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Autonomic nervous control of cervical secretion in the guinea-pig.

Neurogenic influence on cervical secretion was studied in experiments performed on isolated guinea-pig cervix with intact adjacent nerves. The secretion was estimated as carbohydrate efflux. Experiments were undertaken at ovulation time when secretion is most prominent. Hypogastric nerve stimulation significantly increased carbohydrate release. This response was unaffected by phentolamine but inhibited by TTX, scopolamine and hexamethonium, as well as previous destruction of paracervical ganglia. Pelvic nerve stimulation did not affect cervical secretion. Carbachol as well as VIP induced a secretory response of the same extent as did hypogastric nerve stimulation. Scopolamine abolished the VIP-induced secretion. VIP and carbachol in combination had no additive effect. It is concluded that cervical glands of the guinea-pig uterus possess a cholinergic secretomotor innervation. The main pathway is the hypogastric nerve and neurons have a peripheral relay at paracervical ganglia. A more complex peripheral organization cannot be excluded.

Animals↗

Urinary bladder and urethral responses to pelvic and hypogastric nerve stimulation and their relation to vasoactive intestinal polypeptide in the anaesthetized dog.

The effects on the urinary bladder and urethra of pelvic and hypogastric nerve stimulation and their relation to vasoactive intestinal polypeptides (VIP) were investigated in the anaesthetized dog. Both pelvic and hypogastric nerve stimulation elicited a twofold increase in urinary bladder blood flow and a clear-cut increase in bladder venous effluent VIP concentration. Hypogastric nerve stimulation induced an initial, partly alpha-adrenergic and partly non-adrenergic, non-cholinergic, contraction of the urinary bladder followed by a relaxation. The urethra response was a maintained alpha-adrenergic contraction. Pelvic nerve stimulation elicited a bladder contraction with an initial non-cholinergic peak, whereafter the bladder pressure was maintained at a lower level, an effect which was mainly cholinergic in origin. The urethral response was an initial non-adrenergic, non-cholinergic contraction followed by a maintained cholinergic contractile response. Afferent pelvic nerve stimulation led to an efferent activity that seemed to be a combination of activity in pelvic and hypogastric pathways to the urinary bladder and the urethra. VIP (10 nmol) injected i.v. induced a relaxation of the urinary bladder and the urethra, together with a fall in systemic blood pressure. However, despite high plasma concentrations, no vasodilation was elicited in the urinary bladder. Thus, the main target for the VIP release during pelvic and hypogastric nerve stimulation is probably not the bladder vasculature, but instead perhaps the bladder smooth muscle proper.

Afferent Pathways↗

Some effects of pilocarpine and acetylcholine on the responses of the guinea-pig vas deferens to hypogastric nerve stimulation and transmural stimulation and applied (minus)-noradrenaline.

Enhancement by acetylcholine and by pilocarpine of the contractile responses of the guinea-pig isolated vas deferens to hypogastric nerve and transmural stimulation and to exogenous (minus)-noradrenaline was examined. At high concentration (600 nM) acetylcholine enhanced responses in the order:hypogastric nerve greater than transmural greater than exogenous (minus)-noradrenaline. Pilocarpine produced a similar maximal degree of enchancement with all three stimuli. All these effects were reversibly inhibited by 1-4 mum atropine, a concentration which alone did not affected responses to the stimuli. Pilocarpine (50-400 muM) gave a concentration-related inhibition of the enhancement of responses to hypogastric nerve stimulation by 600 nM acetylcholine.

Acetylcholine↗

The activity of post-ganglionic sympathetic nerves to the uterus of the rabbit.

1. The spontaneous activity recorded from the central ends of the uterine nerves of the rabbit is described. The discharges resembled those reported from other post-ganglionic sympathetic nerves; the individual units appeared to discharge remarkably regularly at frequencies between 4/sec and 1 per 3 sec.2. The resting discharge in late pregnant rabbits was usually more intense than in non-pregnant rabbits; otherwise no clear relation existed between variations in the discharges and in the sexual state.3. The discharge was reduced by stimulation of the central end of the depressor nerve and increased by asphyxia.4. Stimulation of either the preganglionic or the post-ganglionic nerve by a single shock evoked a long after-discharge in the post-ganglionic nerve. It is shown that this after-discharge is a property of some of the post-ganglionic neurones supplying the rabbit's uterus which does not depend on the integrity of the preganglionic nerves or of the spinal cord, or on transmission through the ganglia, and that it is not a general property of the sympathetic post-ganglionic neurones of the rabbit.5. Repetitive stimulation of the preganglionic nerve at low frequencies augmented the after-discharge in some cases and depressed it in others; stimulation at high frequencies invariably depressed the after-discharge. Repetitive stimulation of the post-ganglionic nerve at low or high frequencies depressed the after-discharge.6. If the normal preganglionic activity was interrupted abruptly by an anode block, the post-ganglionic discharge also stopped abruptly.7. A ganglion-stimulating action of hexamethonium is described.8. Adrenaline inhibited the post-ganglionic neurones with no sign of excitation. This effect was antagonized by Rogitine without affecting the resting discharge, the action of the depressor nerve or the after-discharge in any way.

