Type of cell death and the role of acetylcholinesterase activity in neurotoxicity induced by paraoxon in cultured rat hippocampal neurons.

Organophosphate (Ops) neurotoxicity is attributed both to its well-known cholinergic and non-cholinergic effects. In the present study we compared enzymatic and morphologic changes in neurons exposed to paraoxon during one day and one week. The effect of exposure time is important in neurotoxicity of Ops. The longer the exposure time is the more damage is observed in neurons, although there are few investigations about the effect in the post-exposure period. Hippocampal cells were obtained from rat neonates and cultured in Neurobasal/B27. Paraoxon at 50 and 100 microM were added. Inverted microscope and electron microscope were used to study cell morphology and Neutral Red staining was used to measure viability. We also assayed caspase-3 and (acetylcholinesterase) AChE activity. Hoechst staining was utilized to determine the type of cell death. Culture medium was replaced after 24 h in one-day group, however, tests were all carried out at the end of the first week in both group. The results indicate that paraoxon reduced the viability in a dose-dependent manner. Our results do not confirm apoptosis in either group; it seems that the cell death in one-day exposure group was not AChE dependent. In conclusion, present data imply that the toxicity of paraoxon is both dose and duration dependent, which may even remain after the cessation of exposure.

Angiotensin-converting enzyme genotype and late respiratory complications of mustard gas exposure.

BACKGROUND: Exposure to mustard gas frequently results in long-term respiratory complications. However the factors which drive the development and progression of these complications remain unclear. The Renin Angiotensin System (RAS) has been implicated in lung inflammatory and fibrotic responses. Genetic variation within the gene coding for the Angiotensin Converting Enzyme (ACE), specifically the Insertion/Deletion polymorphism (I/D), is associated with variable levels of ACE and with the severity of several acute and chronic respiratory diseases. We hypothesized that the ACE genotype might influence the severity of late respiratory complications of mustard gas exposure. METHODS: 208 Kurdish patients who had suffered high exposure to mustard gas, as defined by cutaneous lesions at initial assessment, in Sardasht, Iran on June 29 1987, underwent clinical examination, spirometric evaluation and ACE Insertion/Deletion genotyping in September 2005. RESULTS: ACE genotype was determined in 207 subjects. As a continuous variable, FEV1 % predicted tended to be higher in association with the D allele 68.03 +/- 20.5%, 69.4 +/- 21.4% and 74.8 +/- 20.1% for II, ID and DD genotypes respectively. Median FEV1 % predicted was 73 and this was taken as a cut off between groups defined as having better or worse lung function. The ACE DD genotype was overrepresented in the better spirometry group (Chi2 4.9 p = 0.03). Increasing age at the time of exposure was associated with reduced FEV1 %predicted (p = 0.001), whereas gender was not (p = 0.43). CONCLUSION: The ACE D allele is associated with higher FEV1 % predicted when assessed 18 years after high exposure to mustard gas.

In vivo preconditioning with normobaric hyperoxia induces ischemic tolerance partly by triggering tumor necrosis factor-alpha converting enzyme/tumor necrosis factor-alpha/nuclear factor-kappaB.

Recent studies suggest that intermittent and prolonged normobaric hyperoxia (HO) results in brain ischemic tolerance (BIT), reducing ischemic brain injury. We have attempted to determine the time course of HO-induced BIT, and to explore the putative roles of tumor necrosis factor-alpha (TNF-alpha) converting enzyme (TACE), TNF-alpha, and nuclear factor-kappaB (NF-kappaB) activation in mediating this effect. Two core experimental protocols were applied to rats (experiments 1 [E1] and 2 [E2] respectively). E1 rodents comprised six subgroups, breathing room air (RA; O(2)=21%), or 95% oxygen (HO) for 4, 8, 16 h (4RA, 8RA, 16RA and 4HO, 8HO, 16HO respectively). E2 rodents were divided into subgroups, exposed to 95% inspired HO for 4 h/day for six consecutive days (intermittent hyperoxia, InHO) or for 24 continuous hours (prolonged hyperoxia, PrHO). Each of these had a control group exposed to 21% oxygen in the same chamber. Twenty-four hours after pretreatment, each group was randomly divided to receive 60 min right middle cerebral artery occlusion (MCAO-operated), sham-operation (without MCAO), or no operation (intact). After 24 h reperfusion, neurologic deficit score (NDS), brain water content, Evans Blue extravasation (as a marker of blood-brain barrier permeability), TACE expression, serum TNF-alpha, and phosphor- kappaBalpha levels were assessed in all animals, and infarct volume in the MCAO-operated subgroups. E1: Compared with the control (RA) group, infarct volume was reduced by 58.6% and 64.4% in 16 h and 24 h respectively. NDS and Evans Blue extravasation was also reduced in 16 h and 24 h. There was no statistical difference among 4 h and 8 h. E2: Preconditioning with prolonged and intermittent HO decreased NDS, infarct volume and upregulated TACE and increased phosphor-kappaBalpha and serum TNF-alpha level significantly. Although further studies are needed to clarify the mechanisms of brain ischemic tolerance, InHO and PrHO may partly exert their effects via triggering TACE/TNF-alpha/NF-kappaB.

