RESUMEN
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.
Asunto(s)
Dinorfinas/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Neuropéptidos/metabolismo , Neurotransmisores/metabolismo , Recompensa , Área Tegmental Ventral/metabolismo , Animales , Cocaína/administración & dosificación , Cocaína/farmacología , Neuronas Dopaminérgicas/efectos de los fármacos , Neuronas Dopaminérgicas/metabolismo , Dinorfinas/antagonistas & inhibidores , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Conducta Impulsiva/metabolismo , Péptidos y Proteínas de Señalización Intracelular/antagonistas & inhibidores , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neuropéptidos/antagonistas & inhibidores , Receptores de Orexina/metabolismo , Orexinas , Autoadministración , Transmisión Sináptica/efectos de los fármacos , Área Tegmental Ventral/efectos de los fármacosRESUMEN
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.
Asunto(s)
Aconitum/química , Analgésicos/química , Dinorfinas/metabolismo , Extractos Vegetales/química , Receptores Opioides kappa/metabolismo , Canales Catiónicos TRPV/metabolismo , Administración Oral , Analgésicos Opioides/química , Animales , Temperatura Corporal , Calibración , Supervivencia Celular , Cromatografía Líquida de Alta Presión , Dinorfinas/antagonistas & inhibidores , Dinorfinas/química , Adyuvante de Freund/química , Células HEK293 , Humanos , Inflamación/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos ICR , Raíces de Plantas/químicaRESUMEN
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.
Asunto(s)
Ansiedad/tratamiento farmacológico , Ansiedad/metabolismo , Dinorfinas/metabolismo , Receptores Opioides kappa/metabolismo , Animales , Ansiedad/psicología , Dinorfinas/antagonistas & inhibidores , Humanos , Antagonistas de Narcóticos/farmacología , Antagonistas de Narcóticos/uso terapéutico , Receptores Opioides kappa/antagonistas & inhibidoresRESUMEN
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.
Asunto(s)
Amígdala del Cerebelo/metabolismo , Condicionamiento Operante/fisiología , Dinorfinas/metabolismo , Nicotina/farmacología , Receptores Opioides kappa/metabolismo , Estrés Psicológico/metabolismo , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero/farmacología , Amígdala del Cerebelo/efectos de los fármacos , Animales , Condicionamiento Operante/efectos de los fármacos , Dinorfinas/antagonistas & inhibidores , Masculino , Ratones , Ratones Endogámicos C57BL , Receptores Opioides kappa/agonistas , Receptores Opioides kappa/antagonistas & inhibidores , Estrés Psicológico/psicología , Natación/psicologíaRESUMEN
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.
Asunto(s)
Dinorfinas/metabolismo , Hormona Luteinizante/metabolismo , Neuroquinina B/metabolismo , Ovario/metabolismo , Receptores de Neuroquinina-3/metabolismo , Receptores Opioides kappa/metabolismo , Maduración Sexual , Animales , Implantes de Medicamentos , Dinorfinas/antagonistas & inhibidores , Femenino , Hormona Luteinizante/sangre , Naltrexona/administración & dosificación , Naltrexona/análogos & derivados , Naltrexona/toxicidad , Antagonistas de Narcóticos/administración & dosificación , Antagonistas de Narcóticos/toxicidad , Proteínas del Tejido Nervioso/agonistas , Proteínas del Tejido Nervioso/antagonistas & inhibidores , Proteínas del Tejido Nervioso/metabolismo , Neuroquinina B/agonistas , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Ovario/efectos de los fármacos , Ovario/crecimiento & desarrollo , Fragmentos de Péptidos/administración & dosificación , Fragmentos de Péptidos/toxicidad , Pubertad Precoz/sangre , Pubertad Precoz/inducido químicamente , Pubertad Precoz/metabolismo , Distribución Aleatoria , Ratas , Ratas Wistar , Receptores de Neuroquinina-3/agonistas , Receptores Opioides kappa/antagonistas & inhibidores , Maduración Sexual/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Sustancia P/administración & dosificación , Sustancia P/análogos & derivados , Sustancia P/toxicidad , DesteteRESUMEN
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.
