Reuptake Mechanism Research Articles

Changes in extracellular 5-HIAA concentrations as measured by in vivo microdialysis technique in relation to changes in 5-HT release.

The cerebral microdialysis technique has been widely used to monitor the release of 5-hydroxytryptamine (5-HT). The extracellular concentration of 5-HT has generally been shown to change after pharmacological manipulation as expected. Extracellular levels of the metabolite, 5-hydroxyindoleaceticacid (5-HIAA) does not always change in the same direction as 5-HT and has therefore generally been thought to be of no interest as a marker for 5-HT release. The aim of the present review is to analyse the connection between changes in extracellular levels of 5-HT and 5-HIAA evoked by various pharmacological means. Literature on in vivo microdialysis studies measuring extracellular 5-HT and 5-HIAA has been analysed with special attention to the great importance of the 5-HT re-uptake mechanism in determining their extracellular concentrations. When the 5-HT reuptake mechanism is intact changes in extracellular levels of 5-HT and 5-HIAA go in the same directions, e.g decrease after compounds that decrease 5-HT release and increase after compounds that enhance 5-HT release. Because the extracellular 5-HIAA concentrations is 100-1000 times higher than that of 5-HT similar percentage changes imply that a very small part of the released 5-HT reaches the microdialysis probe under these conditions. When the 5-HT reuptake mechanism is blocked the extracellular 5-HT increases whereas extracellular 5-HIAA decreases mainly because of the 5-HT(1B) receptor-induced decrease in 5-HT release but in part also because of the inhibition of reuptake of 5-HT, both resulting in decreased formation of 5-HIAA. Drug-induced changes in extracellular 5-HIAA levels can give valuable information on the effects of these drugs on the 5-HT release.

Nitric oxide regulation of myocardial contractility and calcium cycling: independent impact of neuronal and endothelial nitric oxide synthases.

The mechanisms by which nitric oxide (NO) influences myocardial Ca2+ cycling remain controversial. Because NO synthases (NOS) have specific spatial localization in cardiac myocytes, we hypothesized that neuronal NOS (NOS1) found in cardiac sarcoplasmic reticulum (SR) preferentially regulates SR Ca2+ release and reuptake resulting in potentiation of the cardiac force-frequency response (FFR). Transesophageal pacing (660 to 840 bpm) in intact C57Bl/6 mice (WT) stimulated both contractility (dP/dtmax normalized to end-diastolic volume; dP/dt-EDV) by 51+/-5% (P<0.001) and lusitropy (tau; tau) by 20.3+/-2.0% (P<0.05). These responses were markedly attenuated in mice lacking NOS1 (NOS1-/-) (15+/-2% increase in dP/dt-EDV; P<0.001 versus WT; and no change in tau; P<0.01 versus WT). Isolated myocytes from NOS1-/- (approximately 2 months of age) also exhibited suppressed frequency-dependent sarcomere shortening and Ca2+ transients ([Ca2+]i) compared with WT. SR Ca2+ stores, a primary determinant of the FFR, increased at higher frequencies in WT (caffeine-induced [Ca2+]i at 4 Hz increased 107+/-23% above 1 Hz response) but not in NOS1-/- (13+/-26%; P<0.01 versus WT). In contrast, mice lacking NOS3 (NOS3-/-) had preserved FFR in vivo, as well as in isolated myocytes with parallel increases in sarcomere shortening, [Ca2+]i, and SR Ca2+ stores. NOS1-/- had increased SR Ca2+ ATPase and decreased phospholamban protein abundance, suggesting compensatory increases in SR reuptake mechanisms. Together these data demonstrate that NOS1 selectively regulates the cardiac FFR via influences over SR Ca2+ cycling. Thus, there is NOS isoform-specific regulation of different facets of rate-dependent excitation-contraction coupling; inactivation of NOS1 has the potential to contribute to the pathophysiology of states characterized by diminished frequency-dependent inotropic responses.

A role for glutamate transporters in neurodegenerative diseases.

