Optimal early development in most species is dependent upon a stable relationship between the mot... more Optimal early development in most species is dependent upon a stable relationship between the mother and her infant. The research described here focuses on the reciprocal nature of this dyad in rodents and humans, with respect to the regulation of responsiveness to stress in both mother and offspring. Dietary influences are critical not only to regulate infant growth but also to modulate the response of the neuroendocrine system to stress and, possibly, to influence some aspects of brain development. In particular, we discuss the role of leptin, a protein produced in the adipose tissue and present in maternal milk, that reduces responses to stress in the infant. We suggest that leptin acts on both central (hypothalamus and hippocampus) and peripheral (pituitary, adrenal gland) targets in the infant to reduce exposure to glucocorticoids and enhance hippocampal development during a sensitive period of brain development. There is also evidence to support the reverse regulatory influenc...
Chronic exposure to high circulating glucocorticoid or ghrelin concentrations increases food inta... more Chronic exposure to high circulating glucocorticoid or ghrelin concentrations increases food intake, weight gain and adiposity, suggesting that ghrelin could contribute to the metabolic effects of chronic glucocorticoids. In male mice, however, blocking ghrelin receptor (GHSR) signaling increased the weight gain and adiposity induced by chronic corticosterone (CORT), rather than attenuating them. In the current study, we investigated the role of GHSR signaling in the metabolic effects of chronic exposure to high circulating CORT in female mice. To do this, female WT and GHSR KO mice were treated with either CORT in a 1% ethanol (EtOH) solution or 1% EtOH alone in their drinking water for 32 days (n = 5–8/group). Body weight, food, and water intake as well as vaginal cyclicity were assessed daily. As expected, CORT treatment-induced significant increases in body weight, food intake, adiposity and also impaired glucose tolerance. In contrast to results observed in male mice, WT and GH...
Most psychiatric disorders are characterized by deficits in the ability to interact socially with... more Most psychiatric disorders are characterized by deficits in the ability to interact socially with others. Ghrelin, a hormone normally associated with the regulation of glucose utilization and appetite, is also implicated in the modulation of motivated behaviors including those associated with food and sex rewards. Here we hypothesized that deficits in ghrelin receptor (growth hormone secretagogue receptor; GHSR) signaling are also associated with deficits in social motivation in male mice. To test this hypothesis, we compared social motivation in male mice lacking GHSR or mice treated with the GHSR antagonist JMV2959 with that of WT or vehicle-treated mice. GHSR signaling in dopamine cells of the ventral tegmental area (VTA) has been implicated in the control of sexual behavior, thus we further hypothesized that GHSR signaling in the VTA is important for social motivation. Thus, we conducted studies where we delivered JMV2959 to block GHSR in the VTA of mice, and studies where we re...
The ability of rat dams to cope with food restriction as well as a concurrent lactation and pregn... more The ability of rat dams to cope with food restriction as well as a concurrent lactation and pregnancy was examined by varying the duration, time of onset and severity of the food restriction schedule. Groups of females impregnated in the postpartum estrus and nursing 8 pups were either allowed to feed ad lib, given 60% of an ad lib ration from days 1-7, 1-14, 8-14 or 15-21 postpartum or 40% of an ad lib ration from days 1-7. Food restriction regimens that extended through the estimated time of implantation for females nursing 8 pups (days 1-14 and 8-14) resulted in a prolongation of pregnancy. Food restriction in the first week postpartum did not result in a prolongation of pregnancy and did not affect the number of females carrying the pregnancy to term. Dams food restricted in the third week postpartum had pregnancies of the same length as ad lib females but a smaller proportion of this group gave birth to a second litter.
