Mothers and offspring: The rabbit as a model system in the study of mammalian maternal behavior and sibling interactions.

UNLABELLED: This article is part of a Special Issue "Parental Care". Jay Rosenblatt effectively promoted research on rabbit maternal behavior through his interaction with colleagues in Mexico. Here we review the activities of pregnant and lactating rabbits (Oryctolagus cuniculus), their neuro-hormonal regulation, and the synchronization of behavior between mother and kits. Changing concentrations of estradiol, progesterone, and prolactin throughout gestation regulate nest-building (digging, straw-carrying, fur-pulling) and prime the mother's brain to respond to the newborn. Nursing is the only mother-young contact throughout lactation. It happens once/day, inside the nest, with ca. 24h periodicity, and lasts around 3min. Periodicity and duration of nursing depend on a threshold of suckling as procedures reducing the amount of nipple stimulation interfere with the temporal aspects of nursing, though not with the doe's maternal motivation. Synchronization between mother and kits, critical for nursing, relies on: a) the production of pheromonal cues which guide the young to the mother's nipples for suckling; b) an endogenous circadian rhythm of anticipatory activity in the young, present since birth. Milk intake entrains the kits' locomotor behavior, corticosterone secretion, and the activity of several brain structures. Sibling interactions within the huddle, largely determined by body mass at birth, are important for: a) maintaining body temperature; b) ensuring normal neuromotor and social development. Suckling maintains nursing behavior past the period of abundant milk production but abrupt and efficient weaning occurs in concurrently pregnant-lactating does by unknown factors. CONCLUSION: female rabbits have evolved a reproductive strategy largely dissociating maternal care from maternal presence, whose multifactorial regulation warrants future investigations.

Ovarian hormones and fasting differentially regulate pituitary receptors for estrogen and gonadotropin-releasing hormone in rabbit female.

To investigate the mechanisms by which caloric restriction affects reproductive function in female rabbits, we measured, in animals intact or ovariectomized (OVX) estrogen-primed and fed ad libitum or fasted for 48 h, the adenohypophysial expression of estrogen receptor-alpha (ESR1) and gonadotropin releasing hormone receptor (GnRHR) and the dynamic secretion of LH following GnRH stimulation. Fasting increased the number of GnRHR-immunoreactive (-IR) cells in intact animals, whereas reduced the density of ESR1-IR cells in OVX rabbits. Estrogen priming decreased the number of ESR1-IR cells in fasted and OVX animals. Ovariectomy increased the number of ESR1-IR cells in fed rabbits, but caused an opposite effect in both fed and fasted animals treated with estrogen. Fasting down regulated the mRNA levels for ESR1 and GnRHR. Estrogen-priming reduced the abundance for ESR1 mRNA in both fed and fasted rabbits, and that for GnRHR in fasted rabbits. Ovariectomy halved ESR1 mRNA levels independently of treatment and feeding condition, whereas increased (P < 001) that for GnRHR in estrogen-primed rabbits. In all rabbits, an LH surge occurred 30 min after GnRH injection but the lowest levels were found in intact fasted rabbits and the highest in fasted, estrogen-primed animals. The LH profile was similar in intact and OVX rabbits and neither fasting nor estrogen priming modified it. In conclusion, fasting differentially modifies the ESR1 and GnRHR expression in the pituitary, depending on the presence of gonadal hormones, indicating complex interactions between metabolic signals and ovarian steroids.

Expression of the cannabinoid receptor type 1 in the pituitary of rabbits and its role in the control of LH secretion.

The aim of this study was to elucidate the possible direct regulatory role of the endocannabinoids in the modulation of LH secretion in rabbits, a reflex ovulator species. The cannabinoid receptor type 1 (CB1) was characterized by RT-PCR techniques in the anterior pituitary of intact and ovariectomized does treated with GnRH and primed with estrogen and CB1 antagonist, rimonabant. Cannabinoid receptor type 1 immune reaction was evidenced by immunohistochemistry in the cytoplasm of approximately 10% of the pituitary cells with a density of 8.5 ± 1.9 (per 0.01 mm(2)), both periodic acid-Schiff positive (30%) and negative (70%). All CB1-immunoreactive cells were also immune reactive for estrogen receptor type 1. Ovariectomy, either alone or combined with estrogen priming, did not modify the relative abundances of pituitary CB1 mRNA, but decreased (P < 0.01) the expression of estrogen receptor type 1 mRNA. Treatment with CB1 antagonist (rimonabant) inhibited (P < 0.01) LH secretory capacity by the pituitary after GnRH injection, and estrogen priming had no effect. The present findings indicate that the endocannabinoid system is a potential candidate for the regulation of the hypothalamic-pituitary-ovarian axis in reflex ovulatory species.

Relevance of ovarian signaling for the early behavioral transition from estrus to pregnancy in the female rabbit.

During estrus, the female domestic rabbit (Oryctolagus cuniculus) displays scent marking behavior (chinning), which is immediately inhibited after mating, temporarily recovers, and then declines and remains inhibited across pregnancy. Chinning is inhibited by progesterone (P) and the activation of the progesterone receptor (PR), but it is unlikely that P participates in the "acute" (immediate) or "early" inhibition of chinning (24 to 96 h post-mating, before plasma P levels rise). Since PR is activated in a ligand-independent manner by a variety of signaling molecules, some of which (e.g., GnRH) are also associated with reflexive ovulation in this species, we hypothesized that neurochemical/neuroendocrine signals associated with mating activate PR, resulting in the inhibition of chinning. In Experiment 1, we tested whether the PR antagonist, RU486 (20 mg, injected s.c. at -1 h, or at -7 h and +3 h relative to mating) prevented the post-mating inhibition of chinning in intact females. RU486 did not prevent the post-mating decline in chinning, indicating that PR activation associated with mating is not necessary for this effect. In Experiment 2, we used ovariectomized (OVX), estradiol benzoate (EB)-treated females to test the hypothesis that ovarian signaling is necessary for the post-mating inhibition of chinning. The acute inhibition of chinning occurred in OVX females, but the early inhibition was absent. We conclude that ovarian signaling is necessary for the early, but not acute, post-mating inhibition of chinning. The PR seems not to participate in either of these phases.

Maternal care of rabbits in the lab and on the farm: endocrine regulation of behavior and productivity.

Maternal behavior in rabbits has been well described in the wild, the laboratory, and the farm. Salient characteristics include: (a) the construction of a nest (inside an underground burrow or a box), composed of straw/grass and body hair and (b) the display of a single, brief (ca. 3 min) nursing bout per day. The onset and decline of nest-building in mid and late pregnancy are controlled by specific combinations of estradiol, progesterone, testosterone, and prolactin. Following parturition (kindling) does can mate and become pregnant again. Management strategies have been used on the farm to enhance productivity, impinging on specific reproductive processes (e.g., use of hormones to synchronize estrus, artificial insemination at kindling, doe-litter separation on specific days of lactation to increase sexual receptivity and fertility). Knowledge about the rabbit's reproductive physiology and behavior will be enriched by integrating research coming from the laboratory (where estrus, pregnant-only, or lactating-only animals are the main categories investigated) with studies performed on the farm or research station (where pregnant-lactating rabbits are prevalent). Similarly, the high productivity demands of modern rabbit farms will benefit from the information obtained in the laboratory where specific issues in reproductive neuroendocrinology are explored with methodologies that are not amenable for farm use.

Progesterone receptor activation signals behavioral transitions across the reproductive cycle of the female rabbit.