In the mammalian central nervous system (CNS) L-glutamic acid acts as a chemical transmitter of excitatory signals. L-glutamate is released in the synaptic cleft and activates a multitude of highly integrated signaling pathways by binding to an array of iono-and metabotropic glutamate receptors. The termination of this chemical neurotransmission occurs via uptake of glutamate by specialized carrier proteins. The reuptake mechanism is capable of maintaining glutamate at levels below 1 p.M in the synaptic cleft. Cells in the CNS possess unique Na+- and K+-coupled transporters with high affinity for glutamate.1 These glutamate transporters are distinct from the recently identified vesicular glutamate transporters, which belong to the family of Na+/phosphate and sulphate transporters.

The cyclo-oxygenase-2 inhibitor celecoxib perturbs intracellular calcium by inhibiting endoplasmic reticulum Ca2+-ATPases: a plausible link with its anti-tumour effect and cardiovascular risks.

Substantial evidence indicates that the cyclo-oxygenase-2 (COX-2) inhibitor celecoxib, a widely prescribed anti-inflammatory agent, displays anti-tumour effect by sensitizing cancer cells to apoptosis. As part of our effort to understand the mechanism by which celecoxib mediates apoptosis in androgen-independent prostate cancer cells, we investigated its effect on intracellular calcium concentration ([Ca(2+)](i)). Digital ratiometric imaging analysis indicates that exposure of PC-3 cells to celecoxib stimulates an immediate [Ca(2+)](i) rise in a dose- and time-dependent manner. Kinetic data show that this Ca(2+) signal arises from internal Ca(2+) release in conjunction with external Ca(2+) influx. Examinations of the biochemical mechanism responsible for this Ca(2+) mobilization indicate that celecoxib blocks endoplasmic reticulum (ER) Ca(2+)-ATPases. Consequently, inhibition of this Ca(2+) reuptake mechanism results in Ca(2+) mobilization from ER stores followed by capacitative calcium entry, leading to [Ca(2+)](i) elevation. In view of the important role of Ca(2+) in apoptosis regulation, this Ca(2+) perturbation may represent part of the signalling mechanism that celecoxib uses to trigger rapid apoptotic death in cancer cells. This Ca(2+)-ATPase inhibitory activity is highly specific for celecoxib, and is not noted with other COX inhibitors tested, including aspirin, ibuprofen, naproxen, rofecoxib (Vioxx), DuP697 and NS398. Moreover, it is noteworthy that this activity is also observed in many other cell lines examined, including A7r5 smooth muscle cells, NIH 3T3 fibroblast cells and Jurkat T cells. Consequently, this Ca(2+)-perturbing effect may provide a plausible link with the reported toxicities of celecoxib such as increased cardiovascular risks in long-term anti-inflammatory therapy.

Influence of central serotonergic mechanisms on lower urinary tract function

The effects of serotonergic drugs on the central nervous system control of the lower urinary tract have revealed possible approaches for the treatment of detrusor overactivity and urinary incontinence. Studies in animals of the distribution of serotonergic nerves and receptors and the changes in voiding function induced by serotonin (5-hydroxytryptamine [5-HT]) receptor agonists and antagonists, as well as 5-HT reuptake inhibitors, form a basis for the development of such treatments. In rats and cats, spinal reflex circuits involved in voiding function exhibit a dense serotonergic innervation, multiple 5-HT receptors, and sensitivity to various drugs that affect serotonergic transmission. Although there is some evidence in the rat for serotonergic facilitation of voiding, most experiments in rats and cats indicate that activation of the central serotonergic system by 5-HT reuptake inhibitors, as well as by 5-HT 1A and 5-HT 2 receptor agonists, depresses reflex bladder contractions and increases the bladder volume threshold for inducing micturition. These data indicate that activation of the central serotonergic system can suppress voiding by enhancing efferent control of the urethral outlet and inhibiting the parasympathetic excitatory input to the urinary bladder. The 5-HT receptors and reuptake mechanisms, therefore, represent targets for the development of new treatments of detrusor overactivity and urinary incontinence.

