Ana Leonor A. Nencioni

Behavioral, Electroencephalographic, and Histopathologic Effects of a Neuropeptide Isolated From Tityus serrulatus Scorpion Venom in Rats

The effects of intrahippocampal administration of a neuropeptide (TS-8F toxin) isolated from Tityus serrulatus scorpion venom have been determined on behavior, limbic seizures, and neuronal degeneration in rats. Behavioral observation showed orofacial automatism, wet dog shakes, and myoclonus. Concomitantly, the electroencephalographic record showed high-frequency and high-voltage spikes that evolved to seizure activity in the hippocampus and cortex. Seven days after TS-8F toxin microinjection, neuronal damage was observed in CA1 and CA2 pyramidal cells and in granular cells of the dentate gyrus. The results suggest that TS-8F toxin may be responsible, at least in part, by the epileptic effects observed with the crude venom. Thus, this toxin may be a useful tool in the study of some neurobiological process.

A microdialysis study of glutamate concentration in the hippocampus of rats after TsTX toxin injection and blockade of toxin effects by glutamate receptor antagonists

Scorpion toxins act on ionic channels changing the release of neurotransmitters. In the present study, we investigated the glutamatergic release evoked by intrahippocampal injection of TsTX toxin isolated from Tityus serrulatus scorpion venom in male Wistar rats and the blockade of the toxin effect by glutamatergic antagonists. Microdialysis for neurotransmitter level quantification, electroencephalographic recording, and histopathological analysis were performed. The microdialysis method revealed enhanced levels of extracellular glutamate in the hippocampal area. The toxin injection preceded by injection of the glutamate receptor antagonists dizolcipine maleate (MK-801), D(-)2-amino-5-phosphonopentanoic acid (AP-5), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), L(+)-2-amino-3-phosphonopropionic acid (AP-3), and (+)-alpha-methyl-4-carboxyphenylglycine (MCPG) demonstrated that MK-801 and AP-5 fully blocked the electrographic alterations and the CA1 cell loss induced by the toxin. CNQX, AP-3, and MCPG partially blocked the epileptiform discharges and no hippocampal damage was observed. Thus, we conclude that the toxin evokes glutamate release and that glutamate receptor antagonists can partially or totally block the toxin effect.

Central effects of Tityus serrulatus and Tityus bahiensis scorpion venoms after intraperitoneal injection in rats.

A great number of studies on scorpion venoms associate their effects to the autonomic nervous system, and few data are available about their action on the central nervous system (CNS). The aim of this work was to evaluate some central effects after intraperitoneal injection of Tityus serrulatus or T. bahiensis scorpion venoms. The hippocampal concentration of some neurotransmitters and their metabolites were determined. Electroencephalographic and behavioral observations were performed, and all brains were removed for histopathological analysis of hippocampal areas. Both venoms induced electrographic and behavioral alterations despite T. bahiensis venom affects less the electrographic activity than T. serrulatus venom. Neurochemical analysis demonstrated no alteration in the extracellular levels of almost all the neurotransmitters evaluated, at least in the hippocampus, and no neuronal loss in this area was observed. Meanwhile, extracellular concentration of HVA increased up to 10 times in approximately 1/3 of the animals of both groups. Scorpion venoms seem to exert a small but important central effect. More studies in this field are necessary because they may be useful in developing new strategies to reduce the damage caused by scorpion stings.

Effects of prenatal exposure to Tityus bahiensis scorpion venom on rat offspring development

Scorpion envenoming is a public health problem. In Brazil, the scorpion Tityus serrulatus is considered the most dangerous, but a large number of exposures also occur with Tityus bahiensis. There are quite a few studies in literature about the toxic effects of this venom but it is not known if the venom causes malformations or behavioral defects to the offspring of mothers exposed to the venom during pregnancy. The objective of this work was to determine, in rats, the possible toxic effects of T. bahiensis venom on offspring when injected into rats during different periods of fetal development. Rats were assigned to one of three groups: one control group and two experimental groups that were subcutaneously injected with venom (2.5mg/kg) on the 10th (GD10) or on 16th day (GD16) of gestation. Pups were evaluated for changes in physical and behavioral development. GD10 treatment group offspring showed an increase in body weight gain, earlier ear unfolding, incisor tooth eruption and vaginal opening. A decrease in the time of palmar grasp and surface-righting reflexes was observed only for males. In GD16 treatment group, earlier ear unfolding, incisor tooth eruption, and delay in eye opening were observed in the offspring. In female pups a decrease in weight gain and in time for palmar grasp reflex, and an increase in time for negative geotaxis were observed. In male pups a delay in the testis descent, decrease in the time of palmar grasp, increase in the time of negative geotaxis reflex and in the general and locomotor activities could be noticed. Therefore, we concluded that a moderate dose of scorpion venom administered to pregnant rats was able to elicit alterations in physical and behavioral development in the offspring during the postnatal period.

