Neuropeptides in the posterodorsal medial amygdala modulate central cardiovascular reflex responses in awake male rats

The rat posterodorsal medial amygdala (MePD) links emotionally charged sensory stimuli to social behavior, and is part of the supramedullary control of the cardiovascular system. We studied the effects of microinjections of neuroactive peptides markedly found in the MePD, namely oxytocin (OT, 10 ng and 25 pg; n=6/group), somatostatin (SST, 1 and 0.05 μM; n=8 and 5, respectively), and angiotensin II (Ang II, 50 pmol and 50 fmol; n=7/group), on basal cardiovascular activity and on baroreflex- and chemoreflex-mediated responses in awake adult male rats. Power spectral and symbolic analyses were applied to pulse interval and systolic arterial pressure series to identify centrally mediated sympathetic/parasympathetic components in the heart rate variability (HRV) and arterial pressure variability (APV). No microinjected substance affected basal parameters. On the other hand, compared with the control data (saline, 0.3 µL; n=7), OT (10 ng) decreased mean AP (MAP50) after baroreflex stimulation and increased both the mean AP response after chemoreflex activation and the high-frequency component of the HRV. OT (25 pg) increased overall HRV but did not affect any parameter of the symbolic analysis. SST (1 μM) decreased MAP50, and SST (0.05 μM) enhanced the sympathovagal cardiac index. Both doses of SST increased HRV and its low-frequency component. Ang II (50 pmol) increased HRV and reduced the two unlike variations pattern of the symbolic analysis (P<0.05 in all cases). These results demonstrate neuropeptidergic actions in the MePD for both the increase in the range of the cardiovascular reflex responses and the involvement of the central sympathetic and parasympathetic systems on HRV and APV.

Neuropeptides in the posterodorsal medial amygdala modulate central cardiovascular reflex responses in awake male rats.

The rat posterodorsal medial amygdala (MePD) links emotionally charged sensory stimuli to social behavior, and is part of the supramedullary control of the cardiovascular system. We studied the effects of microinjections of neuroactive peptides markedly found in the MePD, namely oxytocin (OT, 10 ng and 25 pg; n=6/group), somatostatin (SST, 1 and 0.05 µM; n=8 and 5, respectively), and angiotensin II (Ang II, 50 pmol and 50 fmol; n=7/group), on basal cardiovascular activity and on baroreflex- and chemoreflex-mediated responses in awake adult male rats. Power spectral and symbolic analyses were applied to pulse interval and systolic arterial pressure series to identify centrally mediated sympathetic/parasympathetic components in the heart rate variability (HRV) and arterial pressure variability (APV). No microinjected substance affected basal parameters. On the other hand, compared with the control data (saline, 0.3 µL; n=7), OT (10 ng) decreased mean AP (MAP50) after baroreflex stimulation and increased both the mean AP response after chemoreflex activation and the high-frequency component of the HRV. OT (25 pg) increased overall HRV but did not affect any parameter of the symbolic analysis. SST (1 µM) decreased MAP50, and SST (0.05 µM) enhanced the sympathovagal cardiac index. Both doses of SST increased HRV and its low-frequency component. Ang II (50 pmol) increased HRV and reduced the two unlike variations pattern of the symbolic analysis (P<0.05 in all cases). These results demonstrate neuropeptidergic actions in the MePD for both the increase in the range of the cardiovascular reflex responses and the involvement of the central sympathetic and parasympathetic systems on HRV and APV.

Morphological and functional features of the sex steroid-responsive posterodorsal medial amygdala of adult rats.

The rat posterodorsal medial amygdala (MePD) expresses receptors for gonadal hormones and integrates sex steroid-sensitive subcortical networks. Male-female differences are found in the morphology, connectivity, and local neuropil structure of MePD. For example, dendritic spine density is sexually-dimorphic and changes with the estrous cycle and following gonadal hormones manipulations. Due to its connectivity, the MePD may affect emotionally-loaded social behaviors, according to a former Newman's seminal proposition. Unilateral fiber-sparing ibotenic acid damage of the MePD does not impair male sexual behavior. However, microinjecting glutamate and histamine into the right MePD facilitates ejaculation. Further, MePD-lesioned rats are not different from normal rats in anxiety-like behavior as evaluated by the elevated plus maze test or innate fear test induced by a live cat. In another study, an adapted model for inducing aggressive behavior in rats by a brief period of restraint prior to the resident-intruder paradigm was used to study Fos-immunoreactivity in the MePD. Following stressful stimulation (restraint) or the restraint and fight condition, but not after aggression alone, Fos-immunoreactivity was detected in the MePD. Microinjecting the inhibitory neuropeptide somatostatin into the right MePD notably reduces fighting behavior without affecting locomotion. Overall, these data indicate that sex steroids and local neurochemical stimulatory/inhibitory transmitters modulate the MePD and reinforce the idea that this area is a node for modulating social behavior neural networks.

