Vaginocervical Stimulation Research Articles

Analgesic Synergism Between AP5 (an NMDA Receptor Antagonist) and Vaginocervical Stimulation in the Rat

Vaginocervical stimulation (VS) releases multiple neurotransmitters into superfusates of the spinal cord; these can stimulate both nociceptive (e.g., glutamate, and glycine acting at the NMDA site), and antinociceptive (e.g., GABA, norepinephrine, 5-HT, and glycine acting at the strychnine-sensitive receptor) systems. Although the balance between these two opposing systems can determine the nature, magnitude, and duration of the response to VS, the characteristic prevailing response to VS is analgesia. We hypothesized that by counteracting the nociceptive component of this system, the magnitude and duration of the response to VS would be augmented. In the present study, the NMDA receptor antagonist AP5 [10 μg injected intrathecally (IT)] significantly increased the magnitude and duration of the analgesia (measured as tail flick latency to radiant heat) produced by VS (200 g force). At several time points the analgesic effect of AP5 combined with VS was greater than the sum of the effects of AP5 and VS separately, suggesting that they act synergistically. We propose that AP5 potentiates the analgesic effect of VS by two mechanisms: (a) antagonizing the putative pain-producing action of glutamate and glycine acting jointly at the NMDA receptor, and consequently, (b) permitting the unimpeded expression of the analgesic action of inhibitory neurotransmitters released by VS (e.g., glycine at the strychnine-sensitive receptor, and GABA).

Influence of Male-Related Stimuli on Female Postejaculatory Refractory Period in Rats

Yang, L. Y. and L. G. Clemens. Influence of male-related stimuli on female postejaculatory refractory period in rats. Physiol Behav 63(4) 675–682, 1998. Female rats “pace” their sexual contacts with the male when tested in situations where they can escape from the male during copulation. The type and quality of vaginocervical stimulation that the females receive during copulation influences their pacing behavior. This study investigated the effect of several male-related stimuli on the female’s postejaculatory refractory period (PER). Females were tested in a two-compartment test chamber from which they could escape the male through one of four openings along the bottom of the barrier separating the two compartments. Experiment 1 examined the influence of the seminal plug, the penile cup, and prostate secretions on the female’s PER. Results showed that neither the seminal plug, the penile cup, nor prostate secretions contributed to the female’s PER. Experiment 2 investigated the relation of pre-ejaculatory intromission frequency, ejaculation duration, and the number of pelvic thrusts during ejaculation to the female’s PER. Results indicated that pre-ejaculatory intromission frequency and ejaculation duration but not the number of pelvic thrusts during ejaculation were significantly correlated with the female’s PER. In addition, pre-ejaculatory intromission frequency was significantly correlated with ejaculation duration. Partial correlation analysis suggested that pre-ejaculatory intromission frequency affected ejaculation duration which, in turn, influenced the female’s PER. This finding was further supported by the evidence that ejaculation duration and the female’s PER were significantly shorter in tests in which the male ejaculated on the first or second intromission

The activity of ON and OFF cells at the rostroventromedial medulla is modulated by vagino-cervical stimulation

In anesthetized rats it was tested whether or not the activity of the ON and OFF cells within the rostral ventromedial medulla (RVM) is modulated by the mechanical stimulation of the uterine cervix (VS). ON cells were identified by an abrupt increase in their firing rate before the tail flick in response to a noxious heat. OFF cells were identified by a sudden decrease in their firing rate before the tail flick. All (27 out of 27) identified ON cells decreased their firing rate immediately after VS was applied. The effect of VS on the activity of the cells persisted for the entire stimulation period. On the other hand, all (19 out of 19) identified OFF cells increased their firing rate immediately after VS. The effect of VS on the activity of these cells also persisted for the entire stimulation period. The activity of the neutral cells showed no change, neither during the application of noxious heat, nor during VS. These results suggest that the analgesic-like effect produced by VS can be mediated by the activity of the antinociceptive circuit at the RVM. Alternatively, it can be suggested that the afferent inflow from the genital tract can induce the activity of the antinociceptive circuit at RVM, either by projections to the periaqueductal gray matter or by direct projections to RVM.

