Localization of high-affinity binding sites for oxytocin and vasopressin in the human brain. An autoradiographic study

Sites which bind oxytocin and vasopressin with high affinity were detected in the brain and upper spinal cord of 12 human subjects, using in vitro light microscopic autoradiography. Tissue sections were incubated with tritiated vasopressin, tritiated oxytocin or an iodinated oxytocin antagonist. The ligand specificity of binding was assessed with unlabelled vasopressin or oxytocin in excess, as well as in competition experiments using synthetic structural analogues. The distribution of vasopressin binding sites differed markedly from that of oxytocin binding sites in the forebrain, while there was overlap in the brainstem. Vasopressin binding sites were detected in the dorsal part of the lateral septal nucleus, in midline nuclei and adjacent intralaminar nuclei of the thalamus, in the hilus of the dentate gyrus, the dorsolateral part of the basal amygdaloid nucleus and the brainstem. The distribution of oxytocin binding sites in the brainstem has been recently reported (Loup et al., 1989). Oxytocin binding sites were also observed in the basal nucleus of Meynert, the nucleus of the vertical limb of the diagonal band of Broca, the ventral part of the lateral septal nucleus, the preoptic/anterior hypothalamic area, the posterior hypothalamic area, and variably in the globus pallidus and ventral pallidum. The presence of oxytocin and vasopressin binding sites in limbic and autonomic areas suggests a neurotransmitter or neuromodulator role for these peptides in the human central nervous system. They may also affect cholinergic transmission in the basal forebrain and consequently play a role in Alzheimer's disease.     

Localization of oxytocin binding sites in the human brainstem and upper spinal cord: an autoradiographic study

Two different ligands, tritiated oxytocin and a newly synthesized and monoiodinated oxytocin antagonist, were used to reveal sites which bind oxytocin in the brainstem and upper spinal cord of 12 human subjects. Tissue sections were incubated with either ligand at a concentration close to their respective dissociation constants determined in human uterus and rat brain. Specificity of binding was assessed in presence of unlabelled oxytocin in excess. Comparable results were obtained using tritiated or iodinated ligand. Labelling was most intense in the substantia nigra pars compacta, the substantiae gelatinosae of the caudal spinal trigeminal nucleus and of the dorsal horn of the upper spinal cord, as well as in the medio-dorsal region of the nucleus of the solitary tract. Binding was also detected in the rest of the nucleus of the solitary tract and in other areas, including the oral and interpolar parts of the spinal trigeminal nucleus, the hypoglossal nucleus and the area postrema. Presence of oxytocin binding sites in regions concerned with sensory, autonomic and motor processing suggests that oxytocin could act as a neurotransmitter of neuromodulator in the human central nervous system.     

Localization of Oxytocin Binding Sites in the Thoracic and Upper Lumbar Spinal Cord of the Adult and Postnatal Rat: A Histoautoradiographic Study

Oxytocin binding sites were detected by autoradiography on films and emulsion-coated sections in the spinal cord of adult and postnatal rats from C8 to L2, using a highly selective ¹²⁵l-labelled oxytocin antagonist. Oxytocin binding sites were detected on all transverse sections in the dorsal horn, where labelling was scattered over laminae I and II. The autonomic areas, i.e. the intermediolateral cell column, the central grey (lamina X) and the nucleus intercalatus were labelled. Binding in the intermediolateral cell column was most frequently observed on sections from T9 to T11 in adult and T7 to T8 in postnatal rats. In this location, oxytocin binding sites were highly concentrated on cell bodies of putative sympathetic preganglionic neurons; however, not all of these cells were labelled. Diffuse labelling occurred on the dorsal part of the central grey, mainly between T8 and L2. Isolated labelled cells belonging to the nucleus intercalatus were scattered between the central canal and the intermediolateral cell column. In addition, oxytocin binding sites were found on some motoneurons of the lateral group of T12-T13, but only in postnatal rats. The distribution of oxytocin binding sites in the rat spinal cord coincides with that of the oxytocin innervation and strongly suggests a modulatory role of this peptide in sensory and autonomic functions.     

