Rod precursors in the adult retina of the Austrolebias charrua annual fish.

Rod photoreceptors in the adult teleost retina are produced by rod precursors located in the outer nuclear layer (ONL). Annual fishes of the genus Austrolebias exhibit extensive adult retinal cell proliferation and neurogenesis, as well as surprising adaptive strategies to their extreme and changing environment, including adult retinal plasticity. Thus, here we identify and characterize rod precursors in the ONL of the Austrolebias charrua retina. For this aim we used classical histological techniques, transmission electron microscopy, detection of cell proliferation, and immunohistochemistry. Through these complementary approaches, we describe a cell population clearly distinguishable from photoreceptors in the ONL of the adult retina of A. charrua, which we propose corresponds to the rod precursor population. These cells exhibited particular morphological and ultrastructural characteristics, uptake of cell proliferation markers (BrdU+) and expression of stem cell markers (Sox2+). Determining the existence of the population of rod precursors is crucial to understand the sequence of events related to retinal plasticity and regeneration.

Thermoelectric stack sample cooling modification of a commercial atomic force microscopy.

Enabling temperature dependent experiments in Atomic Force Microscopy is of great interest to study materials and surface properties at the nanoscale. By studying Curie temperature of multiferroic materials, temperature dependent phase transitions on crystalline structures or resistive switching phenomena are only a few examples of applications. We present an equipment capable of cooling samples using a thermoelectric cooling stage down to -61.4 °C in a 15 × 15 mm2 sample plate. The equipment uses a four-unit thermoelectric stack to achieve maximum temperature range, with low electrical and mechanical noise. The equipment is installed into a Keysight 5500LS Atomic Force Microscopy maintaining its compatibility with all Electrical and Mechanical modes of operation. We study the contribution of the liquid cooling pump vibration into the cantilever static deflection noise and the temperature dependence of the cantilever deflection. A La0.7Sr0.3MnO3-y thin film sample is used to demonstrate the performance of the equipment and its usability by analyzing the resistive switching phenomena associated with this oxide perovskite.

Olfacto-retinalis pathway in Austrolebias charrua fishes: a neuronal tracer study.

The olfacto-retinal centrifugal system, a constant component of the central nervous system that appears to exist in all vertebrate groups, is part of the terminal nerve (TN) complex. TN allows the integration of different sensory modalities, and its anatomic variability may have functional and evolutionary significance. We propose that the olfacto-retinal branch of TN is an important anatomical link that allows the functional interaction between olfactory and visual systems in Austrolebias. By injecting three different neuronal tracers (biocytin, horseradish peroxidase, and 1,1'-dioctadecyl-3,3,3',3'tetramethyl-indocarbocyanine perchlorate (DiI)) in the left eye of Austrolebias charrua fishes, we identified the olfacto-retinal branch of TN and related neuronal somas that were differentiable by location, shape, and size. The olfacto-retinal TN branch is composed of numerous thin axons that run ventrally along the olfactory bulb (OB) and telencephalic lobes, and appears to originate from a group of many small monopolar neurons located in the rostral portion of both the ipsi- and contralateral OB (referred to as region 1). Labeled cells were found in two other regions: bipolar and multipolar neurons in the transition between the OB and telencephalic lobes (region 2) and two other groups in the preoptic/pretectal area (region 3). In this last region, the most rostral group is constituted by monopolar pear-shaped neurons and may belong to the septo-preoptic TN complex. The second group, putatively located in the pretectal region, is formed by pseudounipolar neurons and coincides with a conserved vertebrate nucleus of the centrifugal retinal system not involved in the TN complex. The found that connections between the olfactory and visual systems via the olfacto-retinal TN branch suggest an early interaction between these sensory modalities, and contribute to the identification of their currently unknown circuital organization.

Proliferation zones in the brain of adult fish Austrolebias (Cyprinodontiform: Rivulidae): a comparative study.

