Cortical expression of IL1-ß, Bcl-2, Caspase-3 and 9, SEMA-3a, NT-3 and P-glycoprotein as biological markers of intrinsic severity in drug-resistant temporal lobe epilepsy.

Mesial temporal lobe epilepsy (mTLE) is the most common epilepsy induced by previous cerebral injury, and one out of three mTLE patients develops drug resistance (DR). AIM: To assess the expression of Bcl-2, Caspase-3, Caspase-9, IL1-ß, SEMA-3a, NT-3 and P-glycoprotein in the temporal cortex and their relationship with the progression of mTLE-DR clinical features in patients with mTLE-DR. METHOD: Tissue samples from 17 patients were evaluated for protein expression by Western blot and the relationships of the evaluated proteins with the clinical features of the mTLE were assessed through hierarchical cluster analysis. RESULTS: The mTLE-DR group showed significantly higher P-glycoprotein, Bcl-2 and Caspase-9 levels ***p < 0.0001, ****p < 0.0001 and ***p < 0.0002, respectively, than the autopsy control group. Four patient clusters were identified: Clusters 1 and 3 showed relationships among the age of mTLE onset, duration of mTLE-DR, average number of epileptic seizures per week, number of previous antiepileptic drugs (AEDs) and increased expression of Caspase-3, Caspase-9, Neurotrophin-3 and Semaphorin-3a. Clusters 2 and 4 showed relationships among the mTLE onset age, current age, average number of epileptic seizures per week, number of previous AEDs and increased expression of IL1-ß, Bcl-2, P-glycoprotein, Caspase-3 and NT-3. CONCLUSION: The relationships among the clinical data the age of mTLE onset, DR duration, number of previous AEDs, and average number of seizures per week and the expression of proteins involved in neuronal death, neuroinflammation and aberrant connection formation, as which are biological markers in the cerebral temporal cortex, are important factors in the progression and severity of mTLE-DR and support the intrinsic severity hypothesis.

Vaginocervical stimulation enhances social recognition memory in rats via oxytocin release in the olfactory bulb.

The ability of vaginocervical stimulation (VCS) to promote olfactory social recognition memory at different stages of the ovarian cycle was investigated in female rats. A juvenile social recognition paradigm was used and memory retention tested at 30 and 300 min after an adult was exposed to a juvenile during three 4-min trials. Results showed that an intact social recognition memory was present at 30 min in animals with or without VCS and at all stages of the estrus cycle. However, whereas no animals in any stage of the estrus cycle showed retention of the specific recognition memory at 300 min, those in the proestrus/estrus phase that received VCS 10 min before the trial started did. In vivo microdialysis studies showed that there was a significant release of oxytocin after VCS in the olfactory bulb during proestrus. There was also increased oxytocin immunoreactivity within the olfactory bulb after VCS in proestrus animals compared with diestrus ones. Furthermore, when animals received an infusion of an oxytocin antagonist directly into the olfactory bulb, or a systemic administration of alpha or beta noradrenaline-antagonists, they failed to show evidence for maintenance of a selective olfactory recognition memory at 300 min. Animals with vagus or pelvic nerve section also showed no memory retention when tested after 300 min. These results suggest that VCS releases oxytocin in the olfactory bulb to enhance the social recognition memory and that this may be due to modulatory actions on noradrenaline release. The vagus and pelvic nerves are responsible for carrying the information from the pelvic area to the CNS.

Effects of repeated 9 and 30-day exposure to extremely low-frequency electromagnetic fields on social recognition behavior and estrogen receptors expression in olfactory bulb of Wistar female rats.

