Diminished gastric prokinetic response to ghrelin in a rat model of spinal cord injury.

BACKGROUND: Patients with cervical or high-thoracic spinal cord injury (SCI) often present reduced gastric emptying and early satiety. Ghrelin provokes motility via gastric vagal neurocircuitry and ghrelin receptor agonists offer a therapeutic option for gastroparesis. We have previously shown that experimental high-thoracic injury (T3-SCI) diminishes sensitivity to another gastrointestinal peptide, cholecystokinin. This study tests the hypothesis that T3-SCI impairs the vagally mediated response to ghrelin. METHODS: We investigated ghrelin sensitivity in control and T3-SCI rats at 3-days or 3-weeks after injury utilizing: (i) acute (3-day post-injury) fasting and post-prandial serum levels of ghrelin; (ii) in vivo gastric reflex recording following intravenous or central brainstem ghrelin; and (iii) in vitro whole cell recording of neurons within the dorsal motor nucleus of the vagus (DMV). KEY RESULTS: The 2-day food intake of T3-SCI rats was reduced while fasting serum ghrelin levels were higher than in controls. Intravenous and fourth ventricle ghrelin increased in vivo gastric motility in fasted 3-day control rats but not fasted T3-SCI rats. In vitro recording of DMV neurons from 3-day T3-SCI rats were insensitive to exogenous ghrelin. For each measure, vagal responses returned after 3-weeks. CONCLUSIONS AND INFERENCES: Hypophagia accompanying T3-SCI produces a significant and physiologically appropriate elevation in serum ghrelin levels. However, higher ghrelin levels did not translate into increased gastric motility in the acute stage of T3-SCI. We propose that this may reflect diminished sensitivity of peripheral vagal afferents to ghrelin or a reduction in the responsiveness of medullary gastric vagal neurocircuitry following T3-SCI.

Gastric vagal motoneuron function is maintained following experimental spinal cord injury.

BACKGROUND: Clinical reports indicate that spinal cord injury (SCI) initiates profound gastric dysfunction. Gastric reflexes involve stimulation of sensory vagal fibers, which engage brainstem circuits that modulate efferent output back to the stomach, thereby completing the vago-vagal reflex. Our recent studies in a rodent model of experimental high thoracic (T3-) SCI suggest that reduced vagal afferent sensitivity to gastrointestinal (GI) stimuli may be responsible for diminished gastric function. Nevertheless, derangements in efferent signals from the dorsal motor nucleus of the vagus (DMV) to the stomach may also account for reduced motility. METHODS: We assessed the anatomical, neurophysiological, and functional integrity of gastric-projecting DMV neurons in T3-SCI rats using: (i) retrograde labeling of gastric-projecting DMV neurons; (ii) whole cell recordings from gastric-projecting neurons of the DMV; and, (iii) in vivo measurements of gastric contractions following unilateral microinjection of thyrotropin-releasing hormone (TRH) into the DMV. KEY RESULTS: Immunohistochemical analysis of gastric-projecting DMV neurons demonstrated no difference between control and T3-SCI rats. Whole cell in vitro recordings showed no alteration in DMV membrane properties and the neuronal morphology of these same, neurobiotin-labeled, DMV neurons were unchanged after T3-SCI with regard to cell size and dendritic arborization. Central microinjection of TRH induced a significant facilitation of gastric contractions in both control and T3-SCI rats and there were no significant dose-dependent differences between groups. CONCLUSIONS & INFERENCES: Our data suggest that the acute, 3 day to 1 week post-SCI, dysfunction of vagally mediated gastric reflexes do not include derangements in the efferent DMV motoneurons.

Ghrelin increases vagally mediated gastric activity by central sites of action.

