The most disabling aspect of human peripheral nerve injuries, the majority of which affect the upper limbs, is the loss of skilled hand movements. Activity-induced morphological and electrophysiological remodeling of the neuromuscular junction has been shown to influence nerve repair and functional recovery. In the current study, we determined the effects of two different treatments on the functional and morphological recovery after median and ulnar nerve injury. Adult Wistar male rats weighing 280 to 330 g at the time of surgery (N = 8-10 animals/group) were submitted to nerve crush and 1 week later began a 3-week course of motor rehabilitation involving either "skilled" (reaching for small food pellets) or "unskilled" (walking on a motorized treadmill) training. During this period, functional recovery was monitored weekly using staircase and cylinder tests. Histological and morphometric nerve analyses were used to assess nerve regeneration at the end of treatment. The functional evaluation demonstrated benefits of both tasks, but found no difference between them (P > 0.05). The unskilled training, however, induced a greater degree of nerve regeneration as evidenced by histological measurement (P < 0.05). These data provide evidence that both of the forelimb training tasks used in this study can accelerate functional recovery following brachial plexus injury.

Cell transplantation is a promising experimental treatment for spinal cord injury. The aim of the present study was to evaluate the efficacy of mononuclear cells from human umbilical cord blood in promoting functional recovery when transplanted after a contusion spinal cord injury. Female Wistar rats (12 weeks old) were submitted to spinal injury with a MASCIS impactor and divided into 4 groups: control, surgical control, spinal cord injury, and one cell-treated lesion group. Mononuclear cells from umbilical cord blood of human male neonates were transplanted in two experiments: a) 1 h after surgery, into the injury site at a concentration of 5 x 10(6) cells diluted in 10 µL 0.9% NaCl (N = 8-10 per group); b) into the cisterna magna, 9 days after lesion at a concentration of 5 x 10(6) cells diluted in 150 µL 0.9% NaCl (N = 12-14 per group). The transplanted animals were immunosuppressed with cyclosporin-A (10 mg/kg per day). The BBB scale was used to evaluate motor behavior and the injury site was analyzed with immunofluorescent markers to label human transplanted cells, oligodendrocytes, neurons, and astrocytes. Spinal cord injury rats had 25% loss of cord tissue and cell treatment did not affect lesion extension. Transplanted cells survived in the injured area for 6 weeks after the procedure and both transplanted groups showed better motor recovery than the untreated ones (P < 0.05). The transplantation of mononuclear cells from human umbilical cord blood promoted functional recovery with no evidence of cell differentiation.

Foi analisada a ultraestrutura da glândula reprodutiva corpo dorsal (CD) de Megalobulimus abbreviatus. Imunoistoquímica para microscopia eletrônica foi utilizada para detectar peptídeos relacionados ao tetrapeptídeo FMRFamida nas terminações axonais existentes nessa glândula. Foi utilizada marcação neuronal retrógada com o intuito de localizar os neurônios envolvidos nesta inervação. O CD de M. abbreviatus possui um aspecto uniforme em toda sua extensão, tanto na porção supraesofágica como subesofágica. As células do CD possuem várias características de glândulas esteroidogênicas, tais como a presença de inúmeras gotículas lipídicas, numerosas mitocôndrias com cristas tubulares e cisternas bem desenvolvidas de retículo endoplasmático liso. Por toda a extensão do CD de M. abbreviatus foram encontradas numerosas terminações axonais fazendo contatos estruturalmente semelhantes a sinapses com as células do CD. As terminações axonais continham grande número de vesículas eletrodensas e esparsas vesículas eletrolúcidas. As terminações axonais no CD apresentavam vesículas com conteúdo imunorreativo à FMRFamida. A injeção de traçador neural no CD resultou em marcação retrógrada de neurônios no metacérebro dos gânglios cerebrais e nos gânglios parietais do complexo ganglionar subesofágico de M. abbreviatus. É discutida a possibilidade de que estes neurônios identificados por marcação retrógrada possam representar a via de controle neural do CD de M. abbreviatus, cujo mediador químico seria um neuropeptídeo relacionado à FMRFamida.

