexone/analogs & derivatives , Naltrexone/pharmacology , Neurons/cytology , Neurons/drug effects , Neurons/metabolism , Organophosphorus Compounds/pharmacology , Ovariectomy
2.
Peptides ; 91: 40-48, 2017 05.
Article in English | MEDLINE | ID: mdl-28363796

ABSTRACT

We previously reported a series of novel endomorphin analogs with unnatural amino acid modifications. These analogs display good binding affinity and functional activity toward the µ opioid receptor (MOP). In the present study, we further investigated the spinal antinociceptive activity of these compounds. The analogs were potent in several nociceptive models. Opioid antagonists and antibodies against several endogenous opioid peptides were used to determine the mechanisms of action of these peptides. Intrathecal pretreatment with naloxone and ß-funaltrexamine (ß-FNA) effectively inhibited analog-induced analgesia, demonstrating that activity of the analogs is regulated primarily through MOP. Antinociception induced by analog 2 through 4 was not reversed by δ opioid receptor (DOP) or κ opioid receptor (KOP) antagonist; antibodies against dynorphin-A (1-17), dynorphin-B (1-13), and Leu5/Met5-enkephalin had no impact on the antinociceptive effects of these analogs. In contrast, antinociceptive effects induced by a spinal injection of the fluorine substituted analog 1 were significantly reversed by KOP antagonism. Furthermore, intrathecal pretreatment with antibodies against dynorphin-B (1-13) attenuated the antinociceptive effect of analog 1. These results indicate that the antinociceptive activity exerted by intrathecally-administered analog 1 is mediated, in part, through KOP with increased release of dynorphin-B (1-13). The chemical modifications used in the present study may serve as a useful tool to gain insight into the mechanisms of endomorphins activity.


Subject(s)
Analgesics, Opioid/chemistry , Analgesics, Opioid/pharmacology , Oligopeptides/chemistry , Oligopeptides/pharmacology , Opioid Peptides/chemistry , Opioid Peptides/pharmacology , Analgesia , Analgesics, Opioid/administration & dosage , Analgesics, Opioid/antagonists & inhibitors , Analysis of Variance , Animals , Antibodies/immunology , Dynorphins/administration & dosage , Dynorphins/antagonists & inhibitors , Dynorphins/chemistry , Dynorphins/pharmacology , Enkephalin, Leucine/administration & dosage , Enkephalin, Leucine/antagonists & inhibitors , Enkephalin, Leucine/chemistry , Enkephalin, Leucine/pharmacology , Enkephalin, Methionine/administration & dosage , Enkephalin, Methionine/antagonists & inhibitors , Enkephalin, Methionine/chemistry , Enkephalin, Methionine/pharmacology , Fluorine/chemistry , Injections, Spinal , Male , Mice , Naloxone/administration & dosage , Naloxone/pharmacology , Naltrexone/administration & dosage , Naltrexone/analogs & derivatives , Naltrexone/pharmacology , Narcotic Antagonists/pharmacology , Oligopeptides/administration & dosage , Oligopeptides/antagonists & inhibitors , Opioid Peptides/administration & dosage , Opioid Peptides/antagonists & inhibitors , Pain/drug therapy , Pain/metabolism , Pain Measurement , Receptors, Opioid, kappa/antagonists & inhibitors , Receptors, Opioid, mu/antagonists & inhibitors , Receptors, sigma/antagonists & inhibitors
3.
Peptides ; 87: 20-27, 2017 01.
Article in English | MEDLINE | ID: mdl-27840228

ABSTRACT

Brain region-specific expression of proteolytic enzymes can control the biological activity of endogenous neuropeptides and has recently been targeted for the development of novel drugs, for neuropathic pain, cancer, and Parkinson's disease. Rapid and sensitive analytical methods to profile modulators of enzymatic activity are important for finding effective inhibitors with high therapeutic value. Combination of in situ enzyme histochemistry with MALDI imaging mass spectrometry allowed developing a highly sensitive method for analysis of brain-area specific neuropeptide conversion of synthetic and endogenous neuropeptides, and for selection of peptidase inhibitors that differentially target conversion enzymes at specific anatomical sites. Conversion and degradation products of Dynorphin B as model neuropeptide and effects of peptidase inhibitors applied to native brain tissue sections were analyzed at different brain locations. Synthetic dynorphin B (2pmol) was found to be converted to the N-terminal fragments on brain sections whereas fewer C-terminal fragments were detected. N-ethylmaleimide (NEM), a non-selective inhibitor of cysteine peptidases, almost completely blocked the conversion of dynorphin B to dynorphin B(1-6; Leu-Enk-Arg), (1-9), (2-13), and (7-13). Proteinase inhibitor cocktail, and also incubation with acetic acid displayed similar results. Bioconversion of synthetic dynorphin B was region-specific producing dynorphin B(1-7) in the cortex and dynorphin B (2-13) in the striatum. Enzyme inhibitors showed region- and enzyme-specific inhibition of dynorphin bioconversion. Both phosphoramidon (inhibitor of the known dynorphin converting enzyme neprilysin) and opiorphin (inhibitor of neprilysin and aminopeptidase N) blocked cortical bioconversion to dynorphin B(1-7), wheras only opiorphin blocked striatal bioconversion to dynorphin B(2-13). This method may impact the development of novel therapies with aim to strengthen the effects of endogenous neuropeptides under pathological conditions such as chronic pain. Combining histochemistry and MALDI imaging MS is a powerful and sensitive tool for the study of inhibition of enzyme activity directly in native tissue sections.


