Conduction blockade in myelinated fibers by gaseous and volatile substances.

The minimum ambient partial pressure required to reversibly disrupt conducted responses in myelinated nerve fibers (Pblock) was determined for 11 gases and chloroform. For all but one substance, Pblock was inversely proportional to their nonaqueous solubility; large-diameter fibers were less vulnerable than fibers of small diameter. No "anesthetic" effect was displayed by SF6. At the Pblock for three of the agents, the time for completion of their anesthetic action (tb) was proportional to their lipid-to-aqueous solubility ratio. When the ratio was large, tb was longer than when the ratio was small; blockade became complete after the partial pressure of the agent in the lipid or nonaqueous phase of the axon membrane became equal to Pblock. The access of these substances to an nonaqueous site was neither pH nor frequency dependent, but in the case of SF6 access did appear to be limited by its molal volume.

Blockade and recovery of cholinergic transmission in rats treated with hemicholinium 3.

Nerve-induced responses of parotid gland and gastrocnemius muscle were reduced by HC-3 (1 mg kg-1) in proportion to the number of stimuli. Contractions by somatic muscle at 100 Hz were abolished after 6.0 X 10(3) stimuli while 14 X 10(3) were applied at 20 Hz before secretion was blocked. As stimulus rate was decreased, blockade of secretion resulted from fewer stimuli but no difference in ACh content was found between stimulated and unstimulated glands. When stimuli were withheld for 1.5 h transmission recovered temporarily; initial secretory flow rate was only 50% of that in untreated controls when stimulation resumed. In both organs, the time during which responses were sustained, however, was much shorter than when the preparations were stimulated initially. After choline, recovery of transmission was dose-dependent: 150 mg kg-1 were required to restore responsiveness to the muscle and the gland comparable to that in HC-3-treated rats stimulated for the first time. Resting recovery, when stimuli are withheld, probably depends upon stored transmitter becoming mobilized rather than on de novo transmitter synthesis because the endogenous choline in plasma is only 1/1000 of that following exogenous choline.

Micturition in naive and morphine-dependent rats.

Voiding responses were recorded in conscious water-loaded rats. Morphine sulphate (5 mg kg-1) elevated the volume threshold for micturition (MV); the group mean MV of 16 rats after morphine was 40% larger than control. Micturition was nevertheless complete since no urine remained in the bladder afterwards. The implantation of 2 or 4 morphine-base pellets (150 or 300 mg morphine) elevated for 12 days the MV in water-loaded rats. On the 3rd to the 10th day following implantation the group mean was approximately twice that of untreated controls. After micturition was over no residual urine was found in the bladder. Within 3 days the rats became tolerant to the antinociceptive action of the morphine-base pellets but little apparent tolerance developed to their action on micturition. On the 1st day after the pellets were removed, the mean MV was reduced. When withdrawal was precipitated by the administration of naloxone the MV was often too small to measure. This component of a withdrawal syndrome could be elicited in the rats throughout the 12 days of morphine pellet implantation. The administration of 20 mg kg-1 morphine sulphate to anaesthetized rats did not decrease the contractions of the urinary bladder to repetitive stimulation of its motor nerves at 1 and 20 Hz.

Potentiation of nerve-induced bladder contractions after calcium channel blockade.

The potentiation of nerve-induced bladder contractions (NIC) by tetraethylammonium chloride (TEA), K+, or carbachol could result from a greater Ca2+ entry through Ca2+ channels in the muscle or from a greater release of transmitter by nerve terminals. Contractions of equal magnitude by the rat urinary bladder in vitro were initiated by carbachol, K+, or transmural stimulation of urinary bladder motor nerves at 1 Hz. Contractions elicited by K+ or carbachol were drastically reduced by verapamil (0.5 microM), but NICs were unaffected. Thus the role of Ca2+ channels in NICs seems uncertain. NICs are potentiated approximately 50% by K+ (15 mM), carbachol (0.5 microM), or 4-aminopyridine (0.2 mM) and over twofold by TEA (5 mM). Although verapamil (1-5 microM) reduced NICs in a dose-dependent relation, potentiation by each compound was the same. Thus Ca2+ channels probably play no role in potentiation. The resistance of the bladder to distention reflects its viscoelasticity and is Ca2+ sensitive. Because viscoelasticity was decreased by verapamil coincident with the reduction in NICs, both may result from lowered intracellular Ca2+ (Cai2+). However, because the potentiating compounds failed to restore bladder viscoelasticity, they probably did not elevate Cai2+. Therefore, in verapamil-treated preparations potentiation is most probably caused by an enhancement of transmitter release.