Ontogeny of oxytocin receptors in rat forebrain: a quantitative study

The ontogeny of oxytocin receptors in rat forebrain was studied using the selective oxytocin receptor antagonist 125I-d(CH2)5[Tyr(Me)2, Thr4, Tyr-NH29]OVT [( 125I]-OTA). With in vitro receptor autoradiography, binding wa noted on the first postnatal day in dorsal subiculum and thalamus. On postnatal days 5-18, intense labeling was evident in posterior cingulate cortex, dorsal subiculum, lateral septum, and the CA1 subfield of hippocampus. Of these regions only the lateral septum expressed oxytocin receptors in adult brain. Competition studies on coronal sections through posterior cingulate, septum, and dorsal subiculum at P10 demonstrated that transient binding sites in these areas were indeed oxytocin selective (OXY greater than AVP greater tha V1 greater than V2). Result of saturation studies on cingulate membranes from 10-day-old pups agreed favorably with previous reports of the kinetics of [125I]-OTA binding to adult oxytocin receptors (Kd = 0.1 nM in P10 cingulate cortex vs. 0.07 nM for adult ventral subiculum). In contrast to these evanescent developmental sites, oxytocin receptors in the bed nucleus of the stria terminalis and the ventromedial nucleus of the hypothalamus only appeared in adulthood, presumably in response to the surge of gonadal steroids at puberty.     

Morphological and electrophysiological evidence for postsynaptic localization of functional oxytocin receptors in the rat dorsal motor nucleus of the vagus nerve

The vagal complex is innervated by oxytocin immunoreactive axons of hypothalamic origin. The presence of oxytocin binding sites in the dorsal motor nucleus of the vagus nerve of the rat was evidenced by autoradiography with a radioiodinated oxytocin antagonist as ligand. Two weeks following a unilateral vagotomy, distal to the nodose ganglion, binding sites were reduced below the level of detection in the ipsilateral dorsal motor nucleus of the vagus nerve. Choline acetyltransferase immunoreactivity was also markedly reduced in the vagal motoneurons whose axons had been transected. Electrophysiological studies were performed in vitro in brainstem slices from control rats. In antidromically identified vagal motoneurones, oxytocin applied at 0.1-1.0 microM either caused a reversible depolarization or generated, under voltage-clamp conditions, a transient inward current. These responses persisted under the condition of synaptic uncoupling. Taken together these observations favour the notion that oxytocin of hypothalamic origin acts directly on rat vagal motoneurones.     

Early appearance and transient expression of vasopressin receptors in the brain of rat fetus and infant. An autoradiographical and electrophysiological study

The development of vasopressin (AVP) receptors in the rat brain, spinal cord and pituitary gland was studied by in vitro light microscopic autoradiography. AVP binding sites were labeled using [3H]AVP in tissue sections from animals aged between embryonic day 12 (E12) and postnatal day 90 (PN90); the binding of [3H]AVP to oxytocin receptors was prevented by adding in the incubation medium a highly selective oxytocin agonist. Specific binding was first detected at E16 in the ventral pontine reticular formation. Many other brain areas were progressively labeled between E18 and PN5. The distribution of binding sites observed at PN5 remained unchanged until the beginning of the third postnatal week. Thereafter binding was markedly reduced or even disappeared in several areas, in particular in the facial nucleus. The adult distribution of AVP binding sites was established at the time of weaning. The properties of transient AVP binding sites in the facial nucleus were studied both by autoradiography and by electrophysiology. Non-radioactive AVP displaced [3H]AVP binding in this nucleus as efficiently as it did in the lateral septum of the adult. Single-unit extracellular recordings showed that AVP can excite facial motoneurones by interacting with receptors which are pharmacologically indistinguishable from V1 (vasopressor) type. Thus, AVP binding sites transiently expressed in the brain of fetal and infant rat probably represent functional neuronal receptors, having the same ligand selectivity and affinity than AVP binding sites present in the adult. This suggests that AVP acts not only as a neuropeptide in the adult brain but may play a significant role during maturation of the central nervous system.     