Paraoxon-induced ultrastructural growth changes of rat cultured hippocampal cells in neurobasal/B27.

Organophosphates (OPs) neurotoxicity is attributed both to their well-known cholinergic and recently attended non-cholinergic effects. Since parathion has been observed to be responsible for more cases of poisoning than any other OP insecticides, it is vitally important to investigate other mechanisms, besides cholinesterase inhibition, which can potentially contribute to the neurotoxicity of parathion (or its metabolite, paraoxon). In present study, hippocampal cells obtained from Wistar rat neonates were cultured in neurobasal medium supplemented with B27 serum where different doses of paraoxon were also introduced. The neuronal growth in the control group and those exposed to paraoxon was compared. Phase contrast microscopy, cell staining (Neutral Red) and computer assessment morphometric study (Motic) were used to study cell morphology, viability and type of cell death. Statistical analysis was carried out using one-way ANOVA. There was no clear morphologic differences between neurons in the control group and those exposed to 10 microM paraoxon; however, deformity of the soma was clear in pellets containing higher concentration of paraoxon. Ultrastructure of cells was markedly altered at 50 microM dose of paraoxon as evidenced by gradual discontinuation of cytoplasm, appearing of numerous vacuoles and intracytoplasmic myelin figure. The processes (neurites) did not grow in media containing 100 microM paraoxon or more. Viability decreased with increasing paraoxon especially above 100 microM. In conclusion, the present data reveal that paraoxon, in 30 microM or higher concentrations, induces a decrease in cell growth, followed by cell swelling and neuronal death (possibly necrosis).

Involvement of nucleus accumbens in L-arginine-induced conditioned place preference in rats.

In the present study, the effects of intraperitoneal, intra-accumbal and intra-ventral tegmental area administration of L-arginine and N(G)-nitro-L-arginine methyl-ester (L-NAME) on conditioned place preference behavior were studied. Intraperitoneal (i.p.; 0.5, 1 and 5 mg/kg) and intra-accumbal (intra-NAc; 0.3, 1 and 3 microg/rat), but not intra-ventral tegmental area (intra-VTA; 0.3, 1 and 3 microg/rat) administrations of L-arginine produced a significant place conditioning. Similar injections of L-NAME did not produce any response. However, intraperitoneal pretreatment of the animals with L-NAME (5, 10 and 20 mg/kg), 30 min before L-arginine administration, significantly abolished the acquisition of place conditioning induced by either intraperitoneal or intra-accumbal injection of L-arginine. Moreover, injection of L-NAME (5, 10 and 20 mg/kg) on the test day did not alter the L-arginine response. The results may indicate that L-arginine induces conditioned place preference via an increase in nitric oxide (NO) in the nucleus accumbens.

The physiological and pathophysiological modulation of the endocrine function of the heart.