Blocking nitric oxide (NO) production, by 3rd ventricle administration of a nitric oxide synthase... more Blocking nitric oxide (NO) production, by 3rd ventricle administration of a nitric oxide synthase (NOS) inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME; 250 microg/5 microl, postpartum [pp]) decreased milk ejections in Day 10 pp rats. On Day 4 pp, L-NAME treatment eliminated pup retrieval and at both stages of lactation suppressed maternal aggression. Fewer rats treated with L-NAME on Day 10 pp retrieved 4-day-old pups than controls, although all nursed older litters. Following exposure to a mobile intruder, Fos expression was lower in the medial preoptic area and the bed nucleus of the stria terminalis in L-NAME-treated rats than in controls but was lower in the medial amygdala only following exposure to an anaesthetized intruder. Thus, the elevated levels of NO observed in lactation may contribute to the mechanism(s) that mediate maternal behavior and aggression.
The mother-infant interaction that occurs immediately postpartum period has important consequence... more The mother-infant interaction that occurs immediately postpartum period has important consequences including changes in protein expression in the astrocytes within cingulate cortex area 2 (Cg2). Because increased expression of basic fibroblast growth factor (bFGF) and glial fibrillary acidic protein (GFAP) has been associated with increased neuronal activity we used immediate early gene expression to evaluate the effects of parturition plus interaction with pups during and immediately after delivery to investigate patterns of neural activation in a number of brain areas including Cg2. In the first experiment, we compared levels of immunoreactivity of two immediate early genes: c-Fos and egr-1, among three groups of animals: Cycling, Late Pregnant, and rats perfused 1.5 h postpartum. The average number of Fos and egr-1-ir cells/section were calculated for the Parietal Cortex (ParCx), nucleus accumbens (NAc), medial amygdala (MEA), medial preoptic area (MPOA), supraoptic area (SON) an...
The nutritional needs of developing young place a large energetic demand on lactating females. In... more The nutritional needs of developing young place a large energetic demand on lactating females. In this paper some of the mechanisms through which lactating rats meet these demands are described. Emphasis is placed on the mechanisms that drive increased food intake in suckled rats that are independent of milk delivery and negative energy balance. Evidence is presented to suggest that prolactin (PRL), released from the pituitary in response to suckling stimulation, acts centrally to stimulate food intake during lactation. Brain areas in which PRL may act to stimulate food intake as well as its interactions with other hormones and neuropeptides involved in energy balance are discussed.
The hypoalgesic effect of morphine in lactating rats was assessed using the hot-plate test. At mi... more The hypoalgesic effect of morphine in lactating rats was assessed using the hot-plate test. At midlactation (days 12 and 18 postpartum) females nursing litters of 8 pups were less responsive to the hypoalgesic effect of morphine than ovariectomised or cycling females and females on days 6 or 24 of lactation. Subsequent studies showed that the hypoalgesic response to morphine was inhibited in lactating rats at a number of time points after drug administration and across a variety of doses. This effect was not dependent on milk delivery but was dependent on the hormonal state of the female since separation of dams and their litters for 96 h was sufficient to reinstate the response to morphine if it resulted in a reappearance of vaginal estrus.
Oxytocin (OT; Cys-Tyr-Ile-Gln-Asn-Cys-Pro-leu-Gly), a posterior pituitary peptide hormone, is cha... more Oxytocin (OT; Cys-Tyr-Ile-Gln-Asn-Cys-Pro-leu-Gly), a posterior pituitary peptide hormone, is characterized by a Cys1-Cys6 disulfide bond in its stable, isolated state. This paper describes a simple, one-step method for the production of OT in its reduced, dithiol form (OT dithiol), free of reducing agent. The effects of temperature, pH, and metal-ion chelators on the autoxidation of OT dithiol were examined to establish if this form is likely to persist under biological conditions. It was found that OT dithiol has a half-life of 1.8h with respect to reformation of OT disulfide at 37 degrees C and pH 6.9 in the presence of the copper chelators, DTPA and neocuproine. S-Nitrosation of OT dithiol by acidified nitrite at pH 3.0 was examined by absorption spectroscopy and HPLC-UV-MS, which revealed that both singly and doubly S-nitrosated OT are formed. These results suggest novel chemical aspects to OT signaling, the biological implications of which are discussed here.
