Onlay mesh repair for treatment of small umbilical hernias ≤ 2 cm in adults: a single-centre investigation.

PURPOSE: Previous studies on the repair of small umbilical hernias have suggested a lower recurrence rate with mesh compared to suture repair. An important question is in what anatomical position the mesh should be placed. The purpose of this study was to investigate the outcome of using a standardized 4 × 4 cm onlay-mesh for umbilical hernias ≤ 2 cm. METHODS: A retrospective study was conducted at a single centre in Sweden on all umbilical hernia repairs during 2015-2019. The follow-up time was at least four months. Patients were identified using the hospital medical database. Repairs performed with suture or a sublay, ventral patch and laparoscopic mesh positioning were excluded. The patient's demographics, comorbidities, intra-and post-operative details were considered. The primary outcome was surgical site complications within 30 days. The secondary outcome was a recurrence. RESULTS: 80 patients were repaired with a small onlay-mesh for an umbilical hernia ≤ 2 cm. The median (range) follow-up time was 29.0 (4.3-50.1) months. The median age was 46 (26-76) years old. The median body mass index was 28 (19-38) kg/m2. The male to female ratio was 2:1. 4 patients were identified with a surgical site post-operative complication; three with seromas and one with a superficial wound infection. 3 of these were given antibiotics. 2 patients were treated with wound openings bedside. There were no registered cases of recurrence. CONCLUSIONS: Repairing small umbilical hernias with a small onlay-mesh was safe with a low surgical site complication rate. Randomized trials are needed to assess whether mesh can reduce recurrences in umbilical hernia repairs ≤ 2 cm.

Novel dual 'small' vesicle model of ATP- and noradrenaline-mediated sympathetic neuromuscular transmission.

The main question asked was if sympathetic nerves in guinea-pig vas deferens release the co-transmitters ATP and noradrenaline from the same, or different vesicles, i.e. in fixed combinations or independently. The extracellularly recorded excitatory junction current (EJC) and the fractional increase in overflow of tritium (delta T) were used to monitor the per pulse secretion of ATP and [3H]NA, respectively, during electrical stimulation with 1-3000 pulses at 0.1-40 Hz. The frequency- and train length-dependence and alpha 2-adrenoceptor-mediated autoinhibition of these parameters, and of the ATP-mediated twitch contraction, were compared first in the presence of cocaine (to block noradrenaline reuptake), then after brief exposure to phenoxybenzamine (PBA, to irreversibly 'destroy' alpha 2-autoreceptors). Parallel variations of EJC/p(ulse) and delta T/p(ulse) under all conditions would support, non-parallel variations argue against exocytosis of ATP and noradrenaline from the same vesicles. The main findings were that facilitation and alpha 2-autoinhibition of EJC/p and delta T/p were remarkably similar during stimulation at 2 Hz but increasingly dissimilar at higher frequencies. delta T/p remained strongly facilitated and tightly controlled by activation of alpha 2-autoreceptors at 10-40 Hz, but both the facilitation and the sensitivity to alpha 2-autoinhibition of EJC/p were inversely related to frequency. At 40 Hz EJCs were 'small', minimally facilitated and totally unaffected by cocaine or PBA, i.e. insensitive to alpha 2-autoinhibition. Nevertheless, activation of alpha 2-receptors during the 40 Hz train strongly restricted the 'post-tetanic augmentation' (PTA) of the first EJC 10 s after the tetanus. Comparison between the frequency dependence of EJCs and the twitch contraction in the presence of cocaine or after PBA treatment indicates that it is the 'summed EJC per second', i.e. the ATP-driven current injection per unit time into smooth muscle, which triggers the twitch. The working hypothesis is proposed that these nerves use two classes of 'small vesicles' (SVs) to store and release either 'big' or 'small' ATP and noradrenaline 'quanta', and that differences in properties (Ca2+ affinity, capacity) of Ca2+ receptors in the SV membranes enable the nerves to selectively secrete 'big quanta' at low frequency and 'small quanta' during trains at high frequency.

Do sympathetic nerves release noradrenaline in "quanta"?

