Rosana Mattioli

Natural preference of zebrafish (Danio rerio) for a dark environment

The zebrafish (Danio rerio) has been used as a model in neuroscience but knowledge about its behavior is limited. The aim of this study was to determine the preference of this fish species for a dark or light environment. Initially we used a place preference test and in a second experiment we applied an exit latency test. A two-chamber aquarium was used for the preference test. The aquarium consisted of a black chamber and a white chamber. In the first experiment the animal was placed in the aquarium and the time spent in the two compartments was recorded for 10 min. More time was spent in the black compartment (Wilcoxon matched-pairs signed-rank test, T = 7, N1 = N2 = 18, P = 0.0001). In the second experiment the animal was placed in the black or white compartment and the time it took to go from the initial compartment to the opposite one was recorded. The test lasted a maximum of 10 min. The results showed that the animal spent more time to go from the black to the white compartment (Mann-Whitney rank sum test, T = 48, N1 = 9, N2 = 8, P<0.0230). These data suggest that this fish species has a natural preference for a dark environment and this characteristic can be very useful for the development of new behavioral paradigms for fish.

Gait Training Combining Partial Body-Weight Support, a Treadmill, and Functional Electrical Stimulation: Effects on Poststroke Gait

Background and Purpose: Treadmill training with harness support is a promising, task-oriented approach to restoring locomotor function in people with poststroke hemiparesis. Although the combined use of functional electrical stimulation (FES) and treadmill training with body-weight support (BWS) has been studied before, this combined intervention was compared with the Bobath approach as opposed to BWS alone. The purpose of this study was to evaluate the effects of the combined use of FES and treadmill training with BWS on walking functions and voluntary limb control in people with chronic hemiparesis. Subjects: Eight people who were ambulatory after chronic stroke were evaluated. Methods: An A1-B-A2 single-case study design was applied. Phases A1 and A2 included 3 weeks of gait training on a treadmill with BWS, and phase B included 3 weeks of treadmill training plus FES applied to the peroneal nerve. The Stroke Rehabilitation Assessment of Movement was used to assess motor recovery, and a videography analysis was used to assess gait parameters. Results: An improvement (from 54.9% to 71.0%) in motor function was found during phase B. The spatial and temporal variables cycle duration, stance duration, and cadence as well as cycle length symmetry showed improvements when phase B was compared with phases A1 and A2. Discussion and Conclusions: The combined use of FES and treadmill training with BWS led to an improvement in motor recovery and seemed to improve the gait pattern of subjects with hemiparesis, indicating the utility of this combination method during gait rehabilitation. In addition, this single-case series showed that this alternative method of gait training—treadmill training with BWS and FES—may decrease the number of people required to carry out the training.

Conditioned place-preference analysis in the goldfish with the H1 histamine antagonist chlorpheniramine

The aim of this study was to investigate whether the histamine H1-receptor blocker, chlorpheniramine (CPA), has a reinforcing effect in goldfish. We used a place-preference procedure in an aquarium with two chambers colored black and white. On day 1, the animals were placed in the test chamber for 10 min for habituation. On day 2, they were placed in the start compartment for 30 s, the sliding doors were opened, and the time spent in each compartment was recorded over the 10 min to determine the natural compartment preference for each animal. On day 3, they were injected either with 0.1, 0.4, 1.0, or 4.0 mg/kg of CPA or only with vehicle and placed in the less preferred compartment for 25 min. On day 4, the animals were placed in the start compartment and the time spent in each compartment during the 10-min test period was recorded again. The groups treated with 1.0 and 4.0 mg/kg of CPA, spent significantly more time in the compartment in which they experienced the drug effect than the group treated with vehicle. On the other hand, the group treated with 0.4 mg/kg spent significantly less time in the drug-paired compartment. The results indicate a biphasic effect of CPA. Considering that there is evidence that low doses of CPA can also block H3-receptors, we suggest that in goldfish the histaminergic neural system has an inhibitory role in the reinforcing process.