OBJECTIVE: We investigated the short- and long-term effects of extremely low-frequency electromagnetic fields (EMF) on social recognition behavior and expression of α- and ß-estrogen receptors (ER). METHODS: Rats were exposed to 60-Hz electromagnetic fields for 9 or 30 days and tested for social recognition behavior. Immunohistochemistry and western blot assays were performed to evaluate α- and ß-ER expression in the olfactory bulb of intact, ovariectomized (OVX), and ovariectomized+estradiol (E2) replacement (OVX+E2). RESULTS: Ovariectomization showed impairment of social recognition after 9 days of EMF exposure and a complete recovery after E2 replacement and so did those after 30 days. Short EMF exposure increased expression of ß-ER in intact, but not in the others. Longer exposure produced a decrease in intact but an increase in OVX and OVX+E2. DISCUSSION: Our findings suggest a significant role for ß-estrogen receptors and a lack of effect for α-estrogen receptors on a social recognition task. ABBREVIATIONS: EMF: extremely low frequency electromagnetic fields; ERs: estrogen receptors; OB: olfactory bulb; OVX: ovariectomized; OVX + E2: ovariectomized + estradiol replacement; IEI: interexposure interval; ß-ER: beta estrogen receptor; E2: replacement of estradiol; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; WB: Western blot; PBS: phosphate-buffer saline; PB: phosphate-buffer.

Influence of birth and maternal experience on olfactory bulb neurotransmitter release.

The ewe's ability to selectively recognize her lamb depends upon vaginocervical feedback to the brain stimulating an interest in lamb odours. This process is facilitated by previous maternal experience. We have used in vivo microdialysis to measure changes in the release of intrinsic transmitters in the olfactory bulb (glutamate, dopamine and GABA) at parturition to determine if their release profiles differ depending upon the ewe's past maternal olfactory experience. Glutamate and GABA release increased significantly at parturition in multiparous but not primiparous ewes. Dopamine release increased in both groups but mean basal levels of this transmitter were significantly higher in primiparous ewes during the pre-partum period and the first few hours postpartum. The changes in the underlying neural circuitry which determine these differences are established within 6 h of parturition, as revealed by artificial stimulation of the reproductive tract. This procedure renders the system plastic enabling adoption of strange lambs and, contingent, on this, the release of intrinsic transmitters no longer differs between the two groups of ewes. Pharmacological challenges to the olfactory bulb using retrodialysis in nulliparous and multiparous (maternally inexperienced and experienced) ewes produced significant differences between the groups for induction of glutamate and GABA release, but not that of dopamine. K+ challenges produced greater increases in glutamate and GABA release in multiparous than in nulliparous ewes, while dopamine release did not differ with experience. Glutamate receptor blockade produced increases in glutamate ase without changing GABA release.(ABSTRACT TRUNCATED AT 250 WORDS)

The role of oxytocin release in the paraventricular nucleus in the control of maternal behaviour in the sheep.

Oxytocin (OT) release within the brain is thought to play a major role in inducing maternal behaviour in a number of mammalian species but little is known about the sites of release which are important in this respect. We have investigated whether the paraventricular nucleus of the hypothalamus (PVN) is a site of OT action on maternal behaviour in the sheep. In vivo microdialysis and retrodialysis was used to determine whether OT is released in the region of the PVN during the post-partum induction of maternal behaviour and if its release at this site can stimulate maternal behaviour in non-pregnant animals. In vivo sampling showed that OT concentrations increased significantly in the region of PVN at birth. When OT was retrodialysed bilaterally into the PVN (1 or 10 microM) of multiparous ewes treated with progesterone and oestradiol to stimulate lactation, maternal behaviour was induced in a significant number of animals (1 microM, 6/8 and 10 microM, 5/8) compared with controls (0/8 ewes). Similar infusions of the ring structure of OT, tocinoic acid (TOC-10 microM), also induced maternal behaviour in a significant proportion of animals (5/6 ewes) as did intracerebroventricular (ICV) OT (6/8 ewes) and artificial stimulation of the vagina and cervix (VCS, 8/9 ewes). On the other hand, vasopressin (AVP) 1 microM did not induce maternal behaviour in any ewes and a 10 microM dose only induced it in 2/8 animals. The neurochemical changes accompanying the above treatments were also investigated. Noradrenaline concentrations increased in the PVN after the retrodialysis administration of OT 1 microM and 10 microM, TOC 10 microM and AVP 1 microM, OT ICV and VCS. Dopamine concentrations were also increased by OT 10 microM, TOC 10 microM, AVP 1microM and OT ICV. Aspartate and glutamate concentrations were significantly reduced by retrodialysis infusions of OT 1 microM and AVP 1 and 10 microM but not by any other treatment. Finally, the retrodialysis infusion of OT and TOC, as well as ICV OT, significantly increased plasma OT release whereas AVP infusions did not. These results provide evidence that OT is released in the PVN during parturition and is important for the induction of maternal behaviour. It seems probable that OT release at this site has a positive feedback effect on both parvocellular and magnocellular OT neurons to facilitate co-ordinated OT release both in central OT terminal regions (to facilitate maternal behaviour) and peripherally into the blood (to facilitate uterine contractions/milk let down). The potential functional roles for the actions of OT on monoamine and amino acid transmitter release in the PVN are discussed.