Induction of maternal behaviour in non-pregnant, hormone-primed ewes

AbstractIt has been suggested that the induction of maternal behaviour in non-pregnant ewes by treatment with oestrogen and progesterone followed by artificial, mechanical vaginocervical stimulation (VCS), could be used to produce foster mothers for orphan or triplet lambs. To investigate the reliability of the technique, 46 non-pregnant sheep of two breeds (Suffolk (S) and Scottish Blackface (BF)) and two levels of experience (nulliparous (N) and multiparous (M)) were used. All ewes were given 6 weeks of treatment with vaginal sponges containing oestradiol and progesterone, followed by 2 min of VCS and exposure to a neonatal (< 2 days old) foster lamb. Plasma oestrogen and prolactin concentrations were significantly elevated by the hormonal treatment; prolactin was significantly higher in M ewes compared with N ewes (BFN = 91, SN = 107, BFM = 301, SM = 687 μg/l,P< 0·005). Significant udder development was seen in most ewes (proportionately 1·0 Mv.0·62 N, P = 0·01) and more M ewes produced milk than N ewes (0·67v.0·19,P< 0·005). No ewes showed maternal behaviour to the lamb from hormone treatment alone. After VCS, one of 26 N ewes and five of 12 M ewes expressed maternal behaviour. In the M ewes, significantly more S ewes were maternal than BF ewes (proportionately 0·75 SMv.0·25 BFM,P< 0·05). It is concluded that the technique alone is not sufficiently reliable to be a useful fostering aid. It is suggested, however, that previous exposure to neonatal lambs, handling of ewes before testing and using newborn lambs, may be fruitful areas of investigation to improve the technique.

Intromissive Stimulation from the Male Increases Extracellular Dopamine Release from Fluoro-Gold-Identified Neurons within the Midbrain of Female Hamsters

Extracellular concentrations of dopamine in the nucleus accumbens were monitored using microdialysis in ovariectomized female Syrian hamsters hormonally primed with estradiol and progesterone or with a similar regimen of oil injections. Some females in each of these groups had their vaginas occluded with tape, whereas the remaining females' vaginas stayed unoccluded. When exposed to a male, both groups of hormonally primed females showed high levels of lordosis. However, only in the hormone-primed, unoccluded females were there significant elevations of dialysate dopamine during the sexual interactions with the male. There were no significant elevations in dopamine levels in the oil-treated females during interactions with the male. These data suggest that nucleus accumbens dopamine is responsive to stimuli associated with the vaginocervical stimulation received by the female during intromissions by the male. Histological analyses were based on Fluoro-Gold efflux through the probes combined with immunocytochemistry for tyrosine hydroxylase. Probe placements in the rostral accumbens, caudal accumbens, or rostral bed nucleus of the stria terminalis were not distinguishable based on analyses of basal dopamine levels, volume of Fluoro-Gold injection sites, or Fluoro-Gold labeling of midbrain, tyrosine hydroxylase-stained neurons. The number of midbrain neurons containing Fluoro-Gold was positively related to basal dopamine levels, indicating that the amount of dopamine recovered from the nucleus accumbens in microdialysis studies is a function of the number of neurons contributing to the terminal field in the region of the probe.

Cardiovascular responses to vaginocervical stimulation in the spinal cord-transected rat.

The present study ascertained whether increases in heart rate (HR) and systolic blood pressure (SBP) produced by vaginocervical stimulation (VS; 500 g force) persist in the unanesthetized rat after chronic spinal cord transection at selected levels. Three groups were used: spinal cord transection at T7 (n = 10) or L5 (n = 10) or a sham-operated control group (Sh, n = 10). In the Sh group, VS increased significantly both HR, by 95 +/- 14.3 beats/min (bpm) (22 +/- 3.7% above baseline), and BP, by 37 +/- 5.7 mmHg (37 +/- 7.7% above baseline), confirming earlier findings. In the T7 group, VS significantly decreased HR by 107 +/- 21.4 bpm (27 +/- 4.1% below baseline) and increased BP by 41.3 +/- 12.9 mmHg (32 +/- 8.3% above baseline). In response to VS, HR increased in every rat in the Sh group and decreased in every rat in the T7 group. In the L5 group, VS failed to significantly affect HR or BP. In the present study, specific levels of spinal cord transection produced differential HR and BP responses to VS in the rat. A model is presented addressing the component responses of autonomic dysreflexia that can occur, contingent on the level of spinal cord injury, in women during parturition or sexual intercourse.