The discovery of excitatory junction potentials (EJPs) in guinea-pig vas deferens by Burnstock and Holman (1960) showed for the first time that a sympathetic transmitter, now known to be ATP, is secreted in "quanta". As it was assumed at the time that EJPS are triggered by noradrenaline, this discovery led to attempts to use the fractional overflow of noradrenaline from sympathetically innervated tissues to assess, indirectly, the number of noradrenaline molecules in the average "quantum". The basic finding was that each pulse released 1/50000 of the tissue content of noradrenaline, when reuptake was blocked and prejunctional alpha(2)-adrenoceptors were intact. This provided the constraints, two extreme alternatives: (i) each pulse releases 0.2-3% of the content of a vesicle from all varicosities, or (ii) each pulse releases the whole content of a vesicle from 0.2 to 3% of the varicosities. New techniques have made it possible to address questions about the release probability in individual sites, or the "quantal" size, more directly. Results by optical (comparison of the labelling of SV2 and synaptotagmin, proteins in the membrane of transmitter vesicles), electrophysiological (excitatory junction currents, EJCs, at single visualized varicosities) and amperometric (the noradrenaline oxidation current at a carbon fibre electrode) methods reveal that transmitter exocytosis in varicosities is intermittent. The EJC and noradrenaline oxidation current responses (in rat arteries) to a train of single pulses were observed to be similar in intermittency and amplitude fluctuation. This suggests that they are caused by exocytosis of single or very few "quanta" of ATP and noradrenaline, respectively, equal to the contents of single vesicles, from a small population of release sites. These findings support, but do not conclusively prove the validity of the "intermittent" model of noradrenaline release. The question if noradrenaline is always secreted in packets of preset size ("quanta") and if the "quantum" is a subfraction or the whole content of single synaptic vesicles, still remains open.

Catecholaminergic neurotransmission: flagship of all neurobiology.

The paper describes, with focus on the first half of the century, the roles played by study of the sympathoadrenal system for developing modern neurobiology. Adrenaline isolated from extracts of adrenal medulla was the first intercellular messenger to be chemically identified and synthesized. Similarities between effects of adrenaline and sympathetic nerve stimulation led to the first concrete proposal of chemical neurotransmission. That effluent from a sympathetically or parasympathetically stimulated frog heart induced acceleration or slowing of an unstimulated recipient heart was the first conclusive proof of chemical neurotransmission. Acetylcholine (in parasympathetic or somatomotor) and noradrenaline (in sympathetic nerves) are the first identified mammalian neurotransmitters. The existence of a'receptive substance for adrenaline' represents the first proposal that target cells recognize and react to the released transmitter. Deviations for the '-ergic concept, in which sympathetic and parasympathetic nerves are termed 'adrenergic' and 'cholinergic', led to discovery of 'non-adrenergic, non-cholinergic' nerves and a range of other transmitters. That some effects of e.g. sympathetic nerve stimulation are not blocked by any noradrenaline antagonist led to the recognition that some nerves utilize more than one transmitter. Noradrenaline in sympathetic nerves was the first neurotransmitter to be visualized in the light microscope. catecholamines in adrenal medulla and sympathetic nerves were the first messengers to be shown to be stored in vesicles.

Facilitation and depression of ATP and noradrenaline release from sympathetic nerves of rat tail artery.

1. Excitatory junction currents (EJCs) were used to measure ATP release; noradrenaline (NA) oxidation currents and fractional overflow of labelled NA, [3H]NA, were used to monitor the release of endogenous and exogenous NA, respectively, from post-ganglionic sympathetic nerves of rat tail artery. 2. During nerve stimulation with 100 pulses at 5-20 Hz the EJCs initially grew in size (maximally by 23 %, at 2-10 Hz), and then depressed, maximally by 68 % at 20 Hz. 3. The peak amplitude of NA oxidation currents in response to nerve stimulation with 100 pulses at 2-20 Hz grew in size with frequency, while the area was independent of frequency and roughly constant. 4. The size of the NA oxidation currents evoked by nerve stimulation with 4-100 pulses at 20 Hz grew linearly with train length between pulses 4-16. Between pulses 20-100 there was a train length-dependent depression of the signal. 5. Fractional overflow of [3H]NA in response to nerve stimulation with 5-100 pulses at 20 Hz behaved similarly to the EJCs. It initially grew roughly linearly between pulses 5-25, and then showed a dramatic depression similar to that of the EJCs. 6. The alpha2-adrenoceptor antagonists rauwolscine and yohimbine increased the overflow of [3H]NA and the amplitude of NA oxidation currents, but not that of the EJCs. 7. It is concluded that during high-frequency stimulation (i) the release of ATP and NA is first briefly facilitated then markedly depressed, (ii) facilitation and depression of the two transmitters are similar in magnitude and time course, and (iii) alpha2-adrenoceptor antagonists differentially modify EJCs and the NA signals. The results obtained in the absence of drugs are compatible with the hypothesis that ATP and NA are released in parallel, while the effects of alpha2-adrenoceptor antagonists seem to suggest dissociated release.

