Difference in changes of membrane fluidity of polymorphonuclear leukocytes stimulated with phorbol myristate acetate and formyl-methionyl-leucyl-phenylalanine: role of excited oxygen species

Polymorphonuclear leukocytes (PMN) were stimulated with phorbol myristate acetate (PMA) and N-formyl-methionyl-leucyl phenylalanine (FMLP) to clarify the role of excited oxygen species in inducing changes of membrane fluidity. Membrane fluidity was assessed by the excimer-forming lipid technique using pyrenedecanoic acid and flow cytometry. Membrane fluidity of PMN decreased following stimulation with PMA, and the extent of decrease was both time- and dose-dependent. FMLP at 10(-5) M induced a decrease, while FMLP at 10(-7) M induced a rapid increase. On stimulation with 10(-7) M FMLP as well as in a resting condition, the change of membrane fluidity of PMN from patients with chronic granulomatous disease (CGD) was similar to that of normal PMN. However, on stimulation with PMA or 10(-5) M FMLP, CGD PMN did not show a significant decrease. In addition, normal PMN incubated with catalase inhibited the decrease. These findings suggest that the generation of excited oxygen species, particularly of H2O2, is important in inducing a decrease of PMN membrane fluidity.     

Role of Ca2+ and Mg2+ in some human neutrophil functions as indicated by ionophore A23187.

Studies with the divalent cation ionophore A23187 suggest that both Ca2+ and Mg2+ ion influx play a role in human peripheral blood neutrophil function. Degranulation of neutrophils occurred at ionophore concentrations of 10(-5) M and was Ca2+ but not Mg2+ dependent. Modulation of neutrophil chemotaxis was enhanced optimally by 10(-7) M ionophore and was both Ca2+ and Mg2+ dependent. Concentrations of ionophore as low as 10(-12) M seemed to sensitize the cells to a concentration of phorbol myristate acetate which by itself was not chemotactic. These findings also indicate that factors other than Ca2+ or increases in cyclic nucleotides are important to initiation and modulation of neutrophil function.     

Chemotactic peptide enhancement of phorbol ester-induced protein kinase C activity in human neutrophils

Chemotactic factors and phorbol esters may act synergistically to evoke neutrophil responses, but the mechanism of such interaction is not entirely clear. We investigated the combined effects of the chemotactic peptide n-formyl-methionyl-leucyl-phenylalanine (FMLP) and phorbol myristate acetate (PMA) on protein kinase C (PKC) activity in human neutrophils. FMLP had little effect on the sharp decrease in cytosolic PKC activity induced by PMA. However, cells exposed to FMLP and PMA exhibited a synergistic increase in particulate PKC activity (1 +/- 1 pmol 32P/10(7) PMNs/min in unstimulated cells, 53 +/- 12 pmol 32P with 20 ng/ml PMA, 6 +/- 3 pmol 32P with 10(-7) M FMLP, and 191 +/- 17 pmol 32P with FMLP and PMA). FMLP also markedly increased calcium/phospholipid-independent protein kinase activity in particulate fractions of control and PMA-treated cells. Enhancement of PKC activity required the presence of cytochalasin B during cell stimulation. Cellular calcium was crucial to the FMLP effect since enhancement was decreased in cells incubated with EGTA or Quin2. These results suggest that chemotactic factors and phorbol esters may mediate synergistic effects on neutrophil responses through enhancement of particulate PKC activity. The enhancing effect is probably mediated through chemoattractant-mediated increases in intracellular calcium.     

Chemiluminescence by human alveolar macrophages: stimulation with heat-killed bacteria or phorobol myristate acetate.

Chemiluminescence of human alveolar macrophages (AM) was evaluated in vitro. Unstimulated AM generated chemiluminescence that remained constant during incubation. Addition of heat-killed Staphylococcus aureus 502A (HKB) or a chemical agent, phorbol myristate acetate, produced high rates of chemiluminescence that were significantly (P less than 0.05) increased over unstimulated AM. Phorbol myristate acetate-and HKB-stimulated increases in AM chemiluminescence were completely blocked by the enzyme superoxide dismutase. In comparison with unstimulated polymorphonuclear leukocytes, unstimulated AM had significantly (P less than 0.005) greater levels of chemiluminescence. However, after stimulation by phorbol myristate acetate or HKB, AM showed less chemiluminescence than similarly treated polymorphonuclear leukocytes.     