Subject(s)
Brain/metabolism , Dynorphins/metabolism , Endorphins/metabolism , Neuropeptides/metabolism , Animals , Cysteine Endopeptidases/chemistry , Cysteine Endopeptidases/metabolism , Dynorphins/antagonists & inhibitors , Dynorphins/chemistry , Dynorphins/isolation & purification , Endorphins/antagonists & inhibitors , Endorphins/chemistry , Endorphins/isolation & purification , Glycopeptides/administration & dosage , Humans , Neuropeptides/antagonists & inhibitors , Neuropeptides/chemistry , Neuropeptides/isolation & purification , Oligopeptides/administration & dosage , Parkinson Disease/metabolism , Parkinson Disease/pathology , Protease Inhibitors/administration & dosage , Rats , Salivary Proteins and Peptides/administration & dosage , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
4.
J Transl Med ; 13: 284, 2015 Aug 30.
Article in English | MEDLINE | ID: mdl-26320055

ABSTRACT

BACKGROUND: Processed Chuanwu (PCW), the mother root of Aconitum carmichaelii Debeauxv, has been widely used as a classic Traditional Chinese Medicine for pain relieve for over two millennia clinically. However, its action on chronic inflammatory pain has not been clarified. Here, we investigated the antinociceptive effect of PCW in complete freund's adjuvant (CFA)-induced mice and its possible mechanisms associated with opioid system and TRPV1 ion channel. METHODS: Male ICR mice were intraplantarly injected with CFA. PCW (0.34, 0.68 and 1.35 g/kg) was orally given to mice once a day for 7 days. Von frey hairs and planter test were assessed to evaluate the antinociceptive effect of PCW. To investigate the participation of dynorphin/opioid system in PCW antinociception, subtype-specific opioid receptor antagonists or anti-dynorphin A antiserum were used. To eliminate other central mechanisms that contribute to PCW antinociception, hot plate (50 °C) test were performed. Further, involvements of TRPV1 in PCW antinociception were evaluated in CFA-induced TRPV1(-/-) and TRPV1(+/+) C57BL/6 male mice, and in capsaicin-induced nociception ICR naive mice pretreated with nor-BNI. Meanwhile, calcium imaging was performed in HEK293T-TRPV1 cells. Finally, rotarod, open-field tests and body temperature measurement were carried out to assess side effects of PCW. RESULTS: PCW dose-dependently attenuated mechanical and heat hypersensitivities with no tolerance, which could be partially attenuated by coadministration of k-opioid receptor antagonist nor-binaltorphimine (nor-BNI) or anti-dynorphin A (1-13) antiserum. And PCW antinociception was totally erased by pretreatment with nor-BNI in the hot plate test. In addition, PCW antinociception was decreased in TRPV1(-/-) mice compared to TRPV1(+/+) group. And PCW still manifested inhibitory effects in capsaicin-induced nociception with nor-BNI pretreatment. PCW significantly inhibited capsaicin-induced calcium influx in HEK293T-TRPV1 cells. Finally, no detectable side effects were found in naive mice treated with PCW. CONCLUSIONS: This study shows PCW's potent antinociceptive effect in inflammatory conditions without obvious side effects. This effect may result from the activation of κ-opioid receptor via dynorpin release and the inhibition of TRPV1. These findings indicate that PCW might be a potential agent for the management of chronic inflammatory pain.


Subject(s)
Aconitum/chemistry , Analgesics/chemistry , Dynorphins/metabolism , Plant Extracts/chemistry , Receptors, Opioid, kappa/metabolism , TRPV Cation Channels/metabolism , Administration, Oral , Analgesics, Opioid/chemistry , Animals , Body Temperature , Calibration , Cell Survival , Chromatography, High Pressure Liquid , Dynorphins/antagonists & inhibitors , Dynorphins/chemistry , Freund's Adjuvant/chemistry , HEK293 Cells , Humans , Inflammation/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Inbred ICR , Plant Roots/chemistry
5.
Acta Pharmacol Sin ; 36(7): 783-90, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25982631

ABSTRACT

Anxiety disorders are the most common and prevalent forms of psychiatric disease, although the biological basis of anxiety is not well understood. The dynorphin/κ opioid receptor system is widely distributed in the central nervous system and has been shown to play a critical role in modulating mood and emotional behaviors. In the present review, we summarize current literature relating to the role played by the dynorphin/κ opioid receptor system in anxiety and κ opioid receptor antagonists as potential therapeutic agents for the treatment of anxiety disorders.


Subject(s)
Anxiety/drug therapy , Anxiety/metabolism , Dynorphins/metabolism , Receptors, Opioid, kappa/metabolism , Animals , Anxiety/psychology , Dynorphins/antagonists & inhibitors , Humans , Narcotic Antagonists/pharmacology , Narcotic Antagonists/therapeutic use , Receptors, Opioid, kappa/antagonists & inhibitors
6.
Endocrinology ; 156(2): 576-88, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25490143

ABSTRACT

Kisspeptin/neurokinin B/dynorphin (KNDy) neurons, which coexpress kisspeptins (Kps), neurokinin B (NKB), and dynorphin (Dyn), regulate gonadotropin secretion. The KNDy model proposes that NKB (a stimulator, through NK3R) and Dyn (an inhibitor, through κ-opioid receptor) shape Kp secretion onto GnRH neurons. However, some aspects of this paradigm remain ill defined. Here we aimed to characterize the following: 1) the effects of NKB signaling on FSH secretion and 2) the role of Dyn in gonadotropin secretion after NK3R activation; 3) additionally, we explored the roles of other tachykinin receptors, NK1R and NK2R, on gonadotropin release. Thus, the effects of the NK3R agonist, senktide, on FSH release were explored across postnatal development in male and female rats; gonadotropin responses to agonists of NK1R substance P and NK2R [neurokinin A (NKA)] were also monitored. Moreover, the effects of senktide on gonadotropin secretion were assessed after antagonizing Dyn actions by nor-binaltorphimine didydrochloride. Before puberty, rats of both sexes showed increased FSH secretion to senktide (and Kp-10). Conversely, adult female rats were irresponsive to senktide in terms of FSH, despite proven LH responses, whereas the adult males did not display FSH or LH responses to senktide, even at high doses. In turn, substance P and NKA stimulated gonadotropin secretion in prepubertal rats, whereas in adults modest gonadotropin responses to NKA were detected. By pretreatment with a Dyn antagonist, adult males became responsive to senktide in terms of LH secretion and displayed elevated basal LH and FSH levels; nor-binaltorphimine didydrochloride treatment uncovered FSH responses to senktide in adult females. Furthermore, the expression of Pdyn and Opkr1 (encoding Dyn and κ-opioid receptor, respectively) in the mediobasal hypothalamus was greater in males than in females at prepubertal ages. Overall, our data contribute to refining our understanding on how the elements of the KNDy node and related factors (ie, other tachykinins) differentially participate in the control of gonadotropins at different stages of rat postnatal maturation.


