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Electrophysiological study of neural activity in penile nerve of the rat.

Electrophysiological techniques were used to examine the axonal composition and reflex activity of the penile nerve of the rat. Stimulation of either the pelvic nerve, hypogastric nerve, or sympathetic chain elicited synaptic and axonal volleys in the penile nerve. Synaptic responses were suppressed by nicotinic ganglionic blockade, indicating that they were mediated by cholinergic transmission in peripheral ganglia. Axonal volleys represented in part afferent pathways from receptors in the pelvic muscles, perineum, and anus. Stimulation of the penile or pelvic nerves increased intracavernous pressure. Stimulation of the dorsal nerve of the penis elicited central reflexes (50- to 150-ms latencies) in the penile nerve. Those reflexes were not eliminated by acute or chronic spinalization (T8) but were abolished by transection of preganglionic nerves, indicating an origin in the lumbosacral spinal cord. Thus the penile nerves are composed of a heterogenous population of afferent and efferent axons. Reflex activity elicited in these nerves by stimulation of pathways from the penis is probably involved in the initiation of penile erection.

Animals↗

Sympathetic modulation of cholinergic transmission in cat vesical ganglia is mediated by alpha 1- and alpha 2-adrenoceptors.

Modulation of transmission in cat vesical ganglia by exogenous catecholamines or short periods of hypogastric nerve stimulation (HGS) was examined in vivo. HGS (30 Hz, 30 V, 10- to 20-s train) initially caused inhibition of ganglionic transmission, followed by a period of facilitation. The inhibition could be evoked by lower stimulus intensities than the facilitation. The biphasic effect was mimicked by norepinephrine (2-10 micrograms) injected into the inferior mesenteric artery, whereas phenylephrine (5-10 micrograms) usually caused only facilitation. The administration of specific adrenoceptor antagonists showed that the inhibitory responses were mediated by alpha 2-receptors and the facilitatory responses by alpha 1-receptors. The effects of HGS and norepinephrine were also tested on pelvic nerve-evoked bladder contractions. Both caused a brief reduction in contraction amplitude which, for the HGS effect, was mediated mainly by alpha 2-receptors. Noradrenergic inhibition of bladder contractions was mediated by both alpha 2- and beta-adrenoceptors. These studies have shown that sympathetic modulation of ganglionic transmission in the bladder can be either inhibitory or facilitatory, each mediated by a specific receptor type. These two actions may be important during different stages of bladder storage or voiding.

Adrenergic alpha-Agonists↗

Neural regulation of the vas deferens in the rat: an electrophysiological analysis.

Electrophysiological analysis of the neural control of the vas deferens was performed in urethan-anesthetized rats. Intraluminal distension (0.2 ml/min) or electrical stimulation of hypogastric (threshold 1-5 V, 20 Hz) and pelvic nerves (2-5 V, 20 Hz) produced contractions of the vas deferens. Distension-evoked contractile activity was not abolished by nicotinic ganglionic blockade or ipsilateral hypogastric and pelvic nerve transection. Contractions following hypogastric nerve stimulation were abolished by prazosin, while pelvic nerve-evoked responses were partially blocked by atropine. Hypogastric nerve, pelvic nerve, and sympathetic chain stimulation evoked volleys with latencies of 10-30 ms in vasal nerves. Crude estimates for conduction velocities for these responses (less than 0.5 m/s) corresponded to activation of unmyelinated C-fibers. Stimulation of the dorsal nerve of the penis (DNP) (10-16 V, 10-40 Hz) or administration of 5-methoxy-N,N-dimethyltryptamine, both of which produce seminal emission, elicited reflex discharges in nerves to the vas deferens. Hypogastric nerve but not pelvic nerve transection abolished both spontaneous and evoked (105- to 380-ms latency) reflex activity. These experiments provide insight into the organization of afferents in the DNP and efferents conveyed by autonomic pathways that regulate male reproduction.

Animals↗

Reorganization of the innervation of the vas deferens after sympathetic decentralization.

Reorganization of autonomic efferent pathways to the rat vas deferens was noted after chronic (30 days) sympathetic decentralization produced by hypogastric nerve (HGN) transection. In normal rats, electrical stimulation of the HGN elicited an increase in vasal pressure (VP) bilaterally, whereas pelvic nerve (PN) stimulation did not alter VP. However, after unilateral HGN transection, stimulation of the PN on the transected side but not on the normal side increased VP. The decentralized vas exhibited larger VP responses to stimulation of the contralateral HGN in comparison with the normal vas. After bilateral HGN transection, PN-induced VP responses were elicited at lower stimulus intensities than in rats with unilateral transections. PN-induced VP responses were blocked by hexamethonium and prazosin but were not altered by atropine. Distension of the vas lumen occurred after decentralization. PN-induced VP responses were not detectable in extremely distended vas. These data indicate that, after degeneration of sympathetic preganglionic axons, decentralized adrenergic ganglion cells are reinnervated by parasympathetic or sympathetic preganglionic pathways and that the reinnervation influences vasal function.