Histamine in the posterodorsal medial amygdala modulates cardiovascular reflex responses in awake rats.

Centrally injected histamine (HA) affects heart rate (HR), arterial blood pressure (BP), and sympathetic activity in rats. The posterodorsal medial amygdala (MePD) has high levels of histidine decarboxylase, connections with brain areas involved with the modulation of cardiovascular responses, and is relevant for the pathogenesis of hypertension. However, there is no report demonstrating the role of the MePD histaminergic activity on the cardiovascular function in awake rats. The aims of the present work were: 1) to study the effects of two doses (10-100 nM) of HA microinjected in the MePD on basal cardiovascular recordings and on baroreflex- and chemoreflex-mediated responses; 2) to reveal whether cardiovascular reflex responses could be affected by MePD microinjections of (R)-alpha-methylhistamine (AH3), an agonist of the inhibitory autoreceptor H3; and, 3) to carry out a power spectral analysis to evaluate the contribution of the sympathetic and parasympathetic components in the variability of the HR and BP recordings. When compared with the control group (microinjected with saline, 0.3 microl), HA (10 nM) promoted an increase in the MAP50, i.e. the mean value of BP at half of the HR range evoked by the baroreflex response. Histamine (100 nM) did not affect the baroreflex activity, but significantly decreased the parasympathetic component of the HR variability, increased the sympathetic/parasympathetic balance at basal conditions (these two latter evaluated by the power spectral analysis), and promoted an impairment in the chemoreflex bradycardic response. Microinjection of AH3 (10 microM) led to mixed results, which resembled the effects of both doses of HA employed here. Present data suggest that cardiovascular changes induced by baroreceptors and chemoreceptors involve the histaminergic activity in the MePD. This neural regulation of reflex cardiovascular responses can have important implications for homeostatic and allostatic conditions and possibly for the behavioral displays modulated by the rat MePD.

Ultrastructural features of neurons and synaptic contacts in the posterodorsal medial amygdala of adult male rats.

The aim of the present study was to describe the ultrastructure of neurons (from eight animals) and to analyse the synaptic terminal distribution (from two animals) in the posterodorsal subnucleus of the medial amygdala (MePD) of adult male rats. Using transmission electron microscopy, it was possible to identify many spiny and aspiny dendrites, unmyelinated axonal bundles, single axonal processes, a few myelinated axons, blood vessels and glial processes in the neuropil. Axodendritic synapses were the most frequently observed (67.5%), appearing to be of either the inhibitory or the excitatory types. The presynaptic region contained round or flattened vesicles that occurred either singly or with dense-cored vesicles (DCVs). The dendrites often received many synapses on a single shaft, and axon terminals displayed synaptic contacts with one or more postsynaptic structures. Dendritic spines showed different morphologies and the synapses on them (23.1%) formed a single and apparently excitatory synaptic contact with round, electron-lucid vesicles alone or, less frequently, with DCVs. Inhibitory and excitatory axosomatic synapses (8.2%) and excitatory axoaxonic synapses (1.2%) were also identified. The present report provides new findings relevant to the study of the MePD cellular organization and could be combined with other morphological data in order to reveal the functional activity of this area in male rats.

Influence of sex, estrous cycle and motherhood on dendritic spine density in the rat medial amygdala revealed by the Golgi method.

