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RPEP-00073 · 1988

Opioid Peptides Relaxed Blood Vessels by Blocking Nerve Signals

In rabbit ear and saphenous (leg) arteries, both delta-selective opioid agonists (leu-enkephalin and met-enkephalin) and kappa-selective agonists (dynorphin-(1-13) and ethylketocyclazocine) inhibited nerve-stimulation-evoked contractions. Delta agonists showed a vessel-specific difference: their maximum inhibitory effect was significantly less in the saphenous artery than in the ear artery. Kappa agonists worked similarly in both vessels. Yohimbine (an alpha-2 adrenergic blocker) did not enhance opioid inhibitory effects at any stimulation frequency in either vessel. This disproved the hypothesis that opioid receptors work by boosting alpha-2 receptor inhibition of noradrenaline release.

Gan, E A; Duckles, S P · In Vitro

RPEP-00074 · 1988

Mu-Opioid Receptors in the Amygdala Control Feeding Behavior

The mu-selective opioid agonist DAGO (Tyr-D-Ala-Gly-(Me)Phe-Gly-ol) increased food intake when injected into the central nucleus of the amygdala at doses of 1 and 3 nanomoles. Neither DSLET (a delta-selective agonist) nor dynorphin A (a kappa-selective agonist) affected food intake at the same location, even at the highest dose of 3 nanomoles. Dynorphin A did increase food intake when injected into the medial hypothalamus at 2 nanomoles. This shows that different brain regions use different opioid receptor types to control eating. Bilateral injections of DAGO were no more effective than unilateral injections. Naloxone and the long-acting antagonist beta-chlornaltrexamine both blocked DAGO's feeding effect, confirming opioid receptor involvement.

Gosnell, B A · Animal Study

RPEP-00075 · 1988

Opioid Peptides Did Not Directly Affect Fat Cell Metabolism

Six opioid-related substances were tested on isolated rat adipocytes (fat cells): - Beta-endorphin, dynorphin, met-enkephalin, leu-enkephalin, and morphine (agonists) - Naloxone (antagonist) None altered insulin binding to fat cells. None changed 2-deoxy-glucose uptake (a measure of sugar absorption). None affected glucose incorporation into lipids (fat production). None showed lipolytic (fat-breaking) activity. The conclusion: endogenous opioids do not directly affect fat cell metabolism. The glucose problems seen with opioid drugs must happen through other mechanisms, likely in the brain, liver, or pancreas.

Hauner, H; Glatting, G; Ditschuneit, H H; Pfeiffer, E F · In Vitro

RPEP-00076 · 1988

Seizures Trigger a Surge-Then-Rebound Pattern in Brain Opioid Peptides

After seizures (from electroshock, kainic acid, or kindling), both dynorphin and enkephalin initially decrease in the hippocampus, indicating release. But their recovery patterns differ dramatically. Enkephalin shows a rapid, sustained rebound above normal levels after all three seizure types. Dynorphin shows a large, sustained decrease after electroshock and kindling, with slow recovery. Mu-opioid receptor antagonists shortened the kindling process (where repeated mild stimulation eventually produces seizures). This strongly suggests brain opioid peptides are involved in how seizure susceptibility develops. Enkephalin in the hippocampus appears to mediate wet-dog shakes (a specific seizure behavior seen in rodents), connecting it to the opioid withdrawal syndrome.

Hong, J S; McGinty, J F; Grimes, L; Kanamatsu, T; Obie, J; Mitchell, C L · Review

RPEP-00077 · 1988

Cortisol Blocked Opioid-Induced Prolactin Release in Rats

Cortisol at 25 mg/kg given 24 hours before testing decreased prolactin release triggered by all four opioid peptides (dynorphin, beta-endorphin, met-enkephalin, D-Met-Pro-enkephalinamide) injected into the brain. Actinomycin D pretreatment blocked cortisol's inhibitory effect. This means cortisol works by triggering new protein synthesis, not by directly blocking opioid receptors. In adrenalectomized (no adrenal glands) rats, cortisol's inhibitory effect was even stronger. Maximum inhibition was reached at only 5 mg/kg, lower than the 25 mg/kg used in intact rats. Cortisol did not affect opioid-induced corticosterone release. This selectivity means cortisol specifically targets the prolactin pathway, not all opioid effects.