The female rabbit is an exceptional experimental model to define mechanisms by which progesterone (P) controls the expression of reproductive behaviors. In the rabbit, the rise in P levels during pregnancy inhibits estrous scent marking ("chinning"), stimulates the excavation of a nest burrow ("digging"), and primes behaviors later used for nest construction. The pre-parturient fall of P triggers the construction of a straw nest ("straw carrying") that is lined with hair that she pulls from her own body ("hair pulling"). These behaviors can be replicated in ovariectomized (ovx) females given a schedule of estradiol (E) and P that mimics hormone levels during pregnancy (E from days 0 to 4, E + P from days 5 to 17, E from days 18 to 27). We administered PR antagonists RU486 or CDB(VA)2914 to ovx female rabbits during either the initial (days 5-11) or late (days 12-17) phases of P treatment, to determine the role of PR activation in coordinating the expression of these behaviors. Both antiprogestins attenuated the P-mediated decline in chinning and increase in digging when administered during days 5-11. When given across days 12-17, both antiprogestins triggered an early decline in digging, the onset of nest building in some Ss, and the reinstatement of chinning. These results point to a central role of PR activation for establishing and maintaining the behavioral phenotype of pregnancy, and for the behavioral transition from pregnancy to estrus.

Intracerebroventricular injections of prolactin counteract the antagonistic effect of bromocriptine on rabbit maternal behaviour.

To investigate the participation of prolactin in nest-building and maternal behaviour in rabbits, we administered (from pregnancy day 26 to parturition) rabbit prolactin (rbPRL; or vehicle) intracerebroventricularly (i.c.v.) to primiparous animals injected with bromocriptine subcutaneously (s.c.). Control females (given vehicle s.c. and i.c.v.) built a maternal nest (of straw and body hair) in 77% of cases. This proportion decreased to 19% in the bromocriptine-only group (P < 0.05) and increased to 93% in the group given bromocriptine plus rbPRL (P > 0.05). Maternal behaviour (i.e. the adoption of a crouching posture over the litter inside the nest box) was expressed by 77% of control rabbits, 19% of bromocriptine-only animals (P < 0.05) and 57% of females given bromocriptine plus rbPRL (P > 0.05). Values of nonmaternal activities (i.e. scent-marking, ambulation in an open field) were similar among the three studied groups. These results suggest that prolactin, acting in late pregnancy, plays a major role in the stimulation of nest-building and maternal behaviour in rabbits.

Non-ligand activation of estrous behavior in rodents: cross-talk at the progesterone receptor.

Estrous behavior in rodents is triggered by the binding of progesterone (P) to its intracellular receptor (PR). Non-steroidal agents (i.e., gonadotropin-releasing hormone, noradrenaline, dopamine and others), acting at the membrane, can facilitate estrous behavior in estrogen-primed rats. This action is mediated through the generation of second messengers (cyclic AMP, cyclic GMP, calcium) which, in turn, phosphorylate through diverse kinase systems (protein kinases A, G or C) either the PR or associated effector proteins linking the PR to the trans-activation machinery. P or its metabolites also activate cyclic AMP-signaling pathways by acting directly on the membrane or by modulating neurotransmitter release. Molecular processes resulting from second messenger signaling pathways and those from the progesterone-RP interaction synergize to elicit a full behavioral response.

Immunocytochemical detection of progesterone receptor in the female rabbit forebrain: distribution and regulation by oestradiol and progesterone.

There is no information on the neuroanatomical distribution of the progesterone receptor (PR) in the rabbit. Therefore, we mapped the distribution of PR-immunoreactive cells in the forebrain of ovariectomized female rabbits. Vehicle-injected ovariectomized rabbits showed PR-immunoreactive cells only in the infundibular nucleus (IN) and nucleus X (lateral to the ventromedial hypothalamic nucleus). The injection of oestradiol benzoate (EB; 5 micro g/day for 5 days) increased the number of PR-immunoreactive cells in the IN and in three nuclei of the preoptic region (periventricular, medial, and principal). Abundant PR were also found in the paraventricular nucleus and nucleus X. Administration of progesterone (10 mg/day) for 3 days to EB-treated rabbits (a treatment that induces digging behaviour for the maternal nest and suppresses sexual receptivity and scent-marking) eliminated PR-immunoreactivity from all brain areas analysed except the IN. Thus, one-third of the number of cells seen in the ovariectomized + EB condition persisted in this region despite progesterone injections. Withdrawal of progesterone (and continuation of EB) for 5 (but not for 2) days (in a schedule similar to the one that induces straw-carrying and hair-pulling for the maternal nest) increased the number of PR-immunoreactive cells in all regions analysed. These results show that restricted regions of the female rabbit forebrain express abundant PR which are either: (i). up-regulated by oestradiol and down-regulated by progesterone; (ii). oestradiol-insensitive and down-regulated by progesterone; or (iii). insensitive to both oestradiol and progesterone.