Impairment and restoration of rat urinary bladder responsiveness following distension.

Micturition and bladder responsiveness in vitro were impaired in rats fed isotonic sucrose, afflicted with diabetes mellitus or diabetes insipidus. Their urinary output which was seven times control, initiated micturition responses at volumes three times control. Nerve-induced contractions by bladders from these rats developed substantially less pressure than control. Contractions elicited at 1 Hz by control and impaired bladders were potentiated equally by tetraethylammonium chloride (TEA) (5 mM) or by carbachol (2 X 10(-7) M). Contractions elicited at 20 Hz by normal bladders were not potentiated, those by impaired bladders were. TEA, by increasing transmitter release, and carbachol, by a postjunctional action, substantially reversed bladder dysfunction. Because control and impaired bladders were equally enhanced by TEA, prejunctional and contractile element (CE) activity at 1 Hz were probably unaffected by distension. However, postjunctional sensitivity was probably reduced. Impaired bladders, more compliant than controls, became less compliant after carbachol without elevating resting pressure. Whereas the action of carbachol to enhance bladder responsiveness did not involve tension development, there may have been cholinoceptor facilitation and shortening of CE.

Atropine and micturition responses by rats with intact and partially innervated bladder.

1 Micturition responses by a group of 17 rats wee recorded during a water diuresis. During a 2 h period, uniform volumes of urine were passes at regular intervals; the mean of the voiding responses by each animal was consistent from one water loading period to another. Residual urine volumes were physiologically insignificant. 2 Atropine treatment did not compromise seriously micturition by water-loaded rats. Treated animals micturated more frequently; the mean volume was 68% of control. The residual urine volume was equal to that of controls. 3 Several week after the surgical removal of half the motor innervation of the bladder, there was no significant effect on micturition. Mean voiding volumes were not different from those of controls; residual urine volumes were the same as before denervation. 4 After half the innervation of the bladder had been destroyed. The effect of atropine on micturition was enhanced. Volumes passes were 50% of control; large residual volumes remained when micturition was over. Only in this group could bladder distension be found. 5 It is concluded that functional responses of the rat urinary bladder are not only resistant to atropine but also to the sizeable reduction in the number of neuroeffector units in the bladder itself. The functional reserve of the rat bladder musculature is remarkably high when assessed by its ability to empty adequately.

Ventilatory depression in naive and tolerant rats in relation to plasma morphine concentration.

1 The disappearance of morphine from specially formulated pellets containing 75 mg morphine base was measured for 10 days after they were implanted into adult rats; the morphine content decreased at a rate of 5 mg pellet daily.2 From the 2nd to the 6th day of implantation the plasma morphine concentration increased but by the 10th day had declined to only one half the concentration found on day 6.3 Six and 24 h after the pellets were removed from 6 day implanted animals the plasma concentration of morphine amounted to only one quarter to one sixth of the amount in the plasma, respectively, of animals with pellets intact.4 The pulmonary minute volume of naive and implanted rats was depressed by morphine in proportion to the plasma morphine concentration. Less depression was produced by intravenous morphine in the implanted rats than in the naive animals; the greater morphine tolerance displayed by the implanted animals could be shown by the third day of implantation and appeared to be maintained to the 10th day.5 The pulmonary minute volume of implanted rats on the 6th day was much less than the pulmonary minute volume of naive rats. Six and 24 h after the pellets were removed the pulmonary minute volume increased as the plasma morphine concentration decreased.6 The effects on the pulmonary minute volume produced by the slow release of morphine from the implanted pellets was not changed by the development of tolerance while the effects of morphine produced by rapid injection were diminished by the development of tolerance; the different effects of morphine are accordingly linked to the mode of administration.7 We conclude that the action of morphine on the pulmonary minute volume in tolerant rats following rapid injection is fundamentally different from its action following its slow release from implanted pellets, possibly due to differences in access to an undefined neuronal site.