Function of Intromissions on Intromission-Return Latency of Female Rats during Paced Sexual Behavior

Yang, L. Y. and L. G. Clemens. Function of intromissions on intromission-return latency of female rats during paced sexual behavior. Physiol Behav 61(6): 889–894, 1997.—The objectives of this study were to examine how multiple intromissions affect the temporal pattern of the female rat's copulatory behavior; in particular, her latency to return to the male following intromission (intromission-return latency, IRL) and if different hormone replacement regimens affect the temporal aspects of female copulatory behavior. Repeated intromissions alone, without ejaculation, often resulted in prolonged IRLs equal to the postejaculatory refractory period (PER). The first prolonged IRL occurred most frequently between the 24th and 44th intromission. The similar pattern of IRLs around the PER and the prolonged IRLs may indicate that the mechanisms mediating the occurrence of the prolonged IRL are similar to those for the PER. One possible function of the prolonged IRLs may be to facilitate the male's ejaculation after the female has received enough vaginocervical stimulation for the induction of the progestational state of pregnancy. Finally, females receiving a single dose of 50 μg estradiol benzoate (EB) followed by an injection of 0.5 mg progesterone (P) 48 h later showed a significantly longer PER than those receiving 3 daily injections of 0.5 μg EB followed by an injection of 0.5 mg P 24 h after the last EB injection.

Restricting maternal space during parturition in the pig. Effects on oxytocin, vasopressin and cortisol secretion following vagino-cervical stimulation and administration of naloxone.

This experiment studied the effects on endocrine and birth parameters of parturient pigs produced by restricting maternal freedom of movement without otherwise altering environment. Six primiparous pigs (gilts) were each given a jugular catheter under anaesthesia 7 days before parturition and commenced birth in a strawed pen, 2.0 m x 1.5 m in size. Continuous automated blood sampling (3 ml min-1) from unrestrained gilts began following the birth of the first piglet (stage 1) and continued for 2 h. After at least 30 min of blood collection, maternal space was reduced to 2.0 m x 0.55 m by placing rails across the pen (stage 2). The scope for movement in stage 2 was similar to that offered by a farrowing crate. After at least 25 min each gilt was given the opioid antagonist naloxone (1 mg kg-1 i.v.: stage 3). At each stage, vagino-cervical stimulation (VCS) was applied to mimic foetal ejection. Non-cervically stimulated oxytocin (OT) secretion between stages 1 and 2 was unchanged (P > 0.05) but increased significantly relative to both stages 1 and 2 following naloxone treatment for 15-20 min (P < 0.05, paired t-tests on log10 data). Following VCS in all stages plasma OT rose (P < 0.05) for 1-2 min in a similar way to that seen previously following foetal ejection, the increases being proportionally similar irrespective of stage or baseline secretion. Cortisol secretion did not increase as a consequence of space restriction (mean +/- SEM concentrations were 28.6 +/- 8.51 pmol l-1 and 32.3 +/- 11.8 pmol l-1 in stages 1 and 2, respectively). In addition, VCS did not significantly affect cortisol output. Lysine vasopressin concentrations were not affected as a consequence of either stage or VCS. Parturition was not interrupted following space restriction of gilts. These data suggest that reducing maternal space allowance during parturition is not stressful when the process does not involve the movement of animals to novel surroundings.