Appearance and transient expression of oxytocin receptors in fetal, infant, and peripubertal rat brain studied by autoradiography and electrophysiology

The development of oxytocin (OT) receptors in the rat brain and spinal cord was studied by in vitro light microscopic autoradiography and by electrophysiology. OT receptors were labeled using a monoiodinated OT antagonist in tissue sections from animals aged between embryonic day 12 (E12) and postnatal day 90 (PN90); the response of ongoing spike activity to the addition of OT was assessed in neurons located in the dorsal motor nucleus of the vagus nerve of the neonate. Specific binding was detected first at E14 in a region that later differentiated into the dorsal motor nucleus of the vagus nerve. Many other regions were progressively labeled between E20 and PN5. From PN5 to PN16, the distribution of binding sites remained essentially unchanged but differed markedly from that characteristic of the adult. The change-over from the "infant pattern" to the "adult pattern" occurred in 2 stages: the first change took place between PN16 and PN22, a time corresponding to the preweaning period; the second change occurred after PN35 and thus coincided with the onset of puberty. During the first transition period, binding was reduced or disappeared in several areas intensely labeled at earlier stages, in particular, in the cingulate cortex and the dorsal hippocampus. At the same time, binding sites appeared in the ventral hippocampus. At puberty, high densities of OT binding sites appeared in the ventromedial hypothalamic nucleus and the olfactory tubercle. Electrophysiological activity was recorded from vagal neurons in slices obtained from animals sacrificed at PN1-PN12. OT and a selective OT agonist reversibly increased the firing rate of these neurons in a concentration-dependent manner. The neuronal responsiveness was similar to that reported previously in the adult. These results suggest that OT binding sites detected by autoradiography in the developing rat brain represent, at least in some areas, functional neuronal receptors.     

Histoautoradiographic detection of oxytocin- and vasopressin-binding sites in the telencephalon of the rat

Localization of oxytocin- and vasopressin-binding sites has so far been studied in the rat brain by means of film autoradiographs. The disposal of iodinated ligands with high specificity has allowed us to develop histoautoradiography on emulsion-coated sections and to reinvestigate on a microscopic scale the distribution of these sites in the telencephalon (septum, striatopallidal system, amygdala and hippocampus). This technique showed that oxytocin and vasopressin labelling presented distinct distributions and coincided with delimited zones, corresponding to anatomical subdivisions defined on cytoarchitectural and immunocytochemical bases. Vasopressin sites were seen in the dorsal and intermediate parts of the lateral septum and the juxtacapsular nucleus of the bed nucleus of the stria terminalis. Oxytocin sites were located in the ventral and intermediate parts of the lateral septum, the oval and the principal nuclei of the bed nucleus of the stria terminalis and the septofimbrial nucleus. In the striatopallidal system, vasopressin sites were found in the accumbens nucleus and the fundus striati, whereas oxytocin sites were in the accumbens nucleus, the head, and the posterolateral parts of the caudate-putamen, the striatal cell bridges, and the olfactory tubercle. In the amygdala, vasopressin sites were not found, but oxytocin sites were located in the central, medial, and basomedial nuclei. In the hippocampus, vasopressin sites were located in the dentate gyrus (polymorph and molecular layers), and oxytocin sites, in the subiculum (molecular and pyramidal layers) and in the field CA1 of Ammon's horn (lacunosum moleculare and pyramidal layers). The localization of the binding sites at the microscopic level permitted us to reinvestigate whether or not correlation existed in a same area between innervation, electrophysiological effects, and presence of binding sites. © 1993 Wiley-Liss, Inc.     