Regulation of the glutamate uptake by extracellular calcium

To determine whether [Ca 2+] e modulates glutamate re-uptake, we studied the uptake mechanism into rat cerebrocortical synaptosomes. The removal of extracellular Ca 2+ caused a negative modulation in the uptake mechanism. The calculated K 50 value was 0.185±0.019 mM ( n=4). The Michaelis–Menten data analysis indicate that absence of Ca 2+ diminished the V max kinetic parameter by about 60% without changing significantly the K m suggesting a non-competitive mechanism. We also tested the involvement of intracellular Ca 2+ in this phenomenon by trapping BAPTA into the synaptosomal vesicles to control the Ca 2+ concentration. Our results suggest that intracellular Ca 2+ changes have a less predominant role on the glutamate uptake than do extracellular Ca 2+. These findings argue in favor of an important role of extracellular [Ca 2+] in maintaining the l-glutamate re-uptake mechanism in the mammalian central nervous system.

NO as a signalling molecule in the nervous system.

The discovery that nitric oxide (NO) functions as a signalling molecule in the nervous system has radically changed the concept of neural communication. Indeed, the adoption of the term nitrergic for nerves whose transmitter function depends on the release of NO or for transmission mechanisms brought about by NO (Moncada et al., 1997) emphasizes the specific characteristics of this mediator. The physical properties of NO prevent its storage in lipid-lined vesicles and metabolism by hydrolytic degradatory enzymes. Therefore, unlike established neurotransmitters, NO is synthesized on demand and is neither stored in synaptic vesicles nor released by exocytosis, but simply diffuses from nerve terminals. The distance of this NO diffusion (40 – 300 μm in diameter) implies that structures in the vicinity of the producing cell, both neuronal and non-neuronal, are influenced following its release. This suggests that, as well as acting as a neurotransmitter, NO has a neuromodulatory role (Garthwaite & Boulton, 1995). In addition, it diffuses into rather than binds with protein receptors on adjacent cells, and most of its known actions are the consequence of interplay with intracellular targets that would usually be regarded as secondary messengers. The activity of conventional neurotransmitters is terminated either by re-uptake mechanisms or enzymatic degradation while inactivation of NO follows reaction with a substrate. There are multiple points at which biological control can be exerted over the production and activity of conventional neurotransmitters. However, control of the synthesis of NO is the key to regulating its activity. Endothelial NOS (eNOS) and inducible NOS (iNOS) are present in the nervous system and will be duly addressed here. However, neuronal NOS (nNOS) is the principal isoform present in said system and will be the main focus of this review. All nNOS positive neurones exhibit α-nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase activity, which has become the histochemical marker of nitrergic neurones. However, early results demonstrating this may have been limited by inappropriate fixation procedures and should be viewed with caution (Wolf, 1997). The original cloning of full-length nNOS produced what is now designated as nNOSα, and which accounts for the majority of nNOS activity in nervous tissue (Bredt et al., 1991). In addition, four splice variants have recently been identified (nNOSβ, nNOSγ, nNOSμ and nNOS-2) and these appear to exhibit distinct cellular and tissue locations (Gibson, 2001; Nakane et al., 1993; Silvagno et al., 1996; Alderton et al., 2001). In particular, there is growing evidence that nNOS biosynthesis in excitable tissues is not restricted to neurones while substantial amounts of this enzyme have been identified in skeletal muscle, where it is involved in the regulation of metabolism and muscle contractility (Stamler & Meissner, 2001). The magnitude of literature dealing with the role of NO in the nervous system is so great that it would be impossible to include in this review the entirety of the research carried out. For logistical reasons, only groundbreaking references have been quoted, but when necessary, recent reviews dealing with specific areas within the field have been included and are intended to act as a guideline for further reading.