Morphine depression and tolerance of nerve-induced parotid secretion.

1 The nerve-induced secretion produced by the rat parotid gland is proportional to the frequency of stimulation. Morphine decreased the flow rate during stimulation at 2.5 and 5 Hz, but not at 20 Hz. This frequency-dependent action of morphine and was partially reversed by naloxone. 2 The secretion produced by the rat parotid gland during an intravenous infusion of acetylcholine was not diminished by morphine. Therefore, the action of morphine on nerve-induced secretion is most probably on the motor nerve terminals, which release acetylcholine. 3 Animals that had been implanted with morphine base pellets tolerated 4 times as much morphine as controls; after 6 days the minute ventilation was less depressed by graded doses of morphine than non-implanted controls. 4 Nerve-induced secretion in morphine-implanted animals was less depressed by morphine than control animals 6 and 24 h after the pellets were removed. The flow rates in the 6 h group treated with morphine were greater after naloxone than control (precipitated withdrawal) but at 24 h when withdrawal symptoms were no longer evident, naloxone produced only a slight reversal.

Potentiation of nerve-induced bladder responses by tetraethylammonium in relation to junctional and extrajunctional muscarinic receptors.

1 The single stimulus responses elicited in the rat urinary bladder were enhanced up to 3 fold by tetraethylammonium chloride (TEA) in the range 2.5 to 10 mM. Responses elicited by repetitive stimulation at 20 Hz were potentiated much less by 2.5 and 5 mM TEA; 10 mM TEA depressed the responses to less than control levels.2 The responses elicited in preparations treated with tetrodotoxin or botulinum toxin during the 20 Hz stimulus trains were one third to one half of control, while single stimulus responses were abolished altogether. After 10 mM TEA the response to the 20 Hz stimulus trains were near control but the single stimulus responses were not restored at all.3 The single stimulus response of control and of TEA-treated bladder preparations were unaffected by atropine (2 x 10(-6) M) but responses elicited by a 20 Hz stimulus train were reduced more than 40% by atropine. After 5 mM TEA the responses to the 20 Hz stimulus trains that had been partially blocked by atropine were immediately restored to near control levels.4 The responses of bladders to carbachol were dose-dependent in the range 10(-6) to 10(-5) M and were atropine-sensitive. After 5 mM TEA the means of the responses produced by graded doses of carbachol were less than control; muscarinic receptors that were blocked by TEA are probably also atropine-sensitive.5 It is suggested that muscarinic receptors in the rat urinary bladder may be divided into: (1) junctional receptors that are resistant to atropine and may be indirectly affected by TEA and (2) extrajunctional receptors that are blocked by atropine and may be directly affected by TEA.

Motor innervation of the smooth muscle of the rat seminal vesicle.

Frequency-related isovolumetric contractions of the rat seminal vesicle elicited with transmural electrical stimulation were blocked by tetrodotoxin but unaffected by hexamethonium. The postganglionic motor innervation of the rat seminal vesicle is purely excitatory and contains both an adrenergic and a cholinergic component which are excited simultaneously during transmural stimulation. Contractions elicited by adrenergic nerve stimulation were mediated by norepinephrine acting via alpha adrenoceptors, i.e., 1) responses of untreated vesicles to transmural stimulation and to exogenous norepinephrine were antagonized by phentolamine and potentiated by cocaine, 2) pretreatment of animals with reserpine or 6-hydroxydopamine produced a marked depletion of tissue norepinephrine concentration and reduced the responses to transmural stimulation to a level which resembled that of untreated organs in the presence of phentolamine, 3) the residual responses of vesicles from pretreated rats were not modified by phentolamine or cocaine, and 4) responses to tyramine in untreated organs were antagonized by phentolamine but not by cocaine and were observed in organs from reserpine-pretreated rats only after repletion with exogenous norepinephrine. Responses elicited by cholinergic nerve stimulation were mediated by acetylcholine through muscarinic receptors, i.e., 1) responses of untreated vesicles to transmural stimulation and to exogenous acetylcholine were antagonized by atropine, 2) the residual responses to transmural stimulation of vesicles from animals pretreated with reserpine of 6-hydroxydopamine were nearly abolished by atropine and 3) physostigmine potentiated and prolonged the responses of organs from untreated and reserpine-pretreatd animals to transmural stimulation; these effects of physostigmine were abolished by atropine.

