IgG4-positive plasma cells are more often detected in chronic periapical lesions arising from permanent rather than primary teeth.

AIM: To characterize plasma cell subsets in chronic periapical lesions affecting permanent and primary teeth. METHODOLOGY: Only chronic periapical lesions without root canal treatment were selected. Twenty-one radicular cysts and 7 periapical granulomas affecting permanent teeth and 19 radicular cysts and 4 periapical granulomas affecting primary teeth were assessed for immunoglobulin (Ig) light chain (kappa and lambda), Ig heavy chain (IgG, IgG4, IgA, IgM and IgD) and plasma cell immunohistochemical markers (MUM1/IRF4, EMA and CD138). The data acquired were analysed by Student's t test, Mann-Whitney U, Friedman test followed by Dunn's multiple comparison test and Spearman's rank correlation. RESULTS: All cases were polyclonal (having similar kappa/lambda light chain ratios). IgG was most abundant compared to other Ig heavy chains (all, P < 0.001); like Ig light chains, but unlike IgA, there was greater expression of IgG in the primary compared to the permanent dentition, for both radicular cysts (P < 0.001) and periapical granulomas (P = 0.53). Notably, IgG4 expression was greater in the permanent than the primary dentition, for both radicular cyst (P < 0.05) and periapical granuloma (P = 0.65). IgM and IgD expression was scarce and variable, whereas plasma cell populations were detected efficiently through EMA, CD138 and MUM1/IRF4 markers, the latter being more sensitive in both dentitions. CONCLUSIONS: There were slight variations in the Ig light and heavy chain profiles in chronic periapical lesions when comparing the permanent and primary dentitions. The ability of IgG4+ plasma cell infiltration to modulate inflammatory responses in chronic periapical lesions arising from permanent as opposed to primary teeth should be considered in future studies.

Can differences between continental and insular habitats influence the parasites communities associated with the endemic frog Haddadus binotatus?

Habitats characterized by geographic isolation such as islands have been studied using different organisms as models for understanding the dynamic and insular patterns of biodiversity. Determinants of parasite richness in insular host populations have been conducted mainly with mammals and birds, showing that parasite richness decreases in insular areas. In the present study, we predicted that the type of environment (insular or continental) can influence the richness, diversity and abundance of parasites associated with the endemic frog Haddadus binotatus (Spix, 1824). We sampled frogs in two insular and two mainland fragments to survey their helminth parasites. The total richness was composed of 15 taxa of Nematoda and two of Acanthocephala, and the community composition of the two islands had more similarities between them than the two mainland localities. The insular effect was positive for richness and abundance of helminths, and no significant effect was observed on helminth diversity - even the mean diversity presented high numbers for the islands. We presumed that insular hosts could have lost some parasites in the colonization process when these continental islands were separated from the mainland, approximately 11,000 years ago. However, the high richness and abundance on islands can be explained by an epidemiological argument, which considers high population density due to insularity and other features of the host as factors that increase parasite transmission success among individuals.

NLRP12 Attenuates Inflammatory Bone Loss in Experimental Apical Periodontitis.

Apical periodontitis is an inflammatory disorder that results from the host immune response to microbial infection through the dental pulp, leading to alveolar bone destruction. The nod-like receptor 12 (NLRP12) is an atypical intracellular sensor of the NLR family that is involved in the negative regulation of several inflammatory conditions and also osteoclastogenesis. However, the role of NLRP12 in the regulation of immune response and bone loss induced by bacterial infection remains unclear. Here we investigated the development of apical periodontitis in wild-type (WT) and NLRP12 knockout (NLRP12-/-) mice by using micro-computed tomography together with histological, immunohistochemical, and molecular analyses. We found that NLRP12-/- mice are highly susceptible to apical periodontitis induced by bacterial infection, which is associated with an elevated infiltration of neutrophils and macrophages, periapical lesion extension, and alveolar bone destruction. Furthermore, NLRP12-/- mice showed a high expression of inflammatory cytokines ( Il1b, Il6, and Tnfa) and the osteoclastogenic markers ( Rankl and Acp5) in the periapical tissues. Consistent with this observation, NLRP12-/- mice showed an increased number of tartrate-resistant acid phosphatase-positive cells lining the apical periodontitis site, which was associated with augmented expression of the osteoclast effector genes, Ctsk and Mmp9. Mechanistically, NLRP12-deficient preosteoclasts showed elevated IκB-α degradation and p65 phosphorylation when stimulated with receptor activator of nuclear factor (NF)-κB ligand (RANKL). Similarly, increased IκB-α degradation was observed in the periapical tissue of NLRP12-/- mice. Furthermore, our in vitro study showed that preosteoclasts from NLRP12-/- mice exhibited higher RANKL-induced osteoclastogenesis, which was synergistically amplified by interleukin-1ß and tumor necrosis factor α (mimicking an inflammatory periapical milieu). In conclusion, our data show that NLRP12 exhibits a protective role in the periapical bone destruction by attenuating inflammation and osteoclastogenesis through negative regulation of the NF-κB pathway.