Neuroendocrinology of maternal behavior in the rabbit.

Rabbit maternal behavior consists of building an underground nest of straw and body hair during late pregnancy and displaying, with circadian periodicity, a single 3-min nursing bout/day across lactation. Estrogen, androgen, progesterone, and prolactin regulate specific aspects of nest-building and promote the onset of maternal responsiveness. However, the maintenance of this behavior relies on stimuli from the litter: by preventing mother/young contact at parturition or during early lactation maternal responsiveness is altered or abolished. The brain areas controlling the expression of nest-building and nursing were investigated by implanting estradiol, locating the distribution of estrogen and prolactin receptors, quantifying the expression of immediate-early genes, and lesioning structures of the olfactory system. These studies revealed that: (a) estrogen receptor-alpha, alpha, present in the preoptic region, may mediate the stimulation of nest-building by estradiol; (b) prolactin binding sites, located mainly in periventricular structures, are more abundant in late pregnancy and early lactation; (c) the number of FOS-immunoreactive neurons increases in the lateral septum, but not in the mediobasal hypothalamus, following nursing; (d) the accessory olfactory bulb tonically inhibits the expression of maternal behavior because its removal promotes maternal responsiveness in virgins, which are otherwise unresponsive to daily pup exposure. In summary, rabbits rely on the same hormonal and extrahormonal factors that stimulate maternal behavior in other mammals, yet the way in which such factors promote elaborate nest-building and the unfailing display of circadian nursing is unique to rabbits and warrants future investigation.

Pharmacological evidence that prolactin acts from late gestation to promote maternal behaviour in rabbits.

We investigated the role of prolactin and suckling stimulation in the expression of maternal behaviour of primiparous rabbits. Bromocriptine (1 mg/kg/day), given to intact mothers across postpartum days 1-5, decreased serum concentrations of prolactin to undetectable levels, reduced crouching, and increased time inside the nest. Failure of maternal nest-building, provoked by bromocriptine injections from pregnancy day 26 to parturition or to postpartum day 5, correlated with a stronger reduction in crouching and an increased time inside the nest, measures of disturbed maternal behaviour, on postpartum days 3 and 5. Preventing suckling by thelectomy did not prevent prolactin release but reduced crouching incidence and increased the time spent inside the nest on postpartum days 3 and 5. Bromocriptine, injected in thelectomized mothers across postpartum days 1-5, further reduced the incidence of crouching and increased the time spent inside the nest on postpartum days 3 and 5. We conclude that prolactin acting prepartum facilitates maternal behaviour initiation in rabbits and, together with pup stimulation, maintains this behaviour across lactation.

Mating modifies c-fos expression in the brain of male and female rabbits.

Copulation in rabbits provokes behavioral and neuroendocrine changes in both sexes. To investigate if the activity of particular brain regions is modified accordingly we quantified, by the reverse transcription-polymerase chain reaction method, c-fos expression in the preoptic area, hypothalamus, hippocampus, and frontal cortex of male and female rabbits before mating, immediately afterwards, and 1 h later. Mating immediately increased c-fos expression in the hypothalamus of both sexes, the frontal cortex of females, and the preoptic area of males. c-fos expression did not change in the hippocampus after mating in either sex but decreased in the preoptic area of females following mating. Results show that mating provokes changes in brain activity, in a gender- and region-specific manner, which may underlie the behavioral and endocrine consequences of copulation in rabbits.