Paired pulse analysis of ATP and noradrenaline release from sympathetic nerves of rat tail artery and mouse vas deferens: effects of K+ channel blockers.

1. The paired pulse stimulus paradigm - two pulses of equal strength delivered at variable interpulse intervals was used to study the release of ATP and noradrenaline (NA) from post ganglionic sympathetic nerves of rat tail artery and mouse vas deferens. 2. Excitatory junction currents (EJCs) were used to measure the release of ATP, and differential pulse amperometry to measure that of NA. 3. At interpulse intervals of 0.1 - 1 s paired pulse stimulation caused an increase in the size of the second EJC, both in rat tail artery and mouse vas deferens. As the interpulse interval was increased to 10 s or more, the two EJCs became of equal size. 4. In both preparations the K+ channel blockers tetraethylammonium (TEA, 20 mM) and 4-aminopyridine (4-AP, 1 mM) prolonged the duration of the nerve terminal spike and greatly amplified the first EJC of the pair. 5. In the presence of TEA and 4-AP in rat tail artery paired pulse stimulation caused a dramatic depression of the second EJC without markedly affecting the nerve terminal spike. The depression of the second EJC decreased with increasing interpulse intervals, and also when external Ca2+ was reduced to 0.2 mM. In mouse vas deferens, TEA and 4-AP caused only a modest depression of the second EJC. 6. In rat tail artery in the presence of TEA and 4-AP paired pulse stimulation caused a depression of the NA oxidation current evoked by the second pulse, which was similar in magnitude and time course to that of the EJC. Similar TEA and 4-AP induced depression of the second pulse response was also observed when the purinergic and noradrenergic components of the contractile response were investigated. 7. The results show that in rat tail artery K+ channel blockers cause a dramatic paired pulse depression of the release of ATP and NA. The similarity in the depression of the EJC, the NA oxidation current, and the purinergic and noradrenergic components of the contractile response is compatible with the hypothesis that ATP and NA are released in parallel from the same neuronal sources.

Geometry, kinetics and plasticity of release and clearance of ATP and noradrenaline as sympathetic cotransmitters: roles for the neurogenic contraction.

The paper compares the microphysiology of sympathetic neuromuscular transmission in three model preparations: the guinea-pig and mouse vas deferens and rat tail artery. The first section describes the quantal release of ATP and noradrenaline from individual sites. The data are proposed to support a string model in which: (i) most sites (> or = 99%) ignore the nerve impulse and a few (< or = 1%) release a single quantum of ATP and noradrenaline; (ii) the probability of monoquantal release is extremely non-uniform; (iii) high probability varicosities form 'active' strings; and (iv) an impulse train causes repeated quantal release from these sites. Analogy with molecular mechanisms regulating transmitter exocytosis in other systems is proposed to imply that coincidence of at least two factors at the active zone, Ca2+ and specific cytosolic protein(s), may be required to remove a 'fusion clamp', form a 'fusion complex' and trigger exocytosis of a sympathetic transmitter quantum, and that the availability of these proteins may regulate the release probability. The second section shows that clearance of noradrenaline in rat tail artery is basically > or = 30-fold slower than of co-released ATP, and that saturation of local reuptake and binding to local buffering sites maintain the noradrenaline concentration at the receptors, in spite of a profound decline in per pulse release during high frequency trains. The third section describes differences in the strategies by which mouse vas deferens and rat tail artery use ATP and noradrenaline to trigger and maintain the neurogenic contraction.

A novel electrophysiological approach to monitor pulse by pulse the concentration of released noradrenaline at the presynaptic alpha 2-adrenoceptors of sympathetic nerves in rat tail artery.