L-asparaginase isolated from Streptomyces ansochromogenes promotes Th1 profile and activates CD8+ T cells in human PBMC: an in vitro investigation.

AIMS: A new L-asparaginase produced by Streptomyces ansochromogenes UFPEDA 3420 actinobacteria was used in this study against human lymphocyte cultures to evaluate the immunological profile induced by this enzyme. METHODS AND RESULTS: Cultures of lymphocytes were stimulated with S. ansochromogenes L-asparaginase, and cytotoxicity, cell viability, cell stimulation and cytokine production were analysed. This new S. ansochromogenes L-asparaginase induced activation and proliferation of the TCD8+ lymphocyte subset and produced higher TNF-α, IFN-γ, IL-2 and IL-10 levels in a 24-h assay. CONCLUSION: Streptomyces ansochromogenes L-asparaginase is a promising molecule to be used in in vivo models and to deepen preclinical tests against acute lymphoblast leukaemia. SIGNIFICANCE AND IMPACT OF STUDY: L-asparaginase is an indispensable component of the chemotherapeutic treatment of acute lymphoblast leukaemia (ALL) and acute myeloid leukaemia (AML). Currently, drugs such as Asparaginase® , Kidrolase® , and Elspar® and Erwinase® are efficient against leukemic disease, but promote immunosuppression and other side effects in human organisms. Our purified S. ansochromogenes L-asparaginase showed promissory results inducing, in vitro, higher immunostimulation in human PBMC, especially in T CD8+ lymphocyte subsets.

Morphometric and phylogenetic analyses of Serpentirhabdias viperidicus n. sp. (Nematoda: Rhabdiasidae) from the lancehead snake Bothrops moojeni Hoge, 1966 (Reptilia: Serpentes: Viperidae) in Brazil.

Serpentirhabdias viperidicus n. sp. (Nematoda: Rhabdiasidae) is described from the lungs of the 'Brazilian lancehead' Bothrops moojeni (Hoge, 1966) from the savannah in São Paulo State, Brazil. The new species is the eighth species of Serpentirhabdias described in the Neotropical region, and differs from other species mainly by a combination of characters: lips slightly notable, presence of fine striations at posterior ends, presence of two parallel lines with intercalated pores, a pore-shaped phasmid situated at the level of the anal aperture and another two in the posterior half of the tail. It is the first species of Serpentirhabdias reported in this snake host and the second species of this genus found parasitizing South American viperidian snakes. Molecular phylogenetic analysis using ribosomal (ITS and 28S partial) genes confirms Serpentirhabdias viperidicus n. sp. as a new species that clustered in the Serpentirhabdias clade, sister taxon to Serpentirhabdias fuscovenosa and Serpentirhabdias elaphe. This is the first description of Serpentirhabdias species from Brazil using molecular approaches and morphological characters to confirm the monophyly of this recent genus.

One-session root canal treatment with antimicrobial photodynamic therapy (aPDT): an in vivo study.