Substance P facilitates memory in goldfish in an appetitively motivated learning task

The aim of this study was to investigate if the neuropeptide substance P (SP) can improve learning and memory in goldfish (Carassius auratus). Four groups of fish were trained for seven days to find food in one out of two compartments until discrimination was achieved. On the last training day, they were injected (intra-abdominal) with haloperidol or vehicle before the training, and with SP or vehicle immediately after the training session. Each group of fish received either: (1) vehicle+vehicle (n = 18); (2) vehicle + SP, (n = 20, SP 50 mg/kg); (3) haloperidol+ vehicle (n = 15, haloperidol 2 mg/kg); or (4) haloperidol+ SP (n = 14, haloperidol 2 mg/kg, SP 50 mg/kg). Twenty-four hours later, the time spent to find the food was recorded. Reversal training was done for four consecutive days after this post-injection test and the time spent to find the food was recorded again. The results indicate that only the group treated with vehicle + SP needed more time to reach reversal training than control fish (Mann-Whitney U-test, P = 0.0009). It is suggested that SP can enhance memory in fish and that this effect may have a dopaminergic mediation in discrimination learning task.

Post-Trial Administration of H1 Histamine Receptor Blocker Improves Appetitive Reversal Learning and Memory in Goldfish, Carassius auratus

Based on the hypothesis that neuronal histamine exerts an inhibitory influence on learning and reinforcement, goldfish were tested for post-trial effects of the H1 receptor blocker chlorpheniramine (CPA) on learning the location of a food source in one of two compartments, one black the other white, with a feeder located in each compartment. Testing was carried out over 6 days. On the training day a food pellet was placed into the feeder of one of the compartments. After consumption of the food the fish were injected i.p. with either vehicle or CPA either immediately after training or 3 h later. Twenty-four-hours later, food was placed in the same compartment and the time to begin feeding was recorded. On the next day the location of the food pellet was reversed, and testing was continued for 4 days. On the first test day the time to begin feeding was significantly longer for the vehicle injected fish as compared with those injected with CPA. The vehicle group also took longer to begin feeding than the CPA group on the first reversal test day. The results of the 3-h delay groups indicated no significant differences between vehicle and drug for any experimental session. These results suggest that post-trial blockade of the H1 histamine receptor can affect appetitive learning in goldfish either by improving long-term memory consolidation and/or by the additive reinforcing effects of CPA (known from previous studies) on behavior.

Effects of treadmill-walking training with additional body load on quality of life in subjects with Parkinson's disease

BACKGROUND Parkinsons disease (PD) causes motor and non-motor impairments that affect the subjects quality of life. OBJECTIVE To assess the effects of treadmill-walking training with additional body load on the quality of life and motor function of subjects with PD. METHODS Nine subjects with PD, Hoehn and Yahr stages 2-3, not demented and with capability to ambulate independently took part in this study. The training program was divided into three phases (A₁-B-A₂): treadmill training with additional body load (A₁), control condition (conventional physical therapy group; B) and a second period of treadmill training with load (A₂). Each phase lasted six weeks. Quality of life and motor function were assessed by the PDQ-39 and the motor score of the Unified Parkinsons Disease Rating Scale (UPDRS), respectively. The evaluations and the training were performed during the on-phase of the medication cycle. RESULTS There was improvement in the total PDQ-39 score across the training period. The subscores mobility, activities of daily living and cognition subscores significantly improved after the training period. The improvement in the total score was associated with motor and non-motor factors in all of the training phases. The UPDRS motor score also improved, however it did not present any association with the improvement in quality of life. CONCLUSIONS The results showed that the treadmill-walking training with additional body load allowed an improvement in motor and non-motor aspects related to quality of life and motor function in subjects with PD.

Microinjection of histamine into the cerebellar vermis impairs emotional memory consolidation in mice.