The excitatory junction current (EJC) evoked by electrical stimulation of postganglionic sympathetic nerves of rat tail artery with 100 pulses at 2 Hz, at 1.3 mmol/l external Ca2+, was used as a measure of the per pulse release of ATP. In controls the EJCs were initially facilitated, then gradually depressed during the stimulus train. The first EJC was slightly depressed by the alpha 2-adrenoceptor antagonist yohimbine, but starting from the 4th pulse the EJCs were enhanced. Yohimbine increased the early facilitation without markedly modifying the subsequent depression. The yohimbine-induced enhancement of EJCs caused by pulses 11-100 was, thus, constant. The noradrenaline reuptake blocker cocaine depressed the EJCs, abolished the early facilitation and slightly enhanced the depression. These effects of cocaine were reversed by further addition of yohimbine. The alpha 2-adrenoceptor agonist xylazine (1 and 10 mumol/l) dose dependently depressed the EJCs starting from the first pulse. The inhibitory effect of 1 mumol/l xylazine, but not that of 10 mumol/l xylazine, declined with train length. The inhibition of individual EJCs caused by activation of presynaptic alpha 2-adrenoceptors was used to monitor the concentration of released noradrenaline at these receptors. The ratio of individual EJCs in the presence and absence of yohimbine was assumed to reflect, pulse by pulse, the relative concentration of released noradrenaline at the presynaptic alpha 2-adrenoceptors, and hence termed [NA]alpha 2. For comparison, the concentration of endogenous noradrenaline was monitored electrochemically by differential pulse amperometry with a carbon fibre microelectrode; this signal is termed [NA]CF. [NA]alpha 2 and [NA]CF grew during the first 7-10 or 14-16 pulses, respectively, and then remained relatively constant throughout the stimulus train. Cocaine caused [NA]alpha 2 and [NA]CF to continue to grow during the first 35 and 50 pulses, and enhanced their peak levels by 180% and 320%, respectively. For comparison with the effects on the EJCs mediated via presynaptic alpha 2-adrenoceptors, those caused by varying external Ca2+ level were examined. At 0.65 mmol/l Ca2+ the amplitude of the first EJC was smaller than that at 1.3 mmol/l Ca2+, but the facilitation of later EJCs was enhanced and the subsequent depression reduced. An increase in external Ca2+ to 2.6 mmol/l had the opposite effects. All effects on EJCs caused by changes in external Ca2+ were maximal for the first EJC and then declined with the train length.(ABSTRACT TRUNCATED AT 400 WORDS)

Nerve activity-dependent variations in clearance of released noradrenaline: regulatory roles for sympathetic neuromuscular transmission in rat tail artery.

The aim of this study was to find out if clearance of noradrenaline released from sympathetic nerve terminals in rat isolated tail artery is a physiological variable and if so, to determine its role for the noradrenaline-mediated neurogenic contraction. The per pulse release of noradrenaline induced by electrical nerve stimulation and the fluctuations of the level of noradrenaline at the receptors driving the contractions were assessed from the electrochemically determined noradrenaline oxidation current at a carbon fibre electrode at the surface of the artery. Both were compared with the noradrenaline-mediated neurogenic contraction. The effects on these parameters of cocaine or desipramine, or of corticosterone, were used to assess the relative roles of neuronal and extraneuronal uptake, respectively. The effects of cocaine or desipramine, which enhance the noradrenaline level at the receptors by blocking neuronal reuptake, were compared with those of yohimbine, presumed to act exclusively by enhancing the per pulse release of noradrenaline. The results seem to support the following tentative conclusions. Clearance of released noradrenaline occurs by neuronal uptake and diffusion, while extraneuronal uptake is negligible. The noradrenaline-induced neurogenic contraction is mediated via adrenoceptors on cells near the plane of the nerve plexus; the excitation spreads from these cells throughout the syncytium. The contractile response to exogenous noradrenaline may also be mediated via receptors on the innervated key cells. Reuptake of noradrenaline into the releasing varicosities, i.e. in "active junctions", is highly efficient for single quanta but rapidly saturated by repeated release, while reuptake of noradrenaline in the "surround" of active junctions is probably rarely saturated and more independent of nerve activity. Saturation of the transporter by repeated release of quanta from the same varicosity and the consequent accumulation of "residual" noradrenaline and increased diffusion out of the junction and recruitment of noradrenaline receptors in the surround may be the cause of the rapid growth of the contraction during a high frequency train. Diffusion of released noradrenaline away from the postjunctional receptors is restricted by a local nerve activity-dependent buffering mechanism which, in spite of fading of the per pulse release, helps maintain the noradrenaline concentration at the receptors and the contraction during long high-frequency trains. Reactivation of the clearance mechanisms upon cessation of nerve activity accelerates the relaxation.(ABSTRACT TRUNCATED AT 400 WORDS)

Spatiotemporal pattern of quantal release of ATP and noradrenaline from sympathetic nerves: consequences for neuromuscular transmission.