The ultrastructure of the reproductive gland, dorsal body (DB), of Megalobulimus abbreviatus was analysed. Electron microscope immunohistochemistry was used to detect FMRFamide-like peptides in the nerve endings within this gland. Nerve backfilling was used in an attempt to identify the neurons involved in this innervation. In M. abbreviatus, the DB has a uniform appearance throughout their supraesophageal and subesophageal portions. Dorsal body cells have several features in common with steroid-secreting gland cells, such as the presence of many lipid droplets, numerous mitochondria with tubular cristae and a developed smooth endoplasmic reticulum cisternae. Throughout the DB in M. abbreviatus numerous axonal endings were seen to be in contact with the DB cells exhibiting a synaptic-like structure. The axon terminals contained numerous electron-dense and scanty electron-lucid vesicles. In addition, the DB nerve endings exhibited FMRFamide immunoreactive vesicles. Injection of neural tracer into the DB yielded retrograde labelling of neurons in the metacerebrum lobe of the cerebral ganglia and in the parietal ganglia of the subesophageal ganglia complex. The possibility that some of these retrograde-labelled neurons might be FMRFamide-like neurons that may represent a neural control to the DB in M. abbreviatus is discussed.

The native species of amphibians and reptiles of Uruguay were categorized according to the IUCN Red List criteria. Out of 47 amphibian species, seven are listed as Critically Endangered (CR), five as Endangered (EN), one as Vulnerable (VU), three as Near Threatened (NT), and two as Data Deficient (DD); the remaining species are considered to be Least Concern (LC). Among the 64 species of reptiles evaluated, one is listed as Critically Endangered (CR), seven as Endangered (EN), two as Vulnerable (VU), one as Near Threatened (NT) and seven as Data Deficient (DD); the rest are considered to be Least Concern (LC). The use of these results as an additional criterion in the definition of protected areas in Uruguay will contribute towards the conservation of the aforementioned threatened species and their associated ecosystems.

Las especies nativas de anfibios y reptiles de Uruguay fueron categorizadas de acuerdo a los criterios de la Lista Roja de UICN. De las 47 especies de anfibios, siete se encuentran En Peligro Crítico (CR), cinco En Peligro (EN), una Vulnerable (VU), tres Casi Amenazadas (NT), y dos con Datos Deficientes (DD); las especies restantes son consideradas como Preocupación Menor (LC). Entre las 64 especies de reptiles evaluados, una se encuentra en Peligro Crítico (CR), siete En Peligro (EN), dos como Vulnerables (VU), una Casi Amenazadas (NT) y siete con Datos Deficientes (DD); las especies restantes son consideradas como Preocupación Menor (LC). El uso de estos resultados como un criterio adicional en la definición de áreas protegidas en Uruguay contribuirá hacia la conservación de dichas especies amenazadas y sus ecosistemas asociados.

The objective of the present study was to determine whether lesion of the subthalamic nucleus (STN) promoted by N-methyl-D-aspartate (NMDA) would rescue nigrostriatal dopaminergic neurons after unilateral 6-hydroxydopamine (6-OHDA) injection into the medial forebrain bundle (MFB). Initially, 16 mg 6-OHDA (6-OHDA group) or vehicle (artificial cerebrospinal fluid - aCSF; Sham group) was infused into the right MFB of adult male Wistar rats. Fifteen days after surgery, the 6-OHDA and SHAM groups were randomly subdivided and received ipsilateral injection of either 60 mM NMDA or aCSF in the right STN. Additionally, a control group was not submitted to stereotaxic surgery. Five groups of rats were studied: 6-OHDA/NMDA, 6-OHDA/Sham, Sham/NMDA, Sham/Sham, and Control. Fourteen days after injection of 6-OHDA, rats were submitted to the rotational test induced by apomorphine (0.1 mg/kg, ip) and to the open-field test. The same tests were performed again 14 days after NMDA-induced lesion of the STN. The STN lesion reduced the contralateral turns induced by apomorphine and blocked the progression of motor impairment in the open-field test in 6-OHDA-treated rats. However, lesion of the STN did not prevent the reduction of striatal concentrations of dopamine and metabolites or the number of nigrostriatal dopaminergic neurons after 6-OHDA lesion. Therefore, STN lesion is able to reverse motor deficits after severe 6-OHDA-induced lesion of the nigrostriatal pathway, but does not protect or rescue dopaminergic neurons in the substantia nigra pars compacta.

Variações morfológicas nos órgãos do sistema reprodutor dos gastrópodes pulmonados são observadas ao longo do ano e podem ser correlacionadas a fases de seu ciclo reprodutivo. A partir dessa observação, a massa dos órgãos do sistema reprodutor do caracol Megalobulimus abbreviatus foi tomada em cada estação do ano e foram obtidas secções histológicas das gônadas. Os valores de massa foram utilizados para a obtenção do índice organo-somático e as secções, para calcular o diâmetro médio dos ovócitos e o índice de maturação ovocitária. Concluímos que M. abbreviatus é um caracol "iteroparous", apresentando ciclo reprodutivo anual caracterizado por acasalamento e oviposição durante a primavera e o início do verão e pela preparação do sistema reprodutor para um nova fase reprodutiva, que se inicia em janeiro e se estende até o final do inverno.