Effect of the Estrous Cycle on Olfactory Bulb Response to Vaginocervical Stimulation in the Rat: Results from Electrophysiology and Fos Immunocytochemistry Experiments

To determine whether the stage of the estrous cycle modified the response of olfactory bulb neurons to vaginocervical stimulation, (1) vaginocervical stimulation was applied to animals in proestrus-estrus and metestrus-diestrus and the extracellular electrophysiological response of units in the mitral cell layer of the main olfactory bulb was compared, and (2) the effect of vaginocervical or sham stimulation and the effect of the estrous cycle on the number of neurons stained immunocytochemically for Fos in the main and accessory olfactory bulb was examined. Animals in proestrus-estrus had basal firing rates of 21.8 ± 1.8 spikes per 5 s and vaginocervical stimulation produced an increase in firing rate. In contrast, animals in metestrus-diestrus had a slower basal firing rate (14.3 ± 2.3 spikes per 5 s) and vaginocervical stimulation produced a decrease in the firing rate. For animals in proestrus-estrus, vaginocervical stimulation increased the number of Fos-stained cells in the granular cell layer of the accessory olfactory bulb, and in the glomerular and in external plexiform layers of the main olfactory bulb. In contrast, the number of Fos-stained cells decreased in the granular cell layer of the main olfactory bulb after stimulation was applied to animals in proestrus-estrus. The number of Fos-stained cells in the granular layer of the accessory olfactory bulb and the granular and glomerular cell layers of the main olfactory bulb was modulated by the estrous cycle. Therefore, olfactory bulb activity, measured both electrophysiologically and by Fos staining, was affected by the estrous cycle and vaginocervical stimulation, and the two variables interacted. It is likely that integration of interoceptive and environmental stimulation is important for the normal expression of sexual behavior in the female rat.

Differential induction of Fos in the female rat brain following different amounts of vaginocervical stimulation: modulation by steroid hormones

Vaginocervical stimulation (VCS), produced either by copulation with intromission or by manual stimulation of vagina and cervix with a glass rod, induces neuroendocrine and behavioral responses that are critical for female reproduction in many species. We and others have shown that Fos mRNA and protein are induced within different estrogen-concentrating and -non-concentrating regions of the female rat brain following copulation with intromission and manual VCS. In the present study, we investigated the amount of distributed VCS required to induce Fos immunoreactivity within estrogen-concentrating regions of the medial preoptic area, lateral septum, bed nucleus of the stria terminalis, ventromedial hypothalamus, medial amygdala, and mesencephalic central gray, and whether estrogen and progesterone could alter the threshold or pattern of induction. Ovariectomized rats were administered estradiol benzoate (10 μg) 48 h and progesterone (500 μg) 4 h before receiving either 0, 1, 5, 10, 20, 30, 40, or 50 manual VCSs with a lubricated glass rod. Ovariectomized hormone control rats received injections of the sesame oil vehicle 48 and 4 h before VCS. All rats were sacrificed 75 min after the first VCS. Fos immunoreactivity was induced differentially by VCS within the different regions, and the hormones either augmented, inhibited, or had no effect on the induction. These data demonstrate that cells within different estrogen-concentrating regions of the female rat brain are differentially sensitive to VCS, and that steroid hormones can either increase or decrease the amount of Fos induced by different amounts of VCS. Different brain regions may participate in gating the sensory information of VCS into different behavioral and neuroendocrine events.

VAGINOCERVICAL STIMULATION SUPPRESSES THE EXPRESSION OF C-FOS INDUCED BY MATING IN THORACIC, LUMBAR AND SACRAL SEGMENTS OF THE FEMALE RAT