BACKGROUND: Vagally dependent gastric reflexes are mediated through vagal afferent fibers synapsing upon neurons of the nucleus tractus solitarius (NTS) which, in turn modulate the preganglionic parasympathetic dorsal motor nucleus of the vagus (DMV) neurons within the medullary dorsal vagal complex (DVC). The expression and transport of ghrelin receptors has been documented for the afferent vagus nerve, and functional studies have confirmed that vagal pathways are integral to ghrelin-induced stimulation of gastric motility. However, the central actions of ghrelin within the DVC have not been explored fully. METHODS: We assessed the responses to ghrelin in fasted rats using: (i) in vivo measurements of gastric tone and motility following IVth ventricle application or unilateral microinjection of ghrelin into the DVC and (ii) whole cell recordings from gastric-projecting neurons of the DMV. KEY RESULTS: (i) IVth ventricle application or unilateral microinjection of ghrelin into the DVC-elicited contractions of the gastric corpus via excitation of a vagal cholinergic efferent pathway and (ii) ghrelin facilitates excitatory, but not inhibitory, presynaptic transmission to DMV neurons. CONCLUSIONS & INFERENCES: Our data indicate that ghrelin acts centrally by activating excitatory synaptic inputs onto DMV neurons, resulting in increased cholinergic drive by way of vagal motor innervation to the stomach.

A critical re-evaluation of the specificity of action of perivagal capsaicin.

Perivagal application of capsaicin (1% solution) is considered to cause a selective degeneration of vagal afferent C fibres and has been used extensively to examine the site of action of many gastrointestinal (GI) neuropeptides. The actions of both capsaicin and GI neuropeptides may not be restricted to vagal afferent fibres, however, as other non-sensory neurones have displayed sensitivity to capsaicin and brainstem microinjections of these neuropeptides induce GI effects similar to those obtained upon systemic application. The aim of the present study was to test the hypothesis that perivagal capsaicin induces degeneration of vagal efferents controlling GI functions. Experiments were conducted 7-14 days after 30 min unilateral perivagal application of 0.1-1% capsaicin. Immunohistochemical analyses demonstrated that, as following vagotomy, capsaicin induced dendritic degeneration, decreased choline acetyltransferase but increased nitric oxide synthase immunoreactivity in rat dorsal motor nucleus of the vagus (DMV) neurones. Electrophysiological recordings showed a decreased DMV input resistance and excitability due, in part, to the expression of a large conductance calcium-dependent potassium current and the opening of a transient outward potassium window current at resting potential. Furthermore, the number of DMV neurones excited by thyrotrophin-releasing hormone and the gastric motility response to DMV microinjections of TRH were decreased significantly. Our data indicate that perivagal application of capsaicin induced DMV neuronal degeneration and decreased vagal motor responses. Treatment with perivagal capsaicin cannot therefore be considered selective for vagal afferent C fibres and, consequently, care is needed when using perivagal capsaicin to assess the mechanism of action of GI neuropeptides.

Experimental spinal cord injury in rats diminishes vagally-mediated gastric responses to cholecystokinin-8s.

BACKGROUND: We have shown recently that our model of experimental high-thoracic spinal cord injury (T3-SCI) mirrors the gastrointestinal clinical presentation of neurotrauma patients, whereby T3-SCI animals show diminished gastric emptying and dysmotility. In this study we used cholecystokinin as a model peptide to test the hypothesis that the T3-SCI induced gastroparesis is due, in part, to an impaired vagally-mediated response to gastrointestinal peptides. METHODS: We measured the responses to sulfated cholecystokinin (CCK-8s) in control and T3-SCI (3 or 21 days after injury) rats utilizing: (i) c-fos expression in the nucleus tractus solitarius (NTS) following peripherally administered CCK-8s; (ii) in vivo gastric tone and motility following unilateral microinjection of CCK-8s into the dorsal vagal complex (DVC); and (iii) whole cell recordings of glutamatergic synaptic inputs to NTS neurons. KEY RESULTS: Our results show that: (i) medullary c-fos expression in response to peripheral CCK-8s was significantly lower in T3-SCI rats 3 days after the injury, but recovered to control values at 3 weeks post-SCI, (ii) Unilateral microinjection of CCK-8s in the DVC induced a profound gastric relaxation in control animals, but did not induce any response in T3-SCI rats at both 3 and 21 days after SCI, (iii) Perfusion with CCK-8s increased glutamatergic currents in 55% of NTS neurons from control rats, but failed to induce any response in NTS neurons from T3-SCI rats. CONCLUSIONS & INFERENCES: Our data indicate alterations of vagal responses to CCK-8s in T3-SCI rats that may reflect a generalized impairment of gastric vagal neurocircuitry, leading to a reduction of gastric functions after SCI.