Effects of parturition and maternal experience on noradrenaline and acetylcholine release in the olfactory bulb of sheep.

Acetylcholine (ACh) and noradrenaline (NA) release in the olfactory bulb (OB) of ewes was monitored using microdialysis. Both ACh and NA release increased at parturition in multiparous but not in primiparous ewes. However, vaginocervical stimulation performed 6 hr postpartum induced an increase of ACh and NA release in both primiparous and multiparous ewes, indicating that a maturation process had occurred. Finally, pharmacological challenges to the ACh and NA inputs revealed differential responsiveness between nulliparous and multiparous nongestant ewes. These results suggest that the first parturition induces changes in neural circuitry involving ACh and NA inputs to the OB.

Oxytocin and vasopressin release in the olfactory bulb of parturient ewes: changes with maternal experience and effects on acetylcholine, gamma-aminobutyric acid, glutamate and noradrenaline release.

Maternal behaviour and the ewe's ability to recognize her lamb depend on olfactory cues and parturition, and are facilitated by maternal experience. Parturition induces a variety of neurochemical changes in the brain and, in particular, oxytocin (OT) release. This peptide injected centrally induces maternal behaviour. Oxytocin release occurs in the olfactory bulb (OB) at parturition and yet this structure is involved in the process of selective bonding with lamb. The present study therefore investigated the possibility that oxytocin release in the OB might modulate the release of classical transmitters that are known to be important in controlling selective recognition and whether maternal experience has any effect on this. We have first used in vivo microdialysis to measure OT release, as well as that of the related peptide, arginine-vasopressin (AVP), in the OB of maternally experienced and inexperienced ewes during parturition. While OT release significantly increased in both primiparous and multiparous ewes at parturition this increase was significantly greater in multiparous ewes. No significant change of AVP release was observed in either group. However, vagino-cervical stimulation (VCS) performed at 6 h post-partum caused similar increases in OT but not AVP release in both primiparous and multiparous ewes suggesting that the first birth experience potentiates the ability of VCS to evoke OT release within 6 h of parturition. Using retrodialysis, either OT (10 microM) or AVP (10 microM) were infused into the OB of multiparous and nulliparous ewes and their effects on modulating acetylcholine (ACh), noradrenaline (NA), glutamate and gamma-aminobutyric acid (GABA) release were monitored. Both peptides produced an increase of ACh and NA in multiparous animals and this effect was either absent or less pronounced in nulliparous animals. OT, but not AVP, also increased GABA release equivalently in nulliparous and multiparous animals. Glutamate release was not altered in response to OT or AVP infusion. These results suggest that OT release in the OB at parturition may facilitate the recognition of lamb odours by modulating NA, ACh and GABA release which are of primary importance for olfactory memory. The reduced release of OT in the OB of primiparous ewes at parturition, together with its reduced ability to modulate NA and ACh release, might also partly explain why maternally inexperienced animals require a longer period to selectively bond with their lambs.

The estrous cycle affects pseudorabies virus (PRV) infection of the CNS.