Under physiological conditions, the endocrine heart contributes to the maintenance of cardiovascular homeostasis through the polypeptide hormones ANF and BNP, which are members of the natriuretic peptide (NP) family. Given that NPs are of interest from the basic and clinical points of view, the genetic expression and secretion of ANF and BNP as well as the nature of the interaction of these hormones with their receptors has been the subject of extensive studies since the discovery of ANF in 1980. Following hemodynamic overload, increased secretion of NPs by the heart can be seen. This change may occur without an increase in gene expression as observed for atrial NPs following acute volume expansion, or it can occur with an increase in both ANF and BNP gene expression in atria only as seen in mineralocorticoid escape during which it is obvious that a critical decrease in hormone stores must be reached before transcriptional activation occurs. Chronic hemodynamic pressure or volume overload results in increased expression of NPs in atria and ventricles. Under these circumstances, the increased production of BNP by hypertrophic ventricles changes the normal plasma concentration ratio of ANF to BNP, a fact that has clinical diagnostic and prognostic implications. There are exceptions to this rule: chronic, severe L-NAME hypertension, which may occur without left ventricular hypertrophy, does not cause this effect and increased ventricular NP gene expression can occur in mineralocorticoid hypertension before detectable ventricular hypertrophy. Atrial and ventricular NP gene expression appears to be under different transcriptional control because pharmacological treatments such as chronic ACE inhibition or ET(A) receptor blockade can reverse the increased ventricular NP expression but has no detectable effect on atrial NP gene expression. This is not unlike the myosin heavy chain switch that is observed in certain pathologies, and can be pharmacologically reversed in a manner similar to NPs in the ventricles but it does not occur in atrial muscle. These observations made in vivo or using isolated adult atria often differ strikingly from results obtained using the mixed phenotype afforded by cardiocytes in culture, indicating that the kinds of questions addressed by each approach must be judiciously chosen. G-protein coupled receptor-mediated actions of neurohumors such as endothelin and phenylephrine are normally used to stimulate NP gene expression and release in different in vitro models. The main physiological stimulus for increased ANF release, atrial muscle stretch, also appears to rely on G-protein-coupled mechanisms. Alternative agonists and receptor types at play are suggested by the finding that circulating levels of BNP are selectively increased before and during overt cardiac allograft rejection episodes in human patients. The data suggest that enhanced BNP plasma levels could form a basis for a noninvasive test for cardiac allograft rejection. However, the molecular mechanism by which expression of NPs are regulated in the transplanted heart is not well understood. Conditioned medium from mixed lymphocyte reaction cultures, considered an in vitro model of transplantation immunity, induces specific upregulation of BNP as do individual pro-inflammatory cytokines. Findings such as these suggest that the study of NPs will continue to produce a wealth of information relevant to basic and clinical scientists.

Decreased response of rat knee joint blood vessels to phenylephrine in chronic inflammation: involvement of nitric oxide.

The effect of chronic inflammation induced by Freund's Complete Adjuvant (FCA) on rat articular blood vessels and knee joint diameter was investigated. Blood flow changes in response to phenylephrine (an 1-adrenoceptor agonist) in FCA-treated and contralateral knee joints were studied over a 40 day period, using the laser Doppler flowmetery (LDF) technique. Unilateral injection of FCA (0.2 ml) increased the injected knee diameter on all days examined post-injection (P < 0.001) and its maximum increase (53 +/- 2 %) was reached on day 3. After this, the diameter decreased gradually but did not return to its initial value. In control animals, topical application of 10-13-10- 7 mol phenylephrine onto the exposed joint capsule decreased blood flow dose dependently (11. 1 +/- 4.4 to 58.2 +/- 4.5 %, respectively, P < 0.001). Unilateral injection with FCA attenuated the phenylephrine response in both ipsilateral and contralateral knees compared with the response of control animals (5.2 +/- 1.6 to 48.3 +/- 6.1 % and 1.9 +/- 2.2 to 45. 3 +/- 5.6 %, respectively, P < 0.05). The reduction persisted for 3 weeks after FCA injection (ipsilateral for 21 days; contralateral for 30 days, P < 0.001). Subsequently the response returned towards normal. To avoid the influence of 2-adrenoceptors, yohimbine (an 2-adrenoceptor antagonist) was injected (0.5 mg kg-1, I.P.) 30 min before phenylephrine application. Yohimbine blocked the vasoconstrictor effect of 10-10-10-7 mol clonidine (an 2-adrenoceptor agonist, topical application) by 44-67.7 % inhibition, respectively (P < 0.001). Prazosin (an 1-adrenoceptor antagonist, 0.1 mg kg-1, I.P.) blocked the vasoconstrictor effect of phenylephrine (10-10-10-7 mol, topical application) effectively (42 to 69.8 % inhibition, respectively, P < 0.001). To assess the role of nitric oxide (NO) on the observed responses, N G-nitro-L-arginine methyl ester (L-NAME, NO synthase inhibitor) was applied topically (0.2 micromol) 5 min before phenylephrine application. L-NAME application at 7 and 14 days after FCA injection potentiated the vasoconstrictor response in the FCA-treated knee (P < 0.001) but had no significant effect on the contralateral knee. Blood pressure monitoring during phenylephrine, clonidine and L-NAME administration indicated that topical application of the drugs had no significant effect on the systemic blood pressure. These findings indicate that the vasoconstrictor response to phenylephrine was decreased in chronic inflammation and increased NO production could be involved.