Cavernous and systemic plasma levels of norepinephrine and epinephrine during different penile conditions in healthy men and patients with erectile dysfunction

Objectives. To examine and compare the courses of norepinephrine (NE) and epinephrine (E) plasma levels in the systemic and cavernous blood taken during different penile conditions from healthy men and a group of patients with erectile dysfunction (ED). Knowledge concerning the neurophysiology of penile erection has improved tremendously during the past two decades. However, only few in vivo studies on human peripheral neurotransmission have been carried out to date. Methods. Fifty-three healthy adult male subjects and 47 patients with ED of different etiologies were exposed to erotic stimuli to elicit penile tumescence and, in the group of healthy subjects, rigidity. Whole blood was simultaneously aspirated from the corpus cavernosum and the cubital vein during different penile conditions. Plasma levels of NE and E were determined by means of a radioimmunoassay. Results. In the healthy subjects, a significant reduction of NE in cavernous plasma was detected from flaccidity (362 ± 173 pg/mL) to rigidity (248 ± 122 pg/mL), followed by an increase in the detumescence phase (336 ± 199 pg/mL). Changes in NE levels in the peripheral plasma were less pronounced. Cavernous E levels significantly increased from flaccidity (47 ± 41 pg/mL) to tumescence (130 ± 106 pg/mL) and dropped from rigidity (113 ± 67 pg/mL) to detumescence (76 ± 57 pg/mL). The course of systemic E plasma levels was similar to that in the cavernous blood. In contrast, median NE levels in the systemic and cavernous blood of the ED group slightly increased from flaccidity to tumescence (from 199 ± 88 pg/mL to 210 ± 99 pg/mL and from 273 ± 140 pg/mL to 278 ± 118 pg/mL, respectively). Conclusions. In healthy men, penile erection is accompanied by a reduction of NE in the cavernous blood and a rise in E levels in the peripheral and cavernous blood. That NE levels in the cavernous and systemic blood increase during sexual arousal in patients with ED might be an indication of a somatic dysregulation in sympathetic transmission or alterations of NE reuptake mechanisms as a cause of impaired erectile function.

Glutamate transporters alterations in the reorganizing dentate gyrus are associated with progressive seizure activity in chronic epileptic rats.

The expression of glial and neuronal glutamate transporter proteins was investigated in the hippocampal region at different time points after electrically induced status epilepticus (SE) in the rat. This experimental rat model for mesial temporal lobe epilepsy is characterized by cell loss, gliosis, synaptic reorganization, and chronic seizures after a latent period. Despite extensive gliosis, immunocytochemistry revealed only an up-regulation of both glial transporters localized at the outer aspect of the inner molecular layer (iml) in chronic epileptic rats. The neuronal EAAC1 transporter was increased in many somata of individual CA1-3 neurons and granule cells that had survived after SE; this up-regulation was still present in the chronic epileptic phase. In contrast, a permanent decrease of EAAC1 immunoreactivity was observed in the iml of the dentate gyrus. This permanent decrease in EAAC1 expression, which was only observed in rats that experienced progressive spontaneous seizure activity, could lead to abnormal glutamate levels in the iml once new abnormal glutamatergic synaptic contacts are formed by means of sprouted mossy fibers. Considering the steady growth of reorganizing mossy fibers in the iml, the absence of a glutamate reuptake mechanism in this region could contribute to progression of spontaneous seizure activity, which occurs with a similar time course.

Exciting you: signals from another kingdom

Calcium oscillations in the developing brain have been shown to play a crucial role in the development of neuronal networks. In neurons, spontaneous oscillatory electrical activity and changes in the concentration of intracellular calcium have been well described. However, in the human brain, glial cells, not neurons, are the major cell population, comprising ∼90% of brain cells. Glial cells are important companions for neurons; they not only give structural and metabolic support but, in the case of astrocytes, ensheath synapses and are integrated in synaptic transmission through their efficient neurotransmitter reuptake mechanisms. Signaling between neurons and astrocytes has been well characterized, but the astrocytic signals are detected usually in response to neuronal activation. Parri et al. now show that astrocytes are capable of generating signals and transmitting them to neurons spontaneously 1 Parri H.R. et al. Spontaneous astrocytic Ca2+ oscillations in situ drive NMDAR-mediated neuronal excitation. Nat. Neurosci. 2001; 4: 803-812 Crossref PubMed Scopus (478) Google Scholar .

Sympathetic modulation of blood pressure variability.

