Short-term nerve stimulation increases enkephalin production and content in the guinea pig myenteric plexus.

Methionine-enkephalin content in the guinea pig myenteric plexus was determined before and after acute, short-term electrical or chemical stimulation. Stimulation at 20 Hz for 30 s or exposure to high potassium, the calcium channel agonist, CGP28 392, or the narcotic antagonist, (-)-naloxone, resulted in a significant increase in the content of myenteric methionine-enkephalin. The increase produced by electrical stimulation is dependent upon functional sodium channels and the presence of extracellular calcium.

The effects of etorphine on the carboxylmethylation of synaptosomal proteins of rat striatum.

The stimulation of protein carboxylmethyl transferase (PCMT) activity in rat striatal synaptosomes by the dopamine agonist, apomorphine, and a PCMT substrate, calmodulin, was measured in normal and opioid-treated rats to see if inactivation of calmodulin by methylation is a factor in opioid action. Total carboxyl methyl acceptors were measured in preparations from alkaline homogenates, while those already occupied in vivo were measured in acidic homogenates, since the carboxylmethyl group is stable in acid. The administration of etorphine acutely increased the number of already occupied acceptors while chronic morphine treatment decreased this number.

Effects of delta and mu opiopeptides on the turnover and release of dopamine in rat striatum.

Two mu and two delta opiopeptides were administered intracisternally and morphine was administered systemically to rats. The level of dopamine (DA) and its catabolites, homovanillic acid, dihydroxyphenylacetic acid and 3-methoxytramine were measured by high-pressure liquid chromatography with electrical detector in rat striatum to determine: 1) whether opioids alter the release of DA from striatal neuron (which would be indicated by changes in the level of 3-methoxytramine, the extraneuronal catabolite) and 2) whether delta or mu ligands have a greater effect on DA turnover. We found that the levels of 3-methoxytramine did not rise in response to the administration of any opiopeptide or morphine.

A method for dissection of discrete regions of rat brain following microwave irradiation.

A simple microdissection technique for obtaining discrete areas from rat brains exposed to microwave irradiation is described and illustrated. Using the atlas of Pellegrino et al. [10] as a guide, stereotaxically defined areas were removed from coronal sections prepared with sectioning stages constructed from microscope slides.

Is a calmodulin-opiopeptide interaction related to the mechanism of opioid action?

The effect of several opioids: methadone, etorphine, beta-endorphin and D-ala2met enkephalin on Ca++/calmodulin stimulation of enzyme activities either in pure solution (cyclic nucleotide phosphodiesterase) or in striatal membranes (protein kinases in synaptic membranes) were compared to see if a direct opioid/calmodulin interaction could eliminate the stimulation of enzyme activity as part of the mechanism by which opioids alter ion flow and neurotransmitter release. In other experiments, in which endogenous phosphorylation of proteins in striatal synaptic membranes was altered by opioid treatments, the possibility of restoring protein kinase activity to normal levels in the membrane preparation by supplementation with calmodulin at optimal Ca++ concentration was examined. Some opioids (methadone and D-ala2met enkephalin) did not inhibit calmodulin-stimulated phosphodiesterase, which suggests that they were not able to bind to calmodulin.

The effect of opioids on striatal calmodulin levels.

Phosphorylation of proteins in synaptic membranes of rat striatum by endogenous kinases was quantified by scanning the optical density on radioautograms made from gels after electrophoresis of assay samples. Neither the decreased phosphorylation found in samples from morphine-tolerant rats could be raised to control levels, nor could samples from untreated rats be raised to those found after acute opioid administration by the addition of Ca++ and calmodulin to the assay. However, the addition of opioids in vitro to the assay was able to inhibit the stimulation of protein kinase activity by Ca++ and calmodulin.

The role of calmodulin in opioid-induced changes in the phosphorylation of rat striatal synaptic membrane proteins.