In contrast with mammals, adult fish brains exhibit an enormous potential to produce new cells. Proliferation zones, however, have been described in only a few species, hindering comparisons among genuses and orders. Here we analyzed brain cell proliferation in annual teleostean fishes Austrolebias (Cyprinodontiform: Rivulidae). Immunocytochemistry against 5-bromo-2'-deoxyuridine (BrdU) was quantitated and mapped 24 h after injection in three species with different phylogenetic positions or habitats. All species had similar brain anatomy and total volume, but olfactory bulbs, torus longitudinalis and cerebellum were of different sizes in different species. Cell proliferation was found throughout the brain. Three-D reconstructions provided evidence for contiguity along the rostro-caudal axis and concentration in the vicinity of the ventricles. Brain regions analyzed exhibited high mitotic activity, and the torus longitudinalis had the highest volume-normalized proliferation index. A. affinis exhibited the highest normalized proliferation indexes in visual regions but the lowest in olfactory bulb. A. reicherti showed an inverse pattern, suggesting that these species have a different hierarchy of sensorial modalities that could be related to phylogeny or habitat. Double immunostaining against BrdU and cell-type specific markers was performed to determine the fate of proliferating cells. A widespread gliogenesis was evidenced. Few cells positive for both BrdU and the neuronal marker HuC/D were found in the brain of the three species, demonstrating neurogenesis in the adult Austrolebias brain. Summarizing, adult members of the three species showed similar brain anatomy and cell proliferation patterns. Among species, volume-normalized proliferation indexes varied in regions involved in different sensory modalities. To our knowledge, this is the first report showing proliferating cells with neuronal markers as earlier as 24 h after BrdU injection.

Subcortical deafferentation impairs behavioral reinforcement of long-term potentiation in the dentate gyrus of freely moving rats.

Long-term potentiation is a form of neural functional plasticity which has been related with memory formation and recovery of function after brain injury. Previous studies have shown that a transient early-long-term potentiation can be prolonged by direct stimulation of distinct brain areas, or behavioral stimuli with a high motivational content. The basolateral amygdala and other subcortical structures, like the medial septum and the locus coeruleus, are involved in mediating the reinforcing effect. We have previously shown that the lesion of the fimbria-fornix--the main entrance of subcortical afferents to the hippocampus--abolishes the reinforcing basolateral amygdala-effects on long-term potentiation in the dentate gyrus in vivo. It remains to be investigated, however, if such subcortical afferents may also be important for behavioral reinforcement of long-term potentiation. Young-adult (8 weeks) Sprague-Dawley male rats were fimbria-fornix-transected under anesthesia, and electrodes were implanted at the dentate gyrus and the perforant path. One week after surgery the freely moving animals were studied. Fimbria-fornix-lesion reduced the ability of the animals to develop long-term potentiation when a short pulse duration was used for tetanization (0.1 ms per half-wave of a biphasic stimulus), whereas increasing the pulse duration to 0.2 ms per half-wave during tetanization resulted in a transient early-long-term potentiation lasting about 4 h in the lesioned animals, comparable to that obtained in non-lesioned or sham-operated control rats. In water-deprived (24 h) control animals, i.e. in non-lesioned and sham-operated rats, early-long-term potentiation could be behaviorally reinforced by drinking 15 min after tetanization. However, in fimbria-fornix-lesioned animals long-term potentiation-reinforcement by drinking was not detected. This result indicates that the effect of behavioral-motivational stimuli to reinforce long-term potentiation is mediated by subcortical, heterosynaptic afferents.

[Effects of simultaneous transplant of foetal mesencephalic cells in the striatum and the subthalamic nucleus of hemiparkinsonian rats]. / Efectos del trasplante simultáneo de células mesencefálicas fetales en el estriado y el núcleo subtalámico de ratas hemiparkinsonianas.

INTRODUCTION: The main strategy followed in neural transplants as a method of treatment for Parkinson s disease, both experimental and clinical, has been to introduce foetal mesencephalic cells into the target area: the striatum. However, when the dopaminergic cells in the substantia nigra degenerate, not only is the dopaminergic innervation of the striatum affected but also other nuclei: globus pallidus, substantia nigra, substantia nigra pars reticulata and subthalamic nucleus. A series of data from pharmacological and physiological studies offer strong evidence that the dopamine released in these nuclei may play an important role in regulating the output nuclei of the basal ganglia. AIM: To evaluate the effect of transplanting foetal mesencephalic cells on the behaviour of 6 OH DA rats when introduced into the striatum and the subthalamic nucleus. MATERIALS AND METHODS: 6 OH DA was used to induce lesions in the substantia nigra of rats, which were divided into several experimental groups. The rotating activity induced by D amphetamine (5 mg/kg, intraperitoneally) and apomorphine (0.05 mg/kg, subcutaneously) was evaluated before and three months after the transplant in all the experimental groups, except in the control group of healthy rats. The hemiparkinsonian rats received a total of 350,000 foetal ventral mesencephalic cells, which were implanted within small deposits in the striatum (8) and in the subthalamic nucleus (4). RESULTS AND CONCLUSIONS: Rotation induced by both drugs was significantly lower (p= 0.05) in animals that had had dopaminergic cells transplanted into the striatum body. No significant improvement in this behaviour was to be found when transplants were limited to just the subthalamus or, simultaneously, also to the striatum. A significant increase in rotating behaviour induced by apomorphine was observed in the group which received a transplant in just the subthalamus.