Adenosine A(2) receptors inhibit morphine self-administration in rats.

In the present study, the effect of adenosine receptor agonists and antagonists on morphine self-administration was investigated. Intravenous administration of morphine (0.3-3 mg/kg/injection) induced dose-dependent self-administration. The adenosine receptor antagonists, theophylline (2.5, 5, 10 mg/kg) and 3, 7-Dimethyl-1-propargylxanthine (DMPX; 0.25, 0.5, 1 mg/kg), when injected 1 h before the start of the test, reduced the number of self-administered morphine infusions. The adenosine receptor antagonists when administered in the training period (11 days) greatly increased the number of morphine infusions, however, they did not induce any response by themselves. 5'-N-ethylcarboxamido-adenosine (NECA; 0.5, 1 mg/kg) and 4-[2-[[6-Amino-9-(N-ethyl-beta-D-ribofuranuronamidosyl)-9H-purin-2 -yl ]amino] ethyl]benzenepropanoic acid (CGS21680; 0.001, 0.01, 0.025, 0. 05 mg/kg), given 1 h before the start of the test, increased morphine self-administration. Although the adenosine agonists, when injected during training period (11 days), reduced morphine self-administration. Furthermore, NECA, but not CGS21680, induced significant self-administration. The adenosine A(1) receptor agonist, N(6)-cyclohexyladenosine (CHA; 0.01, 0.1, 0.25, 0.5 and 1 mg/kg), and the adenosine A(1) receptor antagonist, 8-phenyletheophylline (2, 4, 6, 8 mg/kg), themselves neither altered morphine infusion nor induced any response. These results indicate a role for adenosine A(2) receptors in the expression and/or development of morphine self-administration.

Nerve-mediated responses of blood vessels in the rabbit knee joint.

Blood vessels in the anterior region of the knee joint of anaesthetised rabbits showed a biphasic response to the electrical stimulation (10 Hz, 1 ms width, 10 V amplitude) of nerve fibres supplying the knee, as measured by laser Doppler flowmetry. The response consisted of vasoconstriction during nerve stimulation followed by a prolonged dilatation. The vasoconstrictor response was mediated by noradrenaline acting mainly via alpha 1-adrenoceptors as it was substantially reduced by close intra-arterial injection of the alpha-adrenergic antagonist phentolamine (approximately 50% reduction) and the selective alpha 1-adrenergic antagonist prazosin (approximately 50% reduction) but not by the alpha 2-antagonist rauwolscine. Further studies involving prolonged (2-hour) close intra-arterial infusion of prazosin gave a approximately 50% reduction of the constrictor response with a concentration of 10(-5) M and approximately 95% reduction when the concentration was raised to 10(-4) M. At the higher prazosin concentration responses to close intra-arterial injection of the alpha 1-agonist phenylephrine were substantially reduced but responses to the alpha 2-agonists clonidine and UK-14304 were not significantly influenced. Infusions of the alpha 2 antagonist CH 38083 failed to inhibit nerve-mediated vasoconstriction at 10(-5) or 10(-4) M. There did not appear to be a purinergic component, as the constrictor response was unaffected by the P2X desensitiser alpha,beta-methylene adenosine 5'-triphosphate. The dilator response appeared to be mediated principally by substance P (presumably released from sensory C fibers) as it was substantially reduced by intraarticular injection of substance P antagonist D-Pro4D-Trp7,9,10-SP(4-11).(ABSTRACT TRUNCATED AT 250 WORDS)

Localized neural control of blood flow in the posterior region of the knee joint in anaesthetized rabbits.

Radiolabelled microspheres were used to measure blood flow of structures in and around the rabbit knee joint and to determine the effect of electrical stimulation of the posterior articular nerve (PAN) supplying the knee. Surgical exposure of the posterior aspect of the knee joint 'capsule' and section of PAN resulted in an increase in flow restricted to this region. Electrical stimulation produced frequency-dependent reductions of blood flow in the posterior region, but no alterations occurred in the anterior region or in any other surrounding structures, suggesting that the neural supply is specific to the posterior joint capsule.

Responses of bone and joint blood vessels in cats and rabbits to electrical stimulation of nerves supplying the knee.