Asunto(s)
Dinorfinas/metabolismo , Hipotálamo/efectos de los fármacos , Hormona Luteinizante/metabolismo , Progesterona/farmacología , Receptores de GABA-A/metabolismo , Transducción de Señal/efectos de los fármacos , Animales , Bicuculina/farmacología , Dinorfinas/antagonistas & inhibidores , Estradiol/farmacología , Femenino , Antagonistas del GABA/farmacología , Hipotálamo/citología , Hipotálamo/metabolismo , Hipotálamo Anterior/citología , Hipotálamo Anterior/efectos de los fármacos , Hipotálamo Anterior/metabolismo , Kisspeptinas/metabolismo , Ratones Endogámicos C57BL , Naltrexona/análogos & derivados , Naltrexona/farmacología , Neuronas/citología , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Compuestos Organofosforados/farmacología , OvariectomíaRESUMEN
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.
Asunto(s)
Proteína de Unión a Elemento de Respuesta al AMP Cíclico/antagonistas & inhibidores , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Desipramina/farmacología , Dinorfinas/antagonistas & inhibidores , Dinorfinas/biosíntesis , Núcleo Accumbens/metabolismo , Estrés Fisiológico/fisiología , Animales , Células Cultivadas , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/genética , Dinorfinas/genética , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/fisiología , Masculino , Núcleo Accumbens/efectos de los fármacos , Fosforilación/efectos de los fármacos , Fosforilación/fisiología , Embarazo , Ratas , Ratas Sprague-Dawley , Estrés Fisiológico/efectos de los fármacosRESUMEN
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.
Asunto(s)
Estimulantes del Sistema Nervioso Central/farmacología , Dopamina/metabolismo , Tolerancia a Medicamentos/fisiología , Etanol/farmacología , Núcleo Accumbens/efectos de los fármacos , Receptores Opioides kappa/efectos de los fármacos , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero/farmacología , Animales , Estimulantes del Sistema Nervioso Central/administración & dosificación , Esquema de Medicación , Interacciones Farmacológicas , Dinorfinas/agonistas , Dinorfinas/antagonistas & inhibidores , Etanol/administración & dosificación , Masculino , Naltrexona/análogos & derivados , Naltrexona/farmacología , Núcleo Accumbens/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
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.
Asunto(s)
Encéfalo/metabolismo , Dinorfinas/metabolismo , Endorfinas/metabolismo , Neuropéptidos/metabolismo , Animales , Cisteína Endopeptidasas/química , Cisteína Endopeptidasas/metabolismo , Dinorfinas/antagonistas & inhibidores , Dinorfinas/química , Dinorfinas/aislamiento & purificación , Endorfinas/antagonistas & inhibidores , Endorfinas/química , Endorfinas/aislamiento & purificación , Glicopéptidos/administración & dosificación , Humanos , Neuropéptidos/antagonistas & inhibidores , Neuropéptidos/química , Neuropéptidos/aislamiento & purificación , Oligopéptidos/administración & dosificación , Enfermedad de Parkinson/metabolismo , Enfermedad de Parkinson/patología , Inhibidores de Proteasas/administración & dosificación , Ratas , Proteínas y Péptidos Salivales/administración & dosificación , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
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.
