Montford F. Piercey

Inhibition of dopamine neuron firing by pramipexole, a dopamine D3 receptor-preferring agonist : comparison to other dopamine receptor agonists

Pramipexole, an amino-benzathiazole [(S)-4,5,6,7-tetrahydro-N-6-propyl-2, 6-benzothiazolediamine dihydrochloride monohydrate] direct-acting dopamine receptor agonist effective in treating Parkinsons disease, bound selectively and with high affinity to dopamine D2-like receptors, with highest affinity at dopamine D3 receptors. Ergot dopamine receptor agonists (bromocriptine, lisuride, pergolide) bound to both dopamine and non-dopamine receptors. Although all agonists depressed dopamine neuron firing, only pramipexole and quinpirole completely silenced firing when administered in slowly-accumulating doses. High-dose pergolide, but not other ergots, completely suppressed firing when given by a prompt bolus i.v. injection, suggesting efficacy limitations may have involved receptor desensitization for pergolide, but not for bromocriptine and lisuride. We conclude that pramipexole differs from ergot dopamine receptor agonists currently used in the treatment of Parkinsons disease by virtue of its selectivity for dopamine receptors, its preferential affinity for the dopamine D3 receptor subtype, and its greater efficacy for stimulating dopamine receptors, as indicated in these electrophysiology assays.

U-50488H, a pure kappa receptor agonist with spinal analgesic loci in the mouse

U-50,488H is a chemically novel analgesic that is a potent opioid-like agent on the mouse tail flick and electrically stimulated guinea pig ileum tests. U-50,488H is a very weak competitor for naloxone binding sites in brain and ileum. However, the drug has high affinity for kappa receptor binding sites revealed by competition for EKC sites in the presence of dihydromorphine. Morphine has both supraspinal and spinal sites of action since it was a potent analgesic after both intracranial and intraspinal injections. However, U-50,488H works predominantly at the spinal level. Dynorphin may be an endogenous ligand at this site. Studies on cat dorsal horn neurons suggest that U-50,488H analgesia may be due to an increase in threshold for neuron excitation.

The dopamine D3 receptor and autoreceptor preferring antagonists (+)-AJ76 and (+)-UH232; a microdialysis study

The in vivo neurochemical profiles of haloperidol, raclopride and the dopamine D3 and autoreceptor preferring dopamine receptor antagonists (+)-UH232 and (+)-AJ76 on dopamine release and metabolism in the dorsal striatum and in the nucleus accumbens are described. It is shown that both (+)-UH232 and especially (+)-AJ76 have different effects on brain dialysate dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC) as compared to haloperidol or raclopride. It is suggested that the relative increase in dialysate dopamine over the relative increase in DOPAC is a neurochemical fingerprint, unique for different dopamine receptor antagonists. As a consequence the increased release and metabolism of dopamine after systemic administration of dopamine receptor antagonists may be controlled by different receptors and different dopamine antagonists can partly distinguish between these receptors. This may be due to their different interactions with different dopamine D2 type receptors. It is finally concluded that (+)-UH232 and especially (+)-AJ76 seem to prefer release regulating autoreceptors at the level of the axon terminals.

Electrophysiological evidence that spiperone is an antagonist of 5-HT1A receptors in the dorsal raphe nucleus.

The neuroleptic spiperone, which binds to 5-HT1A, 5-HT2 and dopamine (DA) receptors, was studied for its effects on serotonin (5-HT) and DA neurons in dorsal raphe nucleus and substantia nigra pars compacta, respectively. We found that 1 mg/kg i.v. spiperone, but not LY53837 (a 5-HT2 antagonist), antagonized the inhibition induced by 5-HT1A agonists 8-hydroxy-dipropylaminotetralin (8-OH-DPAT) and buspirone in the dorsal raphe nucleus. Lower spiperone doses blocked DA receptors in substantia nigra pars compacta, but did not affect 5-HT neurons. Doses of 8-OH-DPAT completely silencing dorsal raphe neurons were ineffective in substantia nigra pars compacta. However, buspirone antagonized DA receptors in substantia nigra pars compacta with doses similar to those depressing dorsal raphe neurons. It is concluded that spiperone is an antagonist of 5-HT1A receptors in the dorsal raphe nucleus.