Gastric emptying of enterally administered liquid meal in conscious rats and during sustained anaesthesia.

BACKGROUND: Gastric motility studies are frequently conducted with anaesthetized animal models. Some studies on the same animal species have reported differences in vagal control of the stomach that could not be explained solely by slightly different experimental conditions. A possible limitation in the comparison between similar studies relates to the use of different anaesthetic agents. Furthermore, anaesthetic effects may also limit generalizations between mechanistic studies of gastric function and the gastric function of conscious animals. In the present study, we used the [(13)C]-breath test following a liquid mixed-nutrient test meal (Ensure), 1 ml) with the aim to investigate the rate of gastric emptying in animals that were either conscious or anaesthetized with either Inactin or urethane. METHODS: One week after determining the maximum (13)CO(2) concentration, time to peak [(13)C] recovery and gastric half emptying time in control, conscious rats, we repeated the experiment in the same rats anaesthetized with Inactin or urethane. KEY RESULTS: Our data show that Inactin anaesthesia prolonged the time to peak [(13)C] recovery but did not significantly reduce the maximum (13)CO(2) concentration nor delay gastric half emptying time. Conversely, urethane anaesthesia resulted in a significant slowing of all parameters of gastric emptying as measured by the maximum (13)CO(2) concentration, time to peak [(13)C] recovery and half emptying time. CONCLUSIONS & INFERENCES: Our data indicate that Inactin(R) anaesthesia does not significantly affect gastric emptying while urethane anaesthesia profoundly impairs gastric emptying. We suggest that Inactin(R), not urethane, is the more suitable anaesthetic for gastrointestinal research.

Time-course of recovery of gastric emptying and motility in rats with experimental spinal cord injury.

We have shown recently that spinal cord injury (SCI) decreases basal gastric contractions 3 days after injury. In the present study we used the [(13)C]-octanoic acid breath test and gastric strain gauges with the aim to investigate the time-course of recovery from postinjury gastric stasis in rats that underwent experimental SCI at the level of the third thoracic (T3) vertebra. Following verification of the [(13)C]-breath test sensitivity in uninjured rats, we conducted our experiments in rats that underwent T3-spinal contusion injury (T3-CI), T3-spinal transection (T3-TX) or laminectomy (control) surgery at 3 days, 1, 3 or 6 weeks postinjury. Our data show that compared to rats that underwent laminectomy, rats that received SCI showed a significant reduction in the cumulative per cent [(13)C] recovery. Although more marked in T3-TX rats, the delayed gastric emptying in T3-CI and T3-TX rats was comparable in the 3 days to 3 weeks period postinjury. At 6 weeks postinjury, the gastric emptying in T3-CI rats recovered to baseline values. Conversely animals in the T3-TX group still show a significantly reduced gastric emptying. Interestingly, the almost complete functional recovery observed in T3-CI rats using the [(13)C]-breath test was not reflected by analysis of spontaneous gastric contractions after SCI. These data indicate that T3-SCI produces a significant reduction in gastric emptying independent of injury severity (T3-CI vs T3-TX) that persists for at least 3 weeks after injury. However, 6 weeks postinjury T3-CI, but not T3-TX, rats begin to demonstrate functional recovery of gastric emptying.

Gastric dysreflexia after acute experimental spinal cord injury in rats.