In female rats, vaginocervical stimulation induces neuroendocrine responses necessary for pregnancy as well as analgesia to a variety of noxious stimuli. In this study, Fos immunocytochemistry was used to detect vaginocervical stimulation-induced changes in the activity of spinal neurons at levels T11-S3, segments known to receive afferent input from nerves which innervate the reproductive tract. Adult ovariectomized estrogen and progesterone-treated rats were killed 1 h after receiving mating stimulation from males, which included five or 15 intromissions, mounts-without-intromission by use of either vaginal masks or genitally-anaesthetized males, or immediately after being removed from their home cages. At all spinal levels, Fos labelling was lowest in the home cage group (50 ± 22 cells), intermediate in the groups receiving intromissions (84 ± 8 and 118 ± 22 cells) and highest in groups receiving mounts-without-intromission stimulation (187 ± 21 and 218 ± 35 cells). Significant increases above control levels following intromissive stimulation were observed at levels L6, S1 and S2. Surprisingly, both groups receiving mounts-without-intromission showed significantly higher numbers of Fos-positive cells than did the fully mated groups at all levels. Analysis of selected spinal segments by Rexed's laminae revealed that intromissive stimulation increased Fos labelling above control levels in laminae II-V and X at L6, and laminae I, II, V and X at S1; vaginocervical stimulation did not increase labelling at L1. The greater Fos responses seen in mounts-without-intromission animals than in control or intromitted animals were apparent at L1, L6 and S1 within the same laminae (II-V and X).These results suggest that stimulation of the uterine cervix initiates activity within L6-S2 neurons which receive pelvic nerve afferents and that such stimulation suppresses activity at all levels within populations of neurons normally activated by cutaneous somatic inputs received from male mounts. As antinociceptive agents are known to suppress c-fos expression, vaginocervical stimulation received during natural mating may be capable of initiating spinal and/or brain mechanisms of analgesia. Copyright © 1996 IBRO Published by Elsevier Science Ltd.

Neural mechanisms accounting for the increase in blood pressure and heart rate during vagino-cervical stimulation

The rise in blood pressure and heart rate produced by the mechanical stimulation of the uterine cervix (VS) was examined after adrenalectomy, after pelvic or hypogastric neurectomies or after spinal cord transection in anesthetized rats. Neither adrenalectomy, nor hypogastric neurectomy prevented the rise in heart rate and blood pressure produced by VS. After the spinal cord transection at T6 level, VS was still able to produce the rise in blood pressure. However, the rise in blood pressure was significantly lower than that produced in the same animals before the transection. No changes in heart rate were produced by VS after spinal cord transection. This result can be explained because this level of transection prevents the reach of the afferent inflow to the superior cervical ganglia. Pelvic neurectomy abolished completely the effects of VS on blood pressure and heart rate. Low intensity (1–2 times the threshold) electrical stimulation of the pelvic nerve produced a rise in blood pressure. Even though heart rate increased during electrical stimulation, the change in heart rate was not statistically different from the pre-stimulation value. These results suggest that the changes in blood pressure and heart rate produced by VS represent a neuronal reflex response mediated by the pelvic nerve. The fact that the effects of VS on blood pressure persist in spinal cord-transected animals suggests that the reflex is integrated at the spinal level. However, the cardiovascular responses to VS were significantly lower than before transection, suggesting that supraspinal centers are also involved in the reflex.

Antinociceptive action of vaginocervical stimulation in rat spinal cord: 2-DG analysis

Using [14C]2-deoxyglucose (2-DG) autoradiography with computerized densitometric analysis, unilateral foot pinch was found to significantly increase the relative optical density in laminae I and II of the ipsilateral, compared to the contralateral, spinal cord at lumbar 5 (L5). However, during vaginocervical mechanostimulation applied concurrently with the unilateral foot pinch, no comparable difference was observed. No response to foot pinch was observed in other laminae of the spinal cord at L5, and no effects comparable to the above were observed at L3. These findings indicate that vaginocervical mechanostimulation suppresses neural responses to noxious foot pinch stimulation selectively at the laminae I and II level of the spinal cord at L5, but not at L3.