Subject(s)
Aging/metabolism , Follicle Stimulating Hormone/metabolism , Kisspeptins/metabolism , Luteinizing Hormone/metabolism , Neurokinin B/metabolism , Animals , Dynorphins/antagonists & inhibitors , Dynorphins/metabolism , Enkephalins/metabolism , Female , Hypothalamus/metabolism , Male , Neurokinin B/agonists , Peptide Fragments , Protein Precursors/metabolism , Rats, Wistar , Receptors, Neurokinin-1/agonists , Receptors, Neurokinin-2/agonists , Substance P/analogs & derivatives
7.
Proc Natl Acad Sci U S A ; 111(16): E1648-55, 2014 Apr 22.
Article in English | MEDLINE | ID: mdl-24706819

ABSTRACT

Hypocretin (orexin) and dynorphin are neuropeptides with opposing actions on motivated behavior. Orexin is implicated in states of arousal and reward, whereas dynorphin is implicated in depressive-like states. We show that, despite their opposing actions, these peptides are packaged in the same synaptic vesicles within the hypothalamus. Disruption of orexin function blunts the rewarding effects of lateral hypothalamic (LH) stimulation, eliminates cocaine-induced impulsivity, and reduces cocaine self-administration. Concomitant disruption of dynorphin function reverses these behavioral changes. We also show that orexin and dynorphin have opposing actions on excitability of ventral tegmental area (VTA) dopamine neurons, a prominent target of orexin-containing neurons, and that intra-VTA orexin antagonism causes decreases in cocaine self-administration and LH self-stimulation that are reversed by dynorphin antagonism. Our findings identify a unique cellular process by which orexin can occlude the reward threshold-elevating effects of coreleased dynorphin and thereby act in a permissive fashion to facilitate reward.


Subject(s)
Dynorphins/metabolism , Intracellular Signaling Peptides and Proteins/metabolism , Neuropeptides/metabolism , Neurotransmitter Agents/metabolism , Reward , Ventral Tegmental Area/metabolism , Animals , Cocaine/administration & dosage , Cocaine/pharmacology , Dopaminergic Neurons/drug effects , Dopaminergic Neurons/metabolism , Dynorphins/antagonists & inhibitors , Hypothalamus/drug effects , Hypothalamus/metabolism , Impulsive Behavior/metabolism , Intracellular Signaling Peptides and Proteins/antagonists & inhibitors , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Neuropeptides/antagonists & inhibitors , Orexin Receptors/metabolism , Orexins , Self Administration , Synaptic Transmission/drug effects , Ventral Tegmental Area/drug effects
8.
J Physiol Pharmacol ; 64(3): 331-40, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23959730

ABSTRACT

We characterized pain behavior and cutaneous blood flow response induced by activation of the spinal transient receptor potential ankyrin 1 (TRPA1) channel using intrathecal drug administrations in the rat. Additionally, we assessed whether the pronociceptive actions induced by intrathecally administered dynorphin A, cholecystokinin or prostaglandin F(2α) are mediated by the spinal TRPA1 channel. Cinnamaldehyde, a TRPA1 agonist, produced a dose-related (3-10 µg) cutaneous blood flow increase and mechanical hypersensitivity effect. These effects at the currently used doses were of short duration and attenuated, although not completely, by pretreatment with A-967079, a TRPA1 antagonist. The cinnamaldehyde-induced hypersensitivity was also reduced by pretreatment with minocycline (an inhibitor of microglial activation), but not by carbenoxolone (a gap junction decoupler). In vitro study, however, indicated that minocycline only poorly blocks the TRPA1 channel. The mechanical hypersensitivity effect induced by dynorphin A, but not that by cholecystokinin or prostaglandin F(2α), was attenuated by a TRPA1 antagonist Chembridge-5861528 as well as A-967079. The cinnamaldehyde-induced cutaneous blood flow increase was not suppressed by MK-801, an NMDA receptor antagonist, or bicuculline, a GABA(A) receptor antagonist. The results indicate that spinal TRPA1 channels promote mechanical pain hypersensitivity and due to antidromic activation of nociceptive nerve fibers increase cutaneous blood flow. The attenuation of the cinnamaldehyde-induced hypersensitivity effect by minocycline may be explained by action other than block of the TRPA1 channel. Moreover, the spinal TRPA1 channel is involved in mediating the pronociceptive action of dynorphin A, but not that of the spinal cholecystokinin or prostaglandin F(2α).


Subject(s)
Analgesics, Non-Narcotic/therapeutic use , Back Pain/drug therapy , Dynorphins/antagonists & inhibitors , Hyperalgesia/drug therapy , Posterior Horn Cells/drug effects , Skin/drug effects , TRPC Cation Channels/antagonists & inhibitors , Acrolein/administration & dosage , Acrolein/adverse effects , Acrolein/analogs & derivatives , Acrolein/antagonists & inhibitors , Analgesics, Non-Narcotic/administration & dosage , Animals , Back Pain/etiology , Back Pain/metabolism , Behavior, Animal/drug effects , Cholecystokinin/administration & dosage , Cholecystokinin/antagonists & inhibitors , Cholecystokinin/metabolism , Dinoprost/administration & dosage , Dinoprost/antagonists & inhibitors , Dinoprost/metabolism , Dose-Response Relationship, Drug , Dynorphins/administration & dosage , Dynorphins/metabolism , Hyperalgesia/chemically induced , Hyperalgesia/metabolism , Injections, Spinal , Male , Minocycline/administration & dosage , Minocycline/therapeutic use , Nerve Tissue Proteins/agonists , Nerve Tissue Proteins/antagonists & inhibitors , Nerve Tissue Proteins/metabolism , Oximes/administration & dosage , Oximes/therapeutic use , Physical Stimulation/adverse effects , Posterior Horn Cells/metabolism , Rats , Rats, Wistar , Regional Blood Flow/drug effects , Skin/blood supply , TRPA1 Cation Channel , TRPC Cation Channels/agonists , TRPC Cation Channels/metabolism
9.
J Reprod Dev ; 59(5): 479-84, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23877505