Morphological changes in the sexual organs of the pulmonates were observed throughout a year and correlated with reproductive-cycle periods. Reproductive-organ weights of the snail Megalobulimus abbreviatus were recorded seasonally and gonad sections were analyzed morphologically. The weights were used to obtain the organosomatic index. Mean oocytic diameter and oocytic maturation index were based on gonad sections. It was concluded that M. abbreviatus is an iteroparous snail whose annual reproductive cycle is characterized by mating and egg laying throughout spring and early summer, and also by reproductive system preparation, occurring over the remainder of the summer until the end of winter, for a new breeding season.

We describe the behavior of the snail Megalobulimus abbreviatus upon receiving thermal stimuli and the effects of pretreatment with morphine and naloxone on behavior after a thermal stimulus, in order to establish a useful model for nociceptive experiments. Snails submitted to non-functional (22ºC) and non-thermal hot-plate stress (30ºC) only displayed exploratory behavior. However, the animals submitted to a thermal stimulus (50ºC) displayed biphasic avoidance behavior. Latency was measured from the time the animal was placed on the hot plate to the time when the animal lifted the head-foot complex 1 cm from the substrate, indicating aversive thermal behavior. Other animals were pretreated with morphine (5, 10, 20 mg/kg) or naloxone (2.5, 5.0, 7.5 mg/kg) 15 min prior to receiving a thermal stimulus (50ºC; N = 9 in each group). The results (means ± SD) showed an extremely significant difference in response latency between the group treated with 20 mg/kg morphine (63.18 ± 14.47 s) and the other experimental groups (P < 0.001). With 2.5 mg/kg (16.26 ± 3.19 s), 5.0 mg/kg (11.53 ± 1.64 s) and 7.5 mg/kg naloxone (7.38 ± 1.6 s), there was a significant, not dose-dependent decrease in latency compared to the control (33.44 ± 8.53 s) and saline groups (29.1 ± 9.91 s). No statistically significant difference was found between the naloxone-treated groups. With naloxone plus morphine, there was a significant decrease in latency when compared to all other groups (minimum 64% in the saline group and maximum 83.2% decrease in the morphine group). These results provide evidence of the involvement of endogenous opioid peptides in the control of thermal withdrawal behavior in this snail, and reveal a stereotyped and reproducible avoidance behavior for this snail species, which could be studied in other pharmacological and neurophysiological studies.

A atividade da fosfatase ácida (AcP) foi estudada em duas espécies de teleósteos em duas estações: verão e inverno. A atividade AcP foi detectada em células de Sertoli de tilápia (Oreochromis niloticus) somente durante o período não reprodutivo de seu ciclo anual, que corresponde aos meses de inverno. Em peixe-rei (Odonthestes perugiae), a reação enzimática foi detectada durante o período não reprodutivo (verão) nas células epiteliais dos dutos eferentes, porém não foi detectada nas células de Sertoli. Esses dados sugerem que essa enzima está envolvida no processo de reabsorção do citoplasma residual das espermátides e também na remoção dos espermatozóides remanescentes do período reprodutivo. Em peixe-rei, essa função heterofágica é realizada pelas células dos dutos eferentes e não pelas células de Sertoli.

Acid phosphatase (AcP) activity was investigated in the testes of two species of teleosts in two seasons: summer and winter. AcP activity was detected in Sertoli cells from tilapia (Oreochromis niloticus) only during the nonreproductive period of its annual cycle, corresponding to the winter months. In kingfish (Odonthestes perugiae), the enzymatic reaction was identified during the non-reproductive period (summer) in epithelial cells of the efferent ducts but not in Sertoli cells. These data suggest that the enzyme is involved in the absorption of residual spermatid cytoplasm and as well as in the removal of spermatozoa remaining after the reproductive period. In kingfish, this heterophagous function is carried out by the efferent duct cells and not by Sertoli cells.