Immunocytochemical detection of estrogen and progesterone receptors in the rabbit submandibular gland.

Rabbit submandibular glands produce secretions involved in olfactory communication. The histology of these glands and their secretory activity are: sexually dimorphic; vary across the female reproductive cycle; and are modified by gonadectomy. This suggests that gonadal steroids regulate the structure and function of such glands. To further support this idea we assessed by immunocytochemistry the presence of estrogen and progesterone receptors in male and female rabbit submandibular glands. Immunoreactivity was detected only in the nucleus of acini cells. The number of estrogen receptor-immunoreactive cells/field varied among estrus (26 +/- 6; mean +/- S.E.), ovariectomized (19 +/- 2), and ovariectomized-estrogen-treated animals (13 +/- 3). Intact males showed a significantly smaller number of estrogen receptor-immunoreactive cells/field (12 +/- 1) than estrous females. Interestingly, progesterone receptor-immunoreactive cells were more abundant in estrous (32 +/- 7) than in ovariectomized animals (7 +/- 1). Estradiol benzoate (5 micrograms daily for 5 days) increased the number of progesterone receptor-immunoreactive cells/field in ovariectomized females (17 +/- 1). Intact males showed fewer progesterone receptor-immunoreactive cells/field (16 +/- 2) than estrous females. Results show that the rabbit submandibular gland is a target for estrogen and progesterone and support the idea that these hormones participate in regulating the physiology of this gland.

Indomethacin inhibits lordosis induced by ring A-reduced progestins: possible role of 3alpha-oxoreduction in progestin-facilitated lordosis.

Progestins with a delta-4-3-keto configuration bind to the progestin receptor (PR) and facilitate estrous behavior in estrogen-primed rats. Some ring A-reduced progestins [5alpha-dihydroprogesterone (alphaDHP), allopregnanolone, and epipregnanolone] are more potent estrus-inducing agents than progesterone when iv injected despite their lower affinity for the PR. Yet the estrus-inducing action of such progestins is reduced by the antiprogestin RU486, suggesting that binding to the PR is required for this effect. Because allo- and epi-pregnanolone are oxidized to alpha- and betaDHP, respectively, by 3alpha-hydroxysteroid oxo-reductase (3alphaHSOR), part of their estrus-inducing action may occur through the binding of such DHPs to the PR. Conversely, because 3alphaHSOR reduces alpha- and betaDHP to allo- or epi-pregnanolone, both of which exert membrane effects, the estrus-inducing effect of DHPs may involve actions independent of the PR. To test these possibilities we assessed the effect of indomethacin, a blocker of 3alphaHSOR, on the estrus-inducing action of such progestins. Because indomethacin also inhibits cyclooxygenases, we selected a dose and treatment schedule that does not interfere with prostaglandin-mediated brain processes (e.g., LHRH release). Indomethacin did not significantly modify the effect of progesterone or megestrol acetate on lordosis. Yet, it significantly reduced the action of all ring A-reduced progestins. Results suggest that: (a) oxidation is essential for lordosis facilitation by 3alpha-pregnanolones and (b) reduction of 3-keto progestins generates 3alpha-hydroxy metabolites which synergize with processes triggered by occupation of the PR by 3-keto progestins. The possible participation in this response of other events influenced by indomethacin (e.g., prostaglandin or melatonin synthesis) is discussed.

Ring A reductions of progestins are not essential for estrous behavior facilitation in estrogen-primed rats.