Animals↗

Receptor-specific influence of endothelin-1 in the erectile response of the rat.

Specific receptor antagonists were used to examine the role of endothelin-1 (ET-1) in the erectile response of the rat. In these studies, intact rats were cannulated to permit the continuous recording of mean arterial pressure (MAP) and intracavernosal pressure (CCP). Erection was induced by electrical stimulation of the autonomic ganglion, which regulates blood flow to the penis. The animals were subjected to intracavernosal injection with vehicle only (Cont) or with an antagonist to the endothelin-A receptor (ET(A)) or to the endothelin-B receptor (ET(B)). Blockade of the ET(A) or the ET(B) had no effect on the erectile response (CCP/MAP) during maximal ganglionic stimulation. When ET-1 was injected into Cont rats, there was a marked vasoconstriction with a sharp rise in MAP and a decline in CCP as the cavernosal arterioles constricted and limited inflow. The injection of the ET(A) antagonist prevented the vasoconstriction after ET-1 injection into Cont rats, whereas blockade of the ET(B) had no effect on the vasoconstrictive effect to ET-1. Similar results were obtained during submaximal ganglionic stimulation. With minimal levels of ganglionic stimulation, ET-1 injection led to a moderated degree of vasodilation in the presence of the ET(A) antagonist. The ET(B) antagonist failed to alter the CCP response during minimal stimulation, but it did have a marked effect on the MAP response to ET-1 injection. The results of these studies confirm that cavernosal tissue of the rat penis is highly responsive to ET-1. However, the failure of the ET-1 antagonists to affect penile erection in response to ganglionic stimulation reflects a minimal role of ET-1 in the erectile response in the rat.

Animals↗

Transepithelial voltage changes during prostatic secretion in the dog.

To define the nature of Na+, K+, and Cl- secretion by the dog prostate, the electrical potential difference between fluid in the prostatic urethra and venous blood was recorded during secretion provoked in pentobarbital-anesthetized dogs by electrically stimulating the hypogastric nerves or by administering pilocarpine intravenously. The resultant prostatic fluid samples as well as plasma samples taken before and/or after secretion were analyzed for these electrolytes. During secretion provoked by either means the transepithelial electrical potential difference, which was essentially zero at rest, became lumen negative, the maximum change (which was related to gland size) being about 4 mV. The time courses of both the potential change and the secretion of fluid were very similar. The Na+ concentration in both nerve- and pilocarpine-induced prostatic fluid was equal to that in plasma, whereas the K+ and Cl- concentrations were higher than in plasma. It is concluded that Na+ may move passively from plasma into prostatic fluid, perhaps via the paracellular route, but the movement of K+ and Cl- probably involves active transcellular transport.

Animals↗

Responses of thalamic neurons to input from the male genitalia.

There have been relatively few electrophysiological studies, in any species, describing the supraspinal processing of inputs from the male genital tract. The thalamus was the focus of the present study. In 11 urethan-anesthetized male rats, subregions of the thalamus were surveyed for neuronal responses to the search stimulus, bilateral electrical stimulation of the dorsal nerve of the penis (DNP). A total of 133 DNP-responsive neurons were found and further tested for degree of somatovisceral convergence from other peripheral structures. Histological reconstruction of the recording sites revealed that the penile-responsive neurons were distributed among various thalamic subregions. These thalamic subregions included the medial-dorsal nuclei and ventral and lateral thalamic subregions (majority of neurons responsive to both tactile and pinch stimulation of the penis) as well as intralaminar, posterior and reticular subregions (majority responsive to pinch only). Taken together, the data demonstrate the existence of thalamic neurons with inputs from the male genitalia with widespread somatovisceral convergence. These neurons likely contribute to the neural circuitries underlying various aspects of penile sensation associated with reproductive and nociceptive events.

Animals↗

Postnatal maturational changes in rat pelvic autonomic ganglion cells: a mixture of steroid-dependent and -independent effects.

