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Spontaneous multiquantal release at synapses in guinea-pig hypogastric ganglia: evidence that release can occur in bursts.

1. A study was made of some properties of the spontaneous synaptic potentials recorded in cells of the hypogastric ganglia of guinea-pigs. 2. The distribution of the amplitudes of the spontaneous synaptic potentials arising from a single preganglionic fibre was found to be multimodal, with peaks at roughly integral multiples of a unit peak. 3. It was found that the amplitudes of the larger spontaneous potentials were consistent with them being the result of synchronous or near-synchronous release of two or more unit-sized quanta (multiquantal release). 4. The proportion of multiquantal potentials observed was found to be dependent on the extracellular calcium ion concentration. 5. When the stochastic properties of the spontaneous potentials were examined, it was found that the spontaneous release process was not random and independent but appeared to be clustered. Indeed, the probability of occurrence of a unit spontaneous synaptic potential was greatly enhanced during the 40--60 msec immediately following any given spontaneous synaptic potential. 6. When unit spontaneous potentials were excluded from the analysis, the multiquantal potentials were still found to be clustered although the clustering was less marked than that seen in the over-all process. 7. These results suggested that the multiquantal spontaneous potentials arose from the release of unit quanta in short high frequency bursts and it was found that a mathematical model incorporating such a mechanism could describe the spontaneous quantal release process.

Animals↗

The influence of visceral mechanoreceptors on sympathetic efferent discharge in the cat.

1. Recordings have been made from eighty-three single sympathetic efferent units in the hypogastric nerve in two types of preparation. In all animals the baroreceptors were denervated to exclude changes in sympathetic discharge resulting from any variations in arterial pressure, and the spinal cord was sectioned at the 6th lumbar segment to exclude changes in efferent discharge that might have been due to pelvic nerve afferents from the bladder or other viscera. In some animals the afferent pathways were sectioned from all pelvic and lower abdominal viscera other than the bladder, so that the vesical afferent pathway was the only neural pathway that might mediate reflex events from these viscera. The hypogastric nerve afferent pathway was excited by bladder distension or by bladder contractions induced by electrical stimulation of the sacral cord.2. Approximately half the units gave an increase in spike rate during distension or during contraction of the bladder. Three-quarters of the units tested also gave an increase in spike rate during colonic distension. In no units that exhibited convergence of afferent input did colonic and vesical stimuli cause responses of opposite sign.3. Approximately 10% of units showed a reduction in discharge rate when the bladder was distended or caused to contract.4. The estimated intravesical pressure thresholds for these reflexes were in the range 8-56 mmHg which extends beyond that of the mechanoreceptors which form the afferent limb of the reflex.5. These reflex studies indicate that the hypogastric nerve afferents from the bladder can elicit sympathetic reflexes within the physiological range of intravesical pressures, and that vesico-sympathetic reflexes can be elicited in the absence of pelvic nerve afferent inputs.In 75% of sympathetic efferent units that respond to bladder distension or contraction, there is evidence for convergent inputs with similar actions from the colon.

Action Potentials↗

Presence and axonal transport of cholinoceptor, but not adrenoceptor sites on a cat noradrenergic neurone.

