Changes in CREB activation in the prefrontal cortex and hippocampus blunt ethanol-induced behavioral sensitization in adolescent mice

Drug dependence is a major health problem in adults and has been recognized as a significant problem in adolescents. We previously demonstrated that repeated treatment with a behaviorally sensitizing dose of ethanol in adult mice induced tolerance or no sensitization in adolescents and that repeated ethanol-treated adolescents expressed lower Fos and Egr-1 expression than adult mice in the prefrontal cortex (PFC). In the present work, we investigated the effects of acute and repeated ethanol administration on cyclic adenosine monophosphate (cAMP) response element-binding protein (CREB) DNA-binding activity using the electrophoretic mobility shift assay (EMSA) and the phosphorylated CREB (pCREB)/CREB ratio using immunoblotting in both the PFC and hippocampus in adolescent and adult mice. Adult mice exhibited typical locomotor sensitization after 15 days of daily treatment with 2.0 g/kg ethanol, whereas adolescent mice did not exhibit sensitization. Overall, adolescent mice displayed lower CREB binding activity in the PFC compared with adult mice, whereas opposite effects were observed in the hippocampus. The present results indicate that ethanol exposure induces significant and differential neuroadaptive changes in CREB DNA-binding activity in the PFC and hippocampus in adolescent mice compared with adult mice. These differential molecular changes may contribute to the blunted ethanol-induced behavioral sensitization observed in adolescent mice.

Daily cycling of nitric oxide synthase (NOS) in the hippocampus of pigeons (C. livia)

Background: Nitric oxide synthase (NOS) is essential for the synthesis of nitric oxide (NO), a non-conventional neurotransmitter with an important role in synaptic plasticity underlying processes of hippocampus-dependent memory and in the regulation of biological clocks and circadian rhythms. Many studies have shown that both the NOS cytosolic protein content and its enzymatic activity present a circadian variation in different regions of the rodent brain, including the hippocampus. The present study investigated the daily variation of NOS enzymatic activity and the cytosolic content of nNOS in the hippocampus of pigeons. Results: Adult pigeons kept under a skeleton photoperiod were assigned to six different groups. Homogenates of the hippocampus obtained at six different times-of-day were used for NOS analyses. Both iNOS activity and nNOS cytosolic protein concentrations were highest during the subjective light phase and lowest in the subjective dark phase of the circadian period. ANOVA showed significant time differences for iNOS enzymatic activity (p < 0.05) and for nNOS protein content (p < 0.05) in the hippocampus. A significant daily rhythm for both iNOS and nNOS was confirmed by analysis with the Cosinor method (p < 0.05). The present findings indicate that the enzymatic activity of iNOS and content of nNOS protein in the hippocampus of pigeons exhibit a daily rhythm, with acrophase values occurring during the behavioral activity phase. Conclusions: The data corroborate the reports on circadian variation of NOS in the mammalian hippocampus and can be considered indicative of a dynamic interaction between hippocampus-dependent processes and circadian clock mechanisms.

Efecto del color del sustrato en el comportamiento de fijación del caballito de mar de hocico corto (Hippocampus hippocampus)

El caballito de mar (Hippocampus hippocampus) presenta una fase planctónica durante los primeros días de su vida. Cuando cambia a la fase bentónica ha de fijarse en un sustrato adecuado con su cola prensil. Este estudio, realizando recuentos diarios de la distribución de los caballitos en el acuario, ha obtenido como resultado que no existe una preferencia entre el color rojo y azul del sustrato a la hora de la fijación, no obstante, se observó una adaptación progresiva de los peces al sustrato artificial.The seahorse (Hippocampus hippocampus) presents a planktonic phase during the first days of life. During settlement, it fixes to a suitable substratum using its prehensile tail. This study, through a daily counts of the seahorse distribution in the aquarium, established that juveniles seahorse do not show a preference between the red or blue fixation substrates. However, it was observed a progressive adaptation of the fish to the artificial handmade substratum.