The biogenic amine histamine is an important neurotransmitter in the central nervous system that has been implicated in learning and memory processes. Experimental evidence indicates that the role of the cerebellum may be more complex than the simple regulation of motor responses, and recent studies have demonstrated significant involvement of the cerebellum in emotional memory consolidation. This study investigated the effect of histamine microinjected into the cerebellar vermis on emotional memory consolidation in mice in the elevated plus-maze (EPM). The cerebellar vermis of male mice (Swiss Albino) were implanted with guide cannulae. The mice weighed between 25 and 30 g. After three days of recovery, behavioral tests in the EPM were performed on two consecutive days; the testing periods were called, Trial 1 and Trial 2. Immediately after Trial 1, the animals received microinjections of histamine in the cerebellar vermis (0.54, 1.36, 2.72, and 4.07 nmol/0.1 μl). On both days, the test sessions were recorded to enable analysis of behavioral measures. The decrease in open arm exploration (% entries and % time spent in the open arms) in Trial 2 relative to Trial 1 was used as a measure of learning and memory. The data were analyzed using One-way Analysis of Variance (ANOVA) and Duncans tests. The percentage of open arm entries (%OAE) and the percentage of time spent in the open arms (%OAT) were reduced in Trial 2 relative to Trial 1 for the control group; the same was true for the group that was microinjected with histamine at doses of 0.54 (%OAE and %OAT) and 1.36 nmol (%OAT). However, when the animals received histamine at doses of 2.72 and 4.07 nmol, their open arm exploration did not decrease. No significant changes were observed in the number of enclosed arm entries (EAE), an EPM index of general exploratory activity. These results suggest that there is a dose-dependent inhibitory effect of histamine microinjected into the cerebellar vermis on emotional memory consolidation.

Emotional memory consolidation impairment induced by histamine is mediated by H1 but not H2 receptors

Histaminergic fibers are present in the molecular and granular layers of the cerebellum and have high density in the vermis and flocculus. Evidence indicates that the cerebellar vermis is involved in memory consolidation. Recently, we demonstrated that when histamine is microinjected into the cerebellar vermis it results in impaired emotional memory consolidation in mice that are submitted to the elevated plus maze (EPM). This study investigated whether histamine impairment was mediated by the H(1) or H(2) receptors. The cerebellar vermis of male mice (Swiss Albino) were implanted using a guide cannula. Three days after recovery, behavioral tests were performed in the EPM on two consecutive days (Trial 1 and Trial 2). Immediately after exposure to the EPM (Trial 1), animals received a microinjection of histaminergic drugs. In Experiment 1, saline (SAL) or histamine (HA, 4.07 nmol/0.1 μl) was microinjected 5 min after pretreatment with the H(1) antagonist chlorpheniramine (CPA, 0.16 nmol/0.1μl) or SAL. In Experiment 2, SAL or HA was microinjected into the mice 5 min after pretreatment with the H(2) antagonist ranitidine (RA, 2.85 nmol/0.1 μl) or SAL. Twenty-four hours later, the mice were re-exposed to the EPM (Trial 2) under the same experimental conditions but did not receive an injection. On both days, the test sessions were recorded to enable analysis of the behavioral measures. The decrease in open arm exploration (% entries and % time spent in the open arms) in Trial 2 relative to Trial 1 was used as a measure of learning and memory. The data were analyzed using the two-way analysis of variance (ANOVA) and Duncans tests. In Experiment 1, the Duncans test indicated that the mice entered the open arms less often (%OAE) and spent less time in the open arms (%OAT) in Trial 2 after being microinjected with SAL+SAL, SAL+CPA and CPA+HA. However, the animals that received SAL+HA did not enter the open arms less frequently or spend less time in them, which was significantly different from the CPA+HA group. The results of Experiment 2 demonstrated that the %OAE and %OAT in Trial 2 were different from Trial 1 for the groups that were microinjected with SAL+SAL and SAL+RA. The animals that were microinjected with RA+HA or with SAL+HA did not show a reduction in %OAE. These results demonstrate that the animals treated with HA did not avoid the open arms less on retesting and indicated that CPA did not alter the behavior parameters but did revert the histamine-induced impairment of memory consolidation. Furthermore, the H(2) antagonist RA, at the dose used in this study, did not affect memory consolidation and failed to revert histamine-induced impairment.

