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Vaginocervical mechanostimulation (VS) was shown previously to release oxytocin within the spinal cord and to induce pupillary dilatation. In the present study, (a) injection of oxytocin directly to the spinal cord (10 or 25 microg intrathecally [i.t.] in 5 microl saline) induced pupillary dilatation when observed 1 min after the end of the injection and (b) injection of an oxytocin receptor antagonist ([d(CH2)5-Tyr (Me)2-Orn8]-Vasotocin [OTA]; 25 microg i.t. in 5 microl saline) significantly attenuated the pupillary dilatation response to VS, when VS was applied 3 min after the end of the injection. Since activation of autonomic sympathetic preganglionic neurons in the thoracic spinal cord produces pupillary dilatation, we propose that oxytocin is a central nervous system neurotransmitter that stimulates these neurons directly, or perhaps indirectly, and thus is a mediator of VS-produced pupillary dilatation.

On the basis of evidence that 14C-2-deoxyglucose (2-DG) autoradiography indicates activity at axonal terminals, whereas c-fos immunocytochemistry indicates activity of neuronal cell bodies, we combined these techniques in adjacent histological brain sections to assess excitatory and disinhibitory synaptic relations in selected sites in female rats in which maternal behavior was elicited by natural parturition, sensitization (7- to 10-day cohabitation with foster pups), or hysterectomy. All individuals in these three groups expressed maternal behavior immediately before 2-DG injection. Controls were non-maternal virgins. Parturient and Hysterectomized groups: elevation (compared with controls) in both 2-DG and c-fos activity in medial preoptic area (MPOA) indicated an increase in its input and output activity, i.e., an excitatory interaction; the MPOA was previously shown to be critical for maternal behavior. Sensitized group: a decrease in 2-DG activity of vomeronasal nuclei (bed nucleus of the accessory olfactory tract, BAOT, and medial amygdala, ME, replicating our previous study) and an elevation in c-fos activity, jointly indicate disinhibition of these nuclei, that were previously shown to modulate pup-chemostimulation-induced sensitization. All other sites showed evidence of excitatory input-output relationships (i.e., joint increase in both 2-DG and c-fos activity), e.g., bed nucleus of the stria terminalis (BNST), lateral habenula (LHAB), central gray (CG), thalamus (THAL), septum (SEPT), and ventral tegmental area (VTA). The present study demonstrates the feasibility of measuring 2-DG and c-fos activity jointly in adjacent sections of the same brain, thereby providing evidence to distinguish between localized excitation and disinhibition.

The present study was designed to ascertain whether the vagus nerves convey functional sensory activity from the reproductive tract in rats. Previously, vaginocervical mechanostimulation (VS) was shown to increase pupil diameter (PD) and the threshold of vocalization to tail shock (Voc-T). These responses were attenuated but not abolished by combined bilateral transection of the 'genito-spinal' nerves (i.e. pelvic, hypogastric and pudendal). Subsequent bilateral vagotomy further reduced or abolished the residual responses. In the present study, spinal cord transection above the known level of entry of the genito-spinal nerves was combined with bilateral vagotomy. In ovariectomized rats, after spinal cord transection at thoracic 7 (T7X), lumbar 5 (L5X) levels, or sham surgery (Sh), responses to VS were measured, the vagus nerves were then transected bilaterally, and responses to VS were again measured. VS significantly increased Voc-T and PD after sham procedure or spinal cord transection at either level. Subsequent bilateral vagotomy abolished the VS-induced increase in PD in the T7X group. Due to low survival rate, the effect of vagotomy on Voc-T could not be determined. Consequently, we performed a second experiment. In non-ovariectomized rats, VS significantly increased PD but reduced Voc-T in the T7X group compared to the Sh group, and subsequent bilateral vagotomy abolished both responses. These findings provide evidence that, in the rat, the vagus nerves provide a functional sensory pathway from the reproductive tract directly to the medulla oblongata of the brain, bypassing the spinal cord.