1. Noradrenaline release and radioligand binding studies were carried out in the cat hypogastric nerve ligated in vito 2 cm distal to the inferior mesenteric ganglion for different time periods, and in different effector organs.2. Large quantities of noradrenaline and dopamine beta-hydroxylase (DBH) accumulated in the segments of nerve immediately proximal (P(1)) and distal (D(1)) to the ligation, with rates of about 100 and 25 mm/24 hr for the orthograde and retrograde transport, respectively.3. Nicotine evoked the release of noradrenaline from P(1) and atrial slices; the secretory response to nicotine was completely antagonized by mecamylamine. [(3)H]alpha-bungarotoxin biding to membranes from P(1) allowed the estimation of a K(D) of 2.97 nm and a B(max) of 1639 f-mole/mg protein.4. Acetylcholine inhibited the release of endogenous noradrenaline evoked by high K(+) stimulation in atrial slices, but not in P(1) segments. Similarly, carbachol decreased [(3)H]noradrenaline release induced by electrical stimulation (twenty-six shocks, 2 Hz, 5 msec) in the atrium but not in P(1).5. [(3)H]Quinuclydinilbenzylate ([(3)H]QNB) specifically binds to membranes from P(1) and vas deferens, following a saturation curve. In the case of P(1) segments taken 48 hr after ligation a K(D) of 0.35 nm and a B(max) of 129 f-mole/mg protein were found.6. The fact that the B(max) in P(1) and D(1) increased with the time of ligation suggests that orthograde and retrograde axonal transports of muscarinic binding sites exist in this nerve, with approximate rates of transport of 15 and 8 mm/24 hr, respectively.7. As far as adrenoceptors are concerned, we observed that yohimbine or phentholamine did not modify transmitter release from P(1), evoked by high K(+) or electrical stimulation. However, yohimbine enhanced the release of [(3)H]noradrenaline induced by electrical stimulation from splenic slices of the same animals.8. [(3)H]Clonidine, [(3)H]dihydroergocryptine or [(3)H]dihydroalprenolol ([(3)H]DHA) did not specifically bind to membranes from P(1), in spite of the fact that they showed typical saturation curves for specific binding in cortex and atrial membranes from the same cats.9. In conclusion, these data (a) further show that the ligated hypogastric nerve is a good model of noradrenergic nerve terminal free of effector cell; (b) provide direct evidence for the neural location of nicotinic receptors whose activation trigger noradrenaline release from noradrenergic neurones; (c) demonstrate the neural location and axonal transport of muscarinic receptor sites, but leave certain doubts about its functional role in this noradrenergic neurone; and (d) do not support the hypothesis that alpha and beta-adrenoceptors which modulate noradrenaline release from peripheral noradrenergic nerve terminals are neurally (or prejunctionally) located.

Animals↗

Autonomic control of penile erection in the dog.

1. In anaesthetized dogs, resting mean penile artery pressure (p.a.p.) and corpus cavernosum pressure (c.c.p.) were 70-100% and 10-15% of mean systemic blood pressure, respectively. 2. Stimulation of the pelvic nerve at 10 Hz produced an immediate drop in p.a.p. and c.c.p., followed 10-30 s later by a rise in c.c.p. to the level of p.a.p. This level was 60-90% of systemic pressure, and was maintained throughout stimulation. 3. The threshold for a rise in c.c.p. was 3-5 Hz. Atropine (1 mg/kg), phentolamine (200 micrograms kg-1) and propranolol (200 micrograms kg-1) had no effect on the response to pelvic nerve stimulation. 4. C.c.p., p.a.p. and their changes in response to pelvic nerve stimulation were not significantly altered by either stimulation or section of the hypogastric nerves. 5. Cutting the sympathetic chain on both sides at L5, or administration of phentolamine, had no effect on resting c.c.p. or p.a.p. However, subsequent responses to pelvic nerve stimulation were enhanced. 6. When the pelvic nerve was stimulated during excitation of the sympathetic chain, there was still an initial drop in p.a.p. and c.c.p. but the subsequent increase in c.c.p. was delayed or abolished. These effects were mimicked by close arterial injection of phenylephrine and blocked by alpha-adrenergic antagonists. 7. This study suggests that erections in response to pelvic nerve stimulation result from an initial increase in volume of the corpus spongiosum, followed 20 s later by a stiffening of the corpus cavernosum as its pressure increases. Only the latter process is inhibited by activity of the sympathetic fibres.

Animals↗

Differentiation of sympathetic neurones projecting in the hypogastric nerves in terms of their discharge patterns in cats.