Blunted neuroendocrine responses to stress are reported in lactating females after exposure to va... more Blunted neuroendocrine responses to stress are reported in lactating females after exposure to various stressors. However, many of the stimuli used in these studies have little ethological relevance for maternal protection of the litter in a threatening environment. The question that arises is whether the relevance of the stressor to the infant is critical in the 'gating' of the neuroendocrine response. We hypothesized that the presence of pups with their mothers at the time of exposure to an intruder or a predator odour is an effective way to increase the emotional salience of the psychological stressor, thus eliminating the stress hyporesponsiveness in lactating females. We first compared neuroendocrine responses [corticotropin-releasing factor (CRF) mRNA in the paraventricular nucleus of the hypothalamus (PVN) and central nucleus of the amygdala (CeA), plasma adrenocorticotropic hormone (ACTH) and corticosterone] between early (EL, PPD3-5), late (LL, PPD 15) lactating and virgin (V) females to a male intruder in the home cage. We next investigated the effect of pups' presence at the time of stressor exposure on the magnitude of the hormonal response to a male intruder in the home cage or to a predator odour (fox urine) in a novel environment. In the male intruder paradigm, levels of CRF mRNA expression in the PVN and CeA were lower in LL compared to EL or V females and plasma ACTH and corticosterone secretion was not as elevated in LL compared to EL females. Aggression towards the intruder was high in EL females in the presence of their pups and a positive correlation was found with the integrated ACTH response. Aggression rapidly declined after pup separation (2.5 h or 48 h) or in LL nursing females. In EL females, the presence of the pups with their mothers (EL + pups) at the time of stress significantly increased plasma ACTH and corticosterone responses to either male intruder or predator odour compared to EL females without their pups for 2.5 h or 48 h (EL - pups). Plasma ACTH response to fox urine in EL + pups females was comparable to that of virgin females, suggesting that increasing the salience of emotionally relevant stimuli by keeping the pups present in the cage could eliminate the hyporesponsiveness detected for EL females without their pups. These studies indicate the critical role of the pups in modulating the maternal response to stressors that represent a threat for the litter. We hypothesize that the amygdala, because of its ability to process olfactory stimuli and stimuli with affective properties, might play an essential role in 'gating' the neuroendocrine response to stress during lactation.
We investigated the contribution of prolactin and oxytocin to the increase in staining for NADPH-... more We investigated the contribution of prolactin and oxytocin to the increase in staining for NADPH-d and oxytocin mRNA in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) observed at the end of pregnancy, or following a steroid-priming regimen that mimics the hormonal profile of late pregnant females. Ovariectomized rats received chronic implants of silastic capsules containing oestrogen and progesterone followed by progesterone removal. In experiment 1, oxytocin antagonist (OTA) was administered to rats to investigate whether intranuclear oxytocin release was necessary for NADPH-d staining. In experiments 2a and b, rats received concurrent treatment with bromocryptine (0.5 mg/day) to suppress endogenous prolactin release, and either systemic prolactin (0.5 mg once daily), or prolactin (2 micro g/ micro l), or vehicle infused twice a day into the third ventricle, or chronic oxytocin infusion (24 ng/day) for 3 days following progesterone removal. Brains were then processed for NADPH-d histochemistry. In experiment 3, the interaction of prolactin and oxytocin on oxytocin mRNA within the SON and PVN was examined. NADPH-d staining in the SON and PVN was reduced by the highest dose of the OTA, and by bromocryptine treatment. Central prolactin and oxytocin replacement completely restored NADPH-d staining in bromocryptine-treated rats. Finally, both bromocryptine and the OTA suppressed oxytocin mRNA expression and prolactin replacement restored expression levels to that of controls. Together, these data suggest that the increased capacity to produce nitric oxide in the SON and PVN during late pregnancy is dependent on prolactin stimulating oxytocin gene mRNA and hence intranuclear oxytocin release.