Animals↗

An analysis of the transmission of excitation from autonomic nerves to smooth muscle.

1. An analysis has been made of the transmission of excitation from the hypogastric nerve to the smooth muscle cells of the guinea-pig vas deferens.2. Depolarization of single muscle cells with current pulses from an intracellular electrode gave local depolarizations of the cell membrane which were not propagated. The total membrane resistance after 100 msec of depolarization was 15 MOmega for depolarizations between 10 and 40 mV.3. Depolarization of some cell membranes with a current pulse during the excitatory junction potential (E.J.P.) decreased the amplitude of the E.J.P. from about 10 mV at 20 mV depolarization, to nearly zero at 60 mV depolarization. In some cells the E.J.P. was unchanged during depolarizations of 50 mV.4. The action of transmitter on the smooth muscle cell membrane continued for the duration of the E.J.P. Action potentials which occurred at various times during the E.J.P. failed to remove the remaining phases of the E.J.P.5. It was shown that the slow time course of the E.J.P. could not be due to the instantaneous and simultaneous release of transmitter from a number of relatively distant sources.6. It was shown that each smooth muscle cell was innervated by several axons. The serial sections examined with the electron microscope showed that a smooth muscle had either a single axon terminating within 200 A of the muscle or no axons terminating on it at all. Therefore transmitter must be released along the length of the axons as well as at the terminations of the axons.

Animals↗

The innervation of the vas deferens of the guinea-pig.

1. The compound action potential of the hypogastric nerve of the guinea-pig contained two main elevations. The low-threshold fibres had a range of conduction velocities from 1.5 to 10 m/sec. The high threshold fibres conducted action potentials at less than 1 m/sec. The hypogastric nerve contained small myelinated fibres and non-myelinated fibres.2. In the preparation in vitro, junctional potentials and contractions were elicited by stimulation of the rapidly conducting fibres alone. Trains of C fibre volleys were ineffective.3. In the preparation in vivo, conduction from the hypogastric nerve to the vas deferens nerve was unidirectional and abolished by hexamethonium. After the administration of hexamethonium, the contraction produced by stimulation of the vas deferens nerve was unaffected.4. The close arterial injection of acetylcholine (ACh) into the pelvic viscera caused centrifugal activity in the motor fibres of the vas deferens nerve, but no impulses were detected in the hypogastric nerve.5. Ganglion cells are present in the last 2 cm of the hypogastric nerve.6. It is concluded that there is a ganglionic relay between the hypogastric and vas deferens nerves.

Acetylcholine↗

Sympathetic inhibition of the urinary bladder and of pelvic ganglionic transmission in the cat.

1. Electrophysiological techniques were utilized to study the mechanisms underlying adrenergic inhibition in the urinary bladder of the cat.2. It has been shown that catecholamines administered by close intraarterial injection or released endogenously by electrical stimulation of the hypogastric nerves elicit two distinct inhibitory responses in the bladder: (1) a direct depression of the vesical smooth muscle and (2) a depression of transmission in vesical parasympathetic ganglia.3. Pharmacological studies revealed that the inhibitory mechanisms were mediated via different adrenergic receptors: beta-receptors on the smooth muscle and alpha-receptors in the parasympathetic ganglia.4. We have been unable, however, to demonstrate that either of these mechanisms is activated by naturally occurring sympathetic firing.

Aminobutyrates↗

Metabolic and ionic requirements for the axoplasmic transport of dopamine beta-hydroxylase.

