The Dorsolateral Periaqueductal Gray and Its Role in Mediating Fear Learning to Life Threatening Events

The dorsolateral column of the periaqueductal gray (dlPAG) integrates aversive emotional experiences and represents an important site responding to life threatening situations, such as hypoxia, cardiac pain and predator threats. Previous studies have shown that the dorsal PAG also supports fear learning; and we have currently explored how the dlPAG influences associative learning. We have first shown that N-methyl-D-aspartate (NMDA) 100 pmol injection in the dlPAG works as a valuable unconditioned stimulus (US) for the acquisition of olfactory fear conditioning (OFC) using amyl acetate odor as conditioned stimulus (CS). Next, we revisited the ascending projections of the dlPAG to the thalamus and hypothalamus to reveal potential paths that could mediate associative learning during OFC. Accordingly, the most important ascending target of the dlPAG is the hypothalamic defensive circuit, and we were able to show that pharmacological inactivation using beta-adrenoceptor blockade of the dorsal premammillary nucleus, the main exit way for the hypothalamic defensive circuit to thalamo-cortical circuits involved in fear learning, impaired the acquisition of the OFC promoted by NMDA stimulation of the dlPAG. Moreover, our tracing study revealed multiple parallel paths from the dlPAG to several thalamic targets linked to cortical-hippocampal-amygdalar circuits involved in fear learning. Overall, the results point to a major role of the dlPAG in the mediation of aversive associative learning via ascending projections to the medial hypothalamic defensive circuit, and perhaps, to other thalamic targets, as well. These results provide interesting perspectives to understand how life threatening events impact on fear learning, and should be useful to understand pathological fear memory encoding in anxiety disorders.

Periaqueductal gray matter modulates the hypercapnic ventilatory response

The periaqueductal gray (PAG) is a midbrain structure directly involved in the modulation of defensive behaviors. It has direct projections to several central nuclei that are involved in cardiorespiratory control. Although PAG stimulation is known to elicit respiratory responses, the role of the PAG in the CO2-drive to breathe is still unknown. The present study assessed the effect of chemical lesion of the dorsolateral and dorsomedial and ventrolateral/lateral PAG (dlPAG, dmPAG, and vPAG, respectively) on cardiorespiratory and thermal responses to hypercapnia. Ibotenic acid (IBO) or vehicle (PBS, Sham group) was injected into the dlPAG, dmPAG, or vPAG of male Wistar rats. Rats with lesions outside the dlPAG, dmPAG, or vPAG were considered as negative controls (NC). Pulmonary ventilation (Ve), mean arterial pressure (MAP), heart rate (HR), and body temperature (Tb) were measured in unanesthetized rats during normocapnia and hypercapnic exposure (5, 15, 30 min, 7 % CO2). IBO lesioning of the dlPAG/dmPAG caused 31 % and 26.5 % reductions of the respiratory response to CO2 (1,094.3 +/- 115 mL/kg/min) compared with Sham (1,589.5 +/- 88.1 mL/kg/min) and NC groups (1,488.2 +/- 47.7 mL/kg/min), respectively. IBO lesioning of the vPAG caused 26.6 % and 21 % reductions of CO2 hyperpnea (1,215.3 +/- 108.6 mL/kg/min) compared with Sham (1,657.3 +/- 173.9 mL/kg/min) and NC groups (1,537.6 +/- 59.3). Basal Ve, MAP, HR, and Tb were not affected by dlPAG, dmPAG, or vPAG lesioning. The results suggest that dlPAG, dmPAG, and vPAG modulate hypercapnic ventilatory responses in rats but do not affect MAP, HR, or Tb regulation in resting conditions or during hypercapnia.

Natural history of an ant-plant-butterfly interaction in a Neotropical savanna

Non-myrmecophilous lepidopteran larvae using plants bearing ant attractants such as extrafloral nectaries are good models for studying morphological and behavioural mechanisms against ant predation. Udranomia spitzi (Hesperiidae) is a butterfly whose larvae feed on leaves of Ouratea spectabilis (Ochnaceae), a plant with extrafloral nectaries. We described the early stages of U. spitzi, and used field observations and experiments to investigate the defensive strategies of caterpillars against predatory ants. Larvae pass through five instars and pupation occurs inside larval leaf shelters. Ant-exclusion experiments revealed that the presence of ants did not affect significantly caterpillar survival. Predation experiments showed that vulnerability to ant predation decreased with increase in larval size. The present study showed that predatory ants are not as relevant as demonstrated for other systems, and also illustrates how observational data and field experiments can contribute to a better understanding of the biology and ecology of a species of interest.