Atropine resistance and muscarinic receptors in the rat urinary bladder.

The action of an anticholinesterase and an antimuscarinic drug upon nerve-induced contractions of the rat urinary bladder were examined during transmural stimulation at 20 Hz. Responses were graded in magnitude by limiting the duration of the stimulus trains. 2 Responses of low magnitude produced by short stimulus trains were unchanged by atropine; however, maximal responses resulting from long stimulus trains were diminished in magnitude and shortened in duration. 3 Responses of small magnitude elicited by short stimulus trains involve muscarinic receptors in close proximity to the neuroeffector junction and are resistant to atropine. 4 Maximal responses elicited by long stimulus trains involve 'junctional' muscarinic receptors as well as receptors located at the periphery of the junction; the 'extrajunctional' receptors are blocked by atropine. 5 Responses of low magnitude produced by short stimulus trains were unaffected by echothiophate; however, the duration of maximal responses resulting from the long stimulus trains was extended. 6 The inhibition of cholinesterase did not increase the occupation of muscarinic receptors by the transmitter; however, after large quantities of transmitter were released by the long stimulus trains the association between the receptors and acetylcholine was prolonged.

The motor innervation of the rat urinary bladder.

1. The post-ganglionic nerves visible in silver-impregnated sections of a normal rat bladder were absent 14 days after both pelvic ganglia had been ablated. After ablation of one ganglion the distribution of nerve trunks in either side of the organ was unchanged. Post-ganglionic axons from either side appear to distribute bilaterally.2. The acetylcholine (ACh) content of the rat bladder was reduced from control by 50% after the post-ganglionic nerves from one ganglion had degenerated. However, the ACh content in the two halves of the bladder sectioned along the mid line was the same after one nerve had degenerated.3. Motor responses of bladder preparations elicited during stimulation of both pelvic nerves were compared with responses elicited when each nerve was stimulated separately. In three-quarters of the animals the sum of the individual responses exceeded the response to combined nerve stimulation by no more than 20%. The functional overlap between the two groups of motor nerves to the bladder was therefore no greater than this amount in most animals.4. Motor responses of normally innervated bladder preparations elicited in vitro by transmural stimulation were compared with responses elicited after the post-ganglionic innervation from one side had degenerated. The mean response of bladders with half their innervation was 50-65% of the mean response of bladders normally innervated. The functional overlap by the two groups of nerves was found to be no greater than 15%.

Motor responses of the urinary bladder and skeletal muscle in botulinum intoxicated rats.

1. Type A or type D botulinum toxin administered to rats did not produce a generalized paralysis of skeletal muscles at the time of ventilatory arrest. However, if survival was extended by artificial ventilation complete blockade of neuromuscular transmission developed 6.5 hr after 100 MLD of type D and 5 hr after 1000 MLD of type A toxin. The onset of paralysis of a muscle was shortened by repetitive stimulation of the motor nerves.2. There was no consistent blockade of parasympathetically innervated viscera in animals dying after type A toxin. Animals given type D toxin displayed mydriasis and urinary retention before death.3. Motor responses to electrical stimulation, of bladder preparations in vitro were more vulnerable to type D than to type A toxin. When somatic paralysis was complete in animals treated with type A or type D toxin the excised bladders produced pressure elevations 45 and 25%, respectively, of control preparations.4. During electrical stimulation of bladder preparations nearly paralysed by either toxin, the ACh release was significantly diminished from controls. In the rat bladder botulinum toxin specifically disrupted the liberation of mediator from post-ganglionic nerve endings.