Androgens have potent effects on the maturation and maintenance of a number of neural pathways involved in reproductive behaviors in males. Most studies in this area have focused on central pathways, but androgen receptors are expressed by many peripheral neurons innervating reproductive organs, and previous studies have demonstrated structural and chemical changes in these neurons at puberty and after castration. We have performed the first electrophysiological comparison of pelvic autonomic ganglion neurons in male rats before and after puberty and following pre- or postpubertal castration. Studies were performed in vitro on intact ganglia with hypogastric and pelvic nerves attached to allow synaptic activation of sympathetic or parasympathetic neurons, respectively. Pelvic ganglion neurons underwent many changes in their passive and active membrane properties over the pubertal period, and some of these changes were dependent on exposure to circulating androgens. The most pronounced steroid-dependent effects were on membrane capacitance (soma size) in sympathetic neurons and duration of the action potential afterhyperpolarization in tonic neurons. Our study also showed that rat pelvic ganglion cells and their synaptic inputs were more diverse than previously reported. In conclusion, this study demonstrated that rat pelvic ganglion neurons undergo considerable postnatal changes in their electrophysiological properties. The steroid dependence of some of these changes indicates that circulating androgens may influence reproductive behaviors at many locations within the nervous system not just in the brain and spinal cord.

Age Factors↗

Afferent fibers supplying the uterus in the rat.

1. In the present three-part study electrophysiological techniques were used to characterize responses of afferent fibers in the rat hypogastric nerve to mechanical or chemical stimulation of the uterus, and anatomical techniques were used to identify the spinal segments through which uterine afferent fibers enter the spinal cord. 2. In an in vivo barbiturate-anesthetized preparation, hypogastric afferent fibers responded in a time-locked manner to mechanical stimulation confined to restricted regions of the uterus and adjacent ligament. Receptive fields were most often located on the uterine body, particularly over the cervix. The few located on the uterine horn were usually near regions irritated during preparative surgery. Effective mechanical stimuli (pressure, stretching, squeezing, probing, rarely contractions) were typically greater than 5 g and simultaneously accompanied by transient ischemia around the probe or contracted area. Distension, unless extreme, was not an effective stimulus. Retrospective analysis of the data indicated that fibers may be more sensitive to uterine stimulation when rats are in vaginal estrus/proestrus than in diestrus/metestrus. 3. In an in vitro preparation, hypogastric afferent fibers responded in a dose-dependent fashion to injections into the uterine artery of the algesic chemicals bradykinin, 5-hydroxytryptamine, and KCl. They also responded to high doses of CO2 (in saline) and NaCN, but rarely to lower doses. Nearly all fibers responded to more than one chemical with response characteristics unique to each chemical (e.g., latency, duration, peak rate). 4. Injections of horseradish peroxidase into the uterine body and small portions of the adjacent horns in rats in vaginal estrus consistently labeled a small number of cells in the L1-S1 dorsal root ganglia, with peaks at L2 and L6. Virtually no cells were labeled in rats whose estrous cycle had been disrupted (by inadvertently keeping them in constant light conditions for several weeks). 5. These results indicate that uterine afferent fibers travel to the central nervous system through both the hypogastric (e.g., L1-L4 ganglia) and pelvic (e.g., L5-S1 ganglia) nerves in the rat, and that hypogastric fibers are capable of conveying fairly precise information about temporal and spatial aspects of uterine mechanical and chemical stimulation. The results also encourage future research into the possibility that the responses of these fibers vary as a function of estrous stage or other aspects of the condition of the uterus (e.g., its irritation).(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

Functional differences between afferent fibers in the hypogastric and pelvic nerves innervating female reproductive organs in the rat.

1. The uterus, cervix, and vaginal canal are innervated by afferent fibers in the hypogastric and pelvic nerves. Four studies compared the innervation territory and sensitivity to peripheral stimuli of the two sets of fibers in adult virgin rats. 2. Innervation territory was studied anatomically by injecting different fluorescent dyes into different parts of the reproductive, lower urinary, and lower digestive tracts and examining retrogradely labeled neurons in dorsal root ganglia. It was also studied electrophysiologically in anesthetized rats by summing potentials evoked in branches of the two nerves by electrical stimulation of different parts of the reproductive tract. 3. In both studies sensory innervation of the reproductive tract shifted from the pelvic to the hypogastric nerve (i.e., shifted entry into the spinal cord from the L6-S1 to the T13-L3 dorsal root ganglia, respectively) as the dye or stimulating electrode shifted from the vaginal entrance to the uterine horns, with fibers from both nerves densely innervating the cervix region (i.e., entering the spinal cord through both sets of ganglia). The anatomic results suggested that the regions innervated by fibers in one nerve might also be innervated by a small component of normally quiescent fibers in the other nerve. 4. Response sensitivity was studied electrophysiologically by simultaneously recording multiunit activity in branches of the hypogastric and pelvic nerves in two ways. First, in intact, anesthetized rats, activity was recorded during mechanical stimulation of the reproductive tract (distension of the vagina and uterus, probing the cervix). Second, in an in vitro organ preparation of the uterus and vagina, activity was recorded during chemical stimulation through the uterine artery with bradykinin, serotonin, NaCN, CO2, and KCl. 5. Pelvic nerve fibers were markedly more sensitive than hypogastric nerve fibers to uterine and cervical mechanostimulation. Similarly, pelvic nerve fibers were more likely to respond or responded more vigorously than hypogastric nerve fibers to all chemical stimuli (except KCl). 6. These results provide strong evidence that afferent fibers in the pelvic and hypogastric nerves of nulliparous adult rats subserve different functions in reproduction and sensation. Pelvic nerve fibers seem closely tied to sensory and behavioral processes associated with mating and conception, whereas hypogastric fibers seem closely tied to pregnancy and nociception, with fibers in both nerves serving functions during parturition.