AIM: To evaluate the response of the apical and periapical tissues of dog teeth with apical periodontitis after one-session root canal treatment with and without antimicrobial photodynamic therapy (aPDT) compared with the use of an intracanal dressing. METHODOLOGY: Sixty root canals with an induced periapical lesion were instrumented and assigned to three groups: I, two-session root canal treatment using antibacterial dressing with calcium hydroxide-based paste; II, one-session root canal treatment using aPDT; and III, one-session root canal treatment in which the root canals were filled immediately after biomechanical preparation. The animals were euthanized after a 90-day experimental period. The maxillas and mandibles with teeth were submitted to histotechnical processing and haematoxylin-eosin staining. Descriptive microscopic analysis of the apical and periapical region characteristics was performed, as well as morphometric assessment of the periapical lesion areas in fluorescence microscopy. Quantitative data were analysed statistically by the nonparametric Kruskal-Wallis test and Dunn's post-test (α = 0.05). RESULTS: Group I was characterized by progressive repair, with the presence of fibres, cells and blood vessels. Group II had periodontal ligaments with the presence of collagen fibres and residual inflammatory cells. Group III had a dense inflammatory infiltrate with extensive oedematous areas and fibrillar dissociation, suggesting a persistent inflammatory and resorptive condition. Regarding periapical lesion size, group I had significantly smaller lesions (P < 0.05) than groups II and III, which did not differ significantly from each other. CONCLUSION: Two-session root canal treatment using a calcium hydroxide-based dressing was associated with significantly smaller periapical lesions at 90 days and characterized by progressive repair.

Colored noise and memory effects on formal spiking neuron models.

Simplified neuronal models capture the essence of the electrical activity of a generic neuron, besides being more interesting from the computational point of view when compared to higher-dimensional models such as the Hodgkin-Huxley one. In this work, we propose a generalized resonate-and-fire model described by a generalized Langevin equation that takes into account memory effects and colored noise. We perform a comprehensive numerical analysis to study the dynamics and the point process statistics of the proposed model, highlighting interesting new features such as (i) nonmonotonic behavior (emergence of peak structures, enhanced by the choice of colored noise characteristic time scale) of the coefficient of variation (CV) as a function of memory characteristic time scale, (ii) colored noise-induced shift in the CV, and (iii) emergence and suppression of multimodality in the interspike interval (ISI) distribution due to memory-induced subthreshold oscillations. Moreover, in the noise-induced spike regime, we study how memory and colored noise affect the coherence resonance (CR) phenomenon. We found that for sufficiently long memory, not only is CR suppressed but also the minimum of the CV-versus-noise intensity curve that characterizes the presence of CR may be replaced by a maximum. The aforementioned features allow to interpret the interplay between memory and colored noise as an effective control mechanism to neuronal variability. Since both variability and nontrivial temporal patterns in the ISI distribution are ubiquitous in biological cells, we hope the present model can be useful in modeling real aspects of neurons.

Absence of interleukin 22 affects the oral microbiota and the progression of induced periapical lesions in murine teeth.

AIM: To evaluate the absence of IL-22 on the progression of periapical lesions in wild-type (WT) and IL-22 knockout (IL-22 KO) mice. METHODOLOGY: The evaluation of the oral microbial profile of mice was performed by Checkerboard DNA-DNA hybridization from saliva samples. Periapical lesions were induced in manbibular first molars by pulpal exposure and evaluated after 7, 21 and 42 days (n = 15). Haematoxylin-eosin-stained sections were analysed under conventional and fluorescence microscopy to evaluate the tissue features and size of periapical lesions and tartrate-resistant acid phosphatase histoenzymology (TRAP), Brown & Brenn staining and immunohistochemistry. The scores of the number of bacterial cells present in the oral cavity were analysed by the Mann-Whitney test, and the results and comparisons for periapical lesion size and number of osteoclasts were subjected to one-way anova and Bonferroni's post-test (α = 0.05). RESULTS: Significant differences were observed for bacterial load between the groups of animals for 6 bacterial species (P < 0.05), with five species found in higher levels in the WT group, and one in the IL-22 KO group. WT mice had significantly larger periapical lesions (P < 0.05) between 7 and 42 days and between 21 and 42 days, with an increase in the mean size and number of osteoclasts. IL-22 KO mice had an increase in periapical lesion size and number of osteoclasts between 7 and 21 days (P < 0.05). No differences were found between bacteria localization in the root canal system between the experimental groups. Small variations related to the location of immunostaining were found between the groups. CONCLUSION: This study revealed differences in the composition of oral microbiota between mice that may be taken into account in the susceptibility to infections and development of periapical lesions. The absence of IL-22 in mice resulted in smaller periapical lesions with fewer osteoclasts at the final experimental period, suggesting the participation of IL-22 in the host immune and inflammatory response to a periradicular infection.