Optimal early development in most species is dependent upon a stable relationship between the mot... more Optimal early development in most species is dependent upon a stable relationship between the mother and her infant. The research described here focuses on the reciprocal nature of this dyad in rodents and humans, with respect to the regulation of responsiveness to stress in both mother and offspring. Dietary influences are critical not only to regulate infant growth but also to modulate the response of the neuroendocrine system to stress and, possibly, to influence some aspects of brain development. In particular, we discuss the role of leptin, a protein produced in the adipose tissue and present in maternal milk, that reduces responses to stress in the infant. We suggest that leptin acts on both central (hypothalamus and hippocampus) and peripheral (pituitary, adrenal gland) targets in the infant to reduce exposure to glucocorticoids and enhance hippocampal development during a sensitive period of brain development. There is also evidence to support the reverse regulatory influenc...
Chronic exposure to high circulating glucocorticoid or ghrelin concentrations increases food inta... more Chronic exposure to high circulating glucocorticoid or ghrelin concentrations increases food intake, weight gain and adiposity, suggesting that ghrelin could contribute to the metabolic effects of chronic glucocorticoids. In male mice, however, blocking ghrelin receptor (GHSR) signaling increased the weight gain and adiposity induced by chronic corticosterone (CORT), rather than attenuating them. In the current study, we investigated the role of GHSR signaling in the metabolic effects of chronic exposure to high circulating CORT in female mice. To do this, female WT and GHSR KO mice were treated with either CORT in a 1% ethanol (EtOH) solution or 1% EtOH alone in their drinking water for 32 days (n = 5–8/group). Body weight, food, and water intake as well as vaginal cyclicity were assessed daily. As expected, CORT treatment-induced significant increases in body weight, food intake, adiposity and also impaired glucose tolerance. In contrast to results observed in male mice, WT and GH...
Most psychiatric disorders are characterized by deficits in the ability to interact socially with... more Most psychiatric disorders are characterized by deficits in the ability to interact socially with others. Ghrelin, a hormone normally associated with the regulation of glucose utilization and appetite, is also implicated in the modulation of motivated behaviors including those associated with food and sex rewards. Here we hypothesized that deficits in ghrelin receptor (growth hormone secretagogue receptor; GHSR) signaling are also associated with deficits in social motivation in male mice. To test this hypothesis, we compared social motivation in male mice lacking GHSR or mice treated with the GHSR antagonist JMV2959 with that of WT or vehicle-treated mice. GHSR signaling in dopamine cells of the ventral tegmental area (VTA) has been implicated in the control of sexual behavior, thus we further hypothesized that GHSR signaling in the VTA is important for social motivation. Thus, we conducted studies where we delivered JMV2959 to block GHSR in the VTA of mice, and studies where we re...
The ability of rat dams to cope with food restriction as well as a concurrent lactation and pregn... more The ability of rat dams to cope with food restriction as well as a concurrent lactation and pregnancy was examined by varying the duration, time of onset and severity of the food restriction schedule. Groups of females impregnated in the postpartum estrus and nursing 8 pups were either allowed to feed ad lib, given 60% of an ad lib ration from days 1-7, 1-14, 8-14 or 15-21 postpartum or 40% of an ad lib ration from days 1-7. Food restriction regimens that extended through the estimated time of implantation for females nursing 8 pups (days 1-14 and 8-14) resulted in a prolongation of pregnancy. Food restriction in the first week postpartum did not result in a prolongation of pregnancy and did not affect the number of females carrying the pregnancy to term. Dams food restricted in the third week postpartum had pregnancies of the same length as ad lib females but a smaller proportion of this group gave birth to a second litter.