ABSTRACT

Puberty in mammals is timed by an increase in gonadotropin-releasing hormone (GnRH) secretion. Previous studies have shown involvement of the two neuropeptides, kisspeptin and neurokinin B (NKB), in controlling puberty onset. Little is known about the role of the other key neuropeptide, dynorphin, in controlling puberty onset, although these three neuropeptides colocalize in the arcuate kisspeptin neurons. The arcuate kisspeptin neuron, which is also referred to as the KNDy neuron, has recently been considered to play a role as an intrinsic source of the GnRH pulse generator. The present study aimed to determine if attenuation of inhibitory dynorphin-kappa-opioid receptor (KOR) signaling triggers the initiation of puberty in normal developing female rats. The present study also determined if stimulatory NKB-neurokinin 3 receptor (NK3R) signaling advances puberty onset. Female Wistar-Imamichi rats were weaned and intraperitoneally implanted with osmotic minipumps filled with nor-binaltorphimine (nor-BNI), a KOR antagonist, or senktide, a NK3R agonist, at 20 days of age. Fourteen days of intraperitoneal infusion of nor-BNI or senktide advanced puberty onset, manifested as vaginal opening and the first vaginal estrus in female rats. Frequent blood sampling showed that nor-BNI significantly increased luteinizing hormone (LH) pulse frequency at 29 days of age compared with vehicle-treated controls. Senktide tended to increase this frequency, but its effect was not statistically significant. The present results suggest that the inhibitory input of dynorphin-KOR signaling plays a role in the prepubertal restraint of GnRH/LH secretion in normal developing female rats and that attenuation of dynorphin-KOR signaling and increase in NKB-NK3R signaling trigger the onset of puberty in female rats.


Subject(s)
Dynorphins/metabolism , Luteinizing Hormone/metabolism , Neurokinin B/metabolism , Ovary/metabolism , Receptors, Neurokinin-3/metabolism , Receptors, Opioid, kappa/metabolism , Sexual Maturation , Animals , Drug Implants , Dynorphins/antagonists & inhibitors , Female , Luteinizing Hormone/blood , Naltrexone/administration & dosage , Naltrexone/analogs & derivatives , Naltrexone/toxicity , Narcotic Antagonists/administration & dosage , Narcotic Antagonists/toxicity , Nerve Tissue Proteins/agonists , Nerve Tissue Proteins/antagonists & inhibitors , Nerve Tissue Proteins/metabolism , Neurokinin B/agonists , Neurons/drug effects , Neurons/metabolism , Ovary/drug effects , Ovary/growth & development , Peptide Fragments/administration & dosage , Peptide Fragments/toxicity , Puberty, Precocious/blood , Puberty, Precocious/chemically induced , Puberty, Precocious/metabolism , Random Allocation , Rats , Rats, Wistar , Receptors, Neurokinin-3/agonists , Receptors, Opioid, kappa/antagonists & inhibitors , Sexual Maturation/drug effects , Signal Transduction/drug effects , Substance P/administration & dosage , Substance P/analogs & derivatives , Substance P/toxicity , Weaning
10.
J Neurosci ; 32(4): 1488-95, 2012 Jan 25.
Article in English | MEDLINE | ID: mdl-22279233

ABSTRACT

Many smokers describe the anxiolytic and stress-reducing effects of nicotine, the primary addictive component of tobacco, as a principal motivation for continued drug use. Recent evidence suggests that activation of the stress circuits, including the dynorphin/κ-opioid receptor system, modulates the rewarding effects of addictive drugs. In the present study, we find that nicotine produced dose-dependent conditioned place preference (CPP) in mice. κ-receptor activation, either by repeated forced swim stress or U50,488 (5 or 10 mg/kg, i.p.) administration, significantly potentiated the magnitude of nicotine CPP. The increase in nicotine CPP was blocked by the κ-receptor antagonist norbinaltorphimine (norBNI) either systemically (10 mg/kg, i.p.) or by local injection in the amygdala (2.5 µg) without affecting nicotine reward in the absence of stress. U50,488 (5 mg/kg, i.p.) produced anxiety-like behaviors in the elevated-plus maze and novel object exploration assays, and the anxiety-like behaviors were attenuated both by systemic nicotine (0.5 mg/kg, s.c.) and local injection of norBNI into the amygdala. Local norBNI injection in the ventral posterior thalamic nucleus (an adjacent brain region) did not block the potentiation of nicotine CPP or the anxiogenic-like effects of κ-receptor activation. These results suggest that the rewarding effects of nicotine may include a reduction in the stress-induced anxiety responses caused by activation of the dynorphin/κ-opioid system. Together, these data implicate the amygdala as a key region modulating the appetitive properties of nicotine, and suggest that κ-opioid antagonists may be useful therapeutic tools to reduce stress-induced nicotine craving.