Anti-Aversive Effects of Cannabidiol on Innate Fear-Induced Behaviors Evoked by an Ethological Model of Panic Attacks Based on a Prey vs the Wild Snake Epicrates cenchria crassus Confrontation Paradigm

Several pharmacological targets have been proposed as modulators of panic-like reactions. However, interest should be given to other potential therapeutic neurochemical agents. Recent attention has been given to the potential anxiolytic properties of cannabidiol, because of its complex actions on the endocannabinoid system together with its effects on other neurotransmitter systems. The aim of this study was to investigate the effects of cannabidiol on innate fear-related behaviors evoked by a prey vs predator paradigm. Male Swiss mice were submitted to habituation in an arena containing a burrow and subsequently pre-treated with intraperitoneal administrations of vehicle or cannabidiol. A constrictor snake was placed inside the arena, and defensive and non-defensive behaviors were recorded. Cannabidiol caused a clear anti-aversive effect, decreasing explosive escape and defensive immobility behaviors outside and inside the burrow. These results show that cannabidiol modulates defensive behaviors evoked by the presence of threatening stimuli, even in a potentially safe environment following a fear response, suggesting a panicolytic effect. Neuropsychopharmacology (2012) 37, 412-421; doi:10.1038/npp.2011.188; published online 14 September 2011

Spatial regimes: dynamics of intentional homicides in the city of Sao Paulo between 2000 and 2008

Objective. To identify the existence of spatial and temporal patterns in the occurrence of intentional homicides in the municipality of Sao Paulo (MSP), Brazil, and to discuss the analytical value of taking such patterns into account when designing studies that address the dynamics and factors associated with the incidence of homicides. Methods. A longitudinal ecological study was conducted, having as units of analysis 13 205 census tracts and the 96 census districts that congregate these sectors in Sao Paulo. All intentional homicides reported in the city between 2000 and 2008 were analyzed. The gross homicide rates per 100 000 population was calculated as well as the global and local Bayesian estimates for each census tract during the study period. To verify the possibility of identifying different patterns of the spatial distribution of homicides, we used BoxMap and Moran's I index. Results. The homicide trends in the city of Sao Paulo in the last decade were not homogeneous and systematic. Instead, seven patterns of spatial distribution were identified; that is, seven spatial regimes for the occurrence of intentional homicides, considering the homicide rates within each census tract as well as the rates in adjacent tracts. These spatial distribution regimes were not contained within the limits of the census tracts and districts. Conclusions. The results show the importance of analyzing the spatial distribution of social phenomena without restriction of political and administrative boundaries.

Risk assessment behaviors associated with corticosterone trigger the defense reaction to social isolation in rats: Role of the anterior cingulate cortex

The extent to which the hypothalamic-pituitary-adrenal axis is activated by short-term and long-term consequences of stress is still open to investigation. This study aimed to determine (i) the correlation between plasma corticosterone and exploratory behavior exhibited by rats subjected to the elevated plus maze (EPM) following different periods of social isolation, (ii) the effects of the corticosterone synthesis blocker, metyrapone, on the behavioral consequences of isolation, and (iii) whether corticosterone produces its effects through an action on the anterior cingulate cortex, area 1 (Cg1). Rats were subjected to 30-min, 2-h, 24-h, or 7-day isolation periods before EPM exposure and plasma corticosterone assessments. Isolation for longer periods of time produced greater anxiogenic-like effects on the EPM. However, stretched attend posture (SAP) and plasma corticosterone concentrations were increased significantly after 30 min of isolation. Among all of the behavioral categories measured in the EPM, only SAP positively correlated with plasma corticosterone. Metyrapone injected prior to the 24 h isolation period reversed the anxiogenic effects of isolation. Moreover, corticosterone injected into the Cg1 produced a selective increase in SAP. These findings indicate that risk assessment behavior induced by the action of corticosterone on Cg1 neurons initiates a cascade of defensive responses during exposure to stressors.