Levels of [14C]2-deoxyglucose (2-DG), measured autoradiographically, in the medial preoptic area (MPOA), were higher during natural parturition with concurrent maternal behavior than in non-pregnant non-maternal controls, whereas levels in the vomeronasal system were lower in virgin rats made maternal by cohabitation with young than in control and parturient rats. Previous studies have shown that lesions of MPOA disrupt maternal behavior, whereas lesions of vomeronasal structures stimulate it, and that an increase in 2-DG levels is indicative of an increase in firing activity in neuron terminals. Consequently, the present findings suggest that maternal behavior can be induced by: (a) an increase in parturition-generated sensory stimulatory input to the MPOA in response to mechanostimulation of the birth canal, and (b) a separate chemosensory vomeronasal pathway whose activity is reduced cohabitation with young, thereby disinhibiting maternal behavior.

In urethane-anesthetized rats, the compound action potential of the pelvic nerve was found to consist of three different waves, two in the A delta fiber and one in the C-fiber range of conduction velocity. Electrical stimulation of the pelvic nerve produced a complete inhibition of the withdrawal response to noxious foot pinch or foot compression. The electromyographic (EMG) activity of the contralateral posterior biceps muscle was used to record the withdrawal response. The withdrawal response inhibition was related to the duration and the frequency of electrical stimulation. Low (5-10 Hz) and high (100-300 Hz) frequencies were ineffective in inhibiting the withdrawal response, whereas intermediate frequencies (20-80 Hz) produced a complete inhibition of the withdrawal response. Short (300 ms) trains of stimulation inhibited the withdrawal response only during the stimulation period. Longer trains of stimulation (500 ms-10 s) produced long-lasting inhibition of the response to noxious stimulation. The inhibition persisted for up to 20 s after the end of electrical stimulation of the pelvic nerve. A delta fiber stimulation was adequate to inhibit the withdrawal response in most (15 out of 17) of the animals. However, A delta plus C-fiber stimulation was needed to inhibit the response to noxious stimulation in two animals. In addition to inhibiting the response to noxious stimulation, pelvic nerve electrical stimulation reflexively activated abdominal muscles. On the basis of present findings using electrical stimulation, it can be suggested that, in the rat, A delta and C-visceral afferents of the pelvic nerve mediate the analgesic effect of vaginocervical probing pelvic and A delta afferents the contraction of abdominal muscles in the fetus-expulsion reflex.

To assess possible changes in nociception during copulation in estrous rats, electric shocks that were 20% suprathreshold for eliciting vocalization in response to tail shock (STS), were applied to the tail before the initiation of copulation and, thereafter, coincident with the onset of mounting bouts by the male (Experiment 1). Females vocalized significantly less during non-intromittive mounts (M; P < 0.001), intromissions (I; P < 0.001), and ejaculation (E; P < 0.01) than before the initiation of copulation. In order to assess the importance of vaginal stimulation (VS) by penile insertion during mating, in Experiment 2 30% STS were applied 300-400 ms after the initiation of mounting to ensure that the stimuli fell within the period of penile insertion occurring during I and E. M failed to significantly inhibit vocalizations to 30% STS. By contrast, both I and E markedly inhibited vocalizations in response to STS. This effect was transitory since subjects (Ss) vocalized to nearly all 30% STS when delivered 15 s after I or E. Copulatory analgesia (CA) was abolished by the bilateral transection of the pelvic and hypogastric nerves but not by the transection of the pudendal nerve (Experiment 3). The magnitude of CA was calibrated by determining the doses of morphine sulfate (MS) required to produce similar decrements in vocalization to STS. The analgesic effects of I and E were equivalent to more than 10 mg/kg and 15 mg/kg, respectively, of MS (Experiment 4). Pelvic-hypogastric neurectomy, but not pudendal neurectomy, also significantly reduced the effect of VS on facilitating lordosis, inducing immobilization and hind leg extension, and blocking the withdrawal reflex to foot pinch (Experiment 5).(ABSTRACT TRUNCATED AT 250 WORDS)

The present study was designed to elucidate the mechanism in the spinal cord by which vaginocervical stimulation (VS) attenuates responses to noxious stimulation. This was accomplished by testing the hypothesis that VS reduces noxious stimulation-induced release of substance P at the level of the spinal cord. Noxious foot shock significantly increased the release of substance P (measured using radioimmunoassay) into superfusates of the lumbosacral spinal cord region in urethane-anesthetized rats. VS applied concurrently with foot shock significantly attenuated the release of substance P compared to the foot shock-only condition. In addition, substance P levels were significantly lower after the VS-only condition than after the no stimulation or foot shock-only conditions. These findings indicate that VS may produce analgesia, at least in part, by suppressing the release of substance P within the spinal cord.