1. Sympathetic neurones that project in the hypogastric nerves (HGNs) were analysed for their discharge patterns in anaesthetized cats. The activity of these neurones was recorded from their axons. Afferents from the pelvic organs (urinary bladder, colon, anal canal), and arterial baro-and chemoreceptors were stimulated. 150 postganglionic and nine preganglionic neurones were analysed. 2. The postganglionic neurones exhibited reflex patterns that were typical of visceral vasoconstrictor neurones and various types of motility-regulating neurones. Most motility-regulating neurones and all visceral vasoconstrictor neurones had ongoing activity. 3. Postganglionic motility-regulating neurones were not influenced by stimulation of arterial baro-and chemoreceptors, but showed distinctive reflexes on stimulation of afferents from pelvic organs. Three subgroups of motility-regulating neurones were identified: type 1 neurones (34% of the sample of postganglionic neurones) were excited from the urinary bladder and inhibited or not influenced from the colon. Type 2 neurones (14%) exhibited a reflex pattern reciprocal to that of the type 1 neurones. Anal motility-regulating neurones (8%) were only influenced from the anal canal. The most powerful reflexes in these types of motility-regulating neurones were elicited by mechanical stimulation of the anal mucosa. 4. Postganglionic visceral vasoconstrictor neurones (16% of the sample) were under powerful inhibitory control from the arterial baroreceptors and weakly excited by stimulation of arterial chemoreceptors. Visceral stimuli had little or no effect on most of these neurones. Some visceral vasoconstrictor neurones exhibited some overlap in their functional properties with motility-regulating neurones. 5. Twenty-eight per cent of our sample of postganglionic neurones showed no reflexes to the afferent stimuli used. About half of these neurones had on-going activity. 6. Nine preganglionic neurones with on-going activity were identified. Most of these neurones behaved like visceral vasoconstrictor or motility-regulating neurones. 7. This study shows that the majority of postganglionic neurones that project in the HGNs can be divided into the same functional types as the lumbar preganglionic neurones that project to the inferior mesenteric ganglion. The proportions of the different types of neurones are similar at pre- and postganglionic levels. Thus the centrally generated patterns of activity are most likely faithfully transmitted from the spinal cord to the target organs in the pelvic cavity in functionally separate pathways.

Anal Canal↗

Haemodynamic effects of distension of the descending colon in anaesthetized dogs.

1. This study was undertaken to determine whether distension of the descending colon in anaesthetized dogs reflexly affects the heart rate, arterial blood pressure or the left ventricular inotropic state. 2. Experiments were performed on twenty-six dogs, which were anaesthetized with sodium pentobarbitone and artificially ventilated. A segment of the distal descending colon was isolated and was distended with warm Ringer solution at a steady intraluminal pressure. 3. In each animal, distension of the colon caused an increase in heart rate and aortic blood pressure. The response of an increase in heart rate was augmented by preventing changes in aortic blood pressure, and was graded in seven dogs by step increments in the distending pressure. In the same animals, distension of the colon always caused a small increase in left ventricular (dP/dt)max at constant heart rate and aortic blood pressure. 4. In four of the twenty-six dogs, cutting the pelvic nerves did not abolish the observed responses to the distension. In seven of the twenty-six dogs, which included the four animals with sectioned pelvic nerves, cutting the hypogastric nerves completely abolished all the observed responses. 5. In thirteen of the twenty-six dogs, propranolol or bretylium tosylate completely abolished the reflex increases in heart rate and left ventricular (dP/dt)max, and phentolamine or bretylium tosylate abolished the reflex increase in aortic blood pressure. 6. These results showed that distension of the colon reflexly increased the heart rate, arterial blood pressure and left ventricular inotropic state. These reflex responses were mediated by sympathetic effects and their afferent limb involved the hypogastric nerves.

Animals↗

Phenotype-specific expression of T-type calcium channels in neurons of the major pelvic ganglion of the adult male rat.