Previous work had suggested that mucosal immunity may be affected by the stage of the estrous cycle. Here, susceptibility to a neurotropic virus infection at different stages of the estrous cycle was assessed in a rodent model after direct injection of the virus into visceral organs. In the first two experiments, female Sprague-Dawley rats were infected with pseudorabies virus (PRV, Bartha's K-strain) by injection into either the cervix or the kidney after monitoring their estrous cycle. After either 4- or 5-day survival period post-infection, the rats were euthanized by transcardially perfusion and peripheral and central nervous system tissues were removed for immunocytochemical staining. The number of infected neurons was counted in various regions. Statistical analysis revealed that: (1) the number of infected cells in the sympathetic or parasympathetic ganglion, or the dorsal root ganglia was not affected regardless of the stage of the estrous cycle after cervix injection with PRV; (2) in contrast, the number of infected neurons in the spinal cord was affected significantly by the stage of the estrous cycle during viral infection of the cervix; (3) after kidney infection, the number of infected neurons found within the spinal cord or dorsal root ganglia varied significantly across the estrous cycle. In both cases, animals infected in proestrus or estrus had fewer infected neurons than animals infected in diestrus I or diestrus II (proestrous and estrous animals had less than 20% of infected cells found in diestrus I or diestrus II rats). In the third experiment, older, persistent estrous or persistent diestrous rats were infected by kidney injection and given a 4-day survival period, prior to virus isolation from lower thoracic spinal cord. Animals in persistent estrous had significantly less virus per gram of tissue than the persistent diestrous rats. These data suggest that the CNS of animals in proestrus or estrus is less susceptible to PRV infection compared to animals in either diestrus I or diestrus II. Because estrogen replacement therapy is known to restore some immune functions during reproductive ageing, it is speculated that plasma estrogen levels modulate the central nervous system's susceptibility to viral infections.

Vaginocervical stimulation-induced release of classical neurotransmitters and nitric oxide in the nucleus of the solitary tract varies as a function of the oestrus cycle.

The effects of vaginocervical stimulation (VCS) on glutamate (GLU), aspartate (ASP), gamma-aminobutyric acid (GABA), noradrenaline (NA), arginine (ARG) and nitric oxide (NO) (citrulline) release in the nucleus of the solitary tract (nTS) were measured in anaesthetised female rats as a function of the oestrus cycle. During pro-oestrus/oestrus (P/E), but not during met-oestrus/di-oestrus (M/D), VCS significantly increased concentrations of NA, ASP, GLU, NO (citrulline) and GABA, but not ARG. Basal NA concentrations were also increased in P/E. These effects were prevented by bilateral section of either the vagus nerve or pelvic and hypogastric nerves. Vagotomy also significantly decreased basal NO concentrations in M/D and P/E while pelvic and hypogastric nerve section significantly increased GABA concentrations. Our results therefore confirm that the nTS is a relay structure for the visceral afferents sending information from the uterus into the central nervous system. The ability of VCS to trigger classical transmitter release and NO in the female is influenced by the stage of the oestrous cycle and is routed both via the vagus and pelvic/hypogastric nerves.

Electrophysiological evidences of a bidirectional communication between the locus coeruleus and the suprachiasmatic nucleus.

To assess a possible relation between the suprachiasmatic nucleus (SCN) and the locus coeruleus (LC), a study with evoked potentials was performed in the rat. An evoked potential was recorded in the SCN area after electrical stimulation of the LC. Also, an evoked potential was recorded in the LC after electrical stimulation of the SCN area. The results indicate specificity of the projecting regions and of the activated regions, suggesting the existence of a possible bidirectional communication between the LC and the SCN. The pathways possibly involved are discussed.

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.

Olfactory bulb neurons respond to cervicovaginal distension.

Mitral cell layer neuronal activity in the olfactory bulb (OB) of the anesthetized rat is modulated by cervicovaginal distension. Data are reported on 22 cells that decreased and 6 that increased in response to the distension. These results provide support for the existence of a functional interaction between the reproductive tract and the olfactory system.

Neuronal regeneration and estrous cycle restoration after locus coeruleus-periventricular gray substance section.