Subject(s)
Amygdala/metabolism , Conditioning, Operant/physiology , Dynorphins/metabolism , Nicotine/pharmacology , Receptors, Opioid, kappa/metabolism , Stress, Psychological/metabolism , 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer/pharmacology , Amygdala/drug effects , Animals , Conditioning, Operant/drug effects , Dynorphins/antagonists & inhibitors , Male , Mice , Mice, Inbred C57BL , Receptors, Opioid, kappa/agonists , Receptors, Opioid, kappa/antagonists & inhibitors , Stress, Psychological/psychology , Swimming/psychology
11.
Mol Pharmacol ; 75(3): 704-12, 2009 Mar.
Article in English | MEDLINE | ID: mdl-19106229

ABSTRACT

The nucleus accumbens (NAc) is a critical brain area for reward and motivated behavior. Accumulating evidence suggests that altered function of the transcription factor cAMP response element binding protein (CREB) within the NAc is involved in depressive behavior. In rats, stress activates CREB within the NAc, and elevation of CREB expression in this region produces depressive-like behaviors that are accompanied by activation of CREB-regulated target genes. The depressive-like behaviors seem to be due, at least in part, to CREB-mediated increases in dynorphin function, because they are mimicked by kappa-opioid receptor (KOR) agonists and attenuated by KOR antagonists. We hypothesized that if CREB-mediated dynorphin expression in the NAc contributes to depressive behavior, then antidepressants might reduce dynorphin function in this region. Here, we demonstrate that desipramine (DMI), a norepinephrine reuptake inhibitor that has been used for decades to treat clinical depression, blocks swim stress-induced activation of prodynorphin (encodes dynorphin) in the NAc. In primary cultures of NAc and striatum, DMI decreases basal and stimulated CREB phosphorylation by causing reductions in intracellular calcium (Ca(2+)) availability that are independent of norepinephrine or other monoaminergic inputs, identifying a potential mechanism for alterations in CREB-mediated gene expression. Fluoxetine (FLX), a selective serotonin reuptake inhibitor, has similar effects in culture, suggesting a common intracellular effect of these antidepressants. These findings raise the possibility that a therapeutically relevant mechanism of action of DMI occurs through attenuation of CREB-mediated gene transcription, which is mediated via previously uncharacterized mechanisms that occur directly within the NAc.


Subject(s)
Cyclic AMP Response Element-Binding Protein/antagonists & inhibitors , Cyclic AMP Response Element-Binding Protein/metabolism , Desipramine/pharmacology , Dynorphins/antagonists & inhibitors , Dynorphins/biosynthesis , Nucleus Accumbens/metabolism , Stress, Physiological/physiology , Animals , Cells, Cultured , Cyclic AMP Response Element-Binding Protein/genetics , Dynorphins/genetics , Female , Gene Expression Regulation/drug effects , Gene Expression Regulation/physiology , Male , Nucleus Accumbens/drug effects , Phosphorylation/drug effects , Phosphorylation/physiology , Pregnancy , Rats , Rats, Sprague-Dawley , Stress, Physiological/drug effects
12.
Eur J Pain ; 12(7): 870-8, 2008 Oct.
Article in English | MEDLINE | ID: mdl-18221900

ABSTRACT

Cancer pain impairs the quality of life of cancer patients, but opioid intervention can cause significant side effects that further decrease quality of life. Although electroacupuncture (EA) has been used to treat cancer pain, its mechanisms are largely unknown. To examine its effects and underlying mechanisms on cancer pain, we injected AT-3.1 prostate cancer cells into the tibia to induce bone cancer in the male Copenhagen rat. The resulting pain was treated with 10Hz/2mA/0.4ms pulse EA for 30min daily at the point equivalent to the human acupoint GB30 (Huantiao) between days 14 and 18 after the injection. For sham control, EA needles were inserted into GB30 without stimulation. Thermal hyperalgesia, a decrease in paw withdrawal latency (PWL) to a noxious thermal stimulus, and mechanical hyperalgesia, a decrease in paw withdrawal pressure threshold (PWPT), was measured at baseline and 20min after the EA treatment. Preprodynorphin mRNA and dynorphin were determined by RT-PCR and immunohistochemistry, respectively. Thermal and mechanical hyperalgesia developed ipsilaterally between days 12 and 18 after cancer cell inoculation. EA significantly (P<0.05) attenuated this hyperalgesia, as shown by increased PWL and PWPT, and inhibited up-regulation of preprodynorphin mRNA and dynorphin compared to sham control. Intrathecal injection of antiserum against dynorphin A (1-17) also significantly inhibited the cancer-induced hyperalgesia. These results suggest that EA alleviates bone cancer pain at least in part by suppressing dynorphin expression, and they support the clinical use of EA in the treatment of cancer pain.


Subject(s)
Acupuncture Analgesia , Adenocarcinoma/secondary , Bone Neoplasms/secondary , Dynorphins/biosynthesis , Electroacupuncture , Hyperalgesia/therapy , Protein Precursors/biosynthesis , Spinal Cord/metabolism , Adenocarcinoma/physiopathology , Animals , Bone Neoplasms/physiopathology , Cell Line, Tumor/transplantation , Down-Regulation , Dynorphins/antagonists & inhibitors , Dynorphins/genetics , Dynorphins/immunology , Hyperalgesia/etiology , Immune Sera , Immunization, Passive , Injections, Spinal , Male , Pain Threshold , Protein Precursors/genetics , Rats , Reaction Time , Tibia
13.
Physiol Behav ; 92(1-2): 167-71, 2007 Sep 10.
Article in English | MEDLINE | ID: mdl-17610914

ABSTRACT

Repeated exposure to ethanol has previously been shown to induce alterations in both midbrain dopamine and dynorphin systems. The aim of this study was to investigate functional changes in the sensitivity of dynorphin/kappa-receptor systems following repeated ethanol administration, using dopamine as an indirect marker. The effects of kappa-opioid receptor ligands on dopamine release in the rat nucleus accumbens were investigated following repeated ethanol administration (2 g/kg body weight, twice daily for 7 days). The selective kappa-receptor agonist U50, 488H reduced dopamine levels in both ethanol- and saline-treated animals, although the decline had a later onset and lasted shorter in the ethanol-treated group. Nor-binaltorphimine, a kappa-antagonist, produced a significant increase of dopamine in ethanol-treated rats, but lacked effect in the saline-treated group. This change in responsiveness of dopamine neurons following repeated ethanol administration could be related to changes in the sensitivity of kappa-receptor systems and/or an increase in dynorphin tone in the nucleus accumbens.