The medial nucleus of the amygdala (MeA), a sexually dimorphic area, contains estrogen and androgen receptors and has an integrative role in behavioral, vegetative and endocrine activities of rats. The density of dendritic spines along the first 40 microm of dendritic length was studied in neurons from the anterodorsal (MeAD), posterodorsal (MePD) and posteroventral (MePV) aspects of the MeA in males, in virgin females during the four phases of the estrous cycle and in multiparous females in diestrus. The single-section Golgi method was employed (N=48 observations per experimental group). In the three MeA subnuclei males showed more dendritic spines than virgin females (P<0.001), with the only exception being the MePD data of females in diestrus (P>0.05). In virgin females, whereas more dendritic spines were found in diestrus, a decline in these values was found during the proestrus, estrus and metaestrus in the MePD and MePV (P<0.05) but not in the MeAD (P>0.05). Compared with virgin females in diestrus, postpartum rats showed more spines in the MeAD (P<0.001) and fewer in the MePD (P<0.001) but no difference was found in the MePV (P>0.05). These data suggest that there are subregion-specific differences in the density of dendritic spines within the MeA and that they appear to be affected by sex, cyclic fluctuations in the levels of ovarian steroids and following pregnancy in rats. These findings may add to the understanding of the MeA neuronal plastic changes that affect the ongoing processing of sensory information and the organization of the neuroendocrine and behavioral basis of reproduction.

Functional activities of the amygdala: an overview.

Research to date into the amygdala shows that it has an integrative role in behavioural, vegetative and endocrine activities of animals in their relation with their environment. Animal studies show that amygdala has a role in emotional response, integrating input signals and initiating activities related to them. Different nuclei seem to have different effects. A complete picture of the functional roles of the amygdala is unavailable, and it has been suggested that the amygdala is functionally and anatomically heterogeneous. Amygdaloid subnuclei appear to have a role in the modulation of fear, in memory and attention, and in some sexual and sex-related behaviour of rats. In humans, functional magnetic resonance imaging shows that the amygdala responds preferentially to emotionally charged stimuli. Bilateral amygdala damage in humans can compromise the recognition of fear in facial expressions, an important ability in social judgement. Future study of the amygdala promises to shed light on emotional disorders in humans.

Effects of gonadal hormones on the morphology of neurons from the medial amygdaloid nucleus of rats.

The medial amygdala (MeA) has receptors for gonadal hormones and is a sexually dimorphic area in rats. The aims of the present work were (1) to look at sex differences and the effect of gonadal hormone withdrawal in males castrated as offspring or at adulthood on neuronal soma area in the anterior and posterior MeA and (2) to study the dendritic branching and the density of dendritic spines in neurons from the MeA of intact males and females. Animals were adult rats, for which the single-section Golgi method was used. Stellate and bitufted cells were found in the MeA. Comparing data among groups, no significant difference in cell body area was found. Dendrites divide sparingly and have very different lengths, and a statistical difference (p < 0.001, males higher than females) in the spine density in the anterior MeA, but not in the posterior MeA, was found. These results suggest that castration does not alter the somal area in males submitted to gonadectomy during the early postnatal period or at adulthood. In addition, the already described sex difference in this nucleus may be more related to the neuropil than the neuronal somal area, which may be relevant for the function of the MeA.

Influence of early postnatal gonadal hormones on anxiety in adult male rats.

Behavioral sex differences have been linked to the presence of testosterone secretion during a critical perinatal period. The present experiment tested whether or not castration at different ages (early postnatal period and adulthood) would alter performance in the plus maze, a behavioral test of anxiety. Intact adult male rats (n = 17) were compared to intact adult females (n = 17); adult castrated males (n = 7) to sham-operated adult male rats (n = 9); and newborn castrated males (n = 7) to sham-operated male offspring (n = 8). When adult, the subjects were left on an elevated plus maze for 5 min. Females made a higher percentage of entries onto the open arms and showed a greater number of scans over the edge of an open arm than males. There were no differences in the percentage of arm entries or time spent on the open arms when adult castrated males were compared to sham-operated rats. On the other hand, newborn castrated males showed a significantly higher number of open arm entries and spent a greater percentage of time on the open arms than sham-operated offspring. The results demonstrate that the absence of male gonadal hormones during the perinatal period decreases anxiety, as assessed in the elevated plus maze, leading to a behavioral pattern that resembles that of females. These data provide evidence for the organizational role of gonadal hormones in the development of behavioral inhibitory systems.

Effects of 8-OH-DPAT on sexual behavior of male rats castrated at different ages.