Based on pharmacological evidence that inhibitory amino acids mediate vaginocervical mechano-stimulation produced analgesia (VSPA), we hypothesized that inhibitory amino acids would be released endogenously in the spinal cord in response to vaginocervical mechano-stimulation (VS). This hypothesis was tested by HPLC analysis of the amino acid content of 5-min superfusates of the spinal cord before, during and after VS (400 g force applied against the cervix) in urethane-anesthetized rats. Utilizing an in vivo push-pull superfusion method, artificial cerebrospinal fluid was continuously superfused over the spinal cord through the intrathecal space surrounding the sacral-lower thoracic region. In addition, concentrations of amino acids in the superfusate were measured in response to KCl stimulation (increasing the superfusion medium from 3.4 to 40.0 mM KCl to produce non-specific depolarization), and noxious hind paw mechano-stimulation (pinching the hind paw to produce a sustained flexor response in ipsilateral hind leg). There was a significant increase in the concentration of Gly, Tau, Asp, Glu and Lys in the superfusate in response to VS (n = 8) and to KCl (n = 8), but not to hind paw stimulation (n = 5). Also, GABA concentrations increased in response to KCl, and the concentration of Ala, Ser, Gln, Thr, Arg and Phe increased in response to VS, however, GABA levels were sometimes below the limits of detection. In contrast, there was no significant change in any amino acid concentration in response to hind paw pinch stimulation, and VS did not significantly affect the concentrations of Tyr, His, Ile, Leu, Met, Trp or Val. The present findings support our hypothesis that VS releases inhibitory amino acids in the spinal cord. Moreover, other amino acids, including 'excitatory' amino acids, are released into the superfusate. The profile of amino acid release in response to VS differs from that in response to paw pinch or KCl administration.

The present study describes several muscular reflexes produced by genital stimulation, the nerves that subserve them, and the visceral and postural effects induced by these reflexes. Electrical stimulation of the iliococcygeus (ic) and pubococcygeus (pc) (striated) muscles produced movement of the vaginal orifice and wall, membranous urethra, tail and pelvis. Electrical stimulation of the psoas major (pm) or iliacus (i) (striated) muscles produced movements of the lumbar vertebrae and extension of the ipsilateral hindlimb. Sensory mechanostimulation elicited responses of these muscles as follows: stimulation of the perineal skin, clitoral sheath or distal vagina produced reflex contraction of the ic and pc muscles. Stimulation of the cervix produced reflex contraction of the pm and i muscles and also blocked the above reflex contraction of the ic and pc muscles. Both the cervical stimulation-induced blockage of the ic and pc reflex response, and the cervical stimulation-induced activation of pm and i muscles was prevented by bilateral transection of the viscerocutaneous branch of the pelvic nerve. Based on the above observations, it is proposed that stimulation of the vaginal surface of the cervix resulting from penile intromission and/or seminal plug deposition during mating behavior in the rat may reflexively active pm and i, thereby contributing to the hindleg postural rigidity and lordotic dorsiflexion that are characteristic of the normal mating posture in female rats.

Afferent activity from the reproductive tract activates intrinsic pain attenuating processes. For example, analgesia results from vaginocervical mechano-stimulation in nonpregnant rats and occurs during pregnancy and parturition. In the present study, the effect of uterocervical mechanostimulation on pain thresholds was investigated in order to determine whether direct stimulation of the uterine cervix could play a role in the analgesia of pregnancy. Uterocervical mechanostimulation was applied to nonpregnant rats via a silastic disc implanted in the uterus. The disc abutted against the cervix and was attached to a thread externalized through the vaginal orifice. Application of a force of 150 g, but not 100 g, produced a significant increase in tail flick latency (110.4 +/- 40.6%, P less than 0.03). This effect was abolished by pelvic neurectomy, but was not altered by hypogastric neurectomy. Stimulation of the uterine cervix in combined pelvic and hypogastric neurectomy rats produced a decrease in tail flick latency. These results indicate that the analgesia that occurs during pregnancy and/or parturition may result, at least in part, from the uterocervical mechanostimulation that occurs during this condition.