Animals↗

Modulation of Ca2+ currents by various G protein-coupled receptors in sympathetic neurons of male rat pelvic ganglia.

The modulation of voltage-gated calcium (Ca2+) channels by various G protein-coupled receptor pathways was investigated in sympathetic neurons of the male rat major pelvic ganglion (MPG). Standard whole cell patch-clamp recording techniques were used to record Ca2+ currents from acutely dissociated neurons. The activation of muscarinic receptors, which uses a G protein pathway that was not blocked by either pertussis toxin (PTX) or cholera toxin (CTX), inhibited both N-type and L-type Ca2+ channels. The activation of alpha2 noradrenergic receptors with the selective agonist UK14304, which used primarily a PTX-sensitive G protein pathway, inhibited only N-type Ca2+ channels. The activation of vasoactive intestinal polypeptide (VIP) receptors, which used a CTX-sensitive G protein pathway, also inhibited only N-type Ca2+ channels. UK14304 and VIP induced a bell-shaped inhibition of the Ca2+ current with a peak inhibition at around +10 mV and decreasing inhibition at more positive potentials. In contrast, the muscarine-induced Ca2+ current inhibition was not bell shaped and was more prominent at more positive potentials. Furthermore, a large depolarization, which relieved the current inhibition by UK14304 and VIP, did not relieve the inhibition by muscarine. Besides inhibiting the Ca2+ current, UK14304 and VIP also slowed the activation kinetics, an effect not seen with muscarine. Replacing external Ca2+ with Ba2+ and replacing internal ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) with high bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA) completely blocked the inhibitory effect of muscarine. However, the inhibitory effects of both UK14304 and VIP were unaffected under these conditions. Surprisingly, the facilitation of the Ca2+ current was eliminated under these strong calcium-buffering conditions. The activation of protein kinase C (PKC) with phorbol 12-myristate 13-acetate (PMA) increases the amplitude of the Ca2+ current, diminishes facilitation, and reduces the inhibition of this current by UK14304 and VIP. However, PKC activation did not reduce the muscarine-induced Ca2+ current inhibition. In summary, our data suggest that muscarine uses a mechanism different from UK14304 and VIP to modulate the N-type Ca2+ channels in sympathetic neurons of the MPG. Although VIP and UK14304 use different G protein pathways, these two different pathways most likely converge downstream to compete for the same target site on the N-type Ca2+ channels.

Adrenergic alpha-Agonists↗

Afferent fibers of the hypogastric nerves are involved in the facilitating effects of chemical bladder irritation in rats.

To evaluate the role of bladder afferent fibers in the hypogastric nerves (HGN) in modulation of the micturition reflex induced by chemical bladder irritation, voiding behavior, continuous cystometry, and spinal c-fos expression following intravesical acetic acid instillation were investigated in rats with or without HGN transection. Voiding behavior and continuous cystometry were examined in unanesthetized conscious rats. Following chemical bladder irritation, a significant increase in urinary frequency associated with a marked decrease in the voided volume per micturition, was noted in control rats with the intact HGN, but not in HGN-transected rats. Continuous infusion of acetic acid in control rats elicited irritative bladder responses characterized by a marked decrease in the intercontraction interval and a marked increase in maximal vesical pressure, both of which were absent in capsaicin-desensitized rats. HGN transection prevented the decrease in the intercontraction interval but not an increase in maximal vesical pressure following chemical bladder irritation. Compared with saline infusion, acetic acid infusion caused a significant increase in c-fos expression at L(1) and L(6) of the spinal cord, and HGN transection significantly reduced c-fos expression in the dorsal horn of the spinal cord at L(1) but not at L(6). These results suggest that capsaicin-sensitive bladder afferent fibers in the HGN, which travel through the rostral lumbar spinal cord, have a role in urinary frequency caused by chemical bladder irritation.

Afferent Pathways↗