Further characterization of preproenkephalin mRNA-containing cells in the rodent globus pallidus

The globus pallidus (external pallidum of primates) is an essential nucleus within basal ganglia circuitry, in part because it receives at least one-half of striatal efferent projections. Neurons of the globus pallidus can be divided into subpopulations based on anatomical, physiological, and chemical features. Globus pallidus neurons project to several structures (the striatum, subthalamic nucleus, entopeduncular nucleus, and substantia nigra pars reticulata), have one of two alternative waveforms (positive/negative versus negative/positive), contain either the calcium binding protein parvalbumin or the neuropeptide precursor preproenkephalin mRNA and show differential immediate early gene responses to dopamine receptor agonists and antagonists. The objective of the present study was to characterize in greater detail the preproenkephalin mRNA-containing pallidal neurons using Sprague-Dawley rats. In situ hybridization for preproenkephalin mRNA was combined with immunocytochemical detection of: (i) the neuron-specific nuclear protein, NeuN, (ii) FluoroGold-labeled pallidostriatal and pallidosubthalamic cells, or (iii) Fos induced by either systemic combined D1-class/D2-class dopamine receptor agonists or a D2-class receptor antagonist. These experiments demonstrated that a substantial population (42%) of globus pallidus neurons contains preproenkephalin mRNA, and that globus pallidus neurons retrogradely labeled after FluoroGold injections into the striatum are more frequently preproenkephalinergic, compared to the population of pallidosubthalamic neurons. Furthermore, systemic administration of a D2 receptor antagonist, eticlopride, induced Fos immunoreactivity predominantly in globus pallidus neurons expressing preproenkephalin mRNA, while combined administration of D1 and D2 receptor agonists induced Fos predominantly in pallidal neurons lacking preproenkephalin mRNA.These results support the conclusion that preproenkephalin mRNA identifies one of the two major subpopulations of pallidal neurons. This preproenkephalin mRNA-expressing pallidal subpopulation preferentially targets the striatum and is more readily activated in its immediate early gene expression by D2 receptor antagonists than by dopamine receptor agonists. This projection provides a pallidal substrate for the dopaminergic regulation of striatal information processing.     

Superoxide generation and its modulation by adenosine in the neutrophils of subjects with asthma

Airway inflammation with neutrophil infiltration may play a role in airway hyperreactivity. Neutrophils may exert their effects through the generation of superoxide O2- anion and other oxygen-derived free radicals. O2- generation by neutrophils has been demonstrated to be modulated by adenosine at physiologic concentrations. Therefore, we have investigated the function of peripheral blood neutrophils with respect to O2- anion generation and its regulation by adenosine in both subjects with asthma and normal subjects and also the relationship between O2- anion generation and airway hyperresponsiveness in subjects with asthma. Purified neutrophils were obtained from eight subjects with stable asthma and seven normal control subjects not taking chronic medications. O2- anion generation in subjects with asthma was significantly higher compared with that of normal subjects after stimulation with either N-formyl-methionyl-leucyl-phenylalanine (mean, 14.8 nmol/10(6) cells for subjects with asthma versus mean, 9.6 nmol/10(6) cells for normal subjects; p less than 0.01) or phorbol myristate acetate (mean, 13.6 nmol/10(6) cells versus mean, 8.1 nmol/10(6) cells; p less than 0.05). Adenosine inhibited N-formyl-methionyl-leucyl-phenylalanine-stimulated O2- anion generation in a dose-related fashion in subjects with asthma and normal subjects to a similar degree. Adenosine had no effect on O2- anion generation after phorbol myristate acetate stimulation. These results indicate that neutrophils from subjects with asthma produce more O2- anion when they are stimulated than do neutrophils from normal subjects and that this difference is not due to adenosine modulation. In subjects with asthma, O2- anion generation correlated with the degree of airway hyperresponsiveness to inhaled methacholine.     