Gastric reflexes are mediated mainly by vago-vagal reflex circuits in the caudal medulla. Despite the fact that brainstem vago-vagal circuitry remains intact after spinal cord injury (SCI), patients with SCI at the cervical level most often present gastric stasis with an increased risk of reflux and aspiration of gastric contents. Using a miniature strain gauge sutured to the gastric surface; we tested gastric motility and reflexive gastric relaxation following oesophageal distension (oesophageal-gastric relaxation reflex) in animals 3 days after a severe spinal contusion at either the third or ninth thoracic spinal segment (acute T3- or T9 SCI, respectively). Both basal gastric motility and the oesophageal-gastric relaxation reflex were significantly diminished in animals with T3 SCI. Conversely, both basal gastric motility and the oesophageal-gastric relaxation reflex were not significantly reduced in T9 SCI animals compared to controls. The reduced gastric motility and oesophageal-gastric reflex in T3 SCI rats was not ameliorated by celiac sympathectomy. Our results show that gastric stasis following acute SCI is independent of altered spinal sympathetic input to the stomach caudal to the lesion. Our data suggest that SCI may alter the sensitivity of vagal reflex function, perhaps by interrupting ascending spinosolitary input to brainstem vagal nuclei.

TNF(alpha) modulation of visceral and spinal sensory processing.

The cytokine tumor necrosis factor(alpha) (TNF(alpha)) is associated with a constellation of physiological and behavioral characteristics that follow in response to infection such as fever, fatigue, listlessness, loss of appetite, malaise, and tactile hypersensitivity. These responses are examples of central nervous system (CNS) functions modified by the activated immune system. Our studies have focused on the involvement of TNF(alpha) in CNS control of gastrointestinal function and "visceral malaise". We have demonstrated that TNF(alpha) can elicit gastric stasis in a dose-dependent fashion via its interaction with vago-vagal neurocircuitry in the brainstem. Sensory elements of the vago-vagal reflex circuit (i.e., neurons of the solitary tract [NST] and area postrema [AP]) are activated by exposure to TNF(alpha), while the efferent elements (i.e., dorsal motor neurons of the vagus [DMN]) cause gastroinhibition. Transient exposure to low doses of TNF(alpha) cause potentiated (exaggerated) NST responses to stimulation. Subsequent studies suggest that TNF(alpha) presynaptically modulates the release of glutamate from primary afferents to the NST. Using immunohistochemical studies, we have observed the constitutive expression of the TNFR1 receptor on central vagal afferents and spinal trigeminal afferents in the medulla, as well as on cells and afferent fibers within the dorsal root ganglia and within laminae I and II of the dorsal horn throughout the spinal cord. The constitutive presence of these receptors on these afferents may explain why inflammatory or infectious processes that generate TNF(alpha) can disrupt gastrointestinal functions and cause tactile hypersensitivity. These receptors may also play a critical role in the chronic allodynia and hyper-reflexia observed after spinal cord injury or peripheral nerve damage.

Dissociation of the effects of nucleus raphe obscurus or rostral ventrolateral medulla lesions on eliminatory and sexual reflexes.

Rat preparations were used to investigate long-term changes in external anal sphincter (EAS) contractions and reflexive penile erection following electrolytic lesions of the nucleus raphe obscurus (nRO) or the rostral ventrolateral medulla. EAS contractions were measured electromyographically (EAS EMG) following distention of the EAS with a 5-mm probe. Penile erections were measured using a standard ex copula reflex testing paradigm. At 48 h postlesion, 100% of nRO-lesioned animals displayed reflexive erections and the magnitude of EAS EMG was significantly greater in lesioned animals than in sham controls. These results suggested EAS hyperreflexia following destruction of the nRO. By 14 days postlesion, EAS responsiveness in nRO-lesioned animals had returned to levels comparable to nonlesioned animals. No measures of penile erection were affected by nRO lesions. In animals with nucleus gigantocellularis (Gi) and lateral nucleus paragigantocellularis (Gi-lPGi) lesions, no significant changes to EAS reflexes were observed at any time point. At 48 h postoperative, Gi-lPGi lesions significantly reduced the latency to first erection and increased the number of erections elicited relative to controls. Similar facilitation of erection latency was observed at 14 days postlesion, while erection number and flip total were no longer significantly different from controls. These and previous studies suggest that the nRO regulates defecatory reflexes in the rat. These data further suggest that the comingled EAS and bulbospongiosus (BS) motoneurons are controlled by discrete and separate brainstem circuits and that increases in EAS and penile reflexes after spinal cord lesions are mediated by loss of different descending inputs.