Analysis of the role op histamine in inhibitory avoidance in goldfish

1. Teleost fish have histaminergic cell bodies on the posterior part of the basal hypothalamus. It was suggested that they are homologous to the tuberomammillary E group in rats. However, unlike in rats, fish have fewer ascending fibers. The main projection runs through the ventral telencephalic area reaching the dorsal telencephalon. This projection is considered homologous to the prosencephalic forebrain bundle. 2. The aim of this study was to verify if the histaminergic system has an inhibitory action on learning and memory in goldfish, as suggested previously for higher vertebrates. 3. A two-compartment aquarium with a central sliding door was used. The animals were placed in one of them, the central door was opened after 30 sec and the time spend for crossing between compartments was recorded. After the fish dorsal fin crossed the line between the compartments a 45 g weight was dropped into the compartment the fish entered. 4. On the training day this procedure was done 3 times. Immediately after the 3rd trial the fish was injected i.p. with either vehicle (2 ml/kg), chlorpheniramine (CPA; 1.0, 4.0 and 8.0 mg/kg) or histidine (500 mg/kg). On the next day, fishes were placed in the start compartment and the latency to cross between compartments was again recorded. 5. The group treated with CPA at the dose of 8 mg/kg, presented a significant increase in the latency to leave the start compartment (Wilcoxon rank sum test, p<0.0232). On the other hand, the vehicle and 1-histidine (500 mg/kg) treated groups, presented a decrease in test latency. 6. Thus, we suggest that also in fish, the histaminergic system has an inhibitory role on learning and memory.

L-histidine provokes a state-dependent memory retrieval deficit in mice re-exposed to the elevated plus-maze

The effects of L-histidine (LH) on anxiety and memory retrieval were investigated in adult male Swiss Albino mice (weight 30-35 g) using the elevated plus-maze. The test was performed on two consecutive days: trial 1 (T1) and trial 2 (T2). In T1, mice received an intraperitoneal injection of saline (SAL) or LH before the test and were then injected again and retested 24 h later. LH had no effect on anxiety at the dose of 200 mg/kg since there was no difference between the SAL-SAL and LH-LH groups at T1 regarding open-arm entries (OAE) and open-arm time (OAT) (mean +/- SEM; OAE: 4.0 +/- 0.71, 4.80 +/- 1.05; OAT: 40.55 +/- 9.90, 51.55 +/- 12.10, respectively; P > 0.05, Kruskal-Wallis test), or at the dose of 500 mg/kg (OAE: 5.27 +/- 0.73, 4.87 +/- 0.66; OAT: 63.93 +/- 11.72, 63.58 +/- 10.22; P > 0.05, Fisher LSD test). At T2, LH-LH animals did not reduce open-arm activity (OAE and OAT) at the dose of 200 mg/kg (T1: 4.87 +/- 0.66, T2: 5.47 +/- 1.05; T1: 63.58 +/- 10.22; T2: 49.01 +/- 8.43 for OAE and OAT, respectively; P > 0.05, Wilcoxon test) or at the dose of 500 mg/kg (T1: 4.80 +/- 1.60, T2: 4.70 +/- 1.04; T1: 51.55 +/- 12.10, T2: 43.88 +/- 10.64 for OAE and OAT, respectively; P > 0.05, Fisher LSD test), showing an inability to evoke memory 24 h later. These data suggest that LH does not act on anxiety but does induce a state-dependent memory retrieval deficit in mice.