The recent explosive development in research concerning the fundamental mechanisms of synaptic transmission helps put the present paper in context. It is now evident that not all transmitter vesicles in a nerve terminal, not even all those docked at its active zones, are immediately available for release (36). We watch, fascinated, the unraveling of the amazingly complex cellular mechanisms and molecular machinery that determine whether or not a vesicle is "exocytosis-competent" (77,78,39,79). Studies on quantal release in different systems show that neurons are fundamentally similar in one respect: that transmitter release from individual active zones is monoquantal (2). But they also show that active zones in different neurons differ drastically in the probability of monoquantal release and in the number of quanta immediately available for release (3). This implies that one should not extrapolate directly from transmitter release in one set of presynaptic terminals (e.g., in neuromuscular endplate or squid giant synapse) to that in other nerve terminals, especially if they have a very different morphology. As shown here, one should not even extrapolate from transmitter release in sympathetic nerves in one tissue (e.g., rat tail artery) to that in other tissues or species (e.g., mouse vas deferens). It is noteworthy that most studies of quantal release are based on electrophysiological analysis and therefore deal with release of fast, ionotropic transmitters from small synaptic vesicles at the active zones, especially in neurons in which these events may be examined with high resolution (49,48,46,33,32). Such data are useful as general models of the release of both fast and slow transmitters from small synaptic vesicles at active zones in other systems, provided that these transmitters are released in parallel, as are apparently ATP and NA in sympathetic nerves. They tell us little or nothing, however, about the release of transmitters (e.g., neuropeptides) from the large vesicles, nor about the spatiotemporal pattern of monoquantal release from small synaptic vesicles in the many neurons that have boutons-en-passent terminals. They show that the time course of effector responses to fast, rapidly inactivated transmitters such as ACh or ATP is necessarily release related. But they do not even address the possibility that the effector responses to slow transmitters such as NA, co-released from the same terminals, may obey completely different rules and perhaps rather be clearance related (7).(ABSTRACT TRUNCATED AT 400 WORDS)

Kinetics of ATP- and noradrenaline-mediated sympathetic neuromuscular transmission in rat tail artery.

Electrophysiological, electrochemical and mechanical recordings were employed to study the kinetics of the release and clearance of adenosine 5'-triphosphate (ATP) and noradrenaline (NA) as sympathetic co-transmitters and of the neurogenic and non-neurogenic contractions in rat isolated tail artery. The life-time of ATP and NA released by a single pulse or 10 pulses at 50 Hz was brief (< 100 ms, or < 3 s, respectively); the neurogenic contractile responses occurred largely after the transmitters had been removed from the extracellular space. The ATP-induced neurogenic contractile responses to a single pulse or 10 pulses at 50 Hz were similar in time-course to the responses to direct muscle stimulation at low voltage; both seemed to be caused by activation of nifedipine-sensitive voltage-gated L-type Ca2+ channels. The alpha 1- and alpha 2-adrenoceptor-mediated components of the NA-induced neurogenic contractile response to 10 pulses at 50 Hz were more delayed and prolonged and determined by properties of the post-receptor mechanisms. The per pulse release of both ATP and NA faded rapidly during long high-frequency trains. So did the ATP level at the receptors and the ATP-induced neurogenic contraction. The NA levels and the contractile responses induced via alpha 1- and alpha 2-adrenoceptors were much better maintained during ongoing stimulation at 20 Hz but relaxed rapidly afterwards, suggesting that nerve activity suppressed, and cessation of nerve activity reactivated NA clearance.

Intermittent release of noradrenaline by single pulses and release during short trains at high frequencies from sympathetic nerves in rat tail artery.