Income inequality and elderly self-rated health in Sao Paulo, Brazil

Purpose: To test the association between income inequality and elderly self-rated health and to propose a pathway to explain the relationship. Methods: We analyzed a sample of 2143 older individuals (60 years of age and over) from 49 distritos of the Municipality of Sao Paulo, Brazil. Bayesian multilevel logistic models were performed with poor self-rated health as the outcome variable. Results: Income inequality (measured by the Gini coefficient) was found to be associated with poor self-rated health after controlling for age, sex, income and education (odds ratio, 1.19; 95% credible interval, 1.01-1.38). When the practice of physical exercise and homicide rate were added to the model, the Gini coefficient lost its statistical significance (P>.05). We fitted a structural equation model in which income inequality affects elderly health by a pathway mediated by violence and practice of physical exercise. Conclusions: The health of older individuals may be highly susceptible to the socioeconomic environment of residence, specifically to the local distribution of income. We propose that this association may be mediated by fear of violence and lack of physical activity. (C) 2012 Elsevier Inc. All rights reserved.

Gregarious behavior of two species of Neotropical harvestmen (Arachnida: Opiliones: Gonyleptidae)

We present the first record and description of the gregarious behavior of the Neotropical harvestmen Serracutisoma proximum (Mello-Leitao 1922) and Serracutisoma spelaeum (Mello-Leitao 1933) (Opiliones: Gonyleptidae: Goniosomatinae) (DaSilva & Gnaspini 2010). We followed and described the pattern of these aggregations over a period of 17 months in a cave in southeastern Brazil. Individuals of the two species aggregated with both conspecifics and heterospecifics during the non-reproductive season (i.e., from October to March, the cool and dry season). Aggregations contained up to 81 individuals, usually with a female-biased adult sex ratio. Multispecific aggregations were usually composed mainly of representatives of one of the two species, suggesting that although these species also aggregate with heterospecifics, there is a preference for aggregating with conspecifics. This study provides novel information on the social behavior of harvestmen, specifically regarding the composition of multispecific aggregations.

Cause-specific mortality and income inequality in Sao Paulo, Brazil

OBJECTIVE: To analyze cause-specifi c mortality rates according to the relative income hypothesis. METHODS: All 96 administrative areas of the city of Sao Paulo, southeastern Brazil, were divided into two groups based on the Gini coefficient of income inequality: high (>= 0.25) and low (<0.25). The propensity score matching method was applied to control for confounders associated with socioeconomic differences among areas. RESULTS: The difference between high and low income inequality areas was statistically significant for homicide (8.57 per 10,000; 95% CI: 2.60; 14.53); ischemic heart disease (5.47 per 10,000 [95% CI 0.76; 10.17]); HIV/AIDS (3.58 per 10,000 [95% CI 0.58; 6.57]); and respiratory diseases (3.56 per 10,000 [95% CI 0.18; 6.94]). The ten most common causes of death accounted for 72.30% of the mortality difference. Infant mortality also had signifi cantly higher age-adjusted rates in high inequality areas (2.80 per 10,000 [95% CI 0.86; 4.74]), as well as among males (27.37 per 10,000 [95% CI 6.19; 48.55]) and females (15.07 per 10,000 [95% CI 3.65; 26.48]). CONCLUSIONS: The study results support the relative income hypothesis. After propensity score matching cause-specifi c mortality rates was higher in more unequal areas. Studies on income inequality in smaller areas should take proper accounting of heterogeneity of social and demographic characteristics.

Associação ecológica entre características dos municípios e o risco de homicídios em homens adultos de 20-39 anos de idade no Brasil, 1999-2010