Effect of environment fluctuations on pattern formation of single species.

System-environment interactions are intrinsically nonlinear and dependent on the interplay between many degrees of freedom. The complexity may be even more pronounced when one aims to describe biologically motivated systems. In that case, it is useful to resort to simplified models relying on effective stochastic equations. A natural consideration is to assume that there is a noisy contribution from the environment, such that the parameters that characterize it are not constant but instead fluctuate around their characteristic values. From this perspective, we propose a stochastic generalization of the nonlocal Fisher-KPP equation where, as a first step, environmental fluctuations are Gaussian white noises, both in space and time. We apply analytical and numerical techniques to study how noise affects stability and pattern formation in this context. Particularly, we investigate noise-induced coherence by means of the complementary information provided by the dispersion relation and the structure function.

Root canal morphology of primary molars: a micro-computed tomography study.

AIM: This was to investigate the root canal morphology of primary molar teeth using micro-computed tomography. METHODS: Primary maxillary (n = 20) and mandibular (n = 20) molars were scanned at a resolution of 16.7 µm and analysed regarding the number, location, volume, area, structured model index (SMI), area, roundness, diameters, and length of canals, as well as the thickness of dentine in the apical third. Data were statistically compared by using paired-sample t test, independent sample t test, and one-way analysis of variance with significance level set as 5%. RESULTS: Overall, no statistical differences were found between the canals with respect to length, SMI, dentine thickness, area, roundness, and diameter (p > 0.05). A double canal system was observed in the mesial and mesio-buccal roots of the mandibular and maxillary molars, respectively. The thickness in the internal aspect of the roots was lower than in the external aspect. Cross-sectional evaluation of the roots in the apical third showed flat-shaped canals in the mandibular molars and ribbon- and oval-shaped canals in the maxillary molars. CONCLUSIONS: External and internal anatomy of the primary first molars closely resemble the primary second molars. The reported data may help clinicians to obtain a thorough understanding of the morphological variations of root canals in primary molars to overcome problems related to shaping and cleaning procedures, allowing appropriate management strategies for root canal treatment.

Sensorial differences according to sex and ages.

OBJECTIVE: To investigate age and sex differences in orofacial sensory detection. METHODS: One hundred and twenty-six (126) healthy subjects were divided into five groups according to their ages. They were assessed with a quantitative sensory testing protocol for gustative, olfactory, thermal (cold/warm), mechanical (tactile/vibration/electric), and pain (deep/superficial) detection thresholds. The corneal reflex was also evaluated. Data were analyzed with the one-way ANOVA, chi-squared, Fisher's exact, Mann-Whitney, and Kruskal-Wallis tests. RESULTS: The groups of subjects over 61 years old had higher olfactory (P < 0.001), gustative (sweet P = 0.004, salty P = 0.007, sour P = 0.006), thermal (warm P < 0.001, cold P < 0.001), and tactile (P < 0.001) detection thresholds than the others. The vibration detection threshold was high only for subjects over 75 years old (P < 0.001). The electric and deep pain detection thresholds were different for the 61-75 years old group (P ≤ 0.001). Women in all age groups had lower gustative (sweet P = 0.020, salty P = 0.002, sour P < 0.001, and bitter P = 0.002), olfactory (P = 0.010), warm (P < 0.001) and deep (P < 0.001), and superficial pain (P = 0.008) detection thresholds than men, and men from all age groups had lower vibratory detection thresholds (P = 0.006) than women. CONCLUSION: High sensory detection thresholds were observed in subjects over the 6th decade of life, and women had a more accurate sensory perception than men.

MyD88 knockout mice develop initial enlarged periapical lesions with increased numbers of neutrophils.