Asunto(s)
Analgésicos Opioides/química , Analgésicos Opioides/farmacología , Oligopéptidos/química , Oligopéptidos/farmacología , Péptidos Opioides/química , Péptidos Opioides/farmacología , Analgesia , Analgésicos Opioides/administración & dosificación , Analgésicos Opioides/antagonistas & inhibidores , Análisis de Varianza , Animales , Anticuerpos/inmunología , Dinorfinas/administración & dosificación , Dinorfinas/antagonistas & inhibidores , Dinorfinas/química , Dinorfinas/farmacología , Encefalina Leucina/administración & dosificación , Encefalina Leucina/antagonistas & inhibidores , Encefalina Leucina/química , Encefalina Leucina/farmacología , Encefalina Metionina/administración & dosificación , Encefalina Metionina/antagonistas & inhibidores , Encefalina Metionina/química , Encefalina Metionina/farmacología , Flúor/química , Inyecciones Espinales , Masculino , Ratones , Naloxona/administración & dosificación , Naloxona/farmacología , Naltrexona/administración & dosificación , Naltrexona/análogos & derivados , Naltrexona/farmacología , Antagonistas de Narcóticos/farmacología , Oligopéptidos/administración & dosificación , Oligopéptidos/antagonistas & inhibidores , Péptidos Opioides/administración & dosificación , Péptidos Opioides/antagonistas & inhibidores , Dolor/tratamiento farmacológico , Dolor/metabolismo , Dimensión del Dolor , Receptores Opioides kappa/antagonistas & inhibidores , Receptores Opioides mu/antagonistas & inhibidores , Receptores sigma/antagonistas & inhibidoresRESUMEN
The cAMP response element-binding protein (CREB) is a critical integrator of neural plasticity that is responsive in a brain region-specific manner to a variety of environmental and pharmacological stimuli, including widely prescribed antidepressant medications. We developed inducible transgenic lines of mice that express either CREB or a dominant-negative mutant of CREB (mCREB) in forebrain regions and used these mice to determine the functional significance of this transcription factor in the learned helplessness paradigm, a behavioral model of depression. We also use a complementary viral-mediated gene transfer approach to directly test the effect of mCREB in the nucleus accumbens, a brain region important for motivation and reward. The results demonstrate that blockade of CREB by overexpression of mCREB in transgenic mice or by viral expression of mCREB in the nucleus accumbens produces an antidepressant-like effect, whereas overexpression of CREB in transgenic mice results in the opposite phenotype. In addition, mCREB expression was colocalized with and decreased the expression of prodynorphin in nucleus accumbens medium spiny neurons, and antagonism of dynorphin in the nucleus accumbens was sufficient to produce an antidepressant-like effect similar to that observed after blockade of CREB. Together, the results demonstrate that nucleus accumbens CREB-dynorphin influence behavior in the learned helplessness model and suggest that this signaling cascade may contribute to symptoms of depression.
Asunto(s)
Proteína de Unión a Elemento de Respuesta al AMP Cíclico/antagonistas & inhibidores , Depresión/etiología , Dinorfinas/antagonistas & inhibidores , Naltrexona/análogos & derivados , Núcleo Accumbens/fisiología , Animales , Antidepresivos , Conducta Animal , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/genética , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/fisiología , Depresión/psicología , Encefalinas/análisis , Encefalinas/inmunología , Encefalinas/metabolismo , Desamparo Adquirido , Inmunohistoquímica , Masculino , Ratones , Ratones Transgénicos , Mutación , Naltrexona/farmacología , Prosencéfalo/metabolismo , Precursores de Proteínas/análisis , Precursores de Proteínas/inmunología , Precursores de Proteínas/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores Opioides kappa/antagonistas & inhibidoresRESUMEN
Whereas tissue injury increases spinal dynorphin expression, the functional relevance of this upregulation to persistent pain is unknown. Here, mice lacking the prodynorphin gene were studied for sensitivity to non-noxious and noxious stimuli, before and after induction of experimental neuropathic pain. Prodynorphin knock-out (KO) mice had normal responses to acute non-noxious stimuli and a mild increased sensitivity to some noxious stimuli. After spinal nerve ligation (SNL), both wild-type (WT) and KO mice demonstrated decreased thresholds to innocuous mechanical and to noxious thermal stimuli, indicating that dynorphin is not required for initiation of neuropathic pain. However, whereas neuropathic pain was sustained in WT mice, KO mice showed a return to baselines by post-SNL day 10. In WT mice, SNL upregulated lumbar dynorphin content on day 10, but not day 2, after injury. Intrathecal dynorphin antiserum reversed neuropathic pain in WT mice at post-SNL day 10 (when dynorphin was upregulated) but not on post-SNL day 2; intrathecal MK-801 reversed SNL-pain at both times. Opioid (mu, delta, and kappa) receptor density and G-protein activation were not different between WT and KO mice and were unchanged by SNL injury. The observations suggest (1) an early, dynorphin-independent phase of neuropathic pain and a later dynorphin-dependent stage, (2) that upregulated spinal dynorphin is pronociceptive and required for the maintenance of persistent neuropathic pain, and (3) that processes required for the initiation and the maintenance of the neuropathic pain state are distinct. Identification of mechanisms that maintain neuropathic pain appears important for strategies to treat neuropathic pain.