Kiem, D T; Kanyicska, B; Stark, E; Fekete, M I · Animal Study

RPEP-00078 · 1988

A Commercial Dynorphin Product Was Contaminated and Gave Misleading Results

A commercially purchased synthetic dynorphin A-(1-13) raised cytosolic calcium in rat pituitary cells. However, this effect was NOT caused by dynorphin itself. When the same preparation was purified by HPLC, the calcium-raising effect disappeared. HPLC combined with LHRH radioimmunoassay revealed contamination with a LHRH-like compound (luteinizing hormone-releasing hormone). The LHRH antagonist blocked the calcium effect, confirming the contamination. Pure dynorphin A-(1-13), dynorphin A-(2-13), leu-enkephalin, beta-endorphin, morphine, and U50,488H all had zero effect on pituitary calcium. LHRH itself raised calcium by about 50 nM, which was blocked by the LHRH antagonist.

Knepel, W; Schöfl, C; Wesemeyer, G; Götz, D M · In Vitro

RPEP-00079 · 1988

How the Hypothalamus Uses Peptide Receptors to Control Hormones

Endogenous opioid peptides inhibit the release of both vasopressin and oxytocin from the posterior pituitary. They also affect anterior pituitary hormones through the hypothalamic portal blood system. Dynorphin mRNA co-exists in the same neurons as vasopressin. Stimuli that increase vasopressin secretion (like salt loading) also increase dynorphin mRNA accumulation. This parallel regulation strongly suggests dynorphin has a genuine role alongside vasopressin. Pro-enkephalin A mRNA co-exists with CRF in a different group of hypothalamic cells. Stresses that increase CRF mRNA also increase pro-enkephalin mRNA in the same area. The co-existence of opioid peptides with established hormones in the same cells, combined with their coordinated regulation, provides powerful evidence that opioids are genuine neuroendocrine regulators, not incidental bystanders.

Lightman, S L · Review

RPEP-00080 · 1988

GHK-Cu Stimulated Collagen Production at Extremely Low Concentrations

GHK-Cu stimulated collagen synthesis in fibroblast cell cultures. The effect began at concentrations between 10 picomolar (10^-12 M) and 100 picomolar (10^-11 M), an extraordinarily low threshold. The stimulation maximized at 1 nanomolar (10^-9 M). Above this concentration, the effect plateaued. The collagen increase was independent of cell number changes. GHK-Cu was not simply making more cells; it was making existing cells produce more collagen. A GHK tripeptide sequence (glycine-histidine-lysine) exists naturally within the alpha 2(I) chain of type I collagen. The authors proposed that when wounds break down collagen, proteases could release GHK fragments. These fragments would then bind copper and stimulate new collagen production at the wound site, creating a self-healing feedback loop.

Maquart, F X; Pickart, L; Laurent, M; Gillery, P; Monboisse, J C; Borel, J P · In Vitro

RPEP-00081 · 1988

Inflammation Raised Spinal Dynorphin and Changed Pain Sensitivity

Within 24 hours of inflammation, dynorphin increased specifically in the ipsilateral (same-side) dorsal horn of the spinal cord. Met-enkephalin, leu-enkephalin, and opioid receptor densities did not change. The inflamed paw showed pronounced supersensitivity to morphine's painkilling effect against pressure pain, starting at 24 hours and increasing at 1 week. Opioid antagonists (naloxone and the kappa-preferring MR 2266) made the pain worse, suggesting endogenous opioids were already partially controlling the pain. By 3-5 weeks, inflammation had spread to the opposite paw. Dynorphin was now elevated bilaterally and in the cervico-thoracic cord. Met-enkephalin and leu-enkephalin also rose bilaterally. Adrenal glands were enlarged and plasma beta-endorphin was elevated, signs of chronic stress. No changes in mu, delta, or kappa receptor density were found at any time point.