As shown by electrophysiological analysis, the release of the sympathetic co-transmitter adenosine 5'-triphosphate (ATP) from individual release sites is monoquantal and intermittent; the average release probability may be as low as 0.01. Indirect evidence from biochemical studies of noradrenaline overflow is compatible with a similar monoquantal, low probability release of noradrenaline as well. In the present study our first aim was to address this issue more directly in rat tail artery, using continuous amperometry to monitor in real time the release of noradrenaline from a relatively small number of sympathetic nerve varicosities. The results seem to provide the first direct evidence that noradrenaline, similarly to ATP, may be released intermittently during nerve stimulation at low frequency. Our second aim was to use the same technique to study the release of noradrenaline caused by nerve stimulation with single pulses or short trains (two to eight pulses) at high frequencies. The results show that during stimulation at 20 Hz the peak amplitude of the noradrenaline oxidation current response grew linearly with the train length, but at 50 Hz the curve describing this growth was sigmoid in shape.

Dual contractile effects of ATP released by field stimulation revealed by effects of alpha,beta-methylene ATP and suramin in rat tail artery.

1. The field stimulation-induced release of endogenous ATP and noradrenaline (NA) and contractile response in rat isolated tail artery were examined. The release of ATP was studied by extracellular electrophysiological recording and that of NA by a novel voltammetrical technique. The effects of the P2-purinceptor antagonist, suramin, on these parameters were compared with those of alpha,beta-methylene ATP, a P2X-purinoceptor desensitizing agent. 2. Neither alpha,beta-methylene ATP (10 microM) nor suramin (100-500 microM) had significant effects on the extracellularly recorded nerve terminal action potential but both abolished the ATP-induced excitatory junction current caused by stimulation at 0.1 Hz. Neither agent affected significantly the voltammetrically measured release of NA induced by 10 or 100 pulses at 20 Hz. 3. Combined blockade of both postjunctional alpha 1- and alpha 2-adrenoceptors by prazosin and yohimbine (both 0.1 microM) profoundly depressed the contractile response to 10 pulses at 20 Hz. The small and fast residual contraction in the presence of these agents was abolished by alpha,beta-methylene ATP (10 microM) and inhibited by suramin in a concentration-dependent manner (10-500 microM; IC50 75 microM) and was hence probably caused by ATP or a related nucleotide. 4. When added first, alpha,beta-methylene ATP (10 microM) or suramin (100-500 microM) delayed the onset and enhanced the amplitude of the neurogenic contraction. This enhanced response was abolished by further addition of prazosin and yohimbine (both 0.1 microM). 5. The K+ channel blocker, tetraethylammonium (10 mM), dramatically enhanced the contractile response to 100 pulses at 1 Hz and caused it to become diphasic. Addition of alpha,beta-methylene ATP (10 microM)or suramin (100-500 microM) abolished the large initial twitch component of this contraction and depressed the tonic phase.6. Like alpha,beta-methylene ATP, suramin (500 microM) had no effect on the contraction caused by exogenous NA (1O nM-l10 microM) or KCI (60 mM); both agents almost abolished the contraction caused by ATP(100 microM).7. In conclusion, (i) the contractile response of rat tail artery to electrical field stimulation is mediated by both ATP and NA, and is thus an expression of ATP-NA co-transmission, (ii) the released ATP exerts two opposite effects via 'P2x-like' purinoceptors, triggering the initial rapid phase of the neurogenic contraction and restricting the NA-mediated component of the contraction; and (iii) the source and possible physiological role of the ATP which causes the inhibitory effect are unknown at present.

Kinetics of noradrenaline released by sympathetic nerves.

At the skeletal neuromuscular junction the released neurotransmitter, acetylcholine, is eliminated within some milliseconds. This time course is known with great precision through the electrical response of target cells. At the sympathetic neuroeffector junction the fast electrical response is not mediated by noradrenaline but by a cotransmitter: ATP. The slow electrical response and the slow component of smooth muscle contraction are principally mediated by noradrenaline. These responses are two orders of magnitude slower than the electrical response to ATP. Therefore, great uncertainty remains regarding the kinetics of noradrenaline appearance and elimination. Here, the local noradrenaline concentration at the surface of the isolated rat tail artery was electrochemically monitored in real time using a carbon fibre electrode. We have shown that the time course of the neurogenically released noradrenaline is at least one order of magnitude faster than the resulting contraction. The kinetics of noradrenaline inactivation by neuronal reuptake were also precisely measured.

Frequency- and train length-dependent variation in the roles of postjunctional alpha 1- and alpha 2-adrenoceptors for the field stimulation-induced neurogenic contraction of rat tail artery.

