Rosario Chirino

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.

Visceral and postural reflexes evoked by genital stimulation in urethane-anesthetized female rats

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

Removal of the accessory olfactory bulbs promotes maternal behavior in virgin rabbits

Virgin rabbits exposed to foster pups for 14 days did not show maternal responsiveness. However, surgical removal of the accessory olfactory bulbs (AOB) activated maternal responsiveness (crouching over the litter inside the nest box for about 3min, which is the normal duration of a nursing bout) in 37% of virgin rabbits (P < 0.007). This behavior appeared abruptly and was first observed between days 3 and 13 of pup exposure. This variation in the latency to respond maternally was not related with the number of sniffings or entrances into the nest box displayed by a female on the days that preceded crouching over the litter. Maternal responsiveness was not observed in any AOB-lesioned animals that were also ovariectomized (P < 0.02 versus AOB-lesioned with ovaries). These results indicate a tonic inhibitory action of the AOB over the expression of maternal behavior in virgin rabbits and a stimulation of maternal responsiveness by ovarian hormones following AOB lesions.

Sex steroid regulation of chin-marking behavior in male New Zealand rabbits

Chin-marking behavior (chinning) was evaluated daily in nine intact adult male rabbits. All subjects (Ss) displayed chinning (mean of means +/- SE = 61 +/- 7 marks/10 min) but the frequency of this behavior varied largely across them (range of mean chinning frequency = 19-84 marks/10 min). Chinning frequency showed abrupt variations at intervals of 2-3 days, but periodogram analysis did not reveal the existence of an endogenous rhythm in this behavior. Castration significantly decreased (mean of means +/- SE = 29 +/- 9 marks/10 min; p < 0.01). but did not suppress chinning. Testosterone propionate (TP; 1 mg/day for 16 days) restored chinning in castrated Ss to slightly below precastration levels (mean +/- S.E. V 53 +/- 13 marks/10 min). The daily administration of 1 microgram estradiol benzoate (EB) plus 1 mg dihydrotestosterone propionate (DHTP) stimulated chinning within 2 days (mean increase = 147%; p < 0.005). DHTP (1 mg/day) given alone stimulated chinning only after 11 days of treatment (mean increase = 475%; p < 0.01). At higher doses, both DHTP (10 mg/day) and EB (10 or 50 micrograms/day) stimulated chinning by 450%, 80%, and 100%, respectively, over baseline values. Results indicate that chinning largely depends on testicular steroids. Androgen receptor occupation by T or DHT, which is enhanced by E, optimally activates chinning.

Forebrain implants of estradiol stimulate maternal nest-building in ovariectomized rabbits.

In rabbits, estradiol and progesterone (P) stimulate digging a maternal burrow while P withdrawal promotes straw-carrying. To investigate where such hormones act to regulate those activities, ovariectomized rabbits were implanted with estradiol benzoate (EB; Experiment 1) in the nucleus accumbens (ACC), the principal nucleus of the medial preoptic area or the dorsal hippocampus. Implants were combined with s.c. P injections. In Experiment 2, P (in crystals or dissolved in oil) was implanted in the same regions as in Experiment 1, combined with s.c. injections of EB. Implants of EB into the ACC or MPOA-bed nucleus of the stria terminalis (BNST) stimulated significant digging across the period of P injections in 72% and 67% of females, respectively. Neither EB implants in the hippocampus nor cholesterol implants in the MPOA-BNST were effective in eliciting digging. P withdrawal provoked a rapid decline of digging in all animals; it also stimulated straw-carrying in 53% of females implanted with EB in the MPOA-BNST. P implants failed to stimulate digging in most females injected with EB. Removal of P crystals did not promote straw-carrying. Results support an action of estradiol on the ACC and MPOA-BNST to promote digging while only the MPOA-BNST is involved in stimulating straw-carrying. The failure of P implants to stimulate digging or straw-carrying in EB-treated females suggests that the stimulation of other or additional brain areas by P is necessary to fully activate maternal nest-building.