In Experiment 1 six dose levels (range 0.66-2000 microg) of progesterone (P) and two synthetic progestins with a double bond at C6: megestrol acetate (MA) and chlormadinone acetate (CA), which cannot be reduced at C5, were injected to estrogen-primed (2 microg estradiol benzoate 42 h earlier) ovariectomized (ovx) rats. The three progestins elicited significant lordosis and proceptive behaviors. Potency analysis showed that MA was the most potent progestin for stimulating estrous behavior, followed by P and CA. These results suggest that ring A reduction of progestins to 5alpha/5beta metabolites is not essential for the facilitation of estrous behavior in ovx estrogen-primed rats. Progestins with the 3-ketone group and a double bond at C4 can also be reduced at C3 to yield 3alpha-hydroxysteroid metabolites potentially capable of stimulating estrous behavior. In Experiment 2, the relevance of the formation of 3alpha-hydroxysteroid metabolites for estrous behavior facilitation was tested by concurrently injecting indomethacin (1.5 mg), a blocker of 3alpha-hydroxysteroid oxidoreductase, with 400 microg of P, MA, or CA to ovx estrogen-primed rats. Indomethacin failed to block the stimulatory effect of these progestins on estrous behavior. These results suggest that 3-ketosteroid reduction is also not essential for estrous behavior facilitation by progestins.

Hormones and external factors: are they "on/off" signals for maternal nest-building in rabbits?

Estradiol, progesterone, and prolactin regulate digging, carrying straw, and plucking hair for maternal nest-building in rabbits. To explore whether external factors also modulate this process, we assessed whether shaved pregnant rabbits with straw nests would collect their own, male, or synthetic hair for nest-building. Pregnant (but not estrous) does collected and used all hair types, indicating that hair-plucking can be bypassed and a nest constructed with "alternative" hair. Unshaved pregnant does with straw nests also collected synthetic hair, indicating that this behavior is not triggered by the absence of maternal hair. Yet, if hair-plucking/nest-building had occurred, hair-collecting was prevented, suggesting that an internally triggered "drive" was turned off by perceiving a built nest. When given only straw or hair, shaved pregnant does collected and used the material provided, indicating that nest-building is internally driven and accomplished by using any available elements. When given both materials, increasingly more shaved does built straw nests across prepartum days 7 to 2. Straw-carrying declined thereafter, suggesting that perceiving a straw nest limits the collection of such material. Hair-collecting was postponed until prepartum day 1 to postpartum day 2, indicating that: (a) mothers distinguish between straw and hair and (b) hormonal factors regulate the sequential selection of straw and then hair and when the change from straw to hair occurs. Maternal behavior was normal at parturition and for the next 4 days in a similar proportion of does among all experimental groups. We conclude that hormones and external factors regulate nest-building by acting as "on/off/on" signals.

Importance of mother/young contact at parturition and across lactation for the expression of maternal behavior in rabbits.

We prevented mother/pup contact at parturition or across early or midlactation to investigate the importance of such interaction for maintaining material behavior in rabbits. When pup contact was prevented across lactation Days 1-7 or 11-17 (by anesthetizing multiparous mothers during the oxytocin-induced milk letdown; Experiment 1), nursing incidence was reduced to 40% and 83%, respectively, on the day following anesthesia withdrawal. Both groups also showed a decreased milk output, long latencies to initiate nursing, and several entrances into the nest box not associated with nursing. In Experiment 2 we prevented mother/litter contact at parturition to determine the specific role of pup contact at this time. We found a reduction in the incidence of nursing on postpartum Day 1 from 80% (in control primiparous mothers) to 33%. By contrast, 100% of both deprived and control multiparous mothers displayed nursing on Day 1. These mothers also showed the unusual behaviors found in Experiment 1 and an extemporaneous nest-building. We conclude that: (a) mother/young contact at parturition is crucial for establishing maternal responsiveness in primiparous does, (b) the experience acquired by raising a previous litter allows the retention of maternal responsiveness despite a lack of pup contact at parturition, (c) maternal responsiveness is maintained across early lactation by daily interaction with pups, and (d) interaction with pups across midlactation allows the finely tuned display of maternal behavior.

Progesterone receptor participates in the stimulatory effect of LHRH, prostaglandin E2, and cyclic AMP on lordosis and proceptive behaviours in rats.