Asunto(s)
Dinorfinas/metabolismo , Neuralgia/metabolismo , Neuralgia/fisiopatología , Nervios Espinales/fisiopatología , Animales , Enfermedad Crónica , Modelos Animales de Enfermedad , Maleato de Dizocilpina/administración & dosificación , Dinorfinas/antagonistas & inhibidores , Dinorfinas/farmacología , Antagonistas de Aminoácidos Excitadores/administración & dosificación , Hiperestesia/metabolismo , Hiperestesia/fisiopatología , Sueros Inmunes/administración & dosificación , Inyecciones Espinales , Ligadura , Región Lumbosacra , Masculino , Ratones , Ratones Noqueados , Neuralgia/tratamiento farmacológico , Dimensión del Dolor/efectos de los fármacos , Umbral del Dolor/efectos de los fármacos , Estimulación Física , Tiempo de Reacción/efectos de los fármacos , Receptores Opioides/análisis , Receptores Opioides/metabolismo , Médula Espinal/química , Médula Espinal/metabolismo , Médula Espinal/fisiopatología , Nervios Espinales/cirugíaRESUMEN
Paradoxical opioid-induced pain has been demonstrated repeatedly in humans and animals. The mechanisms of such pain are unknown but may relate to opioid-induced activation of descending pain facilitatory systems and enhanced expression and pronociceptive actions of spinal dynorphin. Here, the possibility that these opioid-induced central changes might mediate increased excitability to the spinal cord was tested. Tactile and thermal hypersensitivity was observed at 7, but not 1, days after subcutaneous morphine pellet implantation; placebo pellets produced no effects. Basal and capsaicin-evoked release of calcitonin gene-related peptide (CGRP) was measured in minced spinal tissues taken from naive rats or rats on post-pellet days 1 and 7. The content and evoked release of CGRP were significantly increased in tissues from morphine-exposed rats at 7, but not 1, days after implantation. Morphine increased spinal dynorphin content on day 7 in rats with sham bilateral lesions of the dorsolateral funiculus (DLF) but not in rats with DLF lesions. Pharmacological application of dynorphin A(2-13), a non-opioid fragment, to tissues from naive rats enhanced the evoked release of CGRP. Enhanced evoked release of CGRP from morphine-pelleted rats was blocked by dynorphin antiserum or by previous lesions of the DLF. Sustained morphine induces plasticity in both primary afferents and spinal cord, including increased CGRP and dynorphin content. Morphine-induced elevation of spinal dynorphin content depends on descending influences and enhances stimulated CGRP release. Enhanced transmitter release may allow increased stimulus-evoked spinal excitation, which is likely to be critical for opioid-induced paradoxical pain. Such pain may manifest behaviorally as antinociceptive tolerance.