No Brasil, a mortalidade por homicídios persiste como importante problema de saúde pública, principalmente entre homens adultos jovens. O objetivo do presente estudo foi analisar o risco de morte por homicídios entre homens de 20-39 anos de idade e sua associação com características sociodemográficas dos municípios brasileiros. Foi realizado estudo ecológico, tendo como unidades de análise todos os municípios do País. Foram estudadas as tendências temporais entre 1999-2010 e as associações do desfecho com indicadores dos municípios em análise transversal referente ao quatriênio 2007-2010. Entre os quatriênios 1999-2002 e 2007-2010, houve aumento das taxas medianas de mortalidade por homicídios entre homens de 20-39 anos, de 22,7 para 35,5 por 100 mil habitantes. No quatriênio 2007-2010, os riscos de homicídios foram estatisticamente superiores (p<0,001) nos municípios de maior porte populacional, maior taxa de fecundidade, baixa proporção de alfabetizados, maior desigualdade aferida pela renda 20/40 e maior urbanização. Para a proporção da população de baixa renda e renda média per capita, as associações indicam excessos nas estimativas de risco de homicídios nos municípios com valores intermediários desses indicadores. Os achados podem auxiliar na focalização de políticas públicas.

Description of immature stages of Phelypera schuppeli (Boheman, 1834) with comments on natural history (Coleoptera: Curculionidae: Hyperinae)

Immatures of the Phelypera schuppeli (Boheman, 1834) (Curculionidae; Hyperinae; Cepurini) are described, illustrated and compared with available descriptions of larvae and pupae of Hyperini. Immatures and adults from midwest (Dourados, Mato Grosso do Sul; Pirenopolis, Goias) and southeast Brazil (Bauru, Sao Paulo) were found on leaves of the host plant, Pachira aquatica Aubl. (Malvaceae, formerly Bombacaceae), a tree used as an ornamental plant in many Brazilian frost-free cities. Larvae of P. schuppeli are exophytic, brightly colored, eruciform and possess abdominal ambulatory ampullae, resembling larvae of Lepidoptera. Mature larvae can spin globular lattice-like cocoons where pupation takes place. Data in the field and under laboratory conditions confirmed previously published biological observations on P. schuppeli. Additional information about defensive behaviors, process of cocoon construction and natural enemies, such as the larval predator Supputius cinticeps (Stal, 1860) (Hemiptera: Pentatomidae) and the prepupal and pupal parasitoid Jaliscoa nudipennis Boucek, 1993 (Hymenoptera: Pteromalidae), are reported.

Central, but not basolateral, amygdala involvement in the anxiolytic-like effects of midazolam in rats in the elevated plus maze

The role of the amygdala in the mediation of fear and anxiety has been extensively investigated. However, how the amygdala functions during the organization of the anxiety-like behaviors generated in the elevated plus maze (EPM) is still under investigation. The basolateral (BLA) and the central (CeA) nuclei are the main input and output stations of the amygdala. In the present study, we ethopharmacologically analyzed the behavior of rats subjected to the EPM and the tissue content of the monoamines dopamine (DA) and serotonin (5-HT) and their metabolites in the nucleus accumbens (NAc), dorsal hippocampus (DH), and dorsal striatum (DS) of animals injected with saline or midazolam (20 and 30 nmol/0.2 mu L) into the BLA or CeA. Injections of midazolam into the CeA, but not BLA, caused clear anxiolytic-like effects in the EPM. These treatments did not cause significant changes in 5-HT or DA contents in the NAc, DH, or DS of animals tested in the EPM. The data suggest that the anxiolytic-like effects of midazolam in the EPM also appear to rely on GABA-benzodiazepine mechanisms in the CeA, but not BLA, and do not appear to depend on 5-HT and DA mechanisms prevalent in limbic structures.

Serotonin-2C receptors in the basolateral nucleus of the amygdala mediate the anxiogenic effect of acute imipramine and fluoxetine administration