1. Experiments were performed to assess the extent to which knee joint blood flow in cats and rabbits is affected by electrical stimulation of the nerve supply to the knee. 2. Absolute changes in blood flow were measured using the radiolabelled microsphere (approximately 15 microns) technique whilst relative changes in blood flow were assessed using laser Doppler flowmetry. 3. Despite deep general anaesthesia, sympathetic nerve fibres innervating cat knee joint blood vessels showed marked 'tone'. 4. Blood flow to the joint capsule (synovium and overlying fibrous and areolar tissues) was substantially reduced (by approximately 90% in the cat and approximately 45% in the rabbit) during electrical stimulation of the articular nerve supply. 5. The percentage change in the laser Doppler flowmeter signal did not differ significantly from the percentage change in blood flow measured by microsphere technique. 6. Blood vessels in the cancellous bone of the distal femur (condyles) and proximal tibia (plateau) appear to be innervated by vasoconstrictor fibres which reach their effectors via the articular nerves. However, the cortical bone and red marrow of the diaphysis of the femur do not receive such innervation. 7. The potency of the vasoconstrictor influences acting on joint blood vessels could be of relevance in the pathogenesis of inflammatory joint diseases.

Alterations in cat knee joint blood flow induced by electrical stimulation of articular afferents and efferents.

1. Experiments were performed in cats anaesthetized with pentobarbitone. Laser Doppler flowmetry was used to assess the responses of knee joint blood vessels to nerve stimulation under control conditions and in the presence of different adrenoceptor antagonists in order to establish the nature of neurotransmitters released from articular nerve fibres. 2. The posterior articular nerve (PAN) supplying the knee was stimulated at different intensities, and frequency-response curves were obtained. In fourteen animals electrical stimulation of PAN produced an initial vasoconstriction during stimulation which in eight of these was followed by a prolonged dilatation on cessation of stimulation. The constrictor response was increased as a function of frequency but was little altered with increasing intensity beyond a threshold level. 3. The constrictor response to electrical stimulation of PAN was markedly reduced by the alpha-adrenergic antagonist phentolamine (10(-5) M, the alpha 1-blocker prazosin (10(-5) M), and guanethidine (10(-5) M) which inhibits the release of noradrenaline, ATP, and neuropeptide Y from sympathetic nerve endings. 4. The constrictor response to PAN stimulation was unaffected by the alpha 2-blocker rauwolscine and the P2-purinoceptor desensitizer alpha,beta-methylene ATP. 5. The dilator response was due to activation of afferent fibres as it could also be produced by direct electrical stimulation of the L7 dorsal roots. 6. The dilator response to stimulation of PAN or the L7 dorsal root was reduced by prior intra-articular injection of 100 micrograms of the substance P antagonist D-Pro4-D-Trp7,9,10-SP4-11. 7. These results suggest that the vasoconstrictor response to electrical stimulation of PAN is most likely to be mediated via noradrenaline acting mainly upon alpha 1-adrenoceptors. As the dilator response to articular nerve stimulation is reduced by a substance P antagonist, the mediator inducing this response may be substance P or a related neurokinin.

Responses of blood vessels in the rabbit knee to acute joint inflammation.

These experiments examined the responses of articular blood vessels in the rabbit knee to induction of acute joint inflammation by intraarticular injection of 4% kaolin suspension. This produced an inflammatory response, which became evident about four hours after injection. Electrical stimulation of the nerve supply to the knee joint before induction of inflammation produced a biphasic response--an initial vasoconstriction during the stimulation phase followed by dilatation after stimulation stopped. These responses were followed up for eight hours and it was noted that the constrictor response became progressively greater, producing a further 19% decrease in blood flow during nerve stimulation about three hours after the injection of kaolin. The sensitivity of postjunctional alpha adrenoceptors, however, showed still greater increase in the inflamed joint as close intraarterial injection of 10(-6) M adrenaline produced an additional 30% reduction in blood flow four hours after kaolin injection compared with the control response. Possibly, the smaller enhancement of the constrictor response to nerve stimulation in the inflamed joint may reflect sensitisation of prejunctional alpha adrenoceptors in addition to the effects exerted on postjunctional alpha adrenoceptors by the inflammatory process. The dilator response also increased over eight hours, though this rise was less marked. These findings indicate that even over the limited time span of the experiments, significant alterations occurred in factors which influence the calibre of articular blood vessels.

Responses of blood vessels in the rabbit knee to electrical stimulation of the joint capsule.