Inhibition of pudendal reflexes in spinal rats. Reassessing the role of serotonin.

The effects of serotonin (5-HT) and thyrotropin-releasing hormone (TRH) on penile reflexes were investigated in intact and spinally transected male rats. Doses of intrathecal 5-HT (0.0, 1.13, 2.26, 11.3, 22.6, and 113.0 nmol), in a range previously shown to inhibit pudendal reflexes in anesthetized spinal preparations, prolonged the latency to the first penile erection in awake intact rats. However, these doses also provoked hyperreactivity and vocalization. Doses of intrathecal TRH (100 and 500 pmol) that effectively inhibited penile erection in intact animals were less effective in spinalized animals. Finally, a combination of subthreshold doses of TRH (100 pmol) and 5-HT (4.0 nmol) at a ratio known to affect other TRH/5-HT-mediated circuits significantly extended erection latency in animals with spinal transections. These data suggest that 5-HT and TRH are both involved in the inhibitory circuits regulating penile erection, either through corelease onto the same population of cells or through independent release onto different populations of neurons.

Intrathecal methiothepin and thyrotropin-releasing hormone effects on penile erection.

Intrathecal thyrotropin-releasing hormone (TRH) potently inhibits penile erection at all doses (100, 500, 1000 or 5000 pmol) tested so far. Since the serotonin receptor antagonist methiothepin (MT) inhibits TRH responses in other systems, this study tested the hypothesis that MT-sensitive receptors mediate the effect of TRH on penile erection in rats. When compared to controls, the highest doses of IT TRH (0, 10 or 500 pmol) or MT (5 or 50 nmol) significantly altered penile reflex latency. When coadministered (50 nmol MT/500 pmol TRH), the effect of TRH was reversed, suggesting that the high dose of MT antagonized the inhibitory actions of TRH. The low dose of MT (5 nmol) did not block the 500 pmol TRH inhibition of reflex latency. These data further suggest that MT sensitive receptors are important in (1) mediating normal penile reflexes and (2) mediating the inhibitory response to TRH.

Descending projections from the nucleus raphe obscurus to pudendal motoneurons in the male rat.

Previous physiological and behavioral studies have shown that the nucleus raphe obscurus (nRO) modulates pelvic floor reflex function (Yamanouchi and Kakeyama [1992] Physiol. Behav. 51:575-579; Beattie et al. [1996] Soc. Neurosci. Abstr. 22:722.4; Holmes et al. [1997] Brain Res. 759:197-204). In the present study, small injections of fluorescent tracers were used to investigate direct descending projections from the rostral and caudal portions of the brainstem nRO to retrogradely labeled pudendal motoneurons (MN) in the male rat. The caudal nRO projects into the ventral and lateral funiculi of the spinal cord, with arborizations in the thoracic intermediolateral cell column; in laminae VII, IX, and X of the lumbosacral cord; and in the sacral parasympathetic nucleus (SPN). Many identified external anal sphincter and ischiocavernosus MNs appeared to be in direct apposition with fibers originating from the caudal nRO; and more than half of the bulbospongiosus MNs that were identified appeared to receive such descending input. In addition to the nRO spinal autonomic and pudendal motoneuronal targets, projections were observed to regions of the intermediate gray that contain interneurons organizing the pelvic floor reflexes and to MN pools that are involved in functionally related somatic activities. Finally, several neurons in the lumbar enlargement were labeled retrogradely with FluoroRuby after injections into the nRO and the immediately adjacent reticular formation. Thus, the nRO may be in a position to modulate the coordinated actions of autonomic preganglionic and functionally related skeletal MN activity involved in sexual and eliminative reflex functions.

External anal sphincter hyperreflexia following spinal transection in the rat.