Lesion of the main olfactory epithelium facilitates maternal behavior in virgin rabbits

Maternal behavior is induced in virgin female rabbits (normally unresponsive to foster pups) by removing the accessory olfactory bulbs. To determine if the main olfactory system (MOS) plays a similar inhibitory role in the present work we investigated the effect of lesioning the olfactory epithelium with a ZnSO(4) spray on the facilitation of maternal behavior in New Zealand white virgin rabbits. Four days after the chemical lesion 40% of females showed behaviors indistinguishable from those of normal mothers, i.e.: rapid entrance into the nest box containing the pups, adoption of a crouching posture over them, acceptance of suckling, and exit from the nest box after ca. 3min. The proportion of females showing these behaviors rose to 70% by day 14 post-lesion. Ovariectomized rabbits sprayed with ZnSO(4) or animals sprayed with NaCl did not behave maternally. ZnSO(4) also provoked a transient reduction in olfactory perception: before the lesion animals from all groups directed significantly more sniffs to a flask containing male urine than to one containing water. This difference was abolished in ZnSO(4)-sprayed females (intact and ovariectomized) for 3-6 days post-lesion and was re-established by 7-9 days. NaCl did not provoke such transitory hyposmia. ZnSO(4) lesions did not provoke malaise in the animals, as determined by food intake and the frequency of scent-marking and ambulation. Results suggest that olfactory cues from the pups are aversive to virgin rabbits and that a transitory reduction in their perception (accompanied by the action of ovarian secretions) is enough to facilitate maternal responsiveness.

Motherhood and nursing stimulate c-FOS expression in the rabbit forebrain.

Mother rabbits nurse once daily with circadian periodicity. The authors investigated brain structures involved in regulating this activity by quantifying c-FOS-immunoreactive (IR) cells in the forebrain of: (1) mothers killed on postpartum Day 1 (PPD 1) after nursing (Group 1) or not given pups (Group 2); (2) mothers killed on PPD 7 after nursing (Group 3) or not given pups on such day (Group 4); (3) unmated virgins (Group 5). Groups 1 through 4 showed similar numbers of c-FOS-IR cells in the preoptic area, an amount around three to fourfold larger than that found in virgins. Nursing increased, on PPD 1 and 7, c-FOS-IR cell number in the lateral septum and paraventricular and supraoptic nuclei. No differences were seen among Groups 1 through 5 in the suprachiasmatic nucleus. In the ventromedial hypothalamus virgins had more c-FOS-IR cells compared with Groups 1 and 2. Results suggest that specific forebrain structures participate in regulating particular aspects of rabbit maternal behavior: the POA and LS seem associated with the establishment of motherhood and the magnocellular nuclei with the occurrence of milk letdown.

Effect of forebrain implants of testosterone or estradiol on scent-marking and sexual behavior in male and female rabbits

Chinning consists of rubbing the chin against an object, thereby depositing secretions from the submandibular glands. As mating, chinning is stimulated in male and female rabbits by testosterone and estradiol, respectively. To investigate the brain sites where steroids act to stimulate chinning and mating we implanted into the ventromedial hypothalamus (VMH) or the medial preoptic area (MPOA) of gonadectomized male and female rabbits testosterone propionate (TP; males) or estradiol benzoate (EB; females) and quantified chinning and sexual behavior. EB implants into the VMH or MPOA reliably stimulated chinning in females. Most of those implanted into the VMH and around half of the ones receiving EB into MPOA or diagonal band of Broca (DBB) showed lordosis. Chinning, but not sexual behavior, was stimulated in males by TP implants into the MPOA or DBB. Neither chinning nor mounting were reliably displayed by males following TP implants into the VMH. Results indicate that, in females, the VMH is an estrogen-sensitive brain area that stimulates both chinning and lordosis while the MPOA seems to contain subpopulations of neurons involved in either behavior. In males, androgen-sensitive neurons of the MPOA, but not the VMH, are involved in chinning stimulation but it is unclear if these areas also participate in the regulation of copulatory behavior.

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.