We explored the effect of the antiprogestin RU486 on the estrous behaviour (lordosis and proceptivity) induced in sexually experienced ovariectomized oestrogen primed rats by: 5 microg luteinizing hormone-releasing hormone (LHRH), 100 microg prostaglandin E2 (PGE2), or 2 mg dibutyryl cyclic AMP (db cAMP). Pretreatment with 5 mg RU 486 (but not with vehicle) 60 min before the injection of the above-mentioned agents significantly decreased both lordosis and proceptive behaviours normally induced by such agents. Results suggest that the estrus-inducing action of LHRH, PGE2 and db cAMP occurs through the activation of the progesterone receptor.

Estradiol, progesterone, and prolactin regulate maternal nest-building in rabbits.

Maternal nest-building in rabbits, expressed across the last third of pregnancy, consists of: digging a burrow, collecting straw and shaping it into a nest inside the burrow, plucking body hair and lining the straw nest with it. The sequential expression of these activities is correlated with specific changes in the plasma concentration of estradiol, progesterone (P), and prolactin (PRL). To further substantiate the participation of these hormones in the control of maternal nest-building we explored in ovariectomized (ovx) New Zealand white rabbits the capacity of several combinations of such hormones to stimulate digging, straw-carrying, and hair-pulling. Does given estradiol benzoate (EB; 5 micrograms/day from days 3 to 21) plus P (2 or 10 mg/day from days 4 to 16) dug into a substrate from the fourth day of the P treatment until withdrawal of this hormone. The intensity of this effect was greater in the group treated with the high dose of P. Straw-carrying and hair-pulling occurred after P withdrawal in a dose-response way. Food intake, which declines in pregnant females shortly before parturition, decreased to the same extent in both groups of ovx EB-treated does after P withdrawal. A significant increase in PRL plasma levels was observed on day 9 in does given EB plus 2 mg P/day and at two days following P withdrawal in does given EB plus 10 mg P/day. When such ovx EB/P-treated does were given bromocriptine to block PRL release (1 or 3 mg/Kg/day, from days 11 to 21) the expression of digging was unmodified. By contrast, bromocriptine abolished the display of straw-carrying and hair-pulling, and also prevented the decline in food intake normally following P withdrawal. The addition of ovine PRL to ovx EB/P-treated does given bromocriptine reduced the expression of digging, did not restore straw-carrying or hair-pulling, and provoked a sharp decline in food intake. The possible mechanisms of interaction between PRL and steroid hormones for the regulation of specific aspects of the pregnant doe's physiology and behavior are discussed.

Oxytocin and vasopressin immunoreactivity in rabbit hypothalamus during estrus, late pregnancy, and postpartum.

Mother rabbits construct an elaborate maternal nest before parturition and display a single, brief, daily nursing bout throughout lactation. These features present a unique model for investigating the relevance of changes in neuroendocrine secretion associated with pregnancy and parturition for the regulation of maternal behavior. In the present study we analyzed changes in the location, somal size, and number of oxytocin (OT)- and arginine vasopressin (AVP)-immunoreactive (IR) neurons in the hypothalamus of rabbits in estrus, late pregnancy (day 29), and postpartum day 1. From estrus to late pregnancy, the number of OT-IR neurons increased in the scattered cell groups located in the lateral hypothalamic area (LHA), but not in the magnocellular nuclei, i.e., paraventricular nucleus (PVN) and supraoptic nucleus (SON). On postpartum day 1 the increase in the number of OT-IR neurons was sustained in the LHA and became apparent also in the main body of the PVN, in which the number of OT-IR neurons doubled. Increases in the somal size of OT-IR cells were seen in all three nuclei only on postpartum day 1. No OT-IR cells were found in the suprachiasmatic nucleus (SCN). From late pregnancy and into postpartum day 1 increases in the somal size of AVP-IR neurons were detected in the PVN, SON, and LHA but not in the SCN. The number of AVP-IR neurons increased between late pregnancy and postpartum day 1 in the SON only. The changes observed in OT and AVP expression in specific hypothalamic nuclei may be related to specific somatic and behavioral events occurring around the time of parturition, e.g., nest-building, maintenance of homeothermy, elevation of blood volume, and nursing in mother rabbits.