Increased fMet-Leu-Phe receptor expression and altered superoxide production of neutrophil granulocytes in septic and posttraumatic patients

Summary Activation of neutrophils by various inflammatory stimuli has been shown to play a pivotal role in septic and posttraumatic tissue injury. To further elucidate the mechanisms modulating the oxidative metabolism, we assessed superoxide production induced by N-formylmethionyl-leucylphenylalanine (FMLP) and phorbol myristate acetate and the expression of FMLP receptors of human neutrophils on several days during sepsis and after trauma. Neutrophils of septic patients isolated on days 0–4 after the diagnosis of sepsis showed a significant, more than twofold increase in specific binding of [³H]FMLP at 1, 120, and 240 nM. Scatchard plot analyses revealed that this increase in specific binding was due to an increase in the number of low- and high-affinity FMLP receptors with no changes in receptor affinity. On days 5–10 after the onset of sepsis the up-regulation of FMLP receptors on circulating neutrophils was followed by receptor down-regulation. Likewise, neutrophils from patients with trauma that was not complicated by sepsis bound significantly more [³H]FMLP than neutrophils from volunteers. However, the increase in FMLP receptors was less than that in septic neutrophils and returned earlier to normal. In accordance with the up-regulation of FMLP receptors, neutrophils obtained from patients with sepsis or after trauma on days 1–4 and days 1–2, respectively, produced significantly more superoxide anion upon stimulation with FMLP. However, after stimulation with phorbol myristate acetate, a receptor-independent activator of protein kinase C, these cells released less superoxide anion than controls. Our findings suggest that during sepsis and trauma circulating neutrophils become transiently primed for an enhanced oxidative metabolism upon stimulation with FMLP but desensitized to protein kinase C dependent stimulation.Abbreviations PMN neutrophilic granulocytes - FMLP N-formylmethionyl-leucyl-phenylalanine - PMA phorbol myristate acetate - SO oxygen superoxide - PKC protein kinase C - TNF tumor necrosis factor - HBSS Hank's balanced salt solution     

Measurement of cytokine secretion, intracellular protein expression, and mRNA in resting and stimulated peripheral blood mononuclear cells

Quantitation of cytokine production is a valuable adjunct to standard immunologic assays in defining several pathologic processes. Nevertheless, there is little agreement about which tissues should be assayed, which type of assay should be performed, and which stimulation protocol should be used. As these types of assays enter the clinical arena, there is need for standardization. There is also a need to maximize the amount of information which may be derived from a single sample. We compared secreted interleukin 4 (IL-4), IL-2, IL-6, tumor necrosis factor alpha (TNF-alpha), and gamma interferon proteins as measured by enzyme-linked immunosorbent assay with intracellular cytokine production (IL-2 and gamma interferon) as detected by flow cytometry and quantitative competitive PCR for IL-2, IL-4, TNF-alpha, and gamma interferon mRNA and cDNA. Results from unstimulated cells and cells stimulated with phorbol myristate acetate, phytohemagglutinin, and phorbol myristate acetate plus phytohemagglutin were compared. All three methodologies detected significant stimulation of cytokine production. The combination of phytohemagglutinin and phorbol myristate acetate was overall the most-potent stimulus.     

培养脊髓神经元中微管相关蛋白5的表达

目的　探讨初代培养脊髓神经元中微管相关蛋白 (MAP5 )的表达特征及其调节因素。方法　应用荧光免疫细胞化学检测初代培养脊髓神经元中MAP5的表达。结果　MAP5在培养脊髓神经元中呈网状分布在胞浆及突起中。诺考达唑引起微管解聚及MAP5的结构破坏 ,荧光物质分布不均匀 ,突起呈串珠或断裂。PMA(phorbol 12 myristate 13 acetate)能诱发微管及MAP5聚合 ,胞浆中荧光物质呈疏松的网状结构。结论　MAP5在初代培养脊髓神经元胞浆及突起中明显表达 ,且表达受微管解聚剂诺考达唑及微管聚合剂PMA的调控。     

Cytotoxic T Lymphocytes: Blocked Differentiation at the ''Poised'' Stage

The effect of different concentrations of phorbol myristate acetate (PMA) on the development of cytotoxic cells was studied. PMA was selectively able to prevent the development of cytotoxic cells in a mixed leucocyte culture, while allowing the responding cells to proliferate. The higher concentration of PMA (10(-5)M) blocked both direct cytotoxicity and lytic activity in the presence of lectin, while the lower concentration (10(-8) M) only prevented direct lytic function. The removal of PMA and subsequent addition of recombinant interleukin 2 (IL-2) or IL-2-containing supernatants effectively reversed the effect of PMA with recovery of antigen-specific lytic function of cells treated with 10(-8) M, while cells treated with 10(-5)M PMA only recovered lectin-dependent cytotoxic ability.     