Asunto(s)
Dinorfinas/metabolismo , Morfina/administración & dosificación , Fibras Nerviosas/efectos de los fármacos , Neuronas Aferentes/efectos de los fármacos , Neurotransmisores/metabolismo , Médula Espinal/efectos de los fármacos , Animales , Conducta Animal/efectos de los fármacos , Péptido Relacionado con Gen de Calcitonina/metabolismo , Implantes de Medicamentos , Tolerancia a Medicamentos , Dinorfinas/antagonistas & inhibidores , Dinorfinas/farmacología , Calor , Hiperalgesia/inducido químicamente , Hiperalgesia/metabolismo , Sueros Inmunes/farmacología , Técnicas In Vitro , Masculino , Compresión Nerviosa , Fibras Nerviosas/metabolismo , Plasticidad Neuronal/efectos de los fármacos , Neuronas Aferentes/metabolismo , Dimensión del Dolor/efectos de los fármacos , Fragmentos de Péptidos/farmacología , Estimulación Física , Ratas , Ratas Sprague-Dawley , Médula Espinal/fisiología , Factores de TiempoRESUMEN
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.
Asunto(s)
Colecistoquinina/antagonistas & inhibidores , Dinorfinas/antagonistas & inhibidores , Sueros Inmunes/farmacología , Morfina/agonistas , Neuralgia/tratamiento farmacológico , Enfermedades del Sistema Nervioso Periférico/tratamiento farmacológico , Sustancia P/antagonistas & inhibidores , Analgésicos Opioides/administración & dosificación , Analgésicos Opioides/agonistas , Animales , Anticuerpos/farmacología , Anticuerpos/uso terapéutico , Colecistoquinina/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Resistencia a Medicamentos/efectos de los fármacos , Resistencia a Medicamentos/fisiología , Sinergismo Farmacológico , Dinorfinas/metabolismo , Inyecciones Espinales , Ligadura , Masculino , Ratones , Morfina/administración & dosificación , Neuralgia/metabolismo , Neuralgia/fisiopatología , Enfermedades del Sistema Nervioso Periférico/metabolismo , Enfermedades del Sistema Nervioso Periférico/fisiopatología , Nervio Ciático/lesiones , Nervio Ciático/cirugía , Neuropatía Ciática/tratamiento farmacológico , Neuropatía Ciática/metabolismo , Neuropatía Ciática/fisiopatología , Sustancia P/metabolismoRESUMEN
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.
Asunto(s)
Envejecimiento/metabolismo , Hormona Folículo Estimulante/metabolismo , Kisspeptinas/metabolismo , Hormona Luteinizante/metabolismo , Neuroquinina B/metabolismo , Animales , Dinorfinas/antagonistas & inhibidores , Dinorfinas/metabolismo , Encefalinas/metabolismo , Femenino , Hipotálamo/metabolismo , Masculino , Neuroquinina B/agonistas , Fragmentos de Péptidos , Precursores de Proteínas/metabolismo , Ratas Wistar , Receptores de Neuroquinina-1/agonistas , Receptores de Neuroquinina-2/agonistas , Sustancia P/análogos & derivadosRESUMEN
To investigate the involvement of endogenous opioids in acute increases in blood pressure and their functional relationship with atrial natriuretic factor and endothelin-1, we assessed plasma levels of beta-endorphin, met-enkephalin, dynorphin B, catecholamines, atrial natriuretic factor, and endothelin-1 before and after administration of the opioid antagonist naloxone hydrochloride (8 mg i.v.) in 28 hypertensive patients with a stress-induced acute increase in blood pressure. Ten patients with established mild or moderate essential hypertension and 10 normotensive subjects served as control groups. Opioids, atrial natriuretic factor, and endothelin-I were radioimmunoassayed after chromatographic preextraction; catecholamines were determined by high-performance liquid chromatography with electrochemical detection. Patients with an acute increase in blood pressure (systolic, 203.2 +/- 2.2 mm Hg; diastolic, 108.4 +/- 1.3) had plasma opioid, catecholamine, and atrial natriuretic factor levels significantly (P < .01) higher than hypertensive control patients (systolic pressure, 176.4 +/- 1.0 mm Hg; diastolic, 100.0 +/- 1.4), who had a hormonal pattern similar to that of normotensive subjects (systolic pressure, 123.2 +/- 1.5 mm Hg; diastolic, 75.0 +/- 2.0). Endothelin-1 did not differ in any group. In patients with an acute increase in blood pressure, naloxone significantly (P < .01) reduced blood pressure, heart rate, opioids, catecholamines, and atrial natriuretic factor 10 minutes after administration. Naloxone effects on blood pressure, heart rate, opioids, and catecholamines wore off within 20 minutes. In control groups, naloxone failed to modify any of the considered parameters. Our findings suggest that pressor effects of opioid peptides mediated by the autonomic nervous system during stress-induced acute episodes of blood pressure increase in hypertensive patients.