Autoradiographic Detection of Oxytocin- and Vasopressin-Binding Sites in Various Subnuclei of the Bed Nucleus of the Stria Terminalis in the Rat. Effects of Functional and Experimental Sexual Steroid Variations

Oxytocin- and vasopressin-binding sites were detected by autoradiography on films and on emulsion-coated sections of rat brains using highly selective [¹²⁵|]-labelled oxytocin and vasopressin antagonists. Two distinct areas with high concentrations of oxytocin-binding sites were detected in the bed nucleus of the stria terminalis: 1) the principal encapsulated nucleus and the associated cell-sparse zone in the posterior medial part, and 2) the oval nucleus in the anterior lateral part. A weak diffuse labelling was, in addition, detected around the oval nucleus in the anterior lateral and anterior dorsal areas. The vasopressin-binding sites were restricted to the anterior lateral part of the bed nucleus of the stria terminalis where they were highly concentrated in the juxtacapsular nucleus and present with lower density in a discrete cell group dorsal to the oval nucleus. Autoradiographic analyses of the bed nucleus of the stria terminalis from pregnant, lactating and ovariectomized rats (oestradiol treated or not) indicated that only the oxytocin-binding sites in the principal encapsulated nucleus and the associated cell-sparse zone were oestrogen-dependent. These observations are in agreement with earlier data suggesting that the two major divisions of the bed nucleus of the stria terminalis are involved in distinct regulations. The anterior lateral part, including the oval nucleus in which oxytocin receptors are not oestrogen-dependent, is, rather, involved in central autonomie regulations. The posterior medial part, where oestrogen-dependent oxytocin receptors are concentrated in the principal encapsulated nucleus and the associated cell-sparse zone, is implicated in neuroendocrine regulations and in reproductive behaviour.     

Localization and pharmacological characterization of high affinity binding sites for vasopressin and oxytocin in the rat brain by light microscopic autoradiography

Sites which bind tritiated vasopressin (AVP) with high affinity were detected in the brain of male, adult rats, by light microscopic autoradiography. Their anatomical localization differed markedly from that of high affinity binding sites for tritiated oxytocin (OT) determined in the same animal. Co-labelling was minimized by using low concentrations of [3H]AVP and [3H]OT. Binding of the former occurred predominantly in several structures of the limbic system (septum, amygdala, bed nucleus of the stria terminalis, accumbens nucleus), in two hypothalamic nuclei (suprachiasmatic and dorsal tuber) and in the area of the nucleus of the solitary tract. Binding of OT was evidenced in the olfactory tubercle, the ventromedial hypothalamic nucleus, the central amygdaloid nucleus and the ventral hippocampus. The ligand specificity of the binding sites was assessed in competition experiments. Synthetic structural analogues were used, allowing to discriminate OT receptors (OH[Thr4,Gly7]OT) from V2 receptors (dDAVP and d[Tyr(Me)2]VDAVP), V1 receptors ([Phe2,Orn8]VT) and V1b receptors (desGly9d(CH2)5AVP). Our main conclusions are, firstly, that AVP and OT binding sites can be readily distinguished, and that there is virtually no overlap in their distribution in the rat brain. Second, we showed that the sites which bind AVP with high affinity in the brain are V1 receptors, different both from the renal V2 receptors and from the anterior pituitary V1b receptors. Our results support the conjecture that AVP and OT play a role in interneuronal communication in the brain.     

Estrogen-Sensitive Oxytocin Binding Sites are Differently Regulated by Progesterone in the Telencephalon and the Hypothalamus of the Rat