Millan, M J; Członkowski, A; Morris, B; Stein, C; Arendt, R; Huber, A; Höllt, V; Herz, A · Animal Study

RPEP-00083 · 1988

Blocking Stress Hormone Receptors Changed Multiple Opioid and Reproductive Peptides

Alpha-helical CRF9-41 (a CRF receptor blocker) applied to hypothalamic slices in vitro caused beta-endorphin and met-enkephalin to drop significantly within 10 minutes. At the same time, LHRH (luteinizing hormone-releasing hormone) rose significantly. Dynorphin also decreased but the change was not statistically significant. When the antagonist was removed, the pattern reversed within 10 minutes: beta-endorphin, met-enkephalin, and dynorphin rose back up while LHRH fell back down. The same results were obtained in vivo using push-pull perfusion of the arcuate-median eminence region in anesthetized rats. This confirmed the in vitro findings in living animals.

Nikolarakis, K E; Almeida, O F; Sirinathsinghji, D J; Herz, A · Animal Study

RPEP-00084 · 1988

GABA Controls Reproductive Hormone Release Through Opioid Peptide Intermediaries

GABA at concentrations from 10^-8 to 10^-4 M caused a dose-dependent increase in GnRH release from rat hypothalamic slices. The GABA-A receptor agonist isoguvacine replicated this, while the GABA-B agonist baclofen had no effect. The GABA-A antagonist SR95103 blocked the GABA effect. Blockade of nerve conduction with tetrodotoxin abolished GABA's stimulatory effect, meaning it works through neural circuits, not directly on GnRH cells. Naloxone (opioid blocker) prevented GABA-induced GnRH release. The CRF antagonist also blocked it. CRF itself decreased GnRH release, and GABA could not reverse this. GABA stimulated the release of beta-endorphin, dynorphin, and met-enkephalin from the same tissue, connecting opioid peptide release to the control of reproduction.

Nikolarakis, K E; Loeffler, J P; Almeida, O F; Herz, A · In Vitro

RPEP-00085 · 1988

Delta-Opioid Peptide Triggered Hibernation in Active Ground Squirrels

DADLE (D-Ala2-D-Leu5 enkephalin), a delta-opioid receptor agonist, at 1.50 mg/kg/day induced summer hibernation comparable to that caused by natural Hibernation Induction Trigger (HIT). Morphine (1.50 mg/kg/day), morphiceptin (0.82 mg/kg/day), and dynorphin A (0.82 mg/kg/day) did not induce hibernation. Strikingly, morphine, morphiceptin, and dynorphin A actually antagonized HIT-induced hibernation. They blocked the natural hibernation signal. This suggests delta-opioid receptors promote hibernation while mu and kappa opioid receptors promote the arousal (waking) state. Natural hibernation may involve a shift in the balance between these receptor systems.

Oeltgen, P R; Nilekani, S P; Nuchols, P A; Spurrier, W A; Su, T P · Animal Study

RPEP-00086 · 1988

Naloxone Enhanced Delayed Stomach Contractions Triggered by Vagus Nerve

Naloxone (100 to 1000 micrograms/kg) did not affect the initial stomach response to vagus nerve stimulation. But it dose-dependently enhanced the delayed response (from vagal afferent fibers). This means endogenous opioids normally restrain the delayed stomach excitation. Met-enkephalin was markedly more potent than the mu-agonist DAGO and the kappa-agonist dynorphin A(1-13) at inhibiting both types of vagal stomach responses. The delta-selective agonist DPDPE mimicked met-enkephalin's inhibitory effects. The delta-selective antagonist ICI 174,864 blocked DPDPE's effects. This confirmed delta-opioid receptors mediate the gastric inhibition. The conclusion: natural opioid peptides (likely enkephalins) tonically restrain vagus nerve-driven stomach excitation through delta receptors.

Okamoto, T; Kurahashi, K; Fujiwara, M · Animal Study

RPEP-00087 · 1988

Opioid Peptides Had Bidirectional Effects on Human Natural Killer Cells

Met-enkephalin, leu-enkephalin, dynorphin (1-13), DSLET (delta agonist), and DAGO (mu agonist) were tested on human NK cells from healthy donors after 18-hour incubation. The results were bidirectional: populations with low baseline NK activity showed enhancement. Populations with high baseline NK activity showed suppression. This immunoregulatory pattern was consistent across different opioid peptides. The effect was confirmed in a serum-free system with recombinant interferon-alpha, ruling out serum-factor interference. Naloxone displayed both antagonist properties (blocking opioid effects) and direct immunomodulatory effects. This suggested lymphocytes themselves may produce opioid peptides in culture, creating an autocrine signaling loop.