Effects of a toxin isolated from Tityus bahiensis scorpion venom on the hippocampus of rats

AIMS The objective of the present study was to determine the effects of a toxin from T. bahiensis scorpion venom on the hippocampus of rats. This toxin, called Tb V-4, was chosen since it shows remarkable convulsive activity. MAIN METHODS Male Wistar rats weighing 250g were used. The toxin (1.0μg/μl) was injected into the hippocampus. The animals were then submitted to electroencephalographic and behavioral examinations or to microdialysis to determine the levels of neurotransmitters. The location of the implanted guide cannulae and electrodes was checked histologically. The number of cells in the CA1, CA3 and CA4 areas of the hippocampus was determined by light microscopy. Changes in the concentration of cytosolic free calcium were evaluated by confocal microscopy. KEY FINDINGS The toxin evoked behavioral alterations such as wet dog shakes, myoclonus, yawning and orofacial automatisms. Electroencephalographic recordings exhibited alterations such as isolated or grouped spikes and epileptic-like discharges. Injection of the toxin augmented glutamate concentration in the extracellular fluid in some animals. There was also a decrease in the number of pyramidal cells, mainly in the CA1 and CA4 areas for some rats. In some slices of the hippocampus, an increase in intracellular calcium mobilization was seen. SIGNIFICANCE The present results suggest that the Tb V-4 toxin may be responsible for the epileptic and behavioral effects observed with the crude venom. We suggest that the convulsive and degenerative effects induced by the toxin could be due to the enhanced release of excitatory amino acids involved in the most important pathways of the hippocampus.

Intrahippocampal injection of TsTX-I, a beta-scorpion toxin, causes alterations in electroencephalographic recording and behavior in rats

AIMS TsTX-I scorpion toxin, also known as γ-toxin, is a β-toxin which binds to site 4 of the sodium channel, shifting its activation potential. There are few studies about its pharmacological action in the central nervous system. The objective of this work was to determine the electroencephalographic, behavioral and histopathological effects of intrahippocampal injection of TsTX-I. MAIN METHODS Rats were anesthetized and fitted with cannulae for injection into the hippocampus and with electrodes for cerebral recording. The animals were treated with Ringer solution, some doses of TsTX-I, DMSO 0.1% or veratridine. Behavioral and electrographic recordings were observed for 4 hours after the injection. After 7 days, the rats were perfused, and their brains removed for histological analysis. KEY FINDINGS Increasing doses of the toxin evoked epileptic-like discharges, wet dog shakes, and in some cases hind limb paralysis and intense respiratory difficulty followed by death. The histopathological analysis demonstrated no cell loss. Animals injected with veratridine developed epileptiform activity in the electrographic recording and neuronal loss. SIGNIFICANCE The results suggest that TsTX-I toxin may be responsible, at least in part, for the epileptic and behavioral effects observed with the crude venom, and although veratridine and TsTX-I act on Na-channel, the differences between them are remarkable, demonstrating that toxins can have different functional effects depending on the site of action in the channel. Thus, animal neurotoxins are often highly selective and may be useful for the identification of the sequence of events underlying neurotransmission.

Effects of in utero exposure to Tityus bahiensis scorpion venom in adult rats

The toxicity of Tityus bahiensis scorpion venom is well known, but there are little data about the damage in offspring of dams that were exposed to the venom during pregnancy. The objective of this work was to determine the toxic effects of venom in adult offspring of Wistar rats exposed to venom in utero. Dams were divided into a control group, subcutaneously injected with saline solution on the 10th (GD10) and 16th (GD16) days, and two experimental groups, subcutaneously injected with venom (2.5mg/kg) on GD10 or GD16, respectively. Adult offspring were evaluated according to behavioral development and neuronal integrity in the hippocampus. Tests performed in the activity box and in the enriched environment demonstrated that males from GD10 had motor decrease. Females from GD10 showed a depressive-like state and were more anxious, as demonstrated by the forced swimming test and social interaction. The plus-maze discriminative avoidance task demonstrated that GD16 males had lower levels of anxiety. The number of neuronal cells was decreased in CA1, CA3 and CA4 hippocampal areas of males and females from GD10 group and in CA1 of females and CA4 of males from GD16 group. Thus, we conclude that venom exposure in pregnant dams causes subtle alteration in the behavioral and neuronal development of offspring in adult life in a gender-dependent manner.