Reversal of scopolamine-induced amnesia and alterations in energy metabolism by the nootropic piracetam: implications regarding identification of brain structures involved in consolidation of memory traces

Pretreatment with scopolamine, 3 mg/kg, prevented the acquisition of a passive avoidance task in rats. These amnesic effects of scopolamine could largely be overcome by treatment with 100 mg/kg of the nootropic drug piracetam. In order to identify the brain structures involved, the effects of these drugs on regional energy metabolism were measured throughout the brain, utilizing Sokoloffs 2-deoxyglucose autoradiographic procedures. Scopolamine, 3 mg/kg, reduced glucose utilization in several areas of the cerebral cortex. These effects were largest in the parietal and temporal cortices. Other areas affected included the sensorimotor and cingulate cortices, the ventral and lateral thalamus, and the dendritic neuropil of the CA1, CA2, and CA3 regions of the hippocampus. The regional depressions in glucose metabolism observed following scopolamine treatment in the rat had some resemblance to depressions in glucose metabolism reported for Alzheimers disease patients in positron emission tomography studies. Piracetam, 100 mg/kg, did not alter the energy metabolism of any of the 41 brain regions examined. However, this dose of piracetam completely reversed the scopolamine-induced depressions in the hippocampus. Piracetam partially but significantly reversed the scopolamine effects in the cingulate cortex. It is concluded that the data provide support for the hippocampal-cholinergic theory of memory as originally formulated by Meyers and Domino in 1964 and give insight into the mechanisms by which nootropics work.

Spinal and supraspinal sites for morphine and nefopam analgesia in the mouse

Using the tail-flick and hot-plate assays, morphine and nefopam were tested for analgesic activity following intraperitoneal (i.p.), intracranial (i.c.) and intraspinal (i.s.) injection in mice. By the i.p. route, morphine was equipotent on both analgesic tests. Nefopam was one-third as potent as morphine on the hot-plate test, but did not affect the tail-flick. Intracranial morphine was more effective on the hot-plate than on the tail-flick, but i.s. morphine was most potent on the tail-flick. Naloxone, 0.5 mg/kg i.p., totally reversed morphines effects on the tail-flick, but only partially reversed these actions on the hot-plate, suggesting the possibility that morphines effects on the mouse hot-plate test may be mediated via multiple receptor types. Nefopam was more potent by the i.c. route than by the i.p. route, but its was inactive spinally. Nefopam analgesia was unaffected by naloxone treatment. It is concluded that nefopam is a novel, centrally acting, non-narcotic analgesic.

Use of substance P fragments to differentiate substance P receptors of different tissues

The C- and N-terminal fragments of substance P were compared to the parent molecule with respect to their ability to: (a) contract the isolated guinea pig ileum, (b) induce salivation in the rat, (c) excite single cat dorsal horn neurones, and (d) induce scratching by intracranial injections in mice. C-terminal fragments as small as the heptapeptide were potent SP agonists on all assay systems. C-terminal fragments containing five amino acids or less were, at most, only weakly active. The C-terminal hexapeptide was a potent SP receptor stimulant on the isolated guinea pig ileum and, when directly applied by microiontophoresis, on cat dorsal horn neurons. However, the same compound was only 2-5% as potent as substance P in eliciting salivation and scratching in vivo, an indication that this fragment may be especially labile to enzymatic degradation. N-terminal fragments were totally inactive on the isolated guinea pig ileum. On the rat salivation and central nervous system assays, however, N-terminal fragments were capable of weak SP-like activity. It is concluded that SP receptors exist in multiple forms which we have labelled SP1 and SP2 receptors for those insensitive or sensitive to N-terminal fragments, respectively.

Pharmacological characterization of scratching behaviour induced by intracranial injection of substance P and somatostatin.

Abstract Substance P (SP) injected intracranially into mice induced the reciprocal hindlimb scratching syndrome (RHS) of Rackham and Share (1979). The C-terminal SP hexapeptide (SP 6–11) also induced scratching, but the pentapeptide (SP 7–11) caused no scratching at all. Thus, the SP receptor for scratching is similar to those of other tissues. Intracranial SP did not alter tail flick latencies at 0.3 μg. Similar injections of other putative neurotransmitters (both biogenic amines and peptides) failed to mimic the effects of SP precisely, although both kainic acid and somatostatin produced weaker scratching syndromes. Substance P scratching was nonspecifically antagonized by a variety of pharmacological agents. Given intravenously or intraperitoneally, SP did not elicit scratching. Therefore, peripherally administered SP did not reach the receptor for scratching. This contrasts with reports that intraperitoneal SP causes long-lasting analgesia. However, intravenous somatostatin elicited scratching with a potency similar to that of intracranial somatostatin; thus, the site of action for this peptide may be distant from the intracranial injection site.

Morphine does not antagonize the substance P mediated excitation of dorsal horn neurons.

Multibarrelled microelectrodes were used to test the effects of iontophoretically released substance P (SP), morphine, glutamate, and naloxone on spinal cord dorsal horn neurons. Cells excited by SP were also excited by noxious stimuli, a finding consistent with the hypothesis that SP is the neurotransmitter released by primary nociceptor afferents to excite dorsal horn neurons. Iontophoretic morphine failed to depress the SP-induced discharges. Indeed, iontophoretic morphine frequently potentiated the SP responses. In addition to potentiating SP-induced discharges, iontophoretic morphine frequently increased both the spontaneous activity of dorsal horn neurons and the activity evoked in these cells by noxious cutaneous heat and iontophoretic glutamate. Naloxone did not antagonize these excitatory effects. Intravenous morphine only depressed spontaneous discharges. Nevertheless, iontophoretic morphine still produced excitatory effects in spinal animals pretreated with analgesic doses of intravenous morphine. It is concluded that such excitatory effects are toxic actions indicative of supratherapeutic morphine concentrations in the vicinity of the neuron being studied. Intravenously administered morphine depressed the spontaneous activity of dorsal horn neurons of spinal cats, but failed to depress their responses to SP. Morphine also failed to antagonize SPs biological effects in peripheral systems (contraction of isolated guinea pig ileum, rabbit hypotensive effect, rat sialogogic response). It is concluded that morphine is not a substance P receptor antagonist. The results are discussed with respect to the hypotheses that (1) the spinal analgesic effects of systemically administered morphine occur on presynaptic terminals of sensory neurons, and (2) an SP antagonist might be a unique analgesic agent.

Analgesic activities of spinal cord substance P antagonists implicate substance P as a neurotransmitter of pain sensation

Substance P (SP) injected into intraspinal (i.s.) spaces caused mice to vigorously scratch and bite their skins in an apparent reaction to a perceived cutaneous sensation. The scratching behavior was similar to the reciprocal hindlimb-scratching syndrome (RHS) described for intracranial (i.c.) SP injections. Radiotracer experiments, as well as potency and latency measurements, demonstrated that SP-induced scratching, whether induced by the i.c. or i.s. route, was due to SP receptor stimulation in the cervicothoracic cord. Similarly, biting was due to SP stimulation of the lumbosacral spinal cord. Mice coated with capsaicin, an irritant chemical, scratched and bit the coated areas in a manner similar to animals injected with i.s. SP. Standard analgesics depressed this scratching behavior elicited by topical capsaicin. Non-analgesic drugs, with the exception of amphetamine, did not affect capsaicin-induced pain. It is concluded that i.s. SP induces a painful sensory experience. Some piperazinone derivatives of substance Ps C-terminal hexapeptide are shown to specifically antagonize the scratching induced by i.s. SP with little or no effect on motor behavior. These antagonists depressed scratching elicited by topical capsaicin and were analgesic on the hot-plate test. It is concluded that SP is a natural neurotransmitter for pain and that antagonism of endogenous SP systems causes analgesia.