Orlando Ghirardi

Aspects of spatial memory and behavioral disinhibition in Tg2576 transgenic mice as a model of Alzheimer's disease

Transgenic mouse models of Alzheimers disease (AD) have been recently advanced. Tg2576 mice have been shown to develop progressive beta-amyloid (Abeta) neuritic plaques and exhibit impairment of cognitive function. The aim of this study was a better characterization of different aspects of spatial memory performance of transgenic mice, observed at a time when levels of soluble Abeta are elevated and Abeta neuritc plaques start to appear. A general elevation of basal locomotory activity in the home cage was found in Tg2576 mice, which also exhibited an impairment of spontaneous alternation in the Y-maze test. Tg2576 mice were not flexible upon changes in the schedule and failed to codify spatially the testing environment. Consistently, a deficit of spatial memory was also observed when mice were assessed for levels of reactivity to spatial change in the modified open-field test with objects. Compared to controls, Tg2576 mice also exhibited an increased number of explorative approaches to the different objects, and failed to discriminate the displacement of the object. Consistently with the hypothesis of increased disinhibition, a differential behavioural response to the plus-maze paradigm was exhibited by Tg2576 mice. Results clearly indicate that Tg2576 mice are characterized by a number of specific behavioral cognitive alterations, compatible with Alzheimers disease (AD), which make them a suitable animal model for testing of novel anti-AD drugs.

Aging brain: effect of acetyl-l-carnitine treatment on rat brain energy and phospholipid metabolism. A study by31P and1H NMR spectroscopy

The effects of acetyl-L-carnitine (ALCAR) on metabolites involved in energy and phospholipid metabolism have been evaluated by mean of 31P and 1H NMR spectroscopy on adult (6 months) and old (24 months) rat brains. A significant increase of glycerophosphorylcholin (GroPCho) in aged rat brain has been observed as compared with adult rat brain. No variations in ATP, phosphocreatine (PCr), Cr, lactate, ADP and inorganic phosphate (Pi) levels have been found between aged and adult brains. Treatment with ALCAR caused a significant increase in PCr levels and a decrease in lactate and sugar phosphate in adult and aged rat brain. These results are suggestive of treatment with ALCAR being responsible for a reduction in brain glycolytic flow and for enhancing the utilization of alternative energy sources, such as lipid substrates or ketone bodies. Furthermore, the changes in GroPCho levels observed after treatment with ALCAR may be indicative of a modulating effect on the activity of the enzymes involved in the acylation-re-acylation process of membrane phospholipids.

Effect of acetyl-l-carnitine chronic treatment on discrimination models in aged rats

Acetyl-l-carnitine (ALC), a natural component of several biological systems, has been found to modify spontaneous and evoked electrocortical activity in young rats and to improve learning ability in old ones. In clinical application it also improves mood and attention in elderly patients. The present study was aimed at ascertaining the effect of a chronic treatment with ALC added to drinking water on two discrimination models in aged rats. In the first model, simple discrimination learning was tested and was found to be significantly improved by treatment. The second model consisted of a differential reinforcement of low rate of responding. The animals receiving treatment performed significantly better as shown by a lower number of nonrewarded responses. Because impaired learning and memory are related to alterations in hippocampal function, these data indicate that ALC is capable of antagonizing the natural age-dependent deterioration process in the hippocampal structure.

Antioxidant enzyme activities in heart and skeletal muscle of rats of different ages

The activity of antioxidant enzymes was measured in cardiac and skeletal muscle in rats aged either 4, 15, or 27 months. Generally, regardless of age, heart contains a greater content of these enzymes than skeletal muscle. Whereas skeletal muscle showed age-dependent increases in glutathione peroxidase, glutathione reductase, and catalase activities, heart tissue showed increases in only the glutathione peroxidase activity. Neither tissue showed any significant age-dependent change in cytosolic or mitochondrial superoxide dismutase content or in cytochrome oxidase.

Effect of acetyl-l-carnitine on recovery of brain phosphorus metabolites and lactic acid level during reperfusion after cerebral ischemia in the rat — study by 13P- and 1H-NMR spectroscopy

The effects of acetyl-L-carnitine (ALCAR) treatment on brain energy state recovery and lactic acid levels following 20 min ischemia and 2, 24 and 48 h reperfusion were investigated by 31P and 1H-NMR spectroscopy. Transient forebrain ischemia was induced by four-vessel occlusion method in fed 6-month-old Fischer rats. ALCAR or saline was administered by intraperitoneal route immediately after 20 min ischemia and again at 1, 4, 24 and 30 h during reperfusion. Twenty-min severe forebrain ischemia was associated with a marked decrease in phosphocreatine (PCr) and ATP levels and a corresponding increase in lactic acid, inorganic phosphate (Pi), AMP, creatine, glycerol 3-phosphate and alanine levels. Following reperfusion, a general tendency to restore pre-ischemic metabolite levels was observed. However, after 2 h reperfusion in saline-treated rats, lactic acid and Pi levels remained significantly higher, while ATP levels were still significantly lower than in non-ischemic controls. On the contrary, in ALCAR-treated animals a complete recovery of all metabolites including Pi and ATP was observed, while PCr levels were even more elevated compared with those in saline-treated rats. Furthermore lactic acid content was significantly lower than that in both saline-treated and non-ischemic control rats. It is concluded that a potential therapeutic role may be claimed for ALCAR in the treatment of cerebral ischemia through mechanisms that include faster recovery and improvement of brain energy production as well as a decreased lactic acid content during early post-ischemic reperfusion.

The effect of early maternal separation on brain derived neurotrophic factor and monoamine levels in adult heterozygous reeler mice

OBJECTIVE AND METHODS The reeler heterozygous (HZ) mice have provided a model for studying the relationship between reelin (a protein of extracellular matrix) haploinsufficiency and the emergence of neuropsychiatric diseases. In a neurodevelopmental framework, the enduring consequences of early maternal separation (5 h/day during the first postnatal week, or handling controls, H) were studied in reeler HZ and wild type (WT) mice at adulthood. The modulatory effects of a chronic treatment with the atypical antipsychotic olanzapine (OLZ, 1.5 mg/kg for 40 days) were also investigated. RESULTS Early maternal separation had long-term effects on brain plasticity, with a reduction of brain- and glial- derived neurotrophic factor (BDNF and GDNF) in several brain areas of mice, but such a consequence was less marked in the HZ genotype. On the other hand, treatment with OLZ did not affect at all the GDNF but led to an increase of BDNF levels in maternally separated (SEP) mice, