María José Zuluaga

Experimental anxiety in the black and white model in cycling, pregnant and lactating rats

This study demonstrates changes in experimental anxiety assessed in the black and white paradigm during various reproductive states of female rats. Low levels of experimental anxiety were observed during late proestrus and on day 17 of gestation, stages related to high progesterone (P) levels. In estrus, metestrus, diestrus and on day 21 of gestation, stages characterized by low P concentrations, high levels of experimental anxiety, similar to those exhibited by ovariectomized females, were found. No changes in experimental anxiety were observed on day 8 of lactation compared to ovariectomized females. These data are discussed from the standpoint of the putative anxiolytic-like effect of progestins.

Maternal condition reduces fear behaviors but not the endocrine response to an emotional threat in virgin female rats.

Lactating dams and maternal virgin females are less fearful in behavioral tests compared with non-maternal animals, suggesting that maternal condition per se reduces the negative value of threatening stimuli. In addition, lactating females exhibit a diminished hypothalamic-pituitary-adrenal response to potential environmental threats. Can the maternal condition, independently of the endocrine profile of lactation, promote a reduction in the behavioral as well as in the endocrine response to an emotional stressor? To answer this question, anxiety-related and fear behaviors as well as the levels of corticosterone were evaluated in response to a bright-lit open field-loud noise model in maternal and non-maternal non-ovariectomized virgin females and lactating dams in the presence of the pups. Maternal animals, both lactating and virgin, presented an increased exploration of the bright-lit open field and a significant reduction of fear behaviors, indicated by the decreased flight and immobility responses to the subsequent activation of a loud noise, in comparison to non-maternal virgins. Interestingly, maternal virgin females, as non-maternal rats, showed high corticosterone plasma levels, in contrast to the lower endocrine response exhibited by lactating dams when confronted to this threat. Present results suggest that maternal condition allows females to take risks when caring for their young, a behavioral strategy that is independent of the reduced hypothalamic-pituitary-adrenal axis response characteristic of lactation. This evidence points towards a clear dissociation in the mechanisms regulating behavioral and endocrine responses to emotional stressors during motherhood.

Social aversive stimuli presented to the mother produce the precocious expression of fear in rat pups

During the stress hypo-responsive period, rat pups do not display fear responses toward adult males, yet they exhibit distress behavior in isolation. Since the mother modulates her offsprings affective development, we hypothesized that by altering the mothers behavior, a prolonged stressful situation would modify the ontogeny of the fear responses and distress behaviors in pups. Therefore, we repeatedly exposed the mother-litter dyad to different socially stressful stimuli and subsequently evaluated in 8-day-old pups their fear responses toward an anesthetized male, as well as their distress behavior in isolation. Our results show that repeated exposure to unfamiliar males and females, which altered maternal behavior by eliciting aggression in the mother, was associated with the precocious fear responses in pups, though without altering their distress behavior in isolation. We propose that the mother, as the principal mediator of environmental influences, provokes the precocious expression of fear in pups through alterations in her maternal behavior.

Maternal and affective behaviors of lactating rats reared in overlapping litters

Postpartum mating in rats gives rise to complex family units consisting of the mother and two overlapping litters. As a consequence, newborn pups of the second litter, since the moment they are born, acquire experience not only from interaction with the mother and age-matched littermates but also from interaction with older siblings. Newborn pups reared in overlapping litters (OLs) receive a different pattern of maternal stimulation compared to those reared in single litters (SL: one litter of same aged pups), as the mothers reduce some maternal behavior components and juvenile pups from the first litter develop maternal behavior. Since there is strong evidence showing that variations in maternal behavior are transmitted throughout generations, we hypothesized that the altered pattern of maternal stimulation received by OL reared females would modify their behavior during motherhood. To test this hypothesis maternal behavior, maternal aggression and experimental anxiety of dams reared under OL and SL conditions during the first postpartum week were compared. No differences were found between the groups in their maternal behavior and aggression. This result may be explained by the maternal behavior of the juveniles that could compensate for the deficits in the caregiving behaviors received by OL litters. However, a subtle temporal reorganization of the licking behavior was found in OL reared mothers, together with an increased anxiety-related behavior in the plus maze test. These results suggest dissociation in the effects provoked by early environmental alterations on different behavioral systems, and more importantly, that independently of their early family composition, both groups can cope effectively with the changing demands of the pups.

Different chemical fractions of fetal fluids account for their attractiveness at parturition and their repulsiveness during late-gestation in the ewe.

We have investigated whether the chemical components of fetal fluids (FFs), which elicit repulsion in late gestating ewes, are also those responsible for the attractiveness of fetal fluids at parturition. An aqueous fraction of FFs (A1), obtained after extraction with hexane, was tested for repulsion in late-pregnant ewes and for attraction at parturition. We also investigated if the repulsive and attractive characteristics of this A1 fraction were maintained after an additional extraction with dichloromethane (DCM, CH(2)Cl(2)) that produced two more fractions (aqueous/high polarity: A2 and dichloromethane/medium polarity: DCM). Thus, late-pregnant ewes were tested for repulsion of aqueous extracts of FFs (A1, A2 and DCM fractions) in a two-choice test of food preference, whereas parturient ewes were tested for attraction toward these same fractions in a two-choice test of licking warm spongy cloths. The A1 fraction was repulsive to late-pregnant ewes and attractive to parturient females. In contrast, neither the A2 nor the DCM fractions were repulsive to late-pregnant ewes, whereas both fractions were attractive to parturient ones. The discordance between the repulsive and attractive properties of the A2 and DCM fractions suggests that the attractiveness of FFs for parturient ewes and its repulsiveness for females outside the peripartum period depend on mixtures of substances that are at least partially different. Some compounds with high and medium polarity in the A2 and DCM fractions would act synergistically to generate the repulsiveness of FFs, whereas both high and medium polarity compounds can evoke attraction independently of each other.

The Expression of Hypoxia-Induced Gene 1 (Higd1a) in the Central Nervous System of Male and Female Rats Differs According to Age

HIGD1A (hypoxia-induced gene domain protein-1a), a mitochondrial inner membrane protein present in various cell types, has been mainly associated with anti-apoptotic processes in response to stressors. Our previous findings have shown that Higd1a mRNA is widely expressed across the central nervous system (CNS), exhibiting an increasing expression in the spinal cord from postnatal day 1 (P1) to 15 (P15) and changes in the distribution pattern from P1 to P90. During the first weeks of postnatal life, the great plasticity of the CNS is accompanied by cell death/survival decisions. So we first describe HIGD1A expression throughout the brain during early postnatal life in female and male pups. Secondly, based on the fact that in some areas this process is influenced by the sex of individuals, we explore HIGD1A expression in the sexual dimorphic nucleus (SDN) of the medial preoptic area, a region that is several folds larger in male than in female rats, partly due to sex differences in the process of apoptosis during this period. Immunohistochemical analysis revealed that HIGD1A is widely but unevenly expressed throughout the brain. Quantitative Western blot analysis of the parietal cortex, diencephalon, and spinal cord from both sexes at P1, P5, P8, and P15 showed that the expression of this protein is predominantly high and changes with age but not sex. Similarly, in the sexual dimorphic nucleus, the expression of HIGD1A varied according to age, but we were not able to detect significant differences in its expression according to sex. Altogether, these results suggest that HIGD1A protein is expressed in several areas of the central nervous system following a pattern that quantitatively changes with age but does not seem to change according to sex.