Although sympathetic nervous activity (SNA) displays oscillations synchronous with the heart beat and respiration, and between 0.1-0.4 Hz, it is apparent that each of these frequencies does not have the same effect on the vasculature. Frequencies above 1 Hz do not produce oscillations in the vasculature but instead contribute to the mean level of vasoconstriction. Slower oscillations in SNA result in a cycle of vasoconstriction and vasodilation within the vasculature, the amplitude of which, generally decreases with increasing frequency. Some studies indicate that, within the same species, differences exist in the frequency responses between vascular beds, such as the skin and gut. This differential responsiveness is also found between the medullary and cortical vasculature regions of the rabbit kidney. Low-pass filter properties have been described in the iliac circulation of rats, and evidence has been provided that noradrenaline reuptake mechanisms are not the frequency limiting step of the vasculature response. Recent studies on isolated rat vascular smooth muscle cells suggest that sympathetic modulation of vascular tone is limited by the alpha-adrenoceptor signal transduction into the cells and not by an intrinsic inability of the cells to contract and relax at higher rates.

Effect of chronic hypoxia on agonist-induced tone and calcium signaling in rat pulmonary artery.

The effect of chronic hypoxia (CH) for 14 days on Ca2+ signaling and contraction induced by agonists in the rat main pulmonary artery (MPA) was investigated. In MPA myocytes obtained from control (normoxic) rats, endothelin (ET)-1, angiotensin II (ANG II), and ATP induced oscillations in intracellular Ca2+ concentration ([Ca2+]i) in 85-90% of cells, whereas they disappeared in myocytes from chronically hypoxic rats together with a decrease in the percentage of responding cells. However, both the amount of mobilized Ca2+ and the sources of Ca2+ implicated in the agonist-induced response were not changed. Analysis of the transient caffeine-induced [Ca2+]i response revealed that recovery of the resting [Ca2+]i value was delayed in myocytes from chronically hypoxic rats. The maximal contraction induced by ET-1 or ANG II in MPA rings from chronically hypoxic rats was decreased by 30% compared with control values. Moreover, the D-600- and thapsigargin-resistant component of contraction was decreased by 40% in chronically hypoxic rats. These data indicate that CH alters pulmonary arterial reactivity as a consequence of an effect on both Ca2+ signaling and Ca2+ sensitivity of the contractile apparatus. A Ca2+ reuptake mechanism appears as a CH-sensitive phenomenon that may account for the main effect of CH on Ca2+ signaling.