A growing body of evidence indicates that facilitation of serotonin-2C receptor (5-HT2CR)-mediated neurotransmission in the basolateral nucleus of the amygdala (BLA) is involved in anxiety generation. We investigated here whether BLA 5-HT(2C)Rs exert a differential role in the regulation of defensive behaviours related to generalized anxiety (inhibitory avoidance) and panic (escape) disorders. We also evaluated whether activation of BLA 5-HT(2C)Rs accounts for the anxiogenic effect caused by acute systemic administration of the antidepressants imipramine and fluoxetine. Male Wistar rats were tested in the elevated T-maze after intra-BLA injection of the endogenous agonist 5-HT, the 5-HT2CR agonist MK-212 or the 5-HT2CR antagonist SB-242084. This test allows the measurement of inhibitory avoidance acquisition and escape expression. We also investigated whether intra-BLA administration of SB-242084 interferes with the acute anxiogenic effect caused by imipramine and fluoxetine in the Vogel conflict test, and imipramine in the elevated T-maze. While intra-BLA administration of 5-HT and MK-212 facilitated inhibitory avoidance acquisition, suggesting an anxiogenic effect, SB-242084 had the opposite effect. None of these drugs affected escape performance. Intra-BLA injection of a sub-effective dose of SB-242084 fully blocked the anxiogenic effect caused either by the local microinjection of 5-HT or the systemic administration of imipramine and fluoxetine. Our findings indicate that 5-HT(2C)Rs in BLA are selectively involved in the regulation of defensive behaviours associated with generalized anxiety, but not panic. The results also provide the first direct evidence that activation of BLA 5-HT(2C)Rs accounts for the short-term aversive effect of antidepressants.

Behavioral analysis of the interaction between the spitting spider Scytodes globula (Araneae: Scytodidae) and the harvestman Discocyrtus invalidus (Opiliones: Gonyleptidae)

Spitting spiders (Scytodidae) have a distinct predatory strategy in which they eject a sticky secretion from their cheliceral fangs to immobilize prey. This behavior could potentially allow the spider not only to avoid defensive secretions but also to bite specific vulnerable spots of a potential prey such as a harvestman. We used an ethogram, a fluxogram and an experiment to analyze the interaction between the harvestman Discocyrtus invalidus Piza 1938 (Arachnida: Opiliones) and the syntopic spider Scytodes globula (Nicolet 1849) (Arachnida: Araneae). These spiders, while readily taking crickets as prey, seldom spat at and never bit the harvestmen, which apparently did not exude repugnatorial secretions. We therefore tested, by clogging the glands and using appropriate controls, whether non-visible amounts of secretions could cause the rejection, but the harvestmen were still refused. This is the first detailed and quantified description of an interaction between a spitting spider and a harvestman. The general conclusions are that S. globula avoids preying on D. invalidus, S. globula behaves differently when attacking harvestmen and crickets and the scent gland secretions of D. invalidus do not play a direct role in this predator-prey interaction.

Opioidergic, GABAergic and serotonergic neurotransmission in the dorsal raphe nucleus modulates tonic immobility in guinea pigs

Tonic immobility (TI) is an innate defensive behavior that can be elicited by physical restriction and postural inversion and is characterized by a profound and temporary state of akinesis. Our previous studies demonstrated that the stimulation of serotonin receptors in the dorsal raphe nucleus (DRN) appears to be biphasic during TI responses in guinea pigs (Cavia porcellus). Serotonin released by the DRN modulates behavioral responses and its release can occur through the action of different neurotransmitter systems, including the opioidergic and GABAergic systems. This study examines the role of opioidergic, GABAergic and serotonergic signaling in the DRN in TI defensive behavioral responses in guinea pigs. Microinjection of morphine (1.1 nmol) or bicuculline (0.5 nmol) into the DRN increased the duration of TI. The effect of morphine (1.1 nmol) was antagonized by pretreatment with naloxone (0.7 nmol), suggesting that the activation of pi opioid receptors in the DRN facilitates the TI response. By contrast, microinjection of muscimol (0.5 nmol) into the DRN decreased the duration of TI. However, a dose of muscimol (0.26 nmol) that alone did not affect TI, was sufficient to inhibit the effect of morphine (1.1 nmol) on TI, indicating that GABAergic and enkephalinergic neurons interact in the DRN. Microinjection of alpha-methyl-5-HT (1.6 nmol), a 5-HT2 agonist, into the DRN also increased TI. This effect was inhibited by the prior administration of naloxone (0.7 nmol). Microinjection of 8-OH-DPAT (1.3 nmol) also blocked the increase of TI promoted by morphine (1.1 nmol). Our results indicate that the opioidergic, GABAergic and serotonergic systems in the DRN are important for modulation of defensive behavioral responses of TI. Therefore, we suggest that opioid inhibition of GABAergic neurons results in disinhibition of serotonergic neurons and this is the mechanism by which opioids could enhance TI. Conversely, a decrease in TI could occur through the activation of GABAergic interneurons. (C) 2012 Elsevier Inc. All rights reserved.