The effects of the 5-hydroxytryptamine (5-HT) 1A receptor agonist, 8-hydroxy-2(di-n-propylamino)tetralin (8-OH-DPAT), on the sexual behavior of male rats castrated at adulthood or during the early postnatal critical period for brain differentiation are presented. In adult castrated males, two distinct groups of animals, divided according to their copulatory activity impairment following castration, were identified: animals that reached a low intromission frequency around Day 60 after testes removal (early reduction group) and males that retained the behavior until approximately Postcastration Day 180 (late reduction group). 8-OH-DPAT (0.10 mg/kg) increased intromission frequency in both groups, but the effect was more pronounced in rats that had a late reduction of mating behavior after castration. In males castrated at 6 hr or 12 days after birth, an increase in intromission frequency, but not in ejaculatory behavior, was also detected following the injection of 8-OH-DPAT (0.10 or 0.25 mg/kg) at adulthood. In conclusion, 8-OH-DPAT was able to enhance intromission behavior, bypassing the lack of testosterone either at the neonatal critical period or at adult age. The 5-HT1A system would exert a parallel modulatory action, in addition to testosterone, on male sexual behavior. Alternatively, since neonatal castrated males showed no ejaculatory behavior after 8-OH-DPAT injection, 5-HT and testosterone would act in an interactive way for the full expression of copulatory behavior.

Effect of estradiol implanted in the corticomedial amygdala on the sexual behavior of castrated male rats.

1. The purpose of the present investigation was to study the effect of beta-estradiol crystals implanted in the corticomedial area of the amygdaloid body on the sexual behavior of castrated male rats. 2. The animals were divided into the following groups: group I (N = 9), castrated rats; group II (N = 4), rats which had been castrated and stereotaxically implanted with cholesterol, both groups being used as controls; group III (N = 6), castrated rats with estradiol implants. Latency to the first anogenital exploration, latency to the first mount and mount frequency were determined during the pre-castration and post-castration phases and after the material had been implanted in groups II and III in 10-min observation sessions. 3. There was diminished sexual behavior of the animals in group I without spontaneous recurrence within the period observed. Group II animals, who had undergone implantation of cholesterol, an inert substance, maintained low levels of sexual behavior (post-castration 0.8 +/- 0.7 vs 0.0 +/- 0.0 and 0.5 +/- 0.5 on the 6th and 9th day after implantation, respectively). Group III animals presented a gradual increase in the number of mounts (from post-castration 1.2 +/- 0.5 to 6.5 +/- 2.7 and 4.1 +/- 1.0 on the 6th and 9th day after implantation, respectively) and a decrease of mount latency (from post-castration 431.2 +/- 55.9 to 226.1 +/- 119.6 and 51.0 +/- 28.9 on the 6th and 9th day after implantation, respectively) reaching pre-castration levels on the 6th and 9th day after beta-estradiol implantation.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of estradiol implanted in the corticomedial amygdala on the sexual behavior of castrated male rats

1. The purpose of the present investigation was to study the effect of ß-estradiol crystals implanted in the corticomedial area of the amygadaloid body on the sexual behavior of castrated male rats. 2. The animals were divided into the following groups: group I (N=9), castrated rats; group II (N=4), rats which had been castrated and stereotaxically implanted with cholesterol, both groups being used as controls; group III (N=6), castrated rats con estradiol implants. Latency to the first anogenital exploration, latency to the first mount and mount frquency were determined during the pre-castration and post-castration phases and after the material had been implanted in groups II and III in 10-min observation sessions. 3. There was diminished sexual behavior of the animals in group I without spontaneous recurrence within the period observed. Group II animals, who had undergone implantation of cholesterol, an inert substance, maintained low levels of sexual behavior (post-castration 0.8 ñ 7 vs 0.0 ñ 0.0 and 0.5 ñ 0.5 on the 6th and 9th day afther implantation, repectively). Group III animals presented a gradual increase in the number of mounts (from post-castration 1.2 ñ 0.5 to 6.5 ñ 2.7 and 4.1 ñ 1.0 on the 6th and 9th day after implantation, respectively) and a decrease of mount latency (from post-castration 431.2 ñ 55.9 to 226.1 ñ 119.6 and 51.0 ñ 28.9 on the 6th and 9th day after implantation, respectively) reaching pre-castration levels on the 6th and 9th day after ß-estradiol implantation. 4. We conclude that, under the effect of estradiol, the amygdaloid region can modulate male sexual behavior, thus injdicating a physiological role for estradiol receptors present in this area