Subject(s)
Central Nervous System Stimulants/pharmacology , Dopamine/metabolism , Drug Tolerance/physiology , Ethanol/pharmacology , Nucleus Accumbens/drug effects , Receptors, Opioid, kappa/drug effects , 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer/pharmacology , Animals , Central Nervous System Stimulants/administration & dosage , Drug Administration Schedule , Drug Interactions , Dynorphins/agonists , Dynorphins/antagonists & inhibitors , Ethanol/administration & dosage , Male , Naltrexone/analogs & derivatives , Naltrexone/pharmacology , Nucleus Accumbens/metabolism , Rats , Rats, Sprague-Dawley
14.
J Pept Res ; 66(5): 263-76, 2005 Nov.
Article in English | MEDLINE | ID: mdl-16218994

ABSTRACT

Dynorphin A (Dyn A), a 17 amino acid peptide H-Tyr-Gly-Gly-Phe-Leu-Arg-Arg-Ile-Arg-Pro-Lys-Leu-Lys-Trp-Asp-Asn-Gln-OH, is a potent opioid peptide which interacts preferentially with kappa-opioid receptors. Research in the development of selective and potent opioid peptide ligands for the kappa-receptor is important in mediating analgesia. Several cyclic disulphide bridge-containing peptide analogues of Dyn A, which were conformationally constrained in the putative message or address segment of the opioid ligand, were designed, synthesized and assayed. To further investigate the conformational and topographical requirements for the residues in positions 5 and 11 of these analogues, a systematic series of Dyn A(1-11)-NH2 cyclic analogues incorporating the sulphydryl-containing amino acids L- and D-Cys and L- and D-Pen in positions 5 and 11 were synthesized and assayed. Cyclic lactam peptide analogues were also synthesized and assayed. Several of these cyclic analogues, retained the same affinity and selectivity (vs. the mu- and delta-receptors) as the parent Dyn A(1-11)-NH2 peptide in the guinea-pig brain (GPB), but exhibited a much lower activity in the guinea-pig ileum (GPI), thus leading to centrally vs. peripherally selective peptides. Studies of the structure-activity relationship of Dyn A peptide provide new insights into the importance of each amino acid residue (and their configurations) in Dyn A analogues for high potency and good selectivity at kappa-opioid receptors. We report herein the progress towards the development of Dyn A peptide ligands, which can act as agonists or antagonists at cell surface receptors that modulate cell function and animal behaviour using various approaches to rational peptide ligand-based drug design.


Subject(s)
Drug Design , Dynorphins/chemical synthesis , Animals , Biological Assay , Brain/drug effects , Brain/enzymology , Capsaicin/pharmacology , Dynorphins/antagonists & inhibitors , Dynorphins/pharmacology , Guinea Pigs , Ileum/drug effects , Ligands , Macaca mulatta , Mice , Muscle, Smooth/drug effects , Muscle, Smooth/enzymology , Opioid Peptides/agonists , Opioid Peptides/antagonists & inhibitors , Opioid Peptides/chemical synthesis , Pain/drug therapy , Peptides/antagonists & inhibitors , Peptides/chemistry , Protein Conformation , Receptors, Cell Surface/antagonists & inhibitors , Receptors, Cell Surface/drug effects , Species Specificity
15.
Neurosci Lett ; 386(1): 46-51, 2005 Sep 23.
Article in English | MEDLINE | ID: mdl-15982809

ABSTRACT

It is generally accepted that neuropathic pain is resistant to amelioration by morphine in clinical studies and insensitivity to intrathecal (i.t.) administered morphine in experimental models of neuropathic pain has been demonstrated. This study is to determine if endogenous dynorphin, substance P or cholecystokinin is involved in the lack of anti-allodynia of morphine in a partial sciatic nerve ligation (PSL) model of CD-1 mice. Mice exhibited tactile allodynia in the ipsilateral hind paw 1 day after PSL, and reached its maximal allodynic effect at 2 days and remained allodynic for 7 days. Morphine (3.0 nmol) given i.t. did not alter the tactile allodynic threshold in ipsilateral paw of mice pretreated i.t. with normal rabbit serum 2 days after PSL. However, the same dose of morphine (3.0 nmol) given i.t. reduced markedly allodynia in mice pretreated for 2h with antiserum against dynorphin A(1-17) (200 microg); the morphine-produced anti-allodynia developed slowly, reached its peak effect at 30 min and returned to an allodynic state in 60 min. Similarly, i.t. injection of morphine reduced the allodynia in PSL mice pretreated with antiserum against substance P (10 microg) or cholecystokinin (200 microg) for 2h. Intrathecal pretreatment with antiserum against dynorphin A(1-17), substance P or cholecystokinin for 2h injected alone did not affect the baseline mechanical tactile threshold in ipsilateral paw 2 days after PSL. The results indicate that endogenous dynorphin A(1-17), substance P and cholecystokinin are involved in PSL-induced neuropathic allodynia to attenuate the anti-allodynic effect of morphine.