1. Neurons from the major pelvic ganglia (MPG) of adult male rats were enzymatically dissociated and the neurochemical phenotype and Ca2+ current properties examined. 2. Neurons were divided into two subpopulations based on the presence or absence of low threshold T-type Ca2+ channels. The subpopulation of neurons expressing T-type Ca2+ channels was characterized by a mean diameter of 34 microns, a mean membrane capacitance (Cm) of 72 pF, tyrosine hydroxylase immunoreactivity (TH-IR), a lack of NADPH diaphorase (NADPHd) reactivity and a high degree of alpha 2-adrenoceptor-mediated Ca2+ current inhibition (60%). 3. The subpopulation of neurons without overt T-type Ca2+ channels had a mean diameter of 23 microns, a mean Cm of 30 pF, a lack of TH-IR and a moderate degree of alpha 2-adrenoceptor-mediated Ca2+ current inhibition (27%). About 50% of this subpopulation stained positively for NADPHd. 4. The contribution of high threshold N-type Ca2+ channels (60-70%), as determined from omega-conotoxin GVIA inhibition, and L-type Ca2+ channels (< 10%), as determined from nifedipine inhibition, to the whole-cell Ca2+ current was similar for both subpopulations of neurons. 5. These data indicate that the MPG contain at least two subpopulations of postganglionic neurons, i.e. adrenergic and non-adrenergic, with distinct electrophysiological and neurochemical properties. Furthermore, we propose that the presence or absence of T-type Ca2+ channels provides an electrophysiological means of identifying adrenergic and non-adrenergic phenotype, respectively, in neurons of the male rat MPG.

Animals↗

Proportional release of norepinephrine and dopamine- -hydroxylase from sympathetic nerves.

Dopamine-beta-hydroxylase(DBH), the enzyme that catalyzes the conversion of dopamine to norepinephrine, is localized in the vesicles containing catecholamine in sympathetic nerves. This enzyme is released with norepinephrine when the nerves to the guinea pig vas deferens are stimulated in vitro, and the amount of enzyme discharged increases as the length of stimulation periods increases. The amount of DBH released is proportional to the amount of norepinephrine released, and the ratio of norepinephrine to DBH discharged into the incubation medium is similar to that in the soluble portion of the contents of the synaptic vesicles from the vas deferens. These data are compatible with the release of the neurotransmitter norepinephrine and DBH from symnpathetic nerves by a process of exocytosis.

Animals↗

Cutaneous nerve stimulation and motoneuronal excitability. II: Evidence for non-segmental influences.

After stimulation of a sensory nerve, even distant motor nuclei undergo excitability changes which are evidenced by recovery curves. In all, two unequal peaks of facilitation can be identified. They appear when a given motor nucleus is tested after stimulation of various sensory nerves or when the same stimulation (of the sural nerve) is studied in various motor nuclei. The first facilitation is seen earlier in the masseter, then in the arm muscles and finally in lower limb muscles. The existence of a supraspinal centre activated by low threshold exteroceptive afferents and facilitating motor nuclei in a rostro-caudal sequence is postulated to account for certain features of the first and second peaks of facilitation.

Adolescent↗

Regional distribution of autonomic nerves in guinea pig uterus.

Uterine adrenergic and cholinergic nerves were examined histochemically and chemically in combination with surgical denervation. There was a clear regional variation with a greater number of axon terminals in the tubal end of the uterine horn and in the cervix compared to the remainder (major part) of the uterine horn. No cholinergic nerves were found in the uterus but were abundant around the uterine artery and its primary ramifications. Clusters of ganglion cells were located near the cervix: the majority were acetylcholinesterase positive, whereas the remainder stored catecholamines. Unexpectedly, the suspensory ligament (connecting the tubal end of the uterine horn with the two lower ribs) was found to contain an extensive number of adrenergic nerves. The major part of the uterine horns, as well as the cervix, was innervated mainly from two sources contributing almost equally: the paracervical ganglia (short adrenergic neurons) and postganglionic fibers running in the hypogastric nerves. A restricted area at the tubal end of the uterus received most of its via the suspensory ligament.

Acetylcholinesterase↗

Peripheral reflex pathways involving abdominal viscera: transmission of impulses through prevertebral ganglia.