Oleson, D R; Johnson, D R · In Vitro

RPEP-00088 · 1988

Cutting Any Peripheral Nerve Caused Brain-Wide Opioid Peptide Changes

Regardless of which nerve was cut (right or left sciatic, both sciatic, right brachial plexus, saphenous, or sural), the same brain-wide pattern emerged. Beta-endorphin decreased significantly in all brain areas except the striatum. Met-enkephalin increased in all brain areas and in the affected spinal cord segments. Substance P, somatostatin, and dynorphin were unaffected. The changes appeared within 24 hours of surgery and persisted for at least 4 months, indicating a long-lasting reorganization of the opioid system. There was no lateralization: unilateral nerve cuts produced bilateral brain changes. This means local nerve damage triggers a global brain response. Serotonergic drugs normalized beta-endorphin levels, suggesting the serotonin system mediates the opioid changes after nerve injury.

Panerai, A E; Sacerdote, P; Brini, A; Bianchi, M; Mantegazza, P · Animal Study

RPEP-00089 · 1988

Two Mouse Strains Had Opposite Stress Pain Relief Despite Similar Opioid Systems

DBA mice had higher baseline pain thresholds than C57 mice. But C57 mice showed much greater stress-induced analgesia (pain relief from stress). The opioid systems differed genetically: DBA mice had significantly more dynorphin in the hypothalamus and neurointermediate pituitary. C57 mice had lower KD values (better affinity) for spinal mu receptors but higher KD (lower affinity) for cerebral kappa receptors. Under stress, the two strains activated different opioid systems. C57 mice: beta-endorphin decreased in hypothalamus and increased in neurointermediate pituitary lobe. DBA mice: dynorphin decreased in hypothalamus and beta-endorphin increased in the anterior pituitary. Both strains showed spinal cord dynorphin decreases under stress.

Przewłocka, B; Vetulani, J; Lasoń, W; Dziedzicka, M; Silberring, J; Castellano, C; Przewłocki, R · Animal Study

RPEP-00090 · 1988

Low-Dose Enkephalin Blocked Substance P Pain Without Producing Direct Pain Relief

Intrathecal (spinal cord) met-enkephalin at low doses markedly reduced the aversive behavior caused by intrathecal substance P. At these same low doses, met-enkephalin produced no measurable analgesia in the tail-flick pain test. Met-enkephalin needed much higher doses to produce general analgesia. Beta-endorphin and dynorphin-(1-17) were different. They blocked substance P-induced behavior and produced general analgesia at roughly the same doses. There was no separation between the two effects. This suggests met-enkephalin has a specific, preferential interaction with substance P-mediated pain signaling in the spinal cord, separate from its general painkilling effects.

Sakurada, T; Takahashi, K; Sakurada, S; Kisara, K; Folkesson, R; Terenius, L · Animal Study

RPEP-00091 · 1988

Beta-Endorphin Specifically Binds to Lung and Liver in Living Rats

Radioactive beta-endorphin injected intravenously accumulated specifically in the lung and liver but not other tissues. Four lines of evidence confirmed this was specific receptor binding, not passive trapping. First, adding excess unlabeled beta-endorphin reduced the labeled peptide in lung and liver. Second, injecting unlabeled beta-endorphin via the femoral vein (which passes through the lungs first) rapidly increased blood levels of the pre-injected labeled peptide, displacing it from lung binding sites. Injection via the carotid artery (bypassing the lungs) did not have this effect. Third, immunoreactive labeled peptide that had lost its receptor-binding ability did not accumulate in lung or liver. Fourth, dynorphin (1-13) and ethylketocyclazocine (kappa agonist) displaced beta-endorphin from these sites, but DADLE (delta agonist) and naloxone (mu antagonist) did not. This pharmacological profile indicates kappa-type binding sites.

Sato, H; Sugiyama, Y; Sawada, Y; Iga, T; Hanano, M · Animal Study

RPEP-00092 · 1988

Gallbladder Muscle Cells Contracted in Response to Opioid Peptides

Three opioid peptides caused dose-dependent contraction of isolated gallbladder muscle cells: met-enkephalin > dynorphin(1-13) > leu-enkephalin. The contractions were blocked by the opioid antagonists naloxone and MR2266 but not by muscarinic, CCK/gastrin, or tachykinin antagonists. This confirmed the response was specifically through opioid receptors. Differential sensitivity to preferential antagonists of mu (naloxone) and kappa (MR2266) receptors, combined with the different potencies of the three agonists, indicated the presence of mu, delta, and kappa opioid receptors on the gallbladder muscle cells. These opioid peptides are known to be present in myenteric neurons of the gut, providing the natural source of opioid signals for gallbladder muscle.