Subject(s)
Cholecystokinin/antagonists & inhibitors , Dynorphins/antagonists & inhibitors , Immune Sera/pharmacology , Morphine/agonists , Neuralgia/drug therapy , Peripheral Nervous System Diseases/drug therapy , Substance P/antagonists & inhibitors , Analgesics, Opioid/administration & dosage , Analgesics, Opioid/agonists , Animals , Antibodies/pharmacology , Antibodies/therapeutic use , Cholecystokinin/metabolism , Disease Models, Animal , Dose-Response Relationship, Drug , Drug Resistance/drug effects , Drug Resistance/physiology , Drug Synergism , Dynorphins/metabolism , Injections, Spinal , Ligation , Male , Mice , Morphine/administration & dosage , Neuralgia/metabolism , Neuralgia/physiopathology , Peripheral Nervous System Diseases/metabolism , Peripheral Nervous System Diseases/physiopathology , Sciatic Nerve/injuries , Sciatic Nerve/surgery , Sciatic Neuropathy/drug therapy , Sciatic Neuropathy/metabolism , Sciatic Neuropathy/physiopathology , Substance P/metabolism
16.
Biol Pharm Bull ; 28(5): 845-8, 2005 May.
Article in English | MEDLINE | ID: mdl-15863891

ABSTRACT

We previously reported that either (+)-matrine (matridin-15-one) or (+)-allomatrine (the C-6 epimer of matrine)-induced antinociceptive effect was attenuated by s.c. pretreatment with a kappa-opioid receptor (KOR) antagonist nor-binaltorphimine (nor-BNI), indicating the critical role of KORs in antinociceptive effects induced by these alkaloids. In the present study, we found that i.c.v. administration of either (+)-matrine- or (+)-allomatrine induced antinociceptive effects in the mouse tail-flick and warm-plate test, whereas these alkaloids when given spinally failed to induce antinociception. In the guanosine-5'-O-(3-[(35)S]thio)trisphosphate ([(35)S]GTPgammaS) binding assay, we demonstrated that neither (+)-matrine nor (+)-allomatrine produced the stimulation of [(35)S]GTPgammaS binding in the membranes of the spinal cord, indicating that (+)-matrine- and (+)-allomatrine-induced supraspinal antinociceptive actions was not due to a direct stimulation of KORs by these alkaloids. Therefore, we next investigated the involvement of dynorphin A (1-17) release at the spinal or supraspinal site in (+)-matrine- or (+)-allomatrine-induced antinociception. The i.c.v. pretreatment with an antiserum against dynorphin A (1-17) could not affect the antinociceptive effect induced by s.c. treatment of (+)-matrine. In contrast, the s.c.-administered (+)-matrine- and (+)-allomatrine-induced antinociceptive effect was significantly attenuated by i.t. pretreatment of an antiserum against dynorphin A (1-17). The present data suggest that either (+)-matrine or (+)-allomatrine when given i.c.v. may stimulate the descending dynorphinergic neuron, resulting in the stimulation of KORs in the spinal cord, and this phenomenon in turn produces the antinociception in mice.


Subject(s)
Alkaloids/pharmacology , Analgesics/pharmacology , Dynorphins/antagonists & inhibitors , Neurons/drug effects , Pain Measurement/drug effects , Spinal Cord/drug effects , Alkaloids/isolation & purification , Analgesics/isolation & purification , Animals , Dynorphins/metabolism , Male , Mice , Mice, Inbred ICR , Narcotic Antagonists/pharmacology , Neural Pathways/drug effects , Neural Pathways/metabolism , Neurons/metabolism , Plant Extracts/isolation & purification , Plant Extracts/pharmacology , Quinolizines , Spinal Cord/metabolism , Matrines
17.
J Neurochem ; 90(5): 1258-68, 2004 Sep.
Article in English | MEDLINE | ID: mdl-15312181

ABSTRACT

Rats exposed to learned helplessness (LH), an animal model of depression, showed a recovery following an intracerebroventricular injection of nor-binaltorphimine dihydrochloride (norBNI; a kappa-opioid antagonist). To investigate the potential role of dynorphin A and dynorphin B, we examined the effects of different stress/depression models on dynorphin A and dynorphin B immunoreactivity in hippocampus and nucleus accumbens (NAc). Immobilization stress (3 h) caused an increase in levels of dynorphin A and dynorphin B immunoreactivity in the hippocampus and the NAc. Forced swim stress also temporally increased dynorphin A levels in the hippocampus. Furthermore, exposure to LH produced a similar increase in dynorphin A and dynorphin B in the hippocampus and NAc. Infusions of norBNI into the dentate gyrus or CA3 regions of hippocampus and into the shell or core regions of NAc produced antidepressant-like effects in the LH paradigm. The degrees of norBNI's effects were stronger in the CA3 region and NAc shell and less effective in the dentate gyrus of hippocampus and NAc core. These results indicate that both dynorphin A and dynorphin B contribute to the effects of stress, and suggest that blockade of kappa-opioid receptors may have therapeutic potential for the treatment of depression.


Subject(s)
Dynorphins/metabolism , Limbic System/drug effects , Naltrexone/analogs & derivatives , Naltrexone/pharmacology , Stress, Physiological/metabolism , Animals , Behavior, Animal , Cell Count/methods , Disease Models, Animal , Dose-Response Relationship, Drug , Dynorphins/antagonists & inhibitors , Endorphins/metabolism , Escape Reaction/drug effects , Helplessness, Learned , Immobilization/adverse effects , Immunohistochemistry/methods , Limbic System/metabolism , Male , Rats , Rats, Sprague-Dawley
18.
Neurosci Lett ; 359(1-2): 85-8, 2004 Apr 08.
Article in English | MEDLINE | ID: mdl-15050718

ABSTRACT

Glucocorticoids (GC) play important roles in response to stressful stimuli, including pain. This study examined the effects of bilateral adrenalectomy (ADX) and dexamethasone (DEX) replacement on the hyperalgesia and spinal preprodynorphin (PPD) mRNA expression induced by injecting complete Freund's adjuvant (CFA) into rats' hind paws. The results demonstrated that CFA induced more intense hyperalgesia and up-expression of spinal PPD mRNA in ADX rats than in control rats, while both of these intensified reactions could be significantly suppressed by subcutaneous pretreatment with DEX. This leads to the conclusion that both exogenous (pharmacological) and endogenous (physiological) GC suppresses the behavioral hyperalgesia and the up-regulation of spinal PPD mRNA induced by sustained peripheral inflammation. The results also suggest that spinal PPD mRNA suppression may partially underlie the inhibition of behavioral hyperalgesia.