1. Metabolic and ionic requirements for the intra-axonal transport of dopamine beta-hydroxylase (DBH) were investigated in the cat hypogastric nerve-inferior mesenteric ganglion preparation in vitro by monitoring the enzyme accumulation above a crush, 2-2.5 cm distal to the ganglion.2. DBH accumulation in the proximal segment immediately above the crush increased linearly up to 6 h, during incubation in normal Krebs solution at 37 degrees C. The rate of transport of the enzyme was about 4 mm/h.3. Removal of the ganglion, electrical stimulation or reserpine pretreatment (1-6 days before the experiment) did not modify the rate of DBH accumulation.4. Anoxia and glucose deprivation, singly, did not affect accumulation of DBH; however, the combined treatment of anoxia plus glucose deprivation, or dinitrophenol plus glucose deprivation, very markedly interfered with accumulation.5. Removal of sodium or potassium from Krebs solution markedly inhibited the transport of DBH. Preincubation of the nerve in a high-calcium Krebs solution at 4 degrees C, and then reincubation at 37 degrees C, prevented the enzyme accumulation.6. N-ethylmaleimide, ouabain and oligomycin markedly inhibited the transport of DBH.7. The results suggest that transport of DBH, and therefore of noradrenaline storage vesicles, within the hypogastic nerve is dependent on metabolic energy derived from either glycolysis or oxidative phosphorylation. It is also suggested that the sodium-potassium-activated ATPase may play an important role in the intra-axonal transport of storage vesicles.

Adenosine Triphosphatases↗

Two populations or granular vesicles in constricted post-ganglionic sympathetic nerves.

1. A comparative study has been made of the effects of different fixatives on the ultrastructural appearance of granular vesicles accumulated against a constriction in cat hypogastric nerves. 2. After fixation in either osmium tetroxide or in glutaraldehyde followed by post-osmication accumulations of large granular vesicles (65-90 nm in diameter) were observed in profiles of swollen axons. 3. Fixation in acrolein and sodium dichromate revealed a second population of small granular vesicles (mostly 30-50 nm in diameter) in addition to the large vesicles seen after the other fixatives. 4. In reserpinized cats the small granular vesicles were absent. The large granular vesicles were much less numerous, irrespective of the fixative used. 5. It is suggested that either the small vesicles occur in normal noradrenergic axons but are only revealed by catecholamine-sensitive fixatives or they occur as a result of constricting the nerve trunk. Evidence for and against these possibilities is discussed.

Acrolein↗

A correlation between the effects of anti-mitotic drugs on microtubule assembly in vitro and the inhibition of axonal transport in noradrenergic neurones.

1. Podophyllotoxin, colchicine and griseofulvin inhibit the intra-axonal movement of noradrenaline storage vesicles in cat hypogastric nerve/inferior mesenteric ganglion preparations maintained in vitro, cause the disappearance of axonal microtubules and inhibit the assembly of microtubules from tubulin in vitro. The order of potency of the three effects is podophyllotoxin greater than colchicine greater than griseofulvin. 2. Lumicolchicine is without effect on the three parameters and does not interfere with the binding of tritiated colchicine to tubulin. 3. Podophyllotoxin causes a more rapid loss of microtubules from axons than the same concentration of colchicine. 4. The experiments provide strong evidence that microtubules are components of the system responsible for the intra-axonal migration of noradrenaline storage vesicles.

Animals↗

Mechanosensitive afferent units in the hypogastric nerve of the cat.

1. Recordings have been made from ninety single afferent units in the hypogastric and lumbar splanchnic nerves of the cat. 2. The majority of units examined had properties similar to those previously described in the splanchnic nerve: they were slowly adapting mechanoreceptors with one to six punctate mechanosensitive sites distributed mainly along blood vessels as they approached the viscera in peritoneal ligaments; they had a wide range of mechanical thresholds and conduction velocities in the range 0.5-24 m s-1. 3. Receptive fields were found over the bladder base or its peritoneal ligaments or both, on the uterus or broad ligament or both, on the colon or mesocolon or both, and in association with the ureter, vas deferens, prostate or pelvic fat pads. 4. Discharges from afferent units associated with the bladder were investigated during spontaneous (or reflex) bladder contractions, passive distensions, and tetanic contractions induced by electrical stimulation of the sacral spinal cord. The mean spike rates of the adapted (tonic) discharges, observed during distensions and induced tetani, differed over part of the range of intravesical pressures examined. Their behaviour is discussed in relation to the concept of 'in series' tension receptors within the bladder wall.

Action Potentials↗