Epitope focus, clonal composition and Th1 phenotype of the human CD4 response to the secretory mycobacterial antigen Ag85

Lymphoproliferation of healthy donors was tested against mycobacterial antigens (PPD, Ag85, Ag85 peptides). All PPD responders recognized the secretory antigen Ag85 and the peptide specificity for Ag85B was defined. Peptide 91-108 was recognized by 85% of donors. In addition, all CD4 T cell lines generated from 12 donors against PPD or Ag85 responded to 91-108. When this peptide was used to generate T cell lines, the cells responded also to tuberculins from atypical mycobacterial species. Thus the cross-reactive peptide behaved as quasi-universal. The analysis of TCR-BV gene usage by cell lines showed that most Ag85-specific T cells correspond to 91-108-specific clonotypes. Intracytoplasmic staining of cell lines after phorbol myristate acetate stimulation resulted in dominance of interferon-gamma (IFN-gamma)-IL-4 double-positive cells, whereas antigen stimulation resulted in production of IFN-gamma only. The data show that peptide 91-108 is the major focus of the CD4 response to mycobacterial antigens in peripheral blood mononuclear cells and in T cell lines from PPD responders.     

IL-10 Production and CD40L Expression in Patients with Common Variable Immunodeficiency

The authors studied CD40 ligand (CD40L) expression and interleukin-10 (IL-10) production in 16 patients with common variable immunodeficiency (CVI). Mean CD40L expression, determined by using cytofluorimetry, and measured as the mean fluorescence intensity following stimulation of peripheral blood mononuclear cells (PBMC) with phorbol myristate acetate (PMA) and calcium ionophore in 12 patients, was comparable to that of controls. However, three CVI patients showed fluorescence intensity in stimulated cells below 2 standard deviations of normal donors' mean and two other patients had only a slight increase of stimulated versus unstimulated cells (<10 channels). IL-10 production after stimulation of PBMC with both anti-CD3 or anti-CD3 plus PMA gave similar results in CVI patients and normal controls. In vitro stimulation of PBMC with anti-CD40 and various combinations of cytokines (IL-2, IL-4 and IL-10) induced IgG production above 100 ng/ml in one CVI patient out of 13 tested. The data suggest that alterations of IL-10 production are unlikely to play a major role in the pathogenesis of impaired IgG production in most CVI patients. CD40L appears to be normally expressed in two thirds of CVI patients, but it may be functionally defective.     

Measurement of Cytokine Secretion, Intracellular Protein Expression, and mRNA in Resting and Stimulated Peripheral Blood Mononuclear Cells

Quantitation of cytokine production is a valuable adjunct to standard immunologic assays in defining several pathologic processes. Nevertheless, there is little agreement about which tissues should be assayed, which type of assay should be performed, and which stimulation protocol should be used. As these types of assays enter the clinical arena, there is need for standardization. There is also a need to maximize the amount of information which may be derived from a single sample. We compared secreted interleukin 4 (IL-4), IL-2, IL-6, tumor necrosis factor alpha (TNF-α), and gamma interferon proteins as measured by enzyme-linked immunosorbent assay with intracellular cytokine production (IL-2 and gamma interferon) as detected by flow cytometry and quantitative competitive PCR for IL-2, IL-4, TNF-α, and gamma interferon mRNA and cDNA. Results from unstimulated cells and cells stimulated with phorbol myristate acetate, phytohemagglutinin, and phorbol myristate acetate plus phytohemagglutin were compared. All three methodologies detected significant stimulation of cytokine production. The combination of phytohemagglutinin and phorbol myristate acetate was overall the most-potent stimulus.     