The locus coeruleus (LC) was anatomically separated from the periventricular gray substance (PVG) by means of knife cuts in the adult female rat presenting regular estrous cycling. This resulted in a transient suppression of the estrous cycling that lasted 10-13 days after surgery. After this period, irregular or regular cycling activity was observed. The regular cycling was restored 30-45 days after the knife cuts. Golgi impregnation of some of the brains of these rats revealed regenerative elements in the knife-cut-insulted area. Thus, blood vessels, macrophagic-like elements, and glial-like elements were observed in close relation with the knife-cut pathway. Additionally, well-defined stained neurons typical of the LC and PVG were observed in close proximity to the knife-cut pathway. Dendritic and axon projections towards the insulted area were observed. Well defined axons were seen across the knife-cut pathway. These data support, first, that the LC-PVG communication is part of a circuitry for the modulation of gonadotropic activity, and second, that in the restoration of the estrous cyclicity after the knife cut, regenerative processes leading to a LC-PVG functional reconnection occurred after the knife cut.

Vagus nerve afferent and efferent innervation of the rat uterus: an electrophysiological and HRP study.

To determine a possible brainstem connection with the uterus, a study with electrophysiological techniques and horseradish peroxidase (HRP) tracing was performed in the rat. Neurons of the nucleus of the tractus solitarius decreased in discharge frequency during cervicovaginal distension. HRP injections into the uterine walls resulted in the appearance of labelled cells in the nodose ganglion and in the dorsal motor nucleus of the vagus nerve. The results demonstrate a direct bidirectional vagal complex-uterus connection via the vagus nerve. Results are discussed in terms of a complex uterus control system in which the paraventricular nucleus might play an integrative role.

Role of the paraventricular nucleus in the projection from the nucleus of the solitary tract to the olfactory bulb.

Electrophysiological experiments were performed on anesthetized rats to determine the effects of lesions of the paraventricular nucleus on the amplitude of evoked potentials recorded in the periglomerular layer of the olfactory bulb after nucleus of the solitary tract electrical stimulation. Lesions of the paraventricular nucleus enhance the amplitude of both the positive and negative components of the evoked potential in the olfactory bulb. The pathway from the paraventricular nucleus to the olfactory bulb seems to exert a suppressive influence over the projection from the nucleus of the solitary tract to the olfactory bulb under these conditions.

Natriuresis after a water load in humans under different sodium body content.

The present study was aimed at observing the diuretic and natriuretic responses after a water load (2% body weight) in four groups of young consenting volunteers submitted previously, during three days, to a hypersodic (500 mEq Na/day), hyposodic (35 mEq Na/day), and normosodic (200 mEq Na/day) diet, or treated with furosemide (Lasix, 40 mg/day). During the treatment urine was collected each day. On the fourth day, in the morning, the bladder was emptied, the water load was ingested, and the urine collected during 10 periods of 20 min each. The urinary, sodium, and chloride flows were determined. The four groups displayed diuretic curves following a similar pattern. In contrast, the natriuretic curves of the four groups were completely different; totally flat with low values for the furosemide group and a large initial natriuretic curve for the hypersodic group with a gradual decrease but maintaining high values. The results indicate that the way the organism compensates for the excess of water by means of urinary water loss is independent of the body sodium content, whereas the way in which sodium loss is accomplished is determined by its body content and is independent of the way in which the water is lost.

Estradiol prevents ozone-induced increases in brain lipid peroxidation and impaired social recognition memory in female rats.

There is increasing concern about the neurodegenerative and behavioral consequences of ozone pollution in industrialized urban centers throughout the world and that women may be more susceptible to brain neurodegenerative disorders. In the present study we have investigated the effects of chronic (30 or 60 days) exposure to ozone on olfactory perception and memory and on levels of lipid peroxidation, alpha and beta estrogen receptors and dopamine beta-hydroxylase in the olfactory bulb in ovariectomized female rats. The ability of 17beta-estradiol to prevent these effects was then assessed. Results showed that ozone exposure for 30 or 60 days impaired formation/retention of a selective olfactory recognition memory 120 min after exposure to a juvenile stimulus animal with the effect at 60 days being significantly greater than at 30 days. They also showed impaired speed in locating a buried chocolate reward after 60 days of ozone exposure indicating some loss of olfactory perception. These functional impairments could all be prevented by coincident estradiol treatment. In the olfactory bulb, levels of lipid peroxidation were increased at both 30- and 60-day time-points and numbers of cells with immunohistochemical staining for alpha and beta estrogen receptors, and dopamine beta-hydroxylase were reduced as were alpha and beta estrogen receptor protein levels. These effects were prevented by estradiol treatment. Oxidative stress damage caused by chronic exposure to ozone does therefore impair olfactory perception and social recognition memory and may do so by reducing noradrenergic and estrogen receptor activity in the olfactory bulb. That these effects can be prevented by estradiol treatment suggests increased susceptibility to neurodegenerative disorders in aging women may be contributed to by reduced estrogen levels post-menopause.