Evidence for opioidergic inhibition of oxytocin release in periparturient mares

In the horse mare, the onset of parturition is associated with an increase in oxytocin secretion, and it has been suggested that the onset of parturition may be triggered by endogenous oxytocin release. To test the hypothesis that oxytocin secretion is regulated by endogenous opioids in the periparturient period, we have 1) characterized oxytocin secretion in response to vaginocervical stimulation and 2) determined the effect of naloxone, an opioid antagonist, on oxytocin secretion induced by vaginocervical stimulation in prepartum mares and in postpartum mares at estrus and diestrus. During the last 2 months of pregnancy, the first diestrus and subsequent estrus post partum, a total of 66 vaginocervical stimulations were performed. Mares were pretreated with naloxone (0.5 mg/kg iv) or saline, administered 20 min beforevaginocervical stimulation on subsequent days, using a randomized switchback design in which mares served as their own controls. Plasma was collected from 30 min before until 30 min after stimulation and was analyzed for oxytocin concentrations. Vaginocervical stimulation resulted in a significant increase in oxytocin secretion in all mares. Between Days 30 and 20 prepartum, the total amount of oxytocin secreted (calculated as area under the curve for 0 to 10 min after vaginocervical stimulation) was significantly greater in naloxone-treated than in saline-treated mares. From Day 20 prepartum until parturition, the differences between naloxone and saline-treated mares tended to decrease with approaching parturition, and were no longer statistically different. Peak plasma oxytocin concentrations were greater in naloxone-treated mares than in saline-treated mares during the entire prepartum period. During the postpartum period, total amount of oxytocin secreted following vaginocervical stimulation tended to be greater than during the prepartum period, and stimulated oxytocin secretion was significantly greater in naloxone-treated mares than in saline-treated mares. In conclusion, these data suggest that endogenous opioids suppress oxytocin secretion pre and post partum. It appears that opioid inhibition is not limited to the prepartum period, tends to decrease gradually towards parturition and is reinstated after foaling.

Vagotomy blocks responses to vaginocervical stimulation after genitospinal neurectomy in rats

To ascertain whether any effects of vaginocervical stimulation (VS) are mediated by the vagus nerve, all known afferent nerves from the reproductive tract to the spinal cord were transected and the rats were tested for residual responses to VS. After combined bilateral transection of the pelvic, hypogastric, and pudendal nerves (NX), the following responses to VS were greatly reduced or abolished: lordosis to flank-perineum palpation, leg extension, immobilization, and blockage of both tail withdrawal to radiant heat and leg withdrawal to foot pinch. However, after these nerve cuts, the following persisted as significant residual responses to VS: 1) analgesia [measured as increase in vocalization threshold (VOCT) to tailshock], 2) pupil dilatation (PD), and 3) increase in heart rate (HR). Subsequent bilateral subdiaphragmatic vagotomy (VX) significantly reduced the magnitude of PD and abolished the analgesia. By contrast, VX produced no significant effect on the HR increase to VS. The above findings provide evidence that brain-mediated responses to vaginocervical stimulation can be elicited via the vagus nerves.

Infusions of lidocaine into the amygdala, but not the preoptic area, block pseudopregnancy in the rat.

In the rat, vaginocervical stimulation (VCS) received during mating is required for the subsequent expression of 10-12 days of twice-daily prolactin surges that are necessary for pregnancy or pseudopregnancy (PSP). This temporal separation of sensory stimulus and neuroendocrine response suggests that a mnemonic of the vaginocervical stimulation is created in the brain that triggers and sustains the daily prolactin surges. We investigated the possible involvement of the medial preoptic area (mPOA) and the medial amygdala (mAMYG) as potential neural sites involved in the processing of this neuroendocrine arc. Cycling female rats were bilaterally implanted with intracerebral cannulae in either the mPOA or mAMYG. On proestrus, females were manually palpated to confirm sexual receptivity and then received bilateral infusions of either the local anesthetic lidocaine, the Ca(++) channel blocker, verapamil, or phosphate-buffered saline (PBS) into either brain site before or both before and after receipt of 15 intromissions from an experienced male. Unmated control females received comparable infusions of lidocaine or verapamil, and were placed in the empty test arena for 10 min. Infusions consisted of either a single bilateral infusion 15 min before mating (Expt. 1), bilateral infusions both 15 min before and after mating (Expt. 2) or eight bilateral infusions separated by 30 min intervals spanning a period beginning 45 min before and ending 2 h 45 min after mating (Expt. 3). None of the lidocaine infusions into the mPOA prevented the establishment of PSP, and neither verapamil infusions into the mAMYG nor the shorter-term neural block (i.e. single or double lidocaine infusions) of the mAMYG prevented mating-induced PSP. However, the longer-term neural block (i.e. multiple lidocaine infusions) of the mAMYG significantly reduced the incidence of PSP. These data support previous findings that the mAMYG receives sensory input from VCS, and suggest that the mAMYG is a site at which a mnemonic of VCS is established.