Efecto de los misidáceos en la calidad de la puesta en el caballito de mar, Hippocampus hippocampus (Linnaeus, 1758)

[EN]The importance of a suitable feeding in reproduction and spawning quality of teleost fish has been recognized as one of the major ?bottlenecks? in new aquaculture species like seahorses. Mysidacea species has been described as one of the main food for temperate seahorse species (Hippocampus hippocampus and H. guttulatus) in the wild. On the other hand, Artemia has been employed usually as marine food for rearing fish, including seahorses. The aim of this work is to study the effect of two different live preys (Artemia vs Mysis) in spawning quality of H. hippocampus broodstock. The animals were fed two times per day, six times per week. Spawning episodes and larvae quality was recorded. Seahorse fed on mysis showed significantly better results (p<0.05) than Artemia treatment, regarding spawning events, number of offspring?s and size. This fact showed the high potential of mysis as live prey for seahorses or other ornamental species.

Protocolo de anestesia y recuperación para estudio de morfometría de juveniles de caballito de mar, (Hippocampus hippocampus (Linnaeus, 1758)

Anaesthetic and Recovery Protocol for morfometric study of Atlantic seahorse, Hippocampus hippocampus (Linnaeus, 1758), juveniles. Some morphometric studies in marine organism need anaesthetics in order to avoid animal?s sacrifice and allow its recovery. In this study different natural clove essential concentrations has been tested in Atlantic seahorses, Hippocampus hippocampus (Linnaeus, 1758) aged 13 (trunk length 7,48 ± 1,14 mm) and 22 days (trunk length 9,09 ± 1,62 mm). An inversed relation was observed between anaesthetic concentration and effective time. The concentration allowing measures was 25 ppm. This procedure allowed 100% survival rate.

Efecto de la primera alimentación en la supervivencia y el crecimiento de crías de caballito de mar, Hippocampus hippocampus (Linnaeus, 1758)

[EN]Effects of first feeding on survival and growth of Atlantic seahorse juveniles, Hippocampus hippocampus, (Linnaeus, 1758). In this study, two experimental protocols were tested: a first treatment (RA), in which larvae were fed exclusively on rotifers until day 5, and then on Artemia, and a second treatment (A) where larvae were only fed on Artemia. We attempted to improve survival and growth by comparing different diets in the first days of life. The results showed excellent survival (60% average) and growth in juveniles exclusively fed with Artemia, with better values than those obtained by other authors for this species. In comparison, high mortality and poor growth were observed on the first days in larvae fed rotifers.

Experiencias preliminares de cultivo de crías de Hippocampus hippocampus (Linnaeus, 1758) en Gran Canaria

[EN]Seahorses husbandry constitute an innovative aspect on the recovery of wild populations. In this study, two aeration levels were tested (normal and strong) in seahorse juveniles fed on Artemia. Survival and growth were evaluated. The results showed higher survivals (11% average) with strong aerations levels after 90 days. In addition, these juveniles were able to reproduce after 4 months, denoting the early sexual maturation of this species. However, sizes of second-generation larvae at day 0 were smaller compared with those produced by their parents

Caracterización y modulación del color en el caballito de mar (Hippocampus hippocampus)

Máster Oficial en Cultivos Marinos. Trabajo presentado como requisito parcial para la obtención del Título de Máster Oficial en Cultivos Marinos, otorgado por la Universidad de Las Palmas de Gran Canaria (ULPGC), el Instituto Canario de Ciencias Marinas (ICCM), y el Centro Internacional de Altos Estudios Agronómicos Mediterráneos de Zaragoza (CIHEAM)

Modulation of network excitability and epileptiform activity in the hippocampus of immature rats by the activation of GlyRs