The localization at the cellular level and the regulation by progesterone of the estrogen-sensitive oxytocin binding sites was studied in the rat telencephalon and the hypothalamus by using quantitative film-autoradiography and histoautoradiography. Male rats (castrated or not) and ovariectomized females (estradiol supplemented or not) were used to characterize these sites and to precise their localization. They were detected in the striatal cell bridges, the olfactory tubercle, the principal nucleus of the bed nucleus of the stria terminalis and the medial nucleus of the amygdala of the telencephalon and in the medial preoptic, the ventromedial and the ventral premammillary nuclei of the hypothalamus. Estrogen administration in addition induced expression of oxytocin binding sites in the major island of Calleja, the anterior hypothalamic area and the terete nucleus. The density of the estrogen-sensitive oxytocin binding sites varied during the estrous cycle, but differently in the telencephalon and the hypothalamus. in the telencephalon it peaked at proestrus 9 h and was already decreased at proestrus 21 h, whereas in the hypothalamus it was similarly high at proestrus 9h and proestrus 21 h, suggesting the intervention of progesterone in the regulation of the hypothalamic estrogen-sensitive oxytocin binding sites.     

Oxytocin Excites Neurons Located in the Ventromedial Nucleus of the Guinea-Pig Hypothalamus

The area of the ventromedial nucleus of the hypothalamus in the guinea-pig was shown in autoradiographs to contain high affinity binding sites for oxytocin. In order to ascertain whether these sites may represent neuronal receptors, single-cell extracellular recordings were obtained from ventromedial neurons in coronal slices of the hypothalamus of adult guinea-pigs. Oxytocin applied in the nanomolar range excited about half of the neurons tested; none were inhibited. The response to the peptide was reversible and concentration-dependent. It was exerted directly since it persisted under the condition of synaptic isolation. Moreover, the effect was specific since it could be mimicked by a selective oxytocin agonist and since vasopressin was usually at least 10-fold weaker than oxytocin. These findings suggest that the binding sites for oxytocin detected by light microscopic autoradiography in the guinea-pig hypothalamic ventromedial nucleus represent functional receptors.     

Historadioautographic localisation of oxytocin and vasopressin binding sites in the central nervous system of the merione (Meriones shawi)

The distribution of vasopressin and oxytocin binding sites in the central nervous system of the merione (Meriones shawi), a rodent adapted to desert life, was studied by means of conventional film radioautography at macroscopic scale and historadioautography at cellular level using radioiodinated ligands highly selective for either oxytocin or type V1 a vasopressin receptors. Both types of binding sites exhibited the same selectivity for endogenous peptides as in the rat. Distribution of oxytocin binding sites was similar in some structures (limbic system, spinal cord) to that described in the rat and in other rodents. Vasopressin binding sites were much more widely distributed in the merione than in the rat brain. In addition to locations common to most rodents (lateral septum and suprachiasmatic nucleus), in merione vasopressin binding sites occurred in several areas known to express oxytocin binding sites in the rat (olfactory system, hypothalamus). Colocalisation of vasopressin and oxytocin binding sites, which occurred in the CA1 and CA2 fields of Ammon's horns of the hippocampus, the caudate-putamen and the fundus striati of the merione, has so far not been reported in any other rodent.     

Localization and characterization of binding sites for vasopressin and oxytocin in the brain of the guinea pig.

Using autoradiography on film, specific binding sites for arginine-vasopressin (AVP) and for oxytocin (OT) were localized in various areas of the brain of adult male guinea pigs. Vasopressin binding sites were detected with [3H]AVP or with [125I]VPA, a recently synthetized linear vasopressin antagonist radiolabeled with 125I. [125I]VPA and [3H]AVP yielded similar results, thus suggesting that AVP binding sites present in the guinea pig brain are V1 type receptors. Supporting evidence on this was obtained in competing studies using structural analogues allowing to discriminate V1 receptors from V2 and from OT receptors. Oxytocin binding sites were labeled with [3H]OT or with the iodinated OT antagonist [125I]OTA; both ligands yielded similar results. The localization in the guinea pig brain of AVP binding sites differed from that of OT binding sites. AVP binding sites were mainly detected in the olfactory bulb and throughout the cerebral cortex. Oxytocin binding sites were most noticeable in the hypothalamic ventromedial nucleus, in the amygdaloid complex and in restricted areas of the cerebral cortex. A comparison of the present data with those previously described in the rat, the mouse, the human and the hamster brain suggests that similar binding sites are present in these species, but that their anatomical distribution differs markedly. These data are discussed in relation to immunocytochemical and electrophysiological data which suggest that binding sites detected by autoradiography may represent, at least in part, functional neuronal receptors.     