A novel mechanism for the reuptake of CGRP

Unlike classical amine or amino acid transmitters, which are synthesized close to their sites of release, neuropeptides such as the 37-amino-acid calcitonin gene-related peptide (CGRP) are synthesized in the neuronal soma and transported through the axon to sites of release in the nerve terminals. The transport process is time- and energy-consuming, and an intense stimulation could deplete releasable pools of neuropeptides rendering the nerve terminals unresponsive (in terms of release) until this pool has been replenished by newly synthesized and transported peptide. Uptake mechanisms that remove neurotransmitters from the synaptic cleft and spare the synthesis of new molecules of neurotransmitters have been described for small amine or amino acid neurotransmitters. Now, Sams-Nielsen et al. 1xPharmacological evidence for CGRP uptake into perivascular capsaicin sensitive nerve terminals. Sams-Nielsen, A. et al. Br. J. Pharmacol. 2001; 132: 1145–1153Crossref | PubMedSee all References1 provide indirect evidence for a reuptake mechanism for CGRP into capsaicin-sensitive afferent neurones and suggest that this mechanism might have functional relevance for the refilling of a releasable neuropeptide pool in nerve terminals following CGRP release.Isolated segments of the guinea-pig basillary artery, precontracted with prostaglandin F2α (PGF2α), were challenged with capsaicin (10 μm), which caused a CGRP-dependent (blocked by the antagonist CGRP8–37), capsazepine-sensitive relaxation. When the capsaicin challenge was repeated twice again, the relaxation underwent a significant fading (from ∼60% to 20% of PGF2α contraction). At this point, preparations were incubated with CGRP (100 nm) for 20 min and the capsaicin challenges repeated another three times. Following this protocol, the fourth capsaicin challenge (the first after the CGRP incubation) caused a significant relaxation (capsazepine-resistant, but still CGRP-dependent), which showed a significant fading during the fifth and sixth capsaicin challenge. Control experiments showed that repeated challenges with exogenous CGRP induced reproducible relaxations, thus excluding the possibility that desensitization occurs at the smooth-muscle level. In another series of experiments, arterial segments were incubated with [125I]CGRP and [125I] outflow was monitored following relaxation induced by capsaicin in PGF2?-precontracted preparations. Capsaicin induced a significant radioactivity release, which suggests that [125I]CGRP had been taken up by capsaicin-sensitive nerve terminals, even in the absence of a previous capsaicin challenge. Intriguingly, the restoration of capsaicin-induced relaxation was inhibited when CGRP was co-incubated with its antagonist CGRP8–37, suggesting that prejunctional CGRP receptors are involved in the refilling of the neuropeptide in nerve terminals.These results provide indirect evidence for a CGRP reuptake mechanism in capsaicin-sensitive perivascular nerve terminals, possibly mediated through the activation of prejunctional CGRP receptors. Thus, following the activation of prejunctional CGRP receptors and the consequent internalization of the complex peptide ligand–receptor, the peptide ligand might be spared from metabolism and recycled locally for the preservation of physiological functions. In addition, these results highlight a novel role for peptide prejunctional receptors in neurones, as well as in the modulation of neurotransmitter release. Other examples for the transport of relatively large peptides across biological barriers include those mediated by multi-drug resistance protein, other peptide transporters, or specific peptide receptors located at the blood–brain barrier or the choriod plexus epithelium; however, this uptake mechanism for CGRP is the first example of a neuronal uptake of peptides. It will be interesting to assess whether this process represents an exception or similar mechanisms spare other neuropeptides.

Vitreal dihydroxyphenylacetic acid (DOPAC) as an index of retinal dopamine release.

Dopamine is generally accepted as a major neurotransmitter associated with light-adaptive processes in the retina. However, little is known about its precise release pattern in vivo, largely due to the lack of an unambiguous method for the determination of dopamine release. We have found that vitreal levels of dihydroxyphenylacetic acid (DOPAC) reflect the rate of dopamine release in chickens. Blocking re-uptake with nomifensine significantly lowered vitreal DOPAC and retinal dopamine, confirming the retinal origin and reliance of vitreal DOPAC on intact re-uptake mechanisms. Further, inhibition of monoamine oxidase with pargyline reduced vitreal as well as retinal DOPAC levels, confirming that the DOPAC detected is generated by monoamine oxidase. Finally, we found that DOPAC diffused freely into and out of isolated vitreous bodies and we found the vitreous to be metabolically inert with respect to DOPAC, supporting the idea that vitreal levels of DOPAC are consequential to the retinal metabolism of dopamine. Exposure to light, which is known to increase retinal dopamine release, readily increased vitreal DOPAC levels. The accumulation of DOPAC in the vitreous over 6 h light fitted a mathematical model of DOPAC accumulation based on zero-order influx (proportional to dopamine release rates) and diffusion driven, first-order efflux.

Altered cardiac sarcoplasmic reticulum function of intact myocytes of rat ventricle during metabolic inhibition.