In neurophysiological terms, divergence describes the transmission of impulse traffic from a single afferent line, through an integrating nervous system, and out into multiple efferent lines. This feature has been ascribed to the vertebrate central nervous system and invertebrate ganglionic systems but has not yet been associated with the autonomic nervous system in mammals. Therefore, this study investigated the degree of divergence of afferent impulse traffic through a mammalian autonomic ganglion, the inferior mesenteric ganglion (IMG) in guinea pig. Multiunit discharges were recorded extracellularly from the peripheral nerves, which emerge from the IMG, to determine the lines of efferent outflow (i.e., divergence) of impulse traffic generated by stimulating central efferent and peripheral afferent nerves. Pathways interrupted by a cholinergic ganglion synapse were identified by using hexamethonium. Pathways running directly through the IMG were identified by studying the effects of dividing nerves surgically. A complex arrangement of ascending and descending pathways was revealed, showing a neural network that interconnects the upper gastrointestinal tract, distal colon, and pelvic viscera via prevertebral ganglia.

Animals↗

Cross-organ interactions between reproductive, gastrointestinal, and urinary tracts: modulation by estrous stage and involvement of the hypogastric nerve.

Central nervous system neurons process information converging from the uterus, colon, and bladder, partly via the hypogastric nerve. This processing is influenced by the estrous cycle, suggesting the existence of an estrous-modifiable central nervous system substrate by which input from one pelvic organ can influence functioning of other pelvic organs. Here, we tested predictions from this hypothesis that acute inflammation of colon, uterine horn, or bladder would produce signs of inflammation in the other uninflamed organs (increase vascular permeability) and that cross-organ effects would vary with estrous and be eliminated by hypogastric neurectomy (HYPX). Under urethane anesthesia, the colon, uterine horn, or bladder of rats in proestrus or metestrus, with or without prior HYPX, was treated with mustard oil or saline. Two hours later, Evans Blue dye extravasation was measured to assess vascular permeability. Extravasation was increased in all inflamed organs, regardless of estrous stage. For rats in proestrus, but not metestrus, either colon or uterine horn inflammation significantly increased extravasation in the uninflamed bladder. Much smaller cross-organ effects were seen in colon and uterine horn. HYPX reduced extravasation in the inflamed colon and inflamed uterine horn, but not the inflamed bladder. HYPX eliminated the colon-to-bladder and uterine horn-to-bladder effects. These results demonstrate that inflaming one pelvic organ can produce estrous-modifiable signs of inflammation in other pelvic organs, particularly bladder, and suggest that the cross-organ effects involve the hypogastric nerve and are at least partly centrally mediated. Such effects could contribute to co-occurrence and cyclicity of distressing pelvic disorders in women.

Adaptation, Physiological↗

Influence of estradiol on micturition thresholds in the rat: involvement of the hypogastric nerve.

Studies have shown that the severity of bladder hyperreflexia induced by acute bladder inflammation varies with the ovarian cycle. These results suggest that the hyperreflexia is modulated by ovarian hormones. Other studies have suggested that such modulation involves the bladder's sympathetic innervation. These hypotheses were tested by assessing the development of bladder hyperreflexia in urethane-anesthetized rats subjected to different hormonal manipulations with or without bilateral hypogastric neurectomy (HYPX). The groups included sham ovariectomy (sham OVX), ovariectomy (OVX), OVX with estradiol replacement (OVX+E), OVX+HYPX, and OVX+HYPX+E. Assessments were performed using repeated cystometrograms (CMGs) to measure micturition thresholds (MT) before and hourly for 3 h after intravesicular treatment with 50% turpentine oil (or olive oil in an OVX+E control group). In the uninflamed bladder, treatment with estradiol increased MTs in the OVX+E group compared with the OVX group. As expected, bladder inflammation induced bladder hyperreflexia in sham OVX rats (studied in estrus). This hyperreflexia was eliminated by OVX and restored by either estradiol replacement or HYPX. Combining estradiol replacement and HYPX (i.e., OVX+E+HYPX) did not increase the severity of bladder hyperreflexia compared with either manipulation alone. These results indicate that the bladder hyperreflexia that is induced by bladder inflammation requires the presence of estradiol and suggest that this hormonal modulation is exerted via the sympathetic control of the bladder, possibly via an increase of beta-adrenergic inhibitory actions on the detrusor muscle. Similar mechanisms may contribute to bladder disorders in postmenopausal women.

Animals↗