Separation and chemiluminescence properties of human, canine and rat polymorphonuclear cells

Human as well as canine and rat polymorphonuclear cells (PMN) were separated from whole blood by centrifugation. Two-step discontinuous Percoll gradients with distinct densities were used. The purity of the preparations was 99.2%, 98.4% and 97.9%, respectively and the corresponding recoveries were 80.1; 66.3% and 69%. The chemiluminescence properties of the isolated PMN and of phagocytes in small quantities of whole blood were compared in luminol-enhanced assays after stimulation with various agents: non-opsonized zymosan (3.5 g/l), phorbol myristate acetate (PMA, 2.8 X 10(-6) M), calcium ionophore A 23187 (10(-5) M) and N-formyl-methionyl-leucyl-phenylalanine (FMLP, 3.5 X 10(-6) M). The isolated cells of the three species responded to all of the various stimuli. Species-related sensitivity followed the order: human greater than canine greater than rat. Responses to the various agents in the human cells was ranked: PMA greater than or equal to A 23187 greater than zymosan greater than FMLP; for the dog: A 23187 greater than PMA greater than zymosan greater than FMLP; and for the rat: zymosan greater than or equal to PMA greater than FMLP greater than or equal to A 23187. Time courses and peak maximum responses were different following stimulation in the absence or presence of autologous plasma. Distinct soluble stimuli resulted in maximum responses below the baseline in the whole blood assays with canine (FMLP) and rat (FMLP, A 23187) phagocytes.     

An improved nitroblue tetrazolium test using phorbol myristate acetate-coated coverslips.

The endotoxin--nitroblue tetrazolium (NBT) slide test was modified by precoating coverslips with phorbol myristate acetate instead of endotoxin. The percentage of control polymorphonuclear leukocytes reducing NBT was significantly (P less than .05) greater on phorbol myristate acetate-coated coverslips (99 +/- 0.21%) than on endotoxin-coated coverslips (96+/- 1.8%). Polymorphonuclear leukocytes from patients with chronic granulomatous disease did not reduce NBT on phorbol myristate acetate- or endotoxin-treated coverslips. NBT reduction by polymorphonuclear leukocytes from proven heterozygotic carriers of sex-linked chronic granulomatous disease was intermediate between NBT reductions by those from controls and patients. A statistically significant abnormality of NBT reduction was found in polymorphonuclear leukocytes from one carrier of chronic granulomatous disease with phorbol myristate acetate-treated, but not endotoxin-treated coverslips. The phorbol myristate acetate-NBT coverslip technic is a rapid, simple, reliable way to detect deficiencies in polymorphonuclear leukocytes from patients and carriers of chronic granulomatous disease.     

Enkephalin mRNA production by cochlear and vestibular efferent neurons in the gerbil brainstem

Summary Preproenkephalin mRNA production by efferent neurons projecting to the gerbil inner ear was assessed using combined in situ hybridization and retrograde labeling with florescent tracers. Virtually all vestibular efferent neurons were positive for preproenkephalin mRNA. Of the cochlear efferents, one-half of the medial olivocochlear neurons were positive for enkephalin. All lateral olivocochlear neurons were negative for enkephalin. The results suggest that there are two, biochemically distinct subpopulations of medial olivocochlear efferents in the gerbil. Offprint requests to: Division of Otolaryngology, ENT, V-112C     

Functional peculiarities of monocytes isolated from patients with bronchial asthma: Respiratory burst and lipid peroxidation

The capacity of monocytes obtained from patients with bronchial asthma for a “respiratory burst” in response to phorbol myristate acetate and arachidonic acid was studied, the level of lipid peroxidation in the monocytes of asthmatic patients during the disease onset in comparison with healthy donors was determined, and lipid peroxidation activation in the plasma membranes in response to phorbol myristate acetate and arachidonic acid was studied. The generation of by the bronchial asthma patients' and healthy donors' monocytes after stimulation with phorbol myristate acetate was similar. Arachidonic acid induced no generation of by donor monocytes; however, upon incubation of patient monocytes with arachidonic acid the respiratory burst was markedly enhanced, and the quantity of generated during 15 min was almost 6-fold greater than upon stimulation with phorbol myristate acetate. The lipid peroxidation level in donor and patient monocytes differed significantly. A similar potentiation of lipid peroxidation in response to phorbol myristate acetate was observed in both groups; incubation of cells with arachidonic acid induced a greater increase in lipid peroxidation, which was similar for both groups, the increase in lipid peroxidation products in response to arachidonic acid being significantly higher than in response to phorbol myristate acetate. It can be assumed that the considerable enhancement of the oxygen response of the monocytes of the bronchial asthma patients indicates the activation of mononuclear macrophages and their participation in the pathogenesis of the disease. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N ^o 12, pp. 585–588, December, 1994     

Extracellular release of antimicrobial defensins by human polymorphonuclear leukocytes.