Effects of lesions in the hypothalamic paraventricular, supraoptic and suprachiasmatic nuclei on vasopressin and oxytocin in rat brain and spinal cord

The content of arginine vasopressin and oxytocin in various extrahypothalamic sites of the rat brain and spinal cord was determined by specific radioimmunoassays after lesions had been made in either the paraventricular (PVN), supraoptic (SON) or suprachiasmatic nuclei (SCN). In some animals all 3 nuclei were destroyed together. The PVN provided a considerable amount of the vasopressin innervation of the solitary tract nucleus, and most of that in the spinal cord. Oxytocin was removed from some areas after lesions of the PVN and, again, most of this peptide was lost from the spinal cord. Lesions of the SCN did not appear to be followed by significant quantitative changes in either hormone in any of the areas studied. Lesions of the SON resulted in loss of oxytocin, particularly in the periventricular grey and some other areas, suggesting that extrahypothalamic projections from this nucleus may be more important than was previously assumed. Lesions of all 3 nuclei which included destruction of accessory hypothalamic nuclei resulted in a much more widespread loss of vasopressin and oxytocin, but there was preservation of both peptides in the dorsal raphe nucleus and much of those present in the locus coeruleus. It is concluded that the contribution of the classical hypothalamic nuclei to the extrahypothalamic content of vasopressin and oxytocin in rat brain is less than was originally believed, and that there are areas of the brain such as the locus coeruleus and dorsal raphe nucleus in which the source of these peptides may be outside the hypothalamus.     

Localization of atrial natriuretic factor (ANF) binding sites in the central nervous system of the frog

The distribution of atrial natriuretic factor (ANF) binding sites was investigated in the central nervous system of the frog Rana ridibunda using the technique of in vitro receptor autoradiography by means of [125I]-labeled ANF-28. The anatomic distribution of ANF recognition sites was determined on Kodak ARX films apposed onto tissue sections, and their distribution was examined in greater detail by analysis of autoradiograms generated by using emulsion-coated sections. The highest levels of ANF binding sites were found in the olfactory bulb, the dorsal pallium, the septum, the habenular nucleus, the dorsal infundibular nucleus, the interpeduncular nucleus, and in the tectum. Moderate levels of ANF binding sites were observed in the thalamus and throughout the mesencephalon, whereas low levels were detected in the lateral and medial pallium, the medial forebrain bundle, and the nucleus rotondus. In the pituitary gland, the neural and distal lobes were densely loaded with ANF binding sites, whereas no autoradiographic labeling was observed in the pars intermedia. In general, there was a good correlation between the location of ANF receptors and the distribution of ANF-containing neurons, as previously determined by immunocytochemistry. Together these results support the view that ANF may act as a neurotransmitter or a neuromodulator in various regions of the frog brain.     

Vasopressin, oxytocin and neurophysins in the human brain and spinal cord

The concentrations of arginine vasopressin, oxytocin, and their related neurophysins were compared in many areas of postmortem human brain and spinal cord using specific radioimmunoassays. In the hypothalamus the ratio of vasopressin to oxytocin was approximately 3:1, and in the extrahypothalamic areas of the brain the greatest amount of both peptides was present in the locus coeruleus, and to a lesser extent the periaqueductal grey. Vasopressin only was found in the substantia nigra, and globus pallidus. In the medulla, including the nucleus of the solitary tract, the dorsal nucleus of the vagus, and the nucleus of the spinal tract of the trigeminal nerve, the amount of oxytocin was greater than that of vasopressin. In the spinal cord oxytocin predominated over vasopressin to an even greater extent, and reached particularly high values in certain segments of the intermediolateral grey column and dorsal horn. Estrogen-stimulated and nicotine-stimulated neurophysins (ESN and NSN) were both found in large amounts in those areas of the brain and spinal cord where the concentrations of the nonapeptides were greatest, but when the molar ratios of ESN to oxytocin and NSN to vasopressin were compared there was an excess of ESN.     