Modulation of in vivo striatal transmitter release by nitric oxide and cyclic GMP.

The effects of nitric oxide (NO) and cyclic GMP on in vivo transmitter release in the rat striatum were investigated using microdialysis sampling in urethane-anaesthetised animals. The NO release-inducing substances S-nitrosoacetylpenicillamine (SNAP), S-nitrosoglutathione (SNOG), and sodium nitroprusside (SNP) increased extracellular concentrations of aspartate (Asp), glutamate (Glu), gamma-aminobutyric acid (GABA), taurine (Tau), acetylcholine (ACh), and serotonin (5-HT). Dopamine (DA) concentrations were decreased by SNAP but were increased by SNOG and SNP. An NO scavenger, haemoglobin, blocked or reduced the effects of SNAP on transmitter release. However, the control carrier compounds for SNAP, SNOG, and SNAP (penicillamine, glutathione, and potassium ferricyanide, respectively, which do not induce release of NO) also increased GABA, Tau, DA, and 5-HT concentrations. When NO gas was given directly by dissolving it in degassed Ringer's solution, DA concentrations decreased significantly, and those of Asp, Glu, GABA, Tau, ACh, and 5-HT increased. These effects of NO gas were all inhibited by coadministration of haemoglobin and for GABA, Tau, ACh, and DA showed some calcium dependency. The cyclic GMP agonists 8-bromo-cyclic GMP and dibutryl-cyclic GMP stimulated dose-dependent increases in Asp, Glu, GABA, Tau, ACh, DA, and 5-HT concentrations. Increased striatal transmitter release in response to NO may therefore be mediated by its stimulatory action on cyclic GMP formation. NO inhibition of DA release may be mediated indirectly through its stimulation of local cholinergic and GABAergic neurones.

C-fos and c-jun in the paraventricular nucleus play a role in regulating peptide gene expression, oxytocin and glutamate release, and maternal behaviour.

In sheep, birth leads to the induction of maternal behaviour through brain oxytocin release. Associated with these events is an upregulation of oxytocin, opioid and corticotrophin-releasing hormone (CRH) gene expression, as well as that of the immediate early gene c-fos in the paraventricular nucleus (PVN) of the hypothalamus. We investigated the role of c-fos dimerizing with c-jun in controlling the induction of maternal behaviour, altered peptide gene expression, and oxytocin and amino acid release in this region at birth. Fluorescence-labelled antisense oligodeoxyribonucleotides (ODNs) against c-fos/c-jun were infused bilaterally in the PVN, via microdialysis probes with 100 kDa cut-off membranes, and were incorporated into 50-60% of the cells. Compared with the control (scrambled) sequences, they significantly reduced basal concentration of glutamate (to 31.7% of baseline after 10 h) and prevented birth-induced release of aspartate. In addition, antisense treatment reduced the birth-induced increase in oxytocin concentration in the PVN, but not in blood. Although all the animals were fully maternal, the antisense treatment did reduce the peak expression of two components of maternal behaviour: low-pitched bleats; and lamb sniffing. Finally, in situ hybridization histochemistry revealed that the antisense treatment significantly reduced the birth-induced upregulation of c-fos, oxytocin, CRH and preproenkephalin mRNA expression in the PVN, whilst not affecting that of arginine vasopressin. These results suggest that c-fos/c-jun transcription factors play a role in the birth-induced upregulation of oxytocin, CRH and preproenkephalin gene expression, as well as on glutamate and oxytocin release in the sheep PVN.