Human polymorphonuclear leukocytes (PMN) contain three antimicrobial and cytotoxic peptides which belong to a family of mammalian granulocyte peptides named defensins. To determine their potential availability for extracellular microbicidal or cytotoxic events, we quantified the extracellular release of defensins after stimulation of human PMN with phorbol myristate acetate and opsonized zymosan. As determined by enzyme immunoassay and confirmed by polyacrylamide gel electrophoresis and densitometry, 10(6) human PMN contained 4 to 5 micrograms of defensins. After stimulation with a high concentration of phorbol myristate acetate (1 microgram/ml), about 8% of PMN defensins were found in the media. Release of defensins correlated best with the release of azurophil granule marker beta-glucuronidase or elastase and poorly with the release of either the specific granule marker lactoferrin or cytoplasmic lactate dehydrogenase. Phagocytosis of opsonized zymosan resulted in the extracellular release of less than 3% of PMN defensins. The factors responsible for less release of defensins into media relative to the release of other azurophil granule proteins may include heterogeneity of azurophil granules and the affinity of defensins for cellular surfaces and opsonized particles. In vivo, defensins are most likely to reach effective microbicidal or cytotoxic concentrations in PMN-rich exudates (pus), in confined environments of the phagolysosomes, or in intercellular clefts between PMN and their targets.     

Expression and function of AIM, an activation inducer molecule of human lymphocytes, is dependent on the activation of protein kinase C

AIM is an activation inducer molecule selectively expressed by activated lymphocytes through which agonistic proliferative signals can be triggered. The relationship between the expression of AIM with the activation of protein kinase C (PKC) has been studied. Different activators of PKC such as the active phorbol esters, phorbol myristate acetate and phorbol dibutyrate, or the phorbol-related ester mezerein were able to induce AIM expression on peripheral blood lymphocytes as assessed by immunofluorescence flow cytometry. Moreover, the expression of this activation antigen was also induced by treatment of peripheral blood lymphocytes either with dioctanoyl-rac-glycerol, a synthetic analogue of diacylglycerol, the physiological mediator of PKC activation. Further indirect evidence that AIM expression was dependent on the activation of PKC was obtained by blockade of the induction of its expression in cells treated with H7, an inhibitor of PKC. The AIM expression can be detected as early as 3 h after addition of phorbol esters and it requires active RNA and protein synthesis. The activation of PKC appears to be also required in the proliferative response induced by anti-AIM monoclonal antibody (mAb) in conjunction with phorbol esters. Agents such as phorbol myristate acetate, phorbol dibutyrate or mezerein but not the inactive phorbol ester methyl-phorbol myristate acetate induced a high proliferation of peripheral blood lymphocytes in the presence of anti-AIM mAb. In addition, we have demonstrated that the anti-AIM mAb is not sufficient by itself to induce cellular proliferation once the AIM antigen is expressed at the cell surface, requiring the simultaneous stimulation of the PKC to trigger high proliferative responses. Furthermore, the anti-AIM mAb did not appear to exert its effect on proliferation by rapidly increasing the intracytoplasmic Ca2+ levels. Taken together all these results indicate that the expression and function of AIM antigen is dependent on the activation of PKC.     

The relationship between oral dyskinesias produced by long-term haloperidol treatment, the density of striatal preproenkephalin messenger RNA and enkephalin peptide, and the number of striatal neurons expressing preproenkephalin messenger RNA in rats

Neuroleptic-induced oral dyskinesias in rats, a putative analogue to human tardive dyskinesia, may be due to excitotoxic degeneration within the striatum. Haloperidol treatment for 34 weeks increased the optical density of preproenkephalin messenger RNA in individual striatal neurons and enkephalin peptide in the neuropil, regardless of the level of oral dyskinesia produced. However, using unbiased stereological methods, an increased number of striatal neurons expressing preproenkephalin messenger RNA was observed only in rats that did not develop pronounced oral dyskinesias during haloperidol treatment. Said in another manner, the haloperidol-treated animals that developed pronounced oral dyskinesias, failed to produce an increase in the number of neurons expressing preproenkephalin messenger RNA. These results indicate that the mechanism by which neuroleptics induce oral dyskinesias in rats, and perhaps tardive dyskinesia in humans, involves a functional disturbance or even damage to a subpopulation of enkephalinergic neurons in the striatum.