Subject(s)
Dynorphins/biosynthesis , Glucocorticoids/metabolism , Hyperalgesia/metabolism , Protein Precursors/biosynthesis , RNA, Messenger/biosynthesis , Up-Regulation/physiology , Animals , Dynorphins/antagonists & inhibitors , Freund's Adjuvant , Glucocorticoids/pharmacology , Hyperalgesia/chemically induced , Inflammation/chemically induced , Inflammation/metabolism , Male , Protein Precursors/antagonists & inhibitors , RNA, Messenger/antagonists & inhibitors , Rats , Rats, Sprague-Dawley , Spinal Cord/drug effects , Spinal Cord/metabolism , Up-Regulation/drug effects
19.
J Pharmacol Exp Ther ; 307(3): 1135-41, 2003 Dec.
Article in English | MEDLINE | ID: mdl-14557378

ABSTRACT

We have previously demonstrated that both endomorphin-1 (EM-1) and endomorphin-2 (EM-2) at high doses (1.75-35 nmol) given intrathecally (i.t.) or intracerebroventricularly produce antinociception by stimulation of mu-opioid receptors. Now, we report that EM-2 at small doses (0.05-1.75 nmol), which injected alone did not produce antinociception, produces anti-analgesia against opioid agonist-induced antinociception. The tail-flick (TF) response was used to test the antinociception in male CD-1 mice. Intrathecal pretreatment with EM-2 (0.02-1.75 nmol) 45 min before i.t. morphine (3.0 nmol) injection dose dependently attenuated morphine-induced TF inhibition. On the other hand, a similar dose of EM-1 (1.64 nmol) failed to produce any antianalgesic effect. The EM-2 (1.75 nmol)-produced anti-analgesia against morphine-induced TF inhibition was blocked by i.t. pretreatment with the mu-opioid antagonist naloxone or 3-methoxynaltrexone, but not delta-opioid receptor antagonist naltrindole, kappa-opioid receptor antagonist nor-binaltorphimine, or N-methyl-d-aspartate (NMDA) receptor antagonist MK-801. The EM-2-induced antianalgesic effect against morphine-induced TF inhibition was blocked by i.t. pretreatment with antiserum against dynorphin A(1-17), but not beta-endorphin, [Met]-enkephalin, [Leu]-enkephalin, or cholecystokinin antiserum (200 microg each). The i.t. EM-2 pretreatment also attenuated the TF inhibition induced by other mu-opioid agonists, [d-Ala2,N-Me-Phe4,Gly-ol5]-enkephalin, EM-1 and EM-2, delta-opioid agonist deltorphin II, and kappa-opioid agonist (trans)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl]benzeneacetamide methane-sulfonate hydrate (U50,488H). It is concluded that EM-2 at subanalgesic doses presumably stimulates a subtype of mu-opioid receptor and subsequently induces the release of dynorphin A(1-17) to produce antianalgesic effects against mu-, delta-, or kappa-agonists-induced antinociception. The EM-2-induced antianalgesia is not mediated by the release of [Met]-enkephalin, [Leu]-enkephalin, beta-endorphin, or cholecystokinin, nor does it involve kappa- or delta-opioid or NMDA receptors in the spinal cord.


Subject(s)
Analgesics, Opioid/pharmacology , Dynorphins/physiology , Naltrexone/analogs & derivatives , Oligopeptides/pharmacology , Spinal Cord/drug effects , Analgesics, Opioid/administration & dosage , Analgesics, Opioid/antagonists & inhibitors , Animals , Antibodies, Blocking/pharmacology , Dizocilpine Maleate/pharmacology , Dose-Response Relationship, Drug , Dynorphins/antagonists & inhibitors , Excitatory Amino Acid Antagonists/pharmacology , Injections, Spinal , Male , Mice , Morphine/antagonists & inhibitors , Morphine/pharmacology , Naloxone/pharmacology , Naltrexone/pharmacology , Narcotic Antagonists/pharmacology , Oligopeptides/administration & dosage , Pain Measurement/drug effects , Reaction Time/drug effects , Receptors, Opioid/agonists
20.
J Neuroimmunol ; 141(1-2): 30-9, 2003 Aug.
Article in English | MEDLINE | ID: mdl-12965251

ABSTRACT

Neuroimmune interactions control pain through activation of opioid receptors on sensory nerves by immune-derived opioid peptides. Here we evaluate mechanisms of intrinsic pain inhibition at different stages of Freund's adjuvant-induced inflammation of the rat paw. We use immunohistochemistry and paw pressure testing. Our data show that in early (6 h) inflammation leukocyte-derived beta-endorphin, met-enkephalin and dynorphin A activate peripheral mu-, delta- and kappa-receptors to inhibit nociception. In addition, central opioid mechanisms seem to contribute significantly to this effect. At later stages (4 days), antinociception is exclusively produced by leukocyte-derived beta-endorphin acting at peripheral mu and delta receptors. Corticotropin-releasing hormone (CRH) is an endogenous trigger of these effects at both stages. These findings indicate that peripheral opioid mechanisms of pain inhibition gain functional relevance with the chronicity of inflammation.


Subject(s)
Pain/pathology , Pain/prevention & control , Animals , Corticotropin-Releasing Hormone/administration & dosage , Dynorphins/antagonists & inhibitors , Dynorphins/biosynthesis , Dynorphins/physiology , Edema/immunology , Edema/metabolism , Edema/physiopathology , Endorphins/antagonists & inhibitors , Endorphins/biosynthesis , Endorphins/physiology , Enkephalin, Methionine/antagonists & inhibitors , Enkephalin, Methionine/biosynthesis , Enkephalin, Methionine/physiology , Freund's Adjuvant/administration & dosage , Hindlimb , Inflammation/immunology , Inflammation/metabolism , Inflammation/physiopathology , Injections, Subcutaneous , Leukocytes/drug effects , Leukocytes/metabolism , Leukocytes/physiology , Male , Naloxone/administration & dosage , Pain/immunology , Pain Threshold , Rats , Rats, Wistar , Stress, Physiological/immunology , Stress, Physiological/metabolism , Stress, Physiological/physiopathology , Time Factors
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