Changes in the behavior of the sarcoplasmic reticulum (SR) in rat ventricular myocytes were investigated under conditions of metabolic inhibition using laser-scanning confocal microscopy to measure intracellular Ca(2+) and the perforated patch-clamp technique to measure SR Ca(2+) content. Metabolic inhibition had several effects on SR function, including reduced frequency of spontaneous releases of Ca(2+) (sparks and waves of Ca(2+)-induced Ca(2+) release), increased SR Ca(2+) content (79.4+/-5.7 to 115.2+/-6.6 micromol/L cell volume [mean+/-SEM; P:<0.001]), and, after a wave of Ca(2+) release, slower reuptake of Ca(2+) into the SR (rate constant of fall of Ca(2+) reduced from 8.5+/-1.1 s(-)(1) in control to 5.2+/-0.4 s(-)(1) in metabolic inhibition [P:<0.01]). Inhibition of L-type Ca(2+) channels with Cd(2+) (100 micromol/L) did not reproduce the effects of metabolic inhibition on spontaneous Ca(2+) sparks. These results are evidence of inhibition of both Ca(2+) release and reuptake mechanisms. Reduced frequency of release could be attributable to either of these effects, but the increased SR Ca(2+) content at the time of reduced frequency of spontaneous release of Ca(2+) shows that the dominant effect of metabolic inhibition is to inhibit release of Ca(2+) from the SR, allowing the accumulation of greater than normal amounts of Ca(2+). In the context of ischemia, this extra accumulation of Ca(2+) would present a risk of potentially arrhythmogenic, spontaneous release of Ca(2+) on reperfusion of the tissue.

Decreased reuptake of serotonin in human platelets after surgery.

We have documented earlier a decrease in platelet serotonin and a concurrent increase in plasma serotonin, 5-hydroxytryptamin (5-HT) after various forms of stress, suggesting a disturbed platelet 5-HT reuptake mechanism following stress. In order to further elucidate these findings, we have studied platelet 5-HT reuptake kinetics (Vmax and Km) in nine patients before and 4 days after major, uncomplicated abdominal surgery. We found a significant decrease in the maximal 5-HT reuptake velocity (Vmax) after surgery and changes in Km, verifying alterations in the affinity of the platelet 5-HT transport system. The present results thus confirm the hypothesis that 5-HT reuptake kinetics are altered following adrenergic hyperactivity. A decrease in platelet 5-HT reuptake may bear implications for our understanding of poststress adaptive changes in the cardiovascular system as well as in the central nervous system (CNS) serotonergic neurones following stressful stimulation.

Fluvoxamine. An updated review of its use in the management of adults with anxiety disorders.

Fluvoxamine is a potent and selective serotonin reuptake inhibitor (SSRI) that has little or no effect on other monoamine reuptake mechanisms. Relative to other SSRIs, fluvoxamine is a weak inhibitor of cytochrome P450 (CYP) 2D6, a moderate inhibitor of CYP2C19 and CYP3A4 and a potent inhibitor of CYP1A2. In randomised, double-blind trials. fluvoxamine 100 to 300 mg/day for 6 to 10 weeks significantly reduced symptoms of obsessive-compulsive disorder (OCD) compared with placebo. Response rates of 38 to 52% have been reported with fluvoxamine, compared with response rates of 0 to 18% with placebo. In patients with OCD, fluvoxamine had similar efficacy to that of clomipramine and, in smaller trials, the SSRIs paroxetine and citalopram and was significantly more effective than desipramine. Maintenance therapy with fluvoxamine may reduce the likelihood of relapses in up to 67% of patients with OCD. Fluvoxamine < or = 300 mg/day for 6 to 8 weeks was as effective as imipramine in patients with panic disorder, and significantly more effective than placebo. In addition, treatment with fluvoxamine < or = 300 mg/day for > or = 8 weeks improved symptoms of social phobia (social anxiety disorder), post-traumatic stress disorder (PTSD), pathological gambling, compulsive buying, trichotillomania, kleptomania, body dysmorphic disorder, eating disorders and autistic disorder. Large trials comparing the efficacy of fluvoxamine and other SSRIs in patients with anxiety disorders are warranted. Fluvoxamine is generally well tolerated; in postmarketing studies, nausea was the only adverse event occurring in >10% of patients with less commonly reported events including somnolence, asthenia, headache, dry mouth and insomnia. Fluvoxamine is associated with a low risk of suicidal behaviour, sexual dysfunction and withdrawal syndrome. Fewer anticholinergic or cardiovascular events are associated with fluvoxamine than tricyclic antidepressants. Although comparative data are lacking, the tolerability profile of fluvoxamine appears to be broadly similar to those of other SSRIs. Fluvoxamine has demonstrated short term efficacy in the treatment of OCD, panic disorder, social phobia, PTSD and in a range of obsessive-compulsive spectrum disorders. The drug is as effective as clomipramine in patients with OCD but appears to have a better tolerability profile. On the basis of current treatment guidelines, fluvoxamine, like other SSRIs, is recommended as first-line treatment for a number of anxiety disorders. It appears to offer some pharmacokinetic advantages and a different drug interaction profile to the other SSRIs with a broadly similar spectrum of adverse events. However, direct comparisons are required to assess the relative efficacy and tolerability of the different agents of this drug class.