Asunto(s)
Presión Sanguínea/fisiología , Hipertensión/fisiopatología , Péptidos Opioides/fisiología , Factor Natriurético Atrial/antagonistas & inhibidores , Factor Natriurético Atrial/sangre , Factor Natriurético Atrial/fisiología , Presión Sanguínea/efectos de los fármacos , Dinorfinas/antagonistas & inhibidores , Dinorfinas/sangre , Dinorfinas/fisiología , Endorfinas/antagonistas & inhibidores , Endorfinas/sangre , Endorfinas/fisiología , Endotelina-1/antagonistas & inhibidores , Endotelina-1/sangre , Endotelina-1/fisiología , Encefalina Metionina/antagonistas & inhibidores , Encefalina Metionina/sangre , Encefalina Metionina/fisiología , Femenino , Frecuencia Cardíaca/efectos de los fármacos , Frecuencia Cardíaca/fisiología , Humanos , Hipertensión/sangre , Hipertensión/etiología , Infusiones Intravenosas , Masculino , Persona de Mediana Edad , Naloxona/administración & dosificación , Naloxona/farmacología , Antagonistas de Narcóticos/administración & dosificación , Antagonistas de Narcóticos/farmacología , Norepinefrina/antagonistas & inhibidores , Norepinefrina/sangre , Norepinefrina/fisiología , Péptidos Opioides/antagonistas & inhibidores , Péptidos Opioides/sangre , Estrés Fisiológico/complicaciones , Estrés Psicológico/complicaciones , betaendorfina/antagonistas & inhibidores , betaendorfina/sangre , betaendorfina/fisiologíaRESUMEN
Neuropathic pain states are accompanied by increased sensitivity to both noxious and non-noxious sensory stimuli, characterized as hyperalgesia and allodynia, respectively. In animal models of neuropathic pain, the presence of hyperalgesia and allodynia are accompanied by neuroplastic changes including increased spinal levels of substance P, cholecystokinin (CCK), and dynorphin. N-Methyl-D-aspartate (NMDA) receptors appear to be involved in maintaining the central sensitivity which contributes to neuropathic pain. In addition to its opioid activities, dynorphin has been suggested to act at the NMDA receptor complex. In an attempt to mimic the increased levels of spinal dynorphin seen in animal models of neuropathic pain, rats received a single intrathecal (i.t.) injection of dynorphin A(1-17), dynorphin A(1-13), dynorphin A(2-17) or dynorphin A(2-13) through indwelling catheters. Tactile allodynia was determined by measuring response threshold to probing with von Frey filaments. Dynorphin A(1-17) administration evoked significant and long-lasting tactile allodynia (i.e. > 60 days). Likewise, the i.t. administration of dynorphin A(1-13) or dynorphin A(2-17) or dynorphin A(2-13) also produced long-lasting tactile allodynia. Intrathecal pretreatment, but not post-treatment, with MK-801 prevented dynorphin A(1-17)-induced development of allodynia; i.t. administration of MK-801 alone had no effect on responses to tactile stimuli. In contrast, i.t. pretreatment with naloxone did not affect the development of tactile allodynia induced by dynorphin A(1-17) or alter sensory threshold when given alone. These results demonstrate that a single dose of dynorphin A, or its des-Tyr fragments, produces long-lasting allodynia which may be irreversible in the rat. Further, this effect appears to be mediated through activation of NMDA, rather than opioid, receptors. While the precise mechanisms underlying the development and maintenance of the allodynia is unclear, it seems possible that dynorphin may produce changes in the spinal cord, which may contribute to the development of signs reminiscent of a "neuropathic' state. Given that levels of dynorphin are elevated following nerve injury, it seems reasonable to speculate that dynorphin may have a pathologically relevant role in neuropathic pain states.