Gonadal steroids regulate oxytocin receptors but not vasopressin receptors in the brain of male and female rats. An autoradiographical study

The distribution and the amount of [3H]oxytocin binding were studied in the brain of adult rats of either sex, as well as in male and female castrates, some of which received injections of estradiol or testosterone. Intact males were treated with an aromatase inhibitor. Castration and inhibition of aromatase activity reduced, whereas estradiol and testosterone increased oxytocin binding, particularly in regions of the brain assumed to be involved in reproductive functions, such as the ventrolateral part of the hypothalamic ventromedial nucleus and the islands of Calleja and neighbouring cell groups. Binding of oxytocin to the uterus was also estrogen-dependent. In the same animals, we also studied the distribution of [3H]vasopressin binding sites present in the brain. It was similar in males and females, and was not affected by experimentally manipulating gonadal hormone levels. In immunocytochemical studies we noticed, as others had previously, that the vasopressin content of certain areas of the rat brain was affected by castration, whereas the oxytocin innervation was not. These results are discussed in relation to the possible functions of oxytocin in the brain and of the lack of correspondence between the immunocytochemical and the autoradiographic data.     

Differential excitatory responses to oxytocin in sub-divisions of the bed nuclei of the stria terminalis

The lateral dorsal nucleus of the bed nuclei of the stria terminalis (BST-LD) expresses dense oxytocin binding while lower binding is detected in the medial anterior BST (BST-MA) and adjacent ventrolateral septum (VLS). However, in vitro examination of neuronal responses to oxytocin showed that the BST-LD exhibited small, transient responses which desensitized upon repeated challenge. In contrast, the BST-MA and VLS exhibited significantly larger responses with no significant desensitization. This inverse relationship between oxytocin binding density and electrophysiological responsiveness is also seen in the central and medial amygdaloid nuclei, which have respective associations with the lateral and medial divisions of the BST. Thus, excitatory responses to oxytocin vary markedly between BST sub-divisions and may reflect associations within the extended amygdala.     

Regional Changes in Brain Oxytocin Receptors Post-Partum: Time-Course and Relationship to Maternal Behaviour

In a previous report, receptors for oxytocin in rat brain were reported to increase during the early post-partum period. The current study set out to replicate this finding using the novel, highly selective oxytocin receptor ligand, [¹²⁵ l]d(CH₂)₅[Tyr(Me)², Thr⁴, Tyr-NH₂⁹]OVT ([¹²⁵ I]-OTA). Binding was measured using in vitro receptor autoradiography in rat brain on Day 15 of pregnancy, on Days 1 and 6 post-partum, and at least 6 days following the end of lactation. Relative to pregnancy, oxytocin receptor binding was increased on Day 1 (but not Day 6) post-partum in the bed nucleus of the stria terminalis and the ventromedial nucleus of the hypothalamus. In several other regions, including the anterior olfactory nucleus, the central nucleus of the amygdala, and the ventral subiculum binding was equivalent across the groups. Most surprising, binding in the bed nucleus of the stria terminalis was lowest in the post-lactating group (post-partum Day 1 group was 87% higher than post-lactating group). Saturation studies suggested that binding differences reflected changes in number and not affinity of sites. These findings are consistent with earlier studies of changes in brain oxytocin receptors following exogenous gonadal steroid administration. To determine if maternal behaviour elicited by nonhormonal means (i.e. concaveation) was also associated with increased oxytocin receptors, virgin females with extensive pup exposure were studied. No change in oxytocin receptor binding in the bed nucleus of the stria terminalis was noted in virgin, maternal females—demonstrating that the increase observed post-partum was not essential for the onset of maternal behaviour.