Alterations by norepinephrine of cardiac sympathetic nerve terminal function and myocardial beta-adrenergic receptor sensitivity in the ferret: normalization by antioxidant vitamins.

Chronic excessive norepinephrine (NE) causes cardiac sympathetic nerve terminal abnormalities, myocardial beta-adrenergic receptor downregulation, and beta-adrenergic subsensitivity. The present study was carried out to determine whether these changes could be prevented by antioxidants. Ferrets were administered either NE (1.33 mg/d) or vehicle by use of subcutaneous pellets for 4 weeks. Animals were simultaneously assigned to receive either antioxidant vitamins (beta-carotene, ascorbic acid, and alpha-tocopherol) or placebo pellets. NE increased plasma NE 4- to 5-fold but had no effect on heart rate, heart weight, arterial pressure, or left ventricular systolic function. However, myocardial NE uptake activity and NE uptake-1 site density were reduced, as well as cardiac neuronal NE, tyrosine hydroxylase, and neuropeptide Y. In addition, there was a decrease in myocardial beta-adrenergic receptor density with a selective decrease of the beta(1)-receptor subtype, reduction of the high-affinity site for isoproterenol, decreased basal adenylyl cyclase activity, and the adenylyl cyclase responses to isoproterenol, Gpp(NH)p, and forskolin. All of these changes were prevented by antioxidant vitamins. The effects of NE on myocardial beta-adrenergic receptor density, NE uptake-1 carrier site density, and neuronal NE were also prevented by superoxide dismutase or Trolox C. The toxic effects of NE on the sympathetic nerve terminals are mediated via the formation of NE-derived oxygen free radicals. Preservation of the neuronal NE reuptake mechanism is functionally important, because the antioxidants also prevented myocardial beta-adrenergic receptor downregulation and postreceptor abnormalities. Thus, antioxidant therapy may be beneficial in heart failure, in which cardiac NE release is increased.

Neurotransmitter transporters: fruitful targets for CNS drug discovery.

More than 20 members have been identified in the neurotransmitter transporter family. These include the cell surface re-uptake mechanisms for monoamine and amino acid neurotransmitters and vesicular transporter mechanisms involved in neurotransmitter storage. The norepinephrine and serotonin re-uptake transporters are key targets for antidepressant drugs. Clinically effective antidepressants include those with selectivity for either NE or serotonin uptake, and compounds with mixed actions. The dopamine transporter plays a key role in mediating the actions of cocaine and the amphetamines and in conferring selectivity on dopamine neurotoxins. The only clinically used compound to come so far from research on amino acid transporters is the antiepileptic drug tiagabine, a GABA uptake inhibitor.

Reboxetine: tolerability and safety profile in patients with major depression.

Since reboxetine acts selectively on the reuptake of noradrenaline, it has a different side-effects profile than drugs acting predominantly on the serotonin reuptake mechanism. Data from patients treated with reboxetine (n = 1503) or placebo/comparator drugs (n = 1027) show that reboxetine has no significant cardiovascular effects, a low potential for drug interactions, causes no significant impairment of cognitive or motor function and no increase in suicidal ideation. In contrast to certain serotonergic drugs, there is no evidence of any withdrawal syndrome upon abrupt discontinuation or tapering of reboxetine treatment. Sexual dysfunction appears in only a small fraction of the patients and mainly with doses higher than 8 mg daily. The rate of discontinuation due to adverse effects with reboxetine was not significantly different from that observed with placebo in short-term studies. The level of efficacy and apparently favourable tolerability profile makes reboxetine an important alternative in the medical treatment of depressive illness.