Asunto(s)
Dinorfinas/antagonistas & inhibidores , Antagonistas de Aminoácidos Excitadores/uso terapéutico , Antagonistas de Narcóticos/uso terapéutico , Fragmentos de Péptidos/antagonistas & inhibidores , Umbral Sensorial/efectos de los fármacos , Animales , Enfermedad Crónica , Maleato de Dizocilpina/uso terapéutico , Inyecciones Espinales , Masculino , Naloxona/uso terapéutico , Dolor/inducido químicamente , Enfermedades del Sistema Nervioso Periférico/tratamiento farmacológico , Ratas , Ratas Sprague-DawleyRESUMEN
Neuropathic pain is often associated with the appearance of pain in regions not related to the injured nerve. One mechanism that may underlie neuropathic pain is abnormal, spontaneous afferent drive which may contribute to NMDA-mediated central sensitization by the actions of glutamate and by the non-opioid actions of spinal dynorphin. In the present study, injuries to lumbar or sacral spinal nerves elicited elevation in spinal dynorphin content which correlated temporally and spatially with signs of neuropathic pain. The increase in spinal dynorphin content was coincident with the onset of tactile allodynia and thermal hyperalgesia. Injury to the lumbar (L(5)/L(6)) spinal nerves produced elevated spinal dynorphin content in the ipsilateral dorsal spinal quadrant at the L(5) and L(6) spinal segments and in the segments immediately adjacent. Lumbar nerve injury elicited ipsilateral tactile allodynia and thermal hyperalgesia of the hindpaw. In contrast, S(2) spinal nerve ligation elicited elevated dynorphin content in sacral spinal segments and bilaterally in the caudal lumbar spinal cord. The behavioral consequences of S(2) spinal nerve ligation were also bilateral, with tactile allodynia and thermal hyperalgesia seen in both hindpaws. Application of lidocaine to the site of S(2) ligation blocked thermal hyperalgesia and tactile allodynia of the hindpaws suggesting that afferent drive was critical to maintenance of the pain state. Spinal injection of antiserum to dynorphin A((1-17)) and of MK-801 both blocked thermal hyperalgesia, but not tactile allodynia, of the hindpaw after S(2) ligation. These data suggest that the elevated spinal dynorphin content consequent to peripheral nerve injury may drive sensitization of the spinal cord, in part through dynorphin acting directly or indirectly on the NMDA receptor complex. Furthermore, extrasegmental increases in spinal dynorphin content may partly underlie the development of extraterritorial neuropathic pain.
Asunto(s)
Dinorfinas/metabolismo , Dolor/metabolismo , Médula Espinal/metabolismo , Nervios Espinales/lesiones , Anestésicos Locales/administración & dosificación , Anestésicos Locales/farmacología , Animales , Anticuerpos Bloqueadores/farmacología , Maleato de Dizocilpina/farmacología , Dinorfinas/antagonistas & inhibidores , Dinorfinas/inmunología , Antagonistas de Aminoácidos Excitadores/farmacología , Calor , Hiperalgesia/metabolismo , Inmunoensayo , Lidocaína/administración & dosificación , Lidocaína/farmacología , Ligadura , Masculino , Dolor/etiología , Dimensión del Dolor/efectos de los fármacos , Umbral del Dolor/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Receptores de N-Metil-D-Aspartato/efectos de los fármacos , Receptores de N-Metil-D-Aspartato/metabolismo , Médula Espinal/efectos de los fármacos , Nervios Espinales/metabolismoRESUMEN
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.