Behavioral reinforcement of long-term potentiation is impaired in aged rats with cognitive deficiencies.

Behavioral stimuli with emotional/motivational content can reinforce long-term potentiation in the dentate gyrus, if presented within a distinct time window. A similar effect can be obtained by stimulating the basolateral amygdala, a limbic structure related to emotions. We have previously shown that aging impairs amygdala-hippocampus interactions during long-term potentiation. In this report we show that behavioral reinforcement of long-term potentiation is also impaired in aged rats with cognitive deficits. While among young water-deprived animals drinking 15 min after induction of long-term potentiation leads to a significant prolongation of potentiation, cognitively impaired aged rats are devoid of such reinforcing effects. In contrast, a slight but statistically significant depression develops after drinking in this group of animals. We suggest that an impaired mechanism of emotional/motivational reinforcement of synaptic plasticity might be functionally related to the cognitive deficits shown by aged animals.

[Behavior characterization of a model of Huntington's disease in rats, induced by quinolinic acid]. / Caracterización conductual de un modelo de enfermedad de Huntington en ratas inducido por ácido quinolínico.

INTRODUCTION: Huntington's disease (HD) is a progressive neurodegenerative disorder, characterized by severe degeneration of basal ganglia neurons. Behavioral symptoms of HD include abnormal, uncontrollable and constant choreiform movements, impaired cognitive function and emotional disturbance. OBJECTIVE: In order to explore the changes of cognitive and motor functions induced by quinolinate lesion we realized this experiment. MATERIALS AND METHODS: We studied the behavior of rats with unilateral quinolinate induced lesions of the medial striatum. Intact 3 months old male rats (n = 23) were trained in the Morris Water Maze during three consecutive days, eight trials/day (acquisition), and before surgery they were randomly assigned either to intact or lesion groups. Fifteen days after the lesion the rats were tested using retention test (one day/four trials, with the escape platform in the same position as in acquisition test), on the next three days the rats were tested in the transfer test (three days/eight trials-day, with the platform in the new position). The Paw reaching test and the asymmetrical rotational behavior test in respond to amphetamine were also tested in these rats. RESULTS: Lesioned animals exhibited deficient retrieval of stored memories of visuospatial skills and impaired transfer of learning. In relation with motor activity the lesioned rats showed a profound impairment in the skill of the left forelimb for reaching food compared with its right forelimb as well as with the forelimb abilities of intact rats. The lesioned animals showed significant rotational behavior induced by amphetamine agonist, ipsilateral to the lesioned striatum. CONCLUSIONS: These results are consistent with the notion that the striatal degeneration could sufficiently account for the cognitive abnormalities associated with HD, and with the key role played by basal ganglia in enabling voluntary and postural adjustment of the movements.

Caracterización conductual de un modelo de enfermedad de huntington en ratas inducido por ácido quinolínico / Behaviour characterization of a model of huntington's disease in rats, induced by quinolinic acid

Introducción. La enfermedad de Huntington (EH) es una entidad progresiva y neurodegenerativa, caracterizada por una grave degeneración de los ganglios basales. Los síntomas conductuales de la EH incluyen, movimientos coreicos constantes e incontrolables, deterioro de las funciones cognitivas y desequilibrio emocional. Objetivo. Este estudio se realizó con el objetivo de explorar los cambios en las funciones motora y cognitiva inducidos por la lesión estriatal con ácido quinolínico. Material y métodos. Se realizaron estudios conductuales en ratas con lesión unilateral en el estriado medial inducida con ácido quinolínico. Veintitrés ratas macho de 3 meses fueron entrenadas en el laberinto acuático de Morris durante tres días consecutivos, ocho ensayos por día (prueba de adquisición) y antes de la cirugía los animales fueron asignados al azar al grupo control o lesionado. Quince días después de la lesión, se evaluó a los animales mediante la prueba de retención (cuatro ensayos en un día, con la plataforma situada en el mismo lugar que ocupaba durante la adquisición); en los tres días subsiguientes se realizó la prueba de transferencia (ocho ensayos durante tres días, con la plataforma en una nueva localización). También se llevaron a cabo las pruebas de habilidad de las extremidades anteriores y de conducta rotacional inducida por anfetamina. Resultados. Los animales lesionados exhibieron una recuperación de la memoria visuoespacial deficiente y un deterioro de la transferencia del aprendizaje. En relación con la actividad motora, las ratas lesionadas presentaron un déficit marcado en la habilidad del uso de la extremidad izquierda para coger el alimento, en comparación con la extremidad derecha o con la habilidad general desarrollada por las ratas controles. Los animales lesionados también mostraron rotaciones ipsilaterales al estriado lesionado inducidas por el agonista anfetamina. Conclusiones. Los resultados coinciden plenamente con el planteamiento de que la degeneración estriatal debe ser suficiente para causar deterioro en las funciones cognitivas relacionadas con la EH y, además, para afirmar la importancia del papel de los ganglios basales en la ejecución de movimientos voluntarios y de ajuste postural del cuerpo (AU)

[Simultaneous transplantation of fetal mesencephalic cells to the striatum and globus pallidus of rats with lesions induced by 6-hydroxydopamine]. / Trasplante simultáneo de células mesencefálicas fetales en striatum y globus pallidus de ratas con lesión por 6-hidroxidopamina.