AIM: To characterize the formation and progression of experimentally induced periapical lesions in teeth of MyD88 knockout (MyD88 KO) mice compared with wild-type (WT) mice. METHODOLOGY: Periapical lesions were induced in the mandibular first molars of 30 WT and 30 MyD88 KO mice. After 7, 21 and 42 days, the animals were euthanized and the mandibles were subjected to histotechnical processing. Histological sections were stained with haematoxylin and eosin (HE), TRAP histoenzymology, Brown and Brenn staining and immunohistochemistry (RANK, RANKL, OPG). Data were subjected to statistical analysis by the nonparametric Mann-Whitney and Kruskal-Wallis tests and the Dunn post-test, using the SPSS software, version 17.0 (α = 0.05). RESULTS: Regarding the periapical lesion size, the MyD88 KO group had significantly higher values than the WT group in the periods of 7 (P = 0.001) and 21 days (P = 0.05). A larger number of neutrophils in the MyD88 KO group were observed (P = 0.01 at 7 days, P = 0.004 at 21 days and P < 0.001 at 42 days). Regarding the number of osteoclasts, no statistically significant difference was observed between the groups at any of the experimental periods (P = 0.884 at 7 days, P = 0.506 at 21 days and P = 0.211 at 42 days). CONCLUSIONS: In the absence of MyD88, the animals had larger periapical lesions, with a severe inflammatory infiltrate and a significantly larger number of neutrophils.

Environmental factors, familial aggregation and heritability of total cholesterol, low density lipoprotein-cholesterol and high density lipoprotein-cholesterol in a Brazilian population assisted by the Family Doctor Program.

OBJECTIVES: To estimate familial aggregation and the heritability of total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C) and high density lipoprotein-cholesterol (HDL-C) in families assisted by the Family Doctor Program in a Brazilian city, and to evaluate associations between some environmental factors and familial aggregation of these lipids. STUDY DESIGN: Cross-sectional familial study. METHODS: The association of lipids with sociodemographic factors, lifestyle factors and comorbidities (e.g. physical activity, alcohol consumption, smoking, hypertension, impaired glucose tolerance, body mass index) was estimated using linear models and generalized estimating equations. Correlation of TC, LDL-C and HDL-C between pairs of relatives was estimated with the familial correlation procedure, and heritability was estimated with the ASSOC procedure. RESULTS: All associations were statistically significant. There was familial aggregation of TC (parent/offspring, r=0.33; sibling/sibling, r=0.37), LDL-C (parent/offspring, r=0.29; sibling/sibling, r=0.37) and HDL-C (parent/offspring, r=0.25; sibling/sibling, r=0.48), but less than 3%, 6% and 14%, respectively, which was explained by lifestyle factors. Correlation between pairs with genetic sharing (parent/offspring and sibling/sibling) was higher than that observed between father and mother. Heritability estimates ranged between 0.32 (HDL-C) and 0.50 (TC). Similar results were found for the two approaches used to estimate the contribution of genetic and environmental factors in the correlation of TC, LDL-C and HDL-C between the family pairs. CONCLUSION: The results showed that there is familial aggregation of TC, LDL-C and HDL-C, and point to the predominance of genetic factors because little influence of environmental variables was found.

Stochastic Langevin equations: Markovian and non-Markovian dynamics.

Non-Markovian stochastic Langevin-like equations of motion are compared to their corresponding Markovian (local) approximations. The validity of the local approximation for these equations, when contrasted with the fully nonlocal ones, is analyzed in detail. The conditions for when the equation in a local form can be considered a good approximation are then explicitly specified. We study both the cases of additive and multiplicative noises, including system-dependent dissipation terms, according to the fluctuation-dissipation theorem.

Morphometric analysis of experimentally induced periapical lesions: radiographic vs histopathological findings.

OBJECTIVES: To evaluate the accuracy and reliability of conventional (Kodak Ektaspeed Plus film) and digitized radiographic images to detect the presence as well to estimate the size, as measured by an image analysis programme, of periapical radiolucencies induced in dog teeth in comparison with the histomorphometric data obtained from the same lesions by conventional and fluorescence microscopy. METHOD: After the removal of pulp, the root canals of five premolars from the same animal were left exposed for 7 days after which they were sealed for 60 days. At day 53, three more premolars were opened and left exposed to the oral cavity for 7 days. Intact premolars were used as control. Conventional radiographs were taken at day 0, day 7, day 30, day 45 and day 60. Morphometry in digitized radiographic images and histological sections were compared at day 7 and day 60 after setting the experimental series. RESULTS: Radiographically, periapical lesions were only detected 30 days after coronal sealing. A progressively increasing radiolucent lesion area was observed at day 45 and day 60. Histopathologically, 7 days after pulp removal dense inflammatory infiltrate and root resorption in the periapical region was observed. At day 7 and day 60, the lesion sizes were similar when evaluated by both conventional and fluorescence microscopy. Lesion size was about 20% larger in digitized radiographs in comparison with histological measurements. CONCLUSIONS: Although image digitization could not improve the detection of the early stages of periapical lesions, it provides a valuable quantitative assessment of extensive periapical lesions. In addition, fluorescence light microscopy enhances the visualization of the apical and periapical structures and seems to be a highly useful tool for histological evaluation, valuable for both qualitative and quantitative studies of periapical disease.