Asunto(s)
Dolor/patología , Dolor/prevención & control , Animales , Hormona Liberadora de Corticotropina/administración & dosificación , Dinorfinas/antagonistas & inhibidores , Dinorfinas/biosíntesis , Dinorfinas/fisiología , Edema/inmunología , Edema/metabolismo , Edema/fisiopatología , Endorfinas/antagonistas & inhibidores , Endorfinas/biosíntesis , Endorfinas/fisiología , Encefalina Metionina/antagonistas & inhibidores , Encefalina Metionina/biosíntesis , Encefalina Metionina/fisiología , Adyuvante de Freund/administración & dosificación , Miembro Posterior , Inflamación/inmunología , Inflamación/metabolismo , Inflamación/fisiopatología , Inyecciones Subcutáneas , Leucocitos/efectos de los fármacos , Leucocitos/metabolismo , Leucocitos/fisiología , Masculino , Naloxona/administración & dosificación , Dolor/inmunología , Umbral del Dolor , Ratas , Ratas Wistar , Estrés Fisiológico/inmunología , Estrés Fisiológico/metabolismo , Estrés Fisiológico/fisiopatología , Factores de TiempoRESUMEN
The development of chronically painful states following peripheral nerve injury may involve different mechanisms depending on the nature and extent of the nerve lesion. The altered spinal neurochemistry of two substances, the excitatory amino acid glutamate operating via the N-methyl-D-aspartate receptor and the endogenous opioid peptide dynorphin, have been implicated in behavioral sequelae that follow partial peripheral nerve injury. In addition, dynorphin has nonopioid functions which may involve the N-methyl-D-aspartate receptor. We investigated two hypotheses: that the development of mechanical allodynia following complete nerve injury is not greatly influenced by the N-methyl-D-aspartate receptor, and that spinal dynorphin and glutamate expression is interdependent. These studies employed sciatic cryoneurolysis, a complete but transient peripheral nerve injury that results in a delayed mechanical allodynia beginning 21-28 days after injury. Rats were administered dizocipline maleate (MK-801) at 0.25 mg/kg twice per day intraperitoneally from days 0-7 or from days 0-21 post-lesion to pre-emptively block the N-methyl-D-aspartate receptor. In a separate group of rats, an antibody to dynorphin was administered intraperitoneally at 16.6 mg/kg twice per day from days 14 to 21 post-lesion. For all groups, the outcome of allodynia behavior was assessed using von Frey filaments at 42 days post-lesion and the resulting dynorphin and glutamate immunoreactivity in the substantia gelatinosa was measured using proportional area stained and relative optical density, respectively. Only the 0-7 day MK-801 treatment increased the resulting mechanical thresholds significantly (mean +/- S.E.M. 7.0 +/- 1.2 g) when compared to saline-injected animals (3.9 +/- 0.6 g). However, this effect did not prevent allodynia since baseline thresholds were 12 or 15 g for each group. With regard to resulting spinal immunoreactivity, anti-dynorphin antibody treatment significantly increased glutamate immunoreactivity when compared to saline-treated animals (mean relative optical density +/- S.E.M. = 807.2 +/- 3.6 versus 779.6 +/- 8.3, respectively; P = 0.01) at 42 days post-lesion. We conclude that the development of allodynia following sciatic cryoneurolysis peripheral nerve injury involved a minimal contribution from N-methyl-D-aspartate receptor activity. In addition, this study demonstrated that decreasing available dynorphin using antiserum had a significant and lasting effect on spinal glutamate expression without altering the outcome of allodynia. These conclusions suggest that etiological mechanisms leading to pain behaviors are not equal for all nerve injuries, and that altering kappa opioid levels can affect glutaminergic pathways at a substantially later time.