Blocking nitric oxide (NO) production, by 3rd ventricle administration of a nitric oxide synthase... more Blocking nitric oxide (NO) production, by 3rd ventricle administration of a nitric oxide synthase (NOS) inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME; 250 microg/5 microl, postpartum [pp]) decreased milk ejections in Day 10 pp rats. On Day 4 pp, L-NAME treatment eliminated pup retrieval and at both stages of lactation suppressed maternal aggression. Fewer rats treated with L-NAME on Day 10 pp retrieved 4-day-old pups than controls, although all nursed older litters. Following exposure to a mobile intruder, Fos expression was lower in the medial preoptic area and the bed nucleus of the stria terminalis in L-NAME-treated rats than in controls but was lower in the medial amygdala only following exposure to an anaesthetized intruder. Thus, the elevated levels of NO observed in lactation may contribute to the mechanism(s) that mediate maternal behavior and aggression.
The mother-infant interaction that occurs immediately postpartum period has important consequence... more The mother-infant interaction that occurs immediately postpartum period has important consequences including changes in protein expression in the astrocytes within cingulate cortex area 2 (Cg2). Because increased expression of basic fibroblast growth factor (bFGF) and glial fibrillary acidic protein (GFAP) has been associated with increased neuronal activity we used immediate early gene expression to evaluate the effects of parturition plus interaction with pups during and immediately after delivery to investigate patterns of neural activation in a number of brain areas including Cg2. In the first experiment, we compared levels of immunoreactivity of two immediate early genes: c-Fos and egr-1, among three groups of animals: Cycling, Late Pregnant, and rats perfused 1.5 h postpartum. The average number of Fos and egr-1-ir cells/section were calculated for the Parietal Cortex (ParCx), nucleus accumbens (NAc), medial amygdala (MEA), medial preoptic area (MPOA), supraoptic area (SON) an...
The nutritional needs of developing young place a large energetic demand on lactating females. In... more The nutritional needs of developing young place a large energetic demand on lactating females. In this paper some of the mechanisms through which lactating rats meet these demands are described. Emphasis is placed on the mechanisms that drive increased food intake in suckled rats that are independent of milk delivery and negative energy balance. Evidence is presented to suggest that prolactin (PRL), released from the pituitary in response to suckling stimulation, acts centrally to stimulate food intake during lactation. Brain areas in which PRL may act to stimulate food intake as well as its interactions with other hormones and neuropeptides involved in energy balance are discussed.
The hypoalgesic effect of morphine in lactating rats was assessed using the hot-plate test. At mi... more The hypoalgesic effect of morphine in lactating rats was assessed using the hot-plate test. At midlactation (days 12 and 18 postpartum) females nursing litters of 8 pups were less responsive to the hypoalgesic effect of morphine than ovariectomised or cycling females and females on days 6 or 24 of lactation. Subsequent studies showed that the hypoalgesic response to morphine was inhibited in lactating rats at a number of time points after drug administration and across a variety of doses. This effect was not dependent on milk delivery but was dependent on the hormonal state of the female since separation of dams and their litters for 96 h was sufficient to reinstate the response to morphine if it resulted in a reappearance of vaginal estrus.
Oxytocin (OT; Cys-Tyr-Ile-Gln-Asn-Cys-Pro-leu-Gly), a posterior pituitary peptide hormone, is cha... more Oxytocin (OT; Cys-Tyr-Ile-Gln-Asn-Cys-Pro-leu-Gly), a posterior pituitary peptide hormone, is characterized by a Cys1-Cys6 disulfide bond in its stable, isolated state. This paper describes a simple, one-step method for the production of OT in its reduced, dithiol form (OT dithiol), free of reducing agent. The effects of temperature, pH, and metal-ion chelators on the autoxidation of OT dithiol were examined to establish if this form is likely to persist under biological conditions. It was found that OT dithiol has a half-life of 1.8h with respect to reformation of OT disulfide at 37 degrees C and pH 6.9 in the presence of the copper chelators, DTPA and neocuproine. S-Nitrosation of OT dithiol by acidified nitrite at pH 3.0 was examined by absorption spectroscopy and HPLC-UV-MS, which revealed that both singly and doubly S-nitrosated OT are formed. These results suggest novel chemical aspects to OT signaling, the biological implications of which are discussed here.