In the present study, long-term and short-term rat preparations were used to develop a model for investigating external anal sphincter (EAS) reflexes in intact and spinal cord-injured (SCI) rats. In this model, EAS distension with an external probe elicits reflex contractions of the EAS in intact, unanesthetized animals. At 2 h after spinal cord transection, none of the lesioned animals displayed EAS EMG activity. In fact, once distended, the EAS was incapable of maintaining closure of the anal orifice. Over a period of 4 days, spinalized animals developed a hyperreflexia of the EAS response. By 48 h, the rectified, integrated EAS EMG was significantly elevated in comparison with nonlesioned controls (EAS hyperreflexia). In addition, the duration of the EAS EMG bursts in response to sphincter distension had significantly increased. At 6 weeks after injury, the EAS was significantly hyperreflexic as measured by EMG burst duration and burst area. As with intact animals, posttransection EAS reflexes were highly anesthesia sensitive. These studies indicate that (1) brief distension of the anal orifice is sufficient to evoke a physiologically relevant reflexive activation of the EAS in the rat, (2) the 2- to 24-h postinjury areflexia observed in these experiments may be a suitable model for the study of spinal shock, and (3) the observed EAS hyperreflexia after chronic SCI may represent the permanent effects of removing descending inhibitory circuits and segmental plasticity, making this reflex an appropriate measure of defecatory dysfunction after spinal cord injury.

Nucleus raphe obscurus (nRO) regulation of anorectal motility in rats.

Previous research has demonstrated that anorectal contractions in the rat are modulated by activation of spinal autonomic circuits. In the present study, anterograde tracing of descending pathways originating from the caudal nucleus raphe obscurus (nRO) revealed that this nucleus projects to cells within the intermediolateral (IML) cell column of the thoracic cord and the sacral parasympathetic nucleus (SPN). These anatomical studies suggested that the nRO may influence the regulation of spinal reflexes of the pelvic floor. In a second set of experiments, acute rat preparations were used to investigate changes in anorectal motility during electrical stimulation of the nRO. Anorectal contractions were measured by a fluid-filled manometer. Electrical stimulation of the nRO significantly reduced spontaneous anorectal activity when compared to baseline contractions recorded for 1 min prior to stimulation. Stimulation sites outside the nRO did not affect anorectal contractions when compared to either (a) the 1-min pre-stimulation baseline for that site or (b) the 1-min stimulation period for sites within the nRO. Stimulation of caudal portions of the nRO were more likely than the rostral nRO to reduce anorectal contractions. Given that the SPN contains preganglionic neurons which may be involved in control of anorectal contractions (mediated via the pelvic nerve), the studies presented here suggest a functional role for nRO regulation of preganglionic motoneurons innervating the distal gut of the rat.

Differential effects of intrathecal thyrotropin-releasing hormone (TRH) on perineal reflexes in male rats.

The effects of thyrotropin-releasing hormone (TRH) on the sexual and defecatory reflexes regulated by pudendal motoneurons were investigated. Intrathecal TRH (10 microliters volume; 0.0, 0.01, 1.0 or 100 microM concentration) at lumbosacral spinal segments (L4-S1) in acute preparations produced a dose-dependent increase in external anal sphincter (EAS), but not bulbospongiosus (BS), electromyographic (EMG) activity. Intraspinal (L6) injection of 100 microM TRH (1 microliter/micropipette), significantly increased EAS EMG activity in acute preparations. Electromyographic activity of the BS muscle was unchanged. All doses of intrathecal TRH (10 microliters volume; 0, 10, 50, 100, or 500 microM concentration) in awake animals significantly reduced the proportion of responders to a penile reflex test. Subsequently, all measures of penile reflexes were significantly reduced. Glans tumescence and defecation bouts before or during penile reflex testing were unaffected by intrathecal TRH as were indices of behavioral and motor hyper-reactivity analogous to that produced by serotonin. These data indicate that pudendal motoneurons, in the dorsomedial nucleus, are differentially regulated by neuropeptides present in the lumbosacral spinal cord.

Thyrotropin-releasing hormone (TRH) and CNS regulation of anorectal motility in the rat.