1. An in vitro preparation of the rabbit knee joint, perfused with oxygenated Locke's solution, was used to study the response of articular blood vessels to electrical stimulation of the joint capsule. 2. Using trains of stimulus pulses of different durations, frequency-response curves were obtained. Electrical stimulation always produced vasoconstriction of joint blood vessels, which increased as a function of both frequency and pulse width. 3. This vasoconstrictor response was neurally mediated as it was markedly inhibited after addition to both bath and perfusate of tetrodotoxin. In addition, the response to field stimulation of the capsule was virtually abolished in animals pretreated with reserpine which depletes sympathetic nerve endings of noradrenaline. 4. The response to electrical stimulation was substantially reduced by the alpha-adrenergic antagonist phenoxybenzamine (10(-5) M), the alpha 1-blocker prazosin (10(-6) M), and by guanethidine (10(-5) M) which inhibits the release of noradrenaline, ATP and neuropeptide Y from sympathetic nerve endings. 5. The attenuation of the vasoconstrictor response to field stimulation by prazosin (10(-6) M) was little altered by addition of the alpha 2-adrenoceptor blocker rauwolscine (10(-6) M) to the perfusate. 6. alpha, beta-Methylene ATP (10(-6) M), a P2-purinoceptor desensitizer, had no effect on the vasoconstrictor response to electrical stimulation. 7. These results indicate that the vasoconstrictor response to electrical stimulation of the rabbit knee joint capsule is mediated via noradrenaline acting upon alpha 1-adrenoceptors.

The role of the endothelium in mediating the actions of ATP, adenosine and acetylcholine on flow through blood vessels in the rabbit knee joint.

1. An in vitro preparation of the rabbit knee joint, perfused with oxygenated Locke solution, was used to investigate the presence of purinoceptors and the role of endothelium within articular blood vessels. 2. The basal tone of the blood vessels was not affected by adenosine or acetylcholine. Adenosine 5'-triphosphate (ATP) injection produced vasoconstriction which was unaffected by removal of the endothelial layer, but diminished by alpha, beta methylene ATP, a compound which desensitizes P2-purinoceptors. 3. When knee joint blood vessel tone was raised by perfusion with vasopressin (10(-8) M) or 5-hydroxytryptamine (10(-5) M), acetylcholine, ATP and adenosine were all found to induce concentration-dependent relaxation of these vessels. ATP was found to have a dual effect of transient constriction followed by longer-lasting dilatation. 4. 3-Methylxanthine, a P1-purinoceptor antagonist significantly reduced the relaxation response to adenosine but had no effect on the vasodilator effect of ATP. 5. Removal of the endothelial layer virtually abolished the vasodilator effects of acetylcholine and ATP but not adenosine. 6. These results demonstrate that articular blood vessels supplying the rabbit knee contain P1-purinoceptors located on the vascular smooth muscle which mediate vasodilatation. P2-purinoceptors mediating a constrictor effect are also present on this smooth muscle. It is likely that the vasodilator effect of ATP is mediated via P2-purinoceptors located on the endothelial layer.

Adrenoceptor profile of blood vessels in the knee joint of the rabbit.

1. An in vitro preparation of the rabbit knee joint, perfused with oxygenated Locke solution, was used to assess the nature of adrenoceptors within articular blood vessels. 2. Dose/response relationships were obtained to intra-arterial injection of alpha- and beta-adrenoceptor agonists. 3. Adrenaline and noradrenaline produced a similar pattern of increasing constriction of articular vessels with increasing dose of drug. 4. The alpha 1-agonist, phenylephrine, also produced dose-dependent constrictor responses, but the alpha 2-agonist; clonidine, had no effect. The alpha 2-agonist UK-14304 did, however, produce modest vasoconstriction which was not greatly altered by the alpha 1-blocker prazosin. The constrictor effect of noradrenaline was abolished by both the alpha 1,2-blocker phenoxybenzamine and by prazosin but not by the alpha 2-blocker rauwalscine. 5. The beta-adrenoceptor agonist, isoprenaline, had little effect at a dose of 10(-6) M or lower, but gave rise to a constrictor effect at higher concentrations. This response was blocked by phenoxybenzamine but not by the beta 1,2-blocker propranolol, suggesting that the constrictor effect was mediated via alpha-adrenoceptors. 6. The results suggest that alpha 1- and alpha 2-adrenoceptors are present within articular blood vessels, but that beta-receptors are absent. The effects of noradrenaline appear to be mediated principally via alpha 1-adrenoceptors.