In molluscs, the number of peripheral neurons far exceeds those found in the central nervous system. Although previous studies on the morphology of the peripheral nervous system exist, details of its organization remain unknown. Moreover, the foot of the terrestrial species has been studied less than that of the aquatic species. As this knowledge is essential for our experimental model, the pulmonate gastropod Megalobulimus oblongus, the aim of the present study was to investigate monoamines in the pedal plexus of this snail using two procedures: glyoxylic acid histofluorescence to identify monoaminergic structures, and the unlabeled antibody peroxidase anti-peroxidase method using antiserum to detect the serotonergic component of the plexus. Adult land snails weighing 48-80 g, obtained from the counties of Barra do Ribeiro and Charqueadas (RS, Brazil), were utilized. Monoaminergic fibers were detected throughout the pedal musculature. Blue fluorescence (catecholamines, probably dopamine) was observed in nerve branches, pedal and subepithelial plexuses, and in the pedal muscle cells. Yellow fluorescence (serotonin) was only observed in thick nerves and in muscle cells. However, when immunohistochemical methods were used, serotonergic fibers were detected in the pedal nerve branches, the pedal and subepithelial plexuses, the basal and lateral zones of the ventral integument epithelial cells, in the pedal ganglion neurons and beneath the ventral epithelium. These findings suggest catecholaminergic and serotonergic involvement in locomotion and modulation of both the pedal ganglion interneurons and sensory information. Knowledge of monoaminergic distribution in this snail´s foot is important for understanding the pharmacological control of reflexive responses and locomotive behavior.

The vascularization of the central nervous system of the snail Megalobulimus oblongus was studied by injection of carmine-gelatin solution into the arterial system and using a histochemical technique for the detection of alkaline phosphatase. The central nervous system of M. oblongus is irrigated by the anterior aorta, from which a series of small branches emerge that supply the subesophageal nervous ganglia. In turn, these branches give rise to a series of smaller vessels that irrigate the buccal bulb, the anterior portion of the foot, the cerebral ganglia, the dorsal body gland, and the anterior portion of the reproductive system. No hemolymph vessels were detected within nervous tissue although such vessels were found in the periganglionic connective sheath. This connective sheath contains vascular loops and had a series of overlaps and projections that follow the contour of the nervous ganglia. This arrangement permits a larger area of interaction between the surface of the nervous tissue and the hemolymph and reduces the distance between the deepest portion of a given ganglion and the hemolymph vessels.

Immunoreactive substance P was investigated in turtle lumbar spinal cord after sciatic nerve transection. In control animals immunoreactive fibers were densest in synaptic field Ia, where the longest axons invaded synaptic field III. Positive neuronal bodies were identified in the lateral column of the dorsal horn and substance P immunoreactive varicosities were observed in the ventral horn, in close relationship with presumed motoneurons. Other varicosities appeared in the lateral and anterior funiculi. After axotomy, substance P immunoreactive fibers were reduced slightly on the side of the lesion, which was located in long fibers that invaded synaptic field III and in the varicosities of the lateral and anterior funiculus. The changes were observed at 7 days after axonal injury and persisted at 15, 30, 60 and 90 days after the lesion. These findings show that turtles should be considered as a model to study the role of substance P in peripheral axonal injury, since the distribution and temporal changes of substance P were similar to those found in mammals.

O sistema nervoso da ordem Scorpiones parece apresentar um plano comum de organização. Uma abordagem anatômica, utilizando o azul de metileno como meio contrastante para o tecido nervoso, e histológica, por intermédio das técnicas da hematoxilina-eosina e hematoxilina férrica de Heindenhain, permitiu identificar um grande número de nervos em B. bonariensis também presentes em diversas outras espécies de escorpiões, pertencentes a famílias distintas da ordem Scorpiones. Contudo, 2 pares de nervos de B. bonariensis com origem no gânglio queliceral, o par de nervos esofagianos e o par de nervos dos arcos aórticos, parecem divergir desse plano comum, visto não terem sido descritos anteriormente nem possuírem equivalentes em outras espécies de escorpiões pelos critérios de homologia utilizados.

The nervous system of the order Scorpiones appears to have a common organizational structure. The combination of an anatomical study using methylene blue as the contrast medium together with a histological analysis using hematoxylin-eosin and Heindenhain´s iron hematoxylin techniques permitted the identkfication of a large number of nerves in B. bonariensis. Many of these are also present in a variety of other species of scorpions, belonging to distinct families of the order Scorpiones. Nevertheless, two pairs of nerves from B. bonariensis originating in the cheliceral ganglion, one pair of esophageal nerves and one pair of nerves from the aortic arch, appear to diverge from this basic organization. They have not been previously described, nor have any equivalents been reported in research on other scorpion species, in which the current homological criteria have been employed.

Seven days after transection of the sciatic nerve NADPH-diaphorase activity increased in the small and medium neurons of the dorsal root ganglia of the turtle. However, this increase was observed only in medium neurons for up to 90 days. At this time a bilateral increase of NADPH-diaphorase staining was observed in all areas and neuronal types of the dorsal horn, and in positive motoneurons in the lumbar spinal cord, ipsilateral to the lesion. A similar increase was also demonstrable in spinal glial and endothelial cells. These findings are discussed in relation to the role of nitric oxide in hyperalgesia and neuronal regeneration or degeneration.