INTRODUCTION: Studies of neural transplants in experimental models of Parkinson's disease have concentrated their attention on ectopic transplants of foetal mesencephalic cells to denervated striatum. However, the external globus pallidus has recently been shown to play an important part in the physiopathology of this disease. OBJECTIVE: Bearing in mind the importance of loss of extra-striatal dopamine in the genesis of the clinical signs found in parkinsonism, the objective of this study was to evaluate the effect of foetal mesencephalic transplantation to the globus pallidus of hemiparkinsonian rats. MATERIAL AND METHODS: Following conventional transplantation methodolgy, suspensions of cells from the ventral mesencephalum of rat embryos (E-14) were implanted. The tissue was grafted into the striatum, pallidum-striatum and pallidum areas of rats with unilateral lesions of the striatonigral bundle. One, two, three and six months after transplantation, the rotatory activity induced by D-amphetamine was evaluated. The rotatory behaviour induced by apomorphine was evaluated at three months. Motor ability of the front legs was evaluated in all experimental groups three months after transplantation using the 'ladder test'. RESULTS: In the experimental groups in which a transplant was made to the globus pallidus there was a significant reduction (p < 0.01) in rotatory activity induced by D-amphetamine and by apomorphine as compared with the non-transplanted groups. CONCLUSIONS: Transplants of foetal dopaminergic cells survive in the globus pallidus of hemiparkinsonian rats and can improve the rotational activity induced by dopaminergic agonists.

[Neuronal toxicity of human recombinant interleukin-2 in rats. Morphological and behavioral validation]. / Toxicidad neuronal de la interleucina-2 recombinante humana en ratas. Validación morfoconductual.

INTRODUCTION AND OBJECTIVE: The memory impairment which accompanies the aging process is a manifestation of diminished cognitive function. This is intimately related to neuropathological and biochemical changes in cholinergic areas of central nervous system (CNS). Cytokines, first described as immunoregulators, are also implied in defense reactions of the brain. Some studies on the action of IL-2 on the CNS suggest an action blocking the release of acetylcholine in the hippocampus. MATERIAL AND METHODS: We have studied the possible central neurotoxic effect of this soluble factor using the chronic intraperitoneal infusion of human recombinant IL-2 (hr-IL 2) to young and old Sprague Dawley rats. RESULTS AND CONCLUSIONS: The results do not show an in vivo action of IL-2 on the cholinergic function but are consistent with the probable role of this cytokine in the senescent cognitive impairment, in particular the age-related loss of spatial memory and/or during the evolution of neurodegenerative related process.

Toxicidad neuronal de la interleucina-2 recombinante humana en ratas: validación morfoconductual / Neuronal toxicity of human recombinant interleukin 2 in rats: morphoconductual validation

Introducción y objetivo. La afectación de memoria que se observa en el envejecimiento es una manifestación de la disminución de las funciones cognitivas con la edad, la cual está estrechamente asociada a cambios neuropatológicos y bioquímicos en áreas colinérgicas del sistema nervioso central (SNC). Las citoquinas, descritas por primera vez como moléculas inmunoreguladoras, están también implicadas en reacciones defensivas del cerebro. Estudios relacionados con la acción de la IL-2 sobre el SNC le atribuyen un efecto bloqueador sobre la secreción de acetilcolina a nivel hipocampal. Material y métodos. Hemos desarrollado un estudio dirigido a caracterizar los efectos neurotóxicos centrales de esta citoquina mediante la infusión crónica intraperitoneal de IL-2 recombinante humana (IL-2rh) en ratas jóvenes y viejas de la línea Sprague Dawley. Resultados y conclusiones. Los resultados obtenidos, aunque parciales, no parecen referir el posible efecto in vivo de la IL-2 sobre la función colinérgica central, pero si son consistentes con la implicación probable de esta citoquina en el deterioro cognitivo senescente y, de manera particular, en el deterioro de la memoria espacial asociada a la edad y/o en el curso de trastornos neurodegenerativos relacionados