Bilateral transection of the pelvic and/or hypogastric nerves, which convey afferent activity from the reproductive tract, was performed to ascertain the role of these nerves in the analgesic and motoric effects of vaginocervical mechanostimulation (VS) in rats. Two indices of analgesia were used: tail flick latency to radiant heat (TFL) and vocalization threshold to electrical shock of the tail (Voc-T). Nerve cuts were performed at least one week prior to behavioral testing. Bilateral transection of both the pelvic and hypogastric nerves eliminated the analgesic effects of VS on the TFL and Voc-T tests. Bilateral transection of only the pelvic nerves reduced the number of rats showing maximal VS-induced elevation in TFL, without altering the effect of VS on Voc-T. By contrast, bilateral transection of only the hypogastric nerves attenuated the Voc-T-elevating effect of VS, without reducing the effect of VS on elevating TFL. The effects of VS on producing immobility, hindlimb extension and blockage of hindlimb withdrawal to foot pinch were eliminated by combined bilateral pelvic and hypogastric neurectomy. However, bilateral transection of either nerve alone did not significantly alter the efficacy of VS in producing these effects. These findings indicate that the pelvic and hypogastric nerves contribute to the immobility- and extensor-inducing, and flexor-inhibiting effects of VS, and differentially mediate the analgesia-producing effects of VS.

In the present study, a significant increase in pain threshold (current to elicit vocalization to tail shock) was found 15 and 60 min after injection of dibutyryl cyclic AMP (db cAMP) (30 micrograms) into the lateral ventricle in rats bearing a transplant of fetal adrenal medulla (AM). By contrast, no effect on pain threshold was observed in rats bearing an AM transplant but receiving no db cAMP, or in rats receiving db cAMP but not bearing an AM transplant. In primary cultures of rat fetal chromaffin cells, db cAMP increased the number of neuron-like cells that showed both vasoactive intestinal polypeptide (VIP)- and tyrosine hydroxylase (TH)-like immunoreactivity. These findings indicate that db cAMP exerts a pharmacological modulation of the functional activity (i.e. elevation in pain thresholds) of fetal adrenal AM transplants, and induces phenotypic changes in cultured chromaffin cells with expression of a peptide that elevates pain threshold.

In rats, stimulation of the vaginal cervix with a glass rod reliably produces analgesia, as measured by the tail-flick test. The present studies sought to identify the neural substrates underlying this potent pain inhibition by examining the effects of decerebration, spinalization and bilateral dorsolateral funiculus (DLF) lesions on vaginal stimulation-produced analgesia (VSPA). These studies indicate that the neural circuitry mediating VSPA is contained within the caudal brainstem and spinal cord, since decerebration did not reduce VSPA when compared with sham-operated controls. A significant though markedly reduced level of analgesia was induced in spinalized rats, indicating that VSPA involves both intraspinal and descending pathways. This descending pathway, originating within supraspinal nuclei of the caudal brainstem, projects to the spinal cord via the DLF, since DLF lesions and spinalization produced equivalent reductions in VSPA compared to sham-operated controls. These results, considered in the light of previous electrophysiological and anatomical findings, indicate that the ventral medullary region may be the source of the descending DLF projection mediating VSPA.

The efferent innervation of the pelvic and pudendal nerves was characterized in this study by identifying the muscles activated by electrical stimulation of the nerves distal to the point at which they bifurcate from the L6-S1 trunk. Pelvic nerve electrical stimulation produced EMG-monitored contraction of the ipsilateral ilio- and pubococcygeus muscles, which was abolished by cutting one ('muscular') branch of the bifurcated nerve. (This 'muscular' branch receives proprioceptive input activated by tail displacement, whereas the other, 'viscero-cutaneous' branch receives sensory innervation from the midline perineal region.) Pudendal nerve electrical stimulation produced contraction of the coccygeus, external anal sphincter, and ischiocavernosus muscles. Movements of the orifice and wall of the vagina were directly visualized during electrical stimulation of the two nerves. Intravaginal pressure measured by balloon was increased by pelvic nerve stimulation and decreased by pudendal nerve stimulation. Reflexive contraction of the ilio- en pubococcygeus muscles was produced by mechanostimulation of the perineum, clitoral sheath and distal vagina. This response was abolished by gentle cervical mechanostimulation. One implication of this finding is that passage of the fetuses through the cervix during parturition may relax the ilio- and pubococcygeus muscles, thereby facilitating delivery.