Lack of effect of bilateral removal of accessory sex glands on sexual behaviour in the male golden hamster ( Mesocricetus auratus)

In the golden hamster removal of the male accessory sex glands (ASG) may impair fertility because of heavy embryonic wastage. As in the rat, induction of pregnancy and parturition in this species depends on adequate vaginocervical stimulation exerted by the stud at the time of mating. It is possible that embryos fail to develop because of adverse effects on copulatory behaviour consequent upon the absence of accessory sex glands and their associated neural input. The aim of the present study was to test this hypothesis. The results showed that intromission latency, ejaculation latency, intromission frequency, interval of ejaculatory series, postejaculatory pause, ejaculatory frequency and duration of ejaculation were not affected by bilateral section of ampullary glands, coagulating glands, dorsolateral prostate, seminal vesicles, ventral prostate or all of the ASG. It was also concluded that none of the ASG function in the elicitation of sexual behaviour in this species.

Brain-mediated responses to vaginocervical stimulation in spinal cord-transected rats: role of the vagus nerves

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.

Corticotrophin releasing factor mRNA expression in the sheep brain during pregnancy, parturition and lactation and following exogenous progesterone and oestrogen treatment

In the multiparous ewe, intracerebroventricular (i.c.v.) infusions of corticotrophin releasing factor (CRF) act centrally to facilitate the induction of maternal behaviour if administered with vaginocervical stimulation. Changes in CRF mRNA expression in the brains of multiparous ewes were therefore examined as a function of late pregnancy, parturition and lactation using in situ hybridisation histochemistry. As the induction of maternal behaviour in sheep is steroid dependent, a comparable analysis was undertaken in ovariectomised ewes treated with the sex steroids oestrogen and progesterone. Changes in CRF mRNA were quantified in the paraventricular nucleus (PVN) and in the bed nucleus of the stria terminalis (BNST). Expression levels in both the PVN and BNST were unaltered during pregnancy and lactation, but were significantly increased immediately post partum. CRF expression in the BNST, but not in the PVN, was significantly increased in response to treatments with progesterone and oestrogen alone or in combination, although there were no significant differences between treatments. These results indicate that CRF mRNA expression is increased in neuroanatomical locations relevant to the control of maternal behaviour when this behaviour is induced or, in the case of steroid influences on the BNST, is inducible by vaginocervical stimulation. They also indicate, that CRF mRNA expression in the BNST and PVN is differentially influenced by sex steroids.

Neural and Neurochemical Control of Olfactory Recognition of Offspring in Sheep

In sheep, rapid onset of maternal behaviour occurs at parturition and an exclusive bond between a mother and her lambs is established within a few hours after birth. Parturient ewes discriminate between their own lambs and alien lambs: they allow the former to access and suckle the udder, while they reject suckling attempts from the latter, often violently. Olfactory signals from the lambs are the most important cues in this context. Microdialysis measurement of neurochemical substances in the sheep olfactory bulb (OB) reveals increased release of intrinsic (glutamate (GLU), Y-aminobutyric acid (GABA) and dopamine) and centrifugal (noradrenaline (NA) and acetylcholine) neurotransmitters at parturition. Increased release of these transmitters also occurs after artificial vaginocervical stimulation (VCS), which induces ewes to accept alien lambs. Therefore VCS which takes place at parturition plays a key role in triggering the release of these transmitters in the OB and establishing a selective maternal bond. Rapid changes in effectiveness of VCS on release of neurotransmitters occur after the first experience of parturition in primiparous ewes, suggesting that maternal experience with lambs enhances the establishment of a lamb recognition memory at the time of subsequent parturitions. The formation of maternal olfactory memory depends in part on release of nitric oxide (NO) and NA, because inhibition of NO synthase activity or blockade of β-adrenergic receptors or NA release during labour prevents selective bonding and allows ewes t o accept alien lambs. Following selective bonding, the release of GLU and GABA in the OB increases in response to own lamb odours, but not to alien ones, indicating that the importance of these transmitters for mediating the recall of the olfactory memory. Moreover, blockade of GABAA receptors in the OB overcomes a selective bond with own lambs and induces acceptance of alien lambs.