The effect of thyrotropin-releasing hormone (TRH) upon anorectal motility was investigated in acute male rat preparations. Micromolar doses of TRH were intrathecally (i.t.) infused at the L6 spinal level at a rate of 1 microliter/min over 8 min. TRH infusions in 1.0-1000 microM concentrations elicited biphasic, dose-dependent anorectal contractions as measured by a rectal manometer. The 100 microM dose yielded the most significant increase in contractions over the greatest period of time. Atropine, administered as a pretreatment (100 micrograms s.c.), blocked contractions normally produced by i.t. infusion of TRH (1000 microM). Intravenous infusions of atropine (10 micrograms) through a jugular catheter immediately blocked anorectal contractions produced by i.t. infusion of 100 microM TRH. Sectioning of the hypogastric nerve, which supplies sympathetic innervation to the colon and internal anal sphincter, did not significantly affect contractions induced by 100 microM TRH applied intrathecally. Disruption of the major pelvic ganglion fibers, however, completely abolished the contractions induced by 100 microM TRH, either through the interruption of preganglionic parasympathetic fibers in the pelvic nerve, or by disrupting postganglionic fibers. These findings extend the role of TRH in the regulation of defecatory behaviors.

Physiology and mechanics of rat levator ani muscle: evidence for a sexual function.

The levator ani (LA) of male rodents is a classic model tissue for the study of hormone-muscle interactions, although its functions remain unknown. Recordings during copulation from chronic electromyographic (EMG) electrodes in the LA and bulbospongiosus (BS) revealed that EMG activity in the LA and BS was tightly coordinated. The LA was not active during noncopulatory behaviors, including the 1-min interval surrounding defecation. Electrical stimulation of the LA motor nerves increased penile bulb pressure. Increases in penile bulb pressure following BS nerve stimulation were markedly attenuated after LA denervation and were reduced further by LA removal. Stimulation of the LA nerve yielded insignificant changes in rectal pressure. Perineal motion analysis demonstrated that the LA acts upon the penile bulb and the surrounding BS exclusively. Apparently the rodent LA muscle is an active component in a highly coordinated neuromuscular system augmenting penile erection and, contrary to its name, is most unlikely to participate in alimentary function.

Electromyographic activity of rat ischiocavernosus muscles during copulation after treatment with a GABA-transaminase inhibitor.

The administration of GABA-transaminase inhibitors (GABA-TIs) to male rats reduces the proportion of mounts that result in intromissions. Copulatory pelvic thrusting remains normal, despite the fact that animals treated with GABA-TIs show gross deficiencies in other motor acts. In order to determine whether altered sexual behavior produced by GABA-TI could be due to deficiencies in activity of striated penile muscles, we recorded the electromyographic (EMG) activity of the ischiocavernosus (IC) muscle during copulation in male rats treated with sodium valproate. The duration of IC EMG bursts was reduced by sodium valproate in separate tests that allowed or prevented intromission. There was no effect on EMG amplitude or frequency. It is suggested that insufficient activity of the IC muscles reduces the likelihood of vaginal penetration. The actions of GABA may be localized to hypothalamic or brain stem nuclei with GABAergic projections to the spinal motoneurons controlling the IC muscles, or GABA may act directly on these neurons.

Erectile function and bulbospongiosus EMG activity in estrogen-maintained castrated rats vary with behavioral context.

Electromyographic (EMG) activity in the bulbospongiosus muscles (BS) was recorded to monitor potential castration-induced alterations in muscle activity during copulation and reflexive erections. EMG recordings were made from intact male rats and from castrated rats maintained from 7 to 50 days on estradiol benzoate (300 micrograms/day) or testosterone (200 micrograms/day). Despite a 40-50% postcastration reduction in the weight of the BS and accessory sexual glands in estrogen-treated rats, the pattern of EMG activity during copulation was similar across groups. In estradiol-treated males, the EMG burst frequency during mounts and burst duration during intromissions exceeded the parameters of intact males and of castrated males maintained on testosterone. Between intromissions, and following ejaculatory patterns, estrogen-treated males displayed spontaneous muscle bursts accompanied by visually confirmed erection of the glans penis, but these males quickly lost the capacity for reflexive erections. These data demonstrate that despite castration-induced atrophy of the penile muscles and, presumably, their spinal motor nuclei, the motor output to these muscles is maintained following androgen removal. The capacity for substantial penile erection is retained during copulation long after reflexive erections have diminished.