Chronic morphine treatment of rats decreased the level of phosphorylation of synaptic membrane proteins of the striatum assayed in vitro. Although the patterns of phosphorylated proteins separated on SDS-gel electrophoresis from morphine-tolerant rats resembled patterns produced by lowering Ca2+ levels in the assay, supplementation of the protein kinase assay with Ca2+ and its binding protein, calmodulin, did not restore full kinase activity. The addition of methadone or etorphine to the protein kinase in vitro however, was able to block the Ca2+-calmodulin stimulation of phosphorylation in both synaptic membranes and intact synaptosomes.

The effect of acute opioid administration on the phosphorylation of rat striatal synaptic membrane proteins.

The acute administration of morphine, methadone, etorphine, methionine-enkephalin, leucine-enkephalin and some related pharmacologically active peptides to rats produces biphasic effects on the degree of phosphorylation of striatal synaptic plasma membrane (SPM) proteins by endogenous protein kinases in the membranes. At 1 to 5 min after opioid administration, the phosphorylation of several SPM proteins is enhanced, followed at 10 to 20 min by a severe reduction in the rate of phosphorylation of the same proteins. Multiphasic responses follow the initial biphasic response after the injection of some opioids.

The effect of morphine on the endogenous phosphorylation of synaptic plasma membrane proteins of rat striatum.

The synaptic plasma membrane (SPM) fraction was prepared from rat striata and assayed for endogenous phosphorylation in vitro. After the separation of membrane proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the incorporation of phosphate into specific membrane proteins was analyzed by gel slicing and autoradiographic techniques. Phosphate was incorporated into several SPM protein bands, two of which were regulated by cyclic adenosine 3':5' monophosphate.

The effect of morphine tolerance on the incorporation of 3H-leucine into proteins of rat synaptic membranes.

The rate of incorporation of 3H-leucine into proteins of the synaptic junctional and nonjunctional membranes and synaptic vesicles of rat brain has been examined in control and morphine-tolerant rats. There are no discernible differences between control and tolerant animals in amount of protein as measured by densitometric tracings of dye-stained proteins separated by acrylamide gel electrophoresis from the three membrane fractions of whole brain areas. However, there are differences in the turnover of membrane protein: three vesicle protein bands and one junctional-membrane protein band are significantly more highly labeled, and one junctional-membrane protein is significantly less highly labeled by 3H-leucine in samples from tolerant rats.

Endorphins in psychiatry: an overview and a hypothesis.

This article presents an overview of the biochemistry, pharmacology, and physiology of endogenous opioid peptides (endorphins). Clinical psychopharmacology of exogenous opiate agonists and antagonists is reviewed. The evidence presented in the review is compatible with a hypothesis that the level of functional endorphins may be related to psychological events, with a normal level needed for psychological homeostasis.

Effects of morphine and haloperidol on the electrical activity of rat nigrostriatal neurons.

The action of opiates on electrical activity in nigrostriatal neurons was compared with the actions of known dopamine receptor agonists and antagonists in order to clarify the effects of opiates on postsynaptic dopamine receptors in the striatum. The systemic administration of either morphine or haloperidol increased the rate of spontaneous firing of neurons in substantia nigra. The injection of either drug directly into the caudate nucleus of the striatum also increased the firing rates of nigral neurons.

The specificity of binding of the narcotic agonist etorphine in synaptic membranes of rat brain in vivo.

When 3H-etorphine was administered to rats in a pharmacologically effective dose (0.75 mug/kg intracisternally), the labeled drug was concentrated in synaptic membrane fractions isolated from the brains of rats killed 10 min after etorphine injection. Pretreatment of the animals with the narcotic antagonists naloxone, diprenorphine or l-cyclorphan, blocked the pharmacological responses to etorphine and reduced 3H-etorphine binding in the membrane fractions.

Effects of narcotic analgesic drugs on the cyclic adenosine 3',5'-monophosphate-adenylate cyclase system in rat brain.

1. The concentrations of cyclic adenosine 3',5'-monophosphate (cyclic AMP), measured in discrete brain areas removed from rats killed by microwave irradiation, rose transiently in most areas after the administration of morphine. The most pronounced changes, however, were found 2 h after doses of either 10 or 60 mg/kg morphine when cyclic AMP levels declined significantly in the hypothalamus, medulla and cerebellum.