The present paper examines the roles of postjunctional alpha 1- and alpha 2-adrenoceptors for the noradrenaline (NA)-induced neurogenic contractile response to field stimulation mainly with 1-100 pulses at 2 or 20 Hz, in the tail artery of adult normotensive rats. Pharmacological tools were employed to isolate and characterize the alpha 1- and alpha 2-adrenoceptor-mediated components of this response. The degree to which the drugs influenced NA release or reuptake was assessed by their effects on the electrochemically determined, stimulation-induced rise in the NA concentration at the innervated outer surface of the media. This response was unaffected by alpha,beta-methylene ATP (10 microM) or suramin (500 microM), added to desensitize or block P2-purinoceptors, respectively prazosin (0.1 microM) or SK&F 104078 (6-chloro-9-[(3-methyl-2-butenyl)oxyl]-3-methyl- 1H-2,3,4,5-tetrohydro-3-benzazepine, 0.1 microM), used to block postjunctional alpha 1- and alpha 2-adrenoceptors respectively, nifedipine (10 microM), blocker of Ca2+ influx through L-type channels, and ryanodine (10 microM), which blocks mobilization of Ca2+ from intracellular stores; it was moderately enhanced by yohimbine (0.1 microM), blocker of pre- and postjunctional alpha 2-adrenoceptors, and strongly enhanced by cocaine (3 microM) or desipramine (1 microM), blockers of NA reuptake. Judging from their inhibitory effects on the contractile responses to the alpha 1- and alpha 2-adrenoceptor agonists, phenylephrine and xylazine, prazosin (0.1 microM) and SK&F 104078 (0.1 microM) could be used to selectively block alpha 1- and alpha 2-adrenoceptors respectively, while yohimbine (0.1 microM) was less selective, strongly depressing alpha 2- and slightly depressing alpha 1-adrenoceptor-mediated responses. The alpha 1-adrenoceptor-mediated component of the contractile response to short trains at 20 Hz was fast in onset, brief in duration and abolished by ryanodine; that mediated by alpha 2-adrenoceptors was more delayed, prolonged and insensitive to ryanodine. Both components were dose-dependently depressed by nifedipine (0.1-10 microM). The small contractile responses to single pulses, or up to 50 pulses at 2 Hz, or short train (< 4 pulses) at 20 Hz, were more markedly depressed by 0.1 microM yohimbine or SK&F 104078 than by 0.1 microM prazosin and, hence, mediated mainly by alpha 2-adrenoceptors. The reverse was true of the much larger response to longer trains at 20 Hz, which thus probably was mediated mainly by alpha 1-adrenoceptors.(ABSTRACT TRUNCATED AT 400 WORDS)

Sympathetic transmitter release in rat tail artery and mouse vas deferens: facilitation and depression during high frequency stimulation.

Electrophysiological and electrochemical methods were used to study the release of adenosine 5'-triphosphate (ATP) and noradrenaline (NA) from sympathetic nerves during stimulation with trains at 20 Hz (tetanus). In mouse vas deferans ATP release was mainly facilitated during the tetanus, but in rat tail artery progressively and reversibly depressed. In rat tail artery reduction of external calcium attenuated the depression and increased the facilitation during the tetanus, while increased external calcium accentuated the depression. Both ATP and NA release were depressed in parallel during the first 100 pulses of the tetanus. The depression of release was not due to action potential failure, or alpha 2-adrenoceptor-mediated autoinhibition.

Fast and local electrochemical monitoring of noradrenaline release from sympathetic terminals in isolated rat tail artery.

Noradrenaline release from sympathetic nerve terminals was evoked by electrical nerve stimulation of an isolated segment of rat tail artery. This release was recorded by a carbon fiber electrode combined with differential pulse amperometry. The active part of the electrode (one carbon fiber 8 microns in diameter and 50 microns in length) was placed in close contact with the arterial surface. The oxidation current appearing at +120 mV and corresponding to the local noradrenaline concentration at the electrode surface was recorded every 0.5 s. No oxidation current was detected under resting conditions, but electrical stimulation evoked an immediate increase in this current. This response was suppressed when tetrodotoxin was added to the perfusion medium and was enhanced when noradrenaline reuptake was inhibited by cocaine. The amplitude of the response was increased with increasing stimulation frequencies (2-25 Hz) and train lengths (1-16 pulses). Finally, the time resolution of the method (0.5 s) was good enough to show that noradrenaline release precedes the postsynaptic response, i.e., the electrically evoked contraction of the artery.