Neuroanatomical approaches of the tectum-reticular pathways and immunohistochemical evidence for serotonin-positive perikarya on neuronal substrates of the superior colliculus and periaqueductal gray matter involved in the elaboration of the defensive behavior and fear-induced analgesia.

Deep layers of the superior colliculus, the dorsal periaqueductal gray matter and the inferior colliculus are midbrain structures involved in the generation of defensive behavior and fear-induced anti-nociception. Local injections of the GABA(A) antagonist bicuculline into these structures have been used to produce this defense reaction. Serotonin is thought to be the main neurotransmitter to modulate such defense reaction in mammals. This study is the first attempt to employ immunohistochemical techniques to locate serotonergic cells in the same midbrain sites from where defense reaction is evoked by chemical stimulation with bicuculline. The blockade of GABA(A) receptors in the neural substrates of the dorsal mesencephalon was followed by vigorous defensive reactions and increased nociceptive thresholds. Light microscopy immunocytochemistry with streptavidin method was used for the localization of the putative cells of defensive behavior with antibodies to serotonin in the rat's midbrain. Neurons positive to serotonin were found in the midbrain sites where defensive reactions were evoked by microinjection of bicuculline. Serotonin was localized to somata and projections of the neural networks of the mesencephalic tectum. Immunohistochemical studies showed that the sites in which neuronal perikarya positive to serotonin were identified in intermediate and deep layers of the superior colliculus, and in the dorsal and ventral columns of the periaqueductal gray matter are the same which were activated during the generation of defense behaviors, such as alertness, freezing, and escape reactions, induced by bicuculline. These findings support the contention that serotonin and GABAergic neurons may act in concert in the modulation of defense reaction in the midbrain tectum. Our neuroanatomical findings indicate a direct neural pathway connecting the dorsal midbrain and monoaminergic nuclei of the descending pain inhibitory system, with profuse synaptic terminals mainly in the pontine reticular formation, gigantocellularis nucleus, and nucleus raphe magnus. The midbrain tectum-gigantocellularis complex and midbrain tectum-nucleus raphe magnus neural pathways may provide an alternative output allowing the organization of the fear-induced anti-nociception by mesencephalic networks.

Mania mista acompanhada de sintomas psicóticos: um ponto de contato entre a esquizofrenia e a psicose maníaco-depressiva? / Mixed mnia with psychotics symptoms: a common way between schizophrenia and maniac depressive psicosis

No presente artigo os autores fazem o relato do caso de uma paciente de trinta anos de idade que, após manifestar, de início, quadro de transtorno bipolar misto, posteriormente evoluiu de forma desfavorável com sintomas psicóticos e deterioração mental grave, levando ao diagnóstico diferencial e prognóstico entre quadros de mania mista com sintomas psicóticos, transtornos esquizoafetivo e esquizofrenia, através de uma revisão de estudos recentes sobre o espectro bipolar e aquelas outras condições clínicas, enfatizando a necessidade de um diagnóstico preciso desses casos e de uma intervenção terapêutica adequada e precoce.

Neuroanatomical and neuropharmacological study of opioid pathways in the mesencephalic tectum: effect of mu(1)- and kappa-opioid receptor blockade on escape behavior induced by electrical stimulation of the inferior colliculus.