Mild reproductive effects of the Tityus bahiensis scorpion venom in rats

BackgroundScorpion envenoming is a public health problem in Brazil, where Tityus serrulatus and T. bahiensis are considered the most dangerous scorpions. They are well adapted to urbanized environments, and there is an increasing probability of human exposure to these venoms, including during pregnancy. Not much is known about the effects of prenatal exposure to the venom, and no information is available to aid in the rational treatment of victims stung during pregnancy. Thus, this study aimed to investigate whether venom from the scorpion T. bahiensis administered once to pregnant female rats at a dose that causes a moderate envenomation may lead to deleterious effects on the reproductive performance of the dams and on the development of their offspring. This is the first work demonstrating that T. bahiensis venom, when administered experimentally to rats, alters maternal reproductive performance and the morphological development of fetuses. The venom was given to dams on the 5th (GD5) or on the 10th (GD10) gestational day. After laparotomy, on GD21, fetuses and placentas were counted, weighed and externally analyzed. The corpora lutea were counted. The sex and vitality of fetuses were evaluated, and each litter was then randomly divided for visceral or skeletal analyses. Data were analyzed by ANOVA followed by the Tukey-Kramer test and Fisher’s exact test. The significance level for all tests was set at p < 0.05.ResultsGD5 group presented an increased number of pre-implantation losses. Weight gains in fetuses and placentas were observed in the GD5 and GD10 groups. Weights of the heart and lungs were elevated in GD5 and GD10 and liver weight in GD10.ConclusionsModerate envenomation by T. bahiensis scorpion venom alters maternal reproductive performance and fetal development. However, these are preliminary results whose causes should be investigated more carefully in future studies.

Convulsive and Neurodegenerative Effects in Rats of Some Isolated Toxins from the Tityus bahiensis Scorpion Venom

Despite Tityus bahiensis being one of the most dangerous scorpions in Brazil, there are few studies about the effects of its venom, which acts mainly on the central nervous system. Previous studies demonstrated the convulsive ability of this venom. The present work aimed to study the hippocampal effects in rats of some toxins isolated from pool V, which induces a pronounced epileptogenic effect. The pool was separated by reverse-phase HPLC, and the peaks with higher yield (Tb V-1, V-5, V-24, V-27, and V-28) were used in the experiments. Cannulae and electrodes were implanted in the hippocampus of male Wistar rats (240–250 g). The animals were divided into six groups that received intracerebral injection of toxin solution (1 or 2 μg/μL) or Ringer solution (control group), and they were submitted to behavioral, electrographic, and histological analysis. All toxins studied evoked electrographic and behavioral epileptic-like activity to different degrees. Moreover, the toxins V-1, V-24, and V-28 caused significant neuronal loss in CA4 ipsi- and contralateral hippocampal areas. These results suggest that toxins from T. bahiensis scorpion, when injected into the hippocampus, are able to act directly on the central nervous system inducing convulsive and neurodegenerative effects.

Some pharmacological effects of Tityus obscurus venom in rats and mice

ABSTRACT There are a great number of studies about Brazilian scorpions. However, little is known about the venom of scorpions of northern Brazil, mainly about Tityus obscurus, which is responsible for the most number of accidents in the Amazon. Thus, this study aimed to evaluate some pharmacological effects of T. obscurus venom in rats and mice. In rats, the venom (10 mg/kg i.p.) caused hemorrhagic patches in the lung parenchyma but did not lead to pulmonary edema. There was a decrease in general activity, observed in the activity box after venom injection. The venom did not induce changes in the occurrence and intensity of experimentally induced convulsions, nor did it cause hippocampal neuronal loss. In mice, the LD50 obtained was 3.13 mg/kg (i.p.). Different doses of the venom (0.2; 1; 5; 10; 15 &mgr;g/30 &mgr;L per hind paw) induced edematogenic and moderate nociceptive activity in mice. The Tiyus serrulatus venom used as comparison caused more intense symptomatology in mice. Comparing to the venom of other Tityus scorpions of medical importance, that have convulsant and intense nociceptive effects and cause lung edema, as described in the literature, we can conclude that the venom of T. obscurus probably has different characteristics. HIGHLIGHTSTityus obscurus is the main scorpion responsible for accidents in the Amazon region.Tityus obscurus, similar to T. serrulatus venom, exerted edematogenic activity in mice.Tityus obscurus venom caused hemorrhagic patches in the lung of rats.Both nociceptive and lethal activities of T. obscurus venom were lower than those caused by T. serrulatus venom in mice.