This study confirms previous observations of an increase in pain threshold prepartum followed by a decrease postpartum. Moreover hypogastric nerve transection significantly attenuated the analgesia of pregnancy by a factor of 5 but did not abolish it. Thus, the hypogastric nerve plays a major, but not exclusive, role in the analgesia of pregnancy.

The present studies extend previous findings that probing the vaginal cervix of rats blocks withdrawal reflexes and induces immobilization44. In the present studies, we report that this effect is apparently not due to an action on the final motor pathway, for limb or facial movement induced by electrical stimulation of the pyramidal tract was not suppressed by the probing. In contrast, the sensory response of neurons in the ventrobasal complex of the thalamus to noxious pinch stimulation was markedly attenuated by probing the vaginal cervix. However, the response of these neurons to gentle tactile stimulation was not attenuated, indicating a selective antinociceptive effect of the probing. The antinociceptive effect was not necessarily related to changes in arousal. These findings were supported by behavioral studies in which probing the vaginal cervix blocked vocalization in response to tail shock, and elevated the current threshold for eliciting vocalization in response to tail shock. Furthermore, during the probing, the rats were found to be capable of vocalizing in response to presumably non-noxious (lifting) stimulation, even though their vocalization response to noxious tail shock was suppresed. These studies suggest that probing the vaginal cervix rats exerts an analgesic action.

A fine (approx. 9 c/sec) tremor of the jaw and/or vibrissae was observed in normal rats while they were standing still and not showing gross bodily movement. The tremor was distinctly different in frequency, intensity, and behavioral context, from movements involved in gnawing, tooth chattering, or exploratory sniffing. Individual tremor movements (recorded as EMG) occurred in synchrony with individual bursts of multiunit activity (MUA) recorded in the ventrobasal complex of the thalamus and with individual 'spikes' in the cortical (frontal-occipital) EEG. Single trains of this rhythmical activity often lasted more than a minute. The phase relationships between EMG and MUA differed among individuals, but tended to remain consistent within each individual. Movement artifacts were ruled out since (1) the moments of occurrence of individual tremor movements and MUA bursts were interdigitated rather than simultaneous, and (2) during high amplitude EMG bursts accompanying sni ffing (associated with EEG theta rhythm), tooth chattering, eating or licking, no corresponding activity in the MUA was observed. We also ruled out the possibility that the neural activity was generated by reafference, for (1) during vigorous non-tremor sniffing movements of the vibrissae, or chattering or chewing movements of the jaws, the rhythmical MUA was absent (although the units did discharge if the vibrissae contacted an obstacle or were brushed by the experimenter), (2) rhythmical MUA often continued both during brief pauses in the motor tremor, and in its absence, and (3) injection of Xylocaine s.c. into the face abolished sensory responses of the thalamic units, but the rhythmical MUA persisted. We discuss evidence which suggests that (1) the rhythmical cortical EEG waves are the equivalent in the rat of the alpha (mu) rhythm, and (2) the existence of parallels between alpha-tremor and Parkinsonian tremor in terms of their mechanisms and functions.

Using a whole-nerve recording method, the genitalia of the female rat were found to receive afferent innervation as follows. Pelvic nerve: vagina, cervix, and perineal skin; hypogastric nerve: cervix and proximal three fifths of the uterus; pudendal nerve: skin of perineum, inner thigh, and clitoral sheath. It is probable that the pudendal and pelvic nerves are activated during copulation, and that all 3 nerves are activated during parturition.

Suction ablations of the medial or lateral fornix were performed in order to transect selectively the medial corticohypothalamic tract (mcht) which originates in the anteroventral subiculum and travels in the lateral fornix terminating in the basal hypothalamus. The circadian rhythmicity of plasma adrenal corticosteroid levels was assessed in individual animals 1--2 weeks postoperatively. Ablation of the lateral fornix disrupted the periodicity of corticosteroid secretion which is normally synchronized with the light--dark cycle, whereas medial fornix ablation or neocortical ablation caused no such disruption. Group mean levels of plasma adrenal corticoids were higher in the lateral fornix-ablated animals than in the medial fornix-ablated, neocortically ablated, or intact control animals. These findings suggest that the anteroventral subiculum is important in the regulation of adrenal corticosteroid rhythmicity, and that it exerts an inhibitory influence upon corticosteroid release.