Deep layers of the superior colliculus (DLSC), the dorsal and ventral periaqueductal gray matter (PAG), and inferior colliculus (IC) are midbrain structures involved in the generation of defensive behavior. beta-Endorphin and Leu-enkephalin are some neurotransmitters that may modulate such behavior in mammals. Light microscopy immunocytochemistry with streptavidin method was used for the localization of the putative cells of defensive behavior with antibodies for endogenous opioids in rat brainstem. Midbrain structures showed positive neurons to beta-endorphin and Leu-enkephalin in similar distributions in the experimental animals, but we also noted the presence of varicose fibers positive to endogenous opioids in the PAG. Neuroanatomical techniques showed varicose fibers from the central nucleus of the inferior colliculus to ventral aspects of the PAG, at more caudal levels. Naloxonazine and nor-binaltorphimine, competitive antagonists that block mu(1)- and kappa-opioid receptors, were then used in the present work to investigate the involvement of opioid peptide neural system in the control of the fear-induced reactions evoked by electrical stimulation of the neural substrates of the inferior colliculus. The fear-like responses were measured by electrical stimulation of the central nucleus of the inferior colliculus, eliciting the escape behavior, which is characterized by vigorous running and jumping. Central administration of opioid antagonists (2.5 microg/0.2 microl and 5.0 microg/0.2 microl) was performed in non-anesthetized animals (Rattus norvegicus), and the behavioral manifestations of fear were registered after 10 min, 2 h, and 24 h of the pretreatment. Naloxonazine caused an increase of the defensive threshold, as compared to control, suggesting an antiaversive effect of the antagonism on mu(1)-opioid receptor. This finding was corroborated with central administration of nor-binaltorphimine, which also induced a decrease of the fear-like responses evoked by electrical stimulation of the inferior colliculus, since the threshold of the escape behavior was increased 2 and 24 h after the blockade of kappa-opioid receptor. These results indicate that endogenous opioids may be involved in the modulation of fear in the central nucleus of the inferior colliculus. Although the acute treatment (after 10 min) of both naloxonazine and nor-binaltorphimine causes nonspecific effect on opioid receptors, we must consider the involvement of mu(1)- and kappa-opioid receptors in the antiaversive influence of the opioidergic interneurons in the dorsal mesencephalon, at caudal level, after chronic (2-24 h) treatment of these opioid antagonists. The neuroanatomical study of the connections between the central nucleus of the inferior colliculus and the periaqueductal gray matter showed neuronal fibers with varicosities and with terminal bottons, both in the pericentral nucleus of the inferior colliculus and in ventral and dorsal parts of caudal aspects of the periaqueductal gray matter.

Neuroanatomical and psychopharmacological evidence for interaction between opioid and GABAergic neural pathways in the modulation of fear and defense elicited by electrical and chemical stimulation of the deep layers of the superior colliculus and dorsal periaqueductal gray matter.

The effects of central administration of opioid antagonists on the aversive responses elicited by electrical (at the freezing and escape thresholds) or chemical stimulation (crossings, rearings, turnings and jumps, induced by microinjections of bicuculline) of the midbrain tectum were determined. Central microinjections of naloxone and naltrexone in the mesencephalic tectum caused a significant increase in the freezing and escape thresholds elicited by electrical midbrain tectum stimulation. Furthermore, both opioid antagonists caused a significant decrease in the mean incidence of aversive behavioral responses induced by microinjections of bicuculline in the deep layers of the superior colliculus (DLSC) and in dorsal aspects of the periaqueductal gray matter (DPAG), as compared with controls. These findings suggest an opioid modulation of the GABAergic inhibitory inputs controlling the aversive behavior elicited by midbrain tectum stimulation. In fact, immunohistochemical evidence suggests that the dorsal mesencephalon is rich in beta-endorphin-containing neurons and fibers with varicosities. Iontophoretical microinjections of the neurotracer biodextran in the substantia nigra, pars reticulata (SNpr), show nigro-tectal pathways connecting SNpr with the same neural substrate of the DPAG rich in neuronal cells immunoreactive for opioid peptides. Labeled neurons of the DLSC and periaqueductal gray matter send inputs with varsicosities to ipsi- and contralateral DPAG and ipsilateral SNpr. These findings, in addition to the psychopharmacological evidence for the interaction between opioid and GABAergic mechanisms, offer a neuroanatomical basis of a possible presynaptic opioid inhibition of GABAergic nigro-tectal neurons modulating the fear in aversive structures of the cranial mesencephalon, in a short link, and maybe through a major neural circuit, also in GABA-containing perikarya of nigro-tectal neurons.