Blunted neuroendocrine responses to stress are reported in lactating females after exposure to va... more Blunted neuroendocrine responses to stress are reported in lactating females after exposure to various stressors. However, many of the stimuli used in these studies have little ethological relevance for maternal protection of the litter in a threatening environment. The question that arises is whether the relevance of the stressor to the infant is critical in the 'gating' of the neuroendocrine response. We hypothesized that the presence of pups with their mothers at the time of exposure to an intruder or a predator odour is an effective way to increase the emotional salience of the psychological stressor, thus eliminating the stress hyporesponsiveness in lactating females. We first compared neuroendocrine responses [corticotropin-releasing factor (CRF) mRNA in the paraventricular nucleus of the hypothalamus (PVN) and central nucleus of the amygdala (CeA), plasma adrenocorticotropic hormone (ACTH) and corticosterone] between early (EL, PPD3-5), late (LL, PPD 15) lactating and virgin (V) females to a male intruder in the home cage. We next investigated the effect of pups' presence at the time of stressor exposure on the magnitude of the hormonal response to a male intruder in the home cage or to a predator odour (fox urine) in a novel environment. In the male intruder paradigm, levels of CRF mRNA expression in the PVN and CeA were lower in LL compared to EL or V females and plasma ACTH and corticosterone secretion was not as elevated in LL compared to EL females. Aggression towards the intruder was high in EL females in the presence of their pups and a positive correlation was found with the integrated ACTH response. Aggression rapidly declined after pup separation (2.5 h or 48 h) or in LL nursing females. In EL females, the presence of the pups with their mothers (EL + pups) at the time of stress significantly increased plasma ACTH and corticosterone responses to either male intruder or predator odour compared to EL females without their pups for 2.5 h or 48 h (EL - pups). Plasma ACTH response to fox urine in EL + pups females was comparable to that of virgin females, suggesting that increasing the salience of emotionally relevant stimuli by keeping the pups present in the cage could eliminate the hyporesponsiveness detected for EL females without their pups. These studies indicate the critical role of the pups in modulating the maternal response to stressors that represent a threat for the litter. We hypothesize that the amygdala, because of its ability to process olfactory stimuli and stimuli with affective properties, might play an essential role in 'gating' the neuroendocrine response to stress during lactation.
We investigated the contribution of prolactin and oxytocin to the increase in staining for NADPH-... more We investigated the contribution of prolactin and oxytocin to the increase in staining for NADPH-d and oxytocin mRNA in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) observed at the end of pregnancy, or following a steroid-priming regimen that mimics the hormonal profile of late pregnant females. Ovariectomized rats received chronic implants of silastic capsules containing oestrogen and progesterone followed by progesterone removal. In experiment 1, oxytocin antagonist (OTA) was administered to rats to investigate whether intranuclear oxytocin release was necessary for NADPH-d staining. In experiments 2a and b, rats received concurrent treatment with bromocryptine (0.5 mg/day) to suppress endogenous prolactin release, and either systemic prolactin (0.5 mg once daily), or prolactin (2 micro g/ micro l), or vehicle infused twice a day into the third ventricle, or chronic oxytocin infusion (24 ng/day) for 3 days following progesterone removal. Brains were then processed for NADPH-d histochemistry. In experiment 3, the interaction of prolactin and oxytocin on oxytocin mRNA within the SON and PVN was examined. NADPH-d staining in the SON and PVN was reduced by the highest dose of the OTA, and by bromocryptine treatment. Central prolactin and oxytocin replacement completely restored NADPH-d staining in bromocryptine-treated rats. Finally, both bromocryptine and the OTA suppressed oxytocin mRNA expression and prolactin replacement restored expression levels to that of controls. Together, these data suggest that the increased capacity to produce nitric oxide in the SON and PVN during late pregnancy is dependent on prolactin stimulating oxytocin gene mRNA and hence intranuclear oxytocin release.
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Papers by Barbara Woodside