Epileptic seizures are the manifestations of epilepsy, which is a major neurological disorder and occurs with a high incidence during early childhood. A fundamental mechanism underlying epileptic seizures is loss of balance between neural excitation and inhibition toward overexcitation. Glycine receptor (GlyR) is ionotropic neurotransmitter receptor that upon binding of glycine opens an anion pore and mediates in the adult nervous system a consistent inhibitory action. While previously it was assumed that GlyRs mediate inhibition mainly in the brain stem and spinal cord, recent studies reported the abundant expression of GlyRs throughout the brain, in particular during neuronal development. But no information is available regarding whether activation of GlyRs modulates neural network excitability and epileptiform activities in the immature central nervous system (CNS). Therefore the study in this thesis addresses the role of GlyRs in the modulation of neuronal excitability and epileptiform activity in the immature rat brain. By using in vitro intact corticohippocampal formation (CHF) of rats at postnatal days 4-7 and electrophysiological methods, a series of pharmacological examinations reveal that GlyRs are directly implicated in the control of hippocampal excitation levels at this age. In this thesis I am able to show that GlyRs are functionally expressed in the immature hippocampus and exhibit the classical pharmacology of GlyR, which can be activated by both glycine and the presumed endogenous agonist taurine. This study also reveals that high concentration of taurine is anticonvulsive, but lower concentration of taurine is proconvulsive. A substantial fraction of both the pro- and anticonvulsive effects of taurine is mediated via GlyRs, although activation of GABAA receptors also considerably contributes to the taurine effects. Similarly, glycine exerts both pro- and anticonvulsive effects at low and high concentrations, respectively. The proconvulsive effects of taurine and glycine depend on NKCC1-mediated Cl- accumulation, as bath application of NKCC1 inhibitor bumetanide completely abolishes proconvulsive effects of low taurine and glycine concentrations. Inhibition of GlyRs with low concentration of strychnine triggers epileptiform activity in the CA3 region of immature CHF, indicating that intrinsically an inhibitory action of GlyRs overwhelms its depolarizing action in the immature hippocampus. Additionally, my study indicates that blocking taurine transporters to accumulate endogenous taurine reduces epileptiform activity via activation of GABAA receptors, but not GlyRs, while blocking glycine transporters has no observable effect on epileptiform activity. From the main results of this study it can be concluded that in the immature rat hippocampus, activation of GlyRs mediates both pro- and anticonvulsive effects, but that a persistent activation of GlyRs is required to prevent intrinic neuronal overexcitability. In summary, this study uncovers an important role of GlyRs in the modulation of neuronal excitability and epileptiform activity in the immature rat hippocampus, and indicates that glycinergic system can potentially be a new therapeutic target against epileptic seizures of children.

Musical Training Induces Functional Plasticity in Human Hippocampus

Training can change the functional and structural organization of the brain, and animal models demonstrate that the hippocampus formation is particularly susceptible to training-related neuroplasticity. In humans, however, direct evidence for functional plasticity of the adult hippocampus induced by training is still missing. Here, we used musicians' brains as a model to test for plastic capabilities of the adult human hippocampus. By using functional magnetic resonance imaging optimized for the investigation of auditory processing, we examined brain responses induced by temporal novelty in otherwise isochronous sound patterns in musicians and musical laypersons, since the hippocampus has been suggested previously to be crucially involved in various forms of novelty detection. In the first cross-sectional experiment, we identified enhanced neural responses to temporal novelty in the anterior left hippocampus of professional musicians, pointing to expertise-related differences in hippocampal processing. In the second experiment, we evaluated neural responses to acoustic temporal novelty in a longitudinal approach to disentangle training-related changes from predispositional factors. For this purpose, we examined an independent sample of music academy students before and after two semesters of intensive aural skills training. After this training period, hippocampal responses to temporal novelty in sounds were enhanced in musical students, and statistical interaction analysis of brain activity changes over time suggests training rather than predisposition effects. Thus, our results provide direct evidence for functional changes of the adult hippocampus in humans related to musical training.

Grafted neuronal precursor cells differentiate and integrate in injured hippocampus in experimental pneumococcal meningitis

Bacterial meningitis (BM) frequently causes persisting neurofunctional sequelae. Autopsy studies in patients dying from BM show characteristic apoptotic brain injury to the stem cell niche in the subgranular zone of the hippocampal dentate gyrus (DG), and this form of brain damage is associated with learning and memory deficits in experimental BM. With an eye to potential regenerative therapies, the survival, migration, and differentiation of neuronal precursor cells (NPCs) were evaluated after engraftment into the injured hippocampus in vitro and in vivo in an infant rat model of pneumococcal meningitis. Green fluorescent protein (GFP)-expressing NPCs were grafted into the DG of organotypic hippocampal slice cultures injured by challenge with live Streptococcus pneumoniae. Seven days after engraftment, NPCs had migrated from the site of injection into the injured granular layer of the DG and electro-functionally integrated into the hippocampal network. In vivo, GFP-expressing NPCs migrated within 1 week from the injection site in the hilus region to the injured granular layer of the hippocampal DG and showed neuronal differentiation at 2 and 4 weeks after transplantation. Hippocampal injury induced by BM guides grafted NPCs to the area of brain damage and provides a microenvironment for neuronal differentiation and functional integration.