Severi, C; Grider, J R; Makhlouf, G M · In Vitro

RPEP-00093 · 1988

Male and Female Rat Brains Showed Different Enkephalin Patterns in a Reproductive Region

Three opioid peptide families showed distinct, non-overlapping distributions in the preoptic brain region. Beta-endorphin fibers were mainly in the periventricular nucleus. Dynorphin B fibers were more uniformly distributed with few cell bodies. Enkephalin (peptide E) had hundreds of cell bodies, concentrated in specific nuclei. The key sexual dimorphism: male rats had significantly more enkephalin cells in the anteroventral periventricular nucleus (AVPv) than females. This was despite the AVPv being physically larger in female rats. This sex difference was at least partially dependent on perinatal gonadal steroids. Early hormone exposure during the critical developmental window permanently organized the number of enkephalin cells, a process called organizational hormone effects. No sex difference was found in enkephalin cells in the anterodorsal preoptic nucleus, showing the dimorphism was region-specific, not a general property of enkephalin neurons.

Simerly, R B; McCall, L D; Watson, S J · Animal Study

RPEP-00094 · 1988

Different Opioid Peptides Had Opposite Effects on Brain Reward in Different Regions

Leu-enkephalin and leu-enkephalinamide inhibited electrical self-stimulation in the substantia nigra/ventral tegmental area (SN-VTA) but not the medial forebrain bundle/lateral hypothalamus (MFB-LH). However, injection into MFB-LH facilitated self-stimulation of the distant SN-VTA. Ala-leu-enkephalin injected into SN-VTA actually facilitated its self-stimulation (opposite to leu-enkephalin). The same peptide in MFB-LH facilitated SN-VTA stimulation like the other enkephalins. Dynorphin A(1-13) injected into SN-VTA facilitated its self-stimulation. Injection into MFB-LH had no effect. Met-enkephalin had no direct or indirect effects in either region. These differences likely reflect different receptor preferences in the SN-VTA neural organization.

Singh, J; Desiraju, T · Animal Study

RPEP-00096 · 1988

Seizure-Induced Opioid Peptides in Spinal Fluid Raised Seizure Thresholds

CSF from rats that had just experienced an electroshock seizure significantly raised seizure thresholds in recipient rats when injected into their brain ventricles. This anticonvulsant activity was blocked by high-dose naloxone and by the selective delta-opioid antagonist ICI 174,864. This identified delta-opioid receptors as the mediators. The active substance was destroyed by heat (90°C) and by trypsin (a protein-digesting enzyme), confirming it is a peptide. It passed through 10,000 dalton membranes but not 5,000 dalton membranes, putting its size between 5,000-10,000 daltons. Post-seizure CSF had increased beta-endorphin (31 amino acids, ~3,500 daltons) but not dynorphin A, leu-enkephalin, or met-enkephalin. However, hidden met-enkephalin sequences (released by trypsin digestion of larger peptides) were found. The size (5,000-10,000 Da) and delta-receptor specificity do not match any known opioid peptide, suggesting an undiscovered anticonvulsant opioid peptide.

Tortella, F C; Long, J B · Animal Study

RPEP-00118 · 1989

Enkephalin Nerve Fibers in the Prostate Decline With Age

Proenkephalin-derived peptide nerve fibers in human prostate were concentrated in the dorsolateral stroma and decreased in density with age. No prodynorphin or POMC-derived peptides were detected.

Jungblut, T; Aumüller, G; Malek, B; Melchior, H · Cross Sectional

RPEP-00123 · 1989

Beta-Endorphin and Dynorphin Mimicked Melatonin Immune-Boosting Effects

Beta-endorphin and dynorphin replicated melatonin's immunoenhancing and anti-stress effects, but with complementary roles: beta-endorphin for normal conditions, dynorphin for stress states. Effects were circadian and opioid-receptor-mediated.

Maestroni, G J; Conti, A · Animal Study