Endocannabinoid signaling mechanisms in the substantia nigra pars reticulata modulate GABAergic nigrotectal pathways in mice threatened by urutu-cruzeiro venomous pit viper.

The substantia nigra pars reticulata (SNpr) is rich in γ-aminobutyric acid (GABA)-ergic neurons and connected to the mesencephalic tectum (MT) structures, such as the superior colliculus and dorsal periaqueductal gray matter. The SNpr presents a high density of cannabinoid receptors (CBRs), suggesting a possible regulatory role that is played by endocannabinoids (eCBs) in the ventral mesencephalon. The present study investigated the involvement of SNpr eCB mechanisms in nigrotectal pathways in the expression of defensive behavior associated with instinctive fear and panic reactions in mice that are confronted with the venomous Viperidae snake Bothrops alternatus. The localization of CB1 receptors (CB1RS) and synaptophysin glycoprotein in the SNpr was also evaluated. Administration of the GABAA receptor antagonist bicuculline in the MT increased defensive responses to the snake that are related to panic, such as freezing and non-oriented escape reactions, sometimes toward the snake itself. Mice that were pretreated with anandamide (5 or 50pmol) in the SNpr, followed by an injection of physiological saline or bicuculline in the MT, exhibited significant decreases in the expression of alertness, freezing, and escape responses. Immunofluorescence showed the presence of fibers that were rich in CB1RS and synaptophysin in the SNpr, indicating that these receptors appear to be located mainly in presynaptic terminals in the striatonigral pathway. These findings suggest that eCB mechanisms in the SNpr facilitate the activity of nigrotectal GABAergic pathways, modulating the activity of striatonigral links during the elaboration and organization of innate fear and panic-like responses in threatening situations.

Effect of chronic intake of sweet substance on nociceptive thresholds and feeding behavior of Rattus norvegicus (Rodentia, Muridae).

The investigation of the influence of sweetened food on feeding behavior targeted to non-sucrose nutrients as well as the sensitivity to painful stimuli in isolated and grouped animals is the aim of the present work. The tail withdrawal latencies in the tail-flick test (a spinal reflex) were measured before and immediately after the treatment with tap water or sucrose (62, 125 or 250 g/l). Our findings suggest that: (a) The analgesic effect of sucrose intake depends on the concentration of sucrose solution and on the time during which the solution is consumed; (b) the most effective concentration of sucrose followed by antinociceptive effect was the one of 250 g/l in both isolated and grouped animals; (c) considering the individually caged rats, the intake of sucrose in the highest concentration (250 g/l) was the smallest as compared with the consumption of sucrose in more diluted solutions (62.5 and 125 g/l), but this higher sweetened solution was followed by antinociception; (d) animals treated with concentrated sucrose solution ate smaller quantities of pellets than animals treated with tap water; (e) tonic intake of highly concentrated sweet substance seems to be crucial for the increase of the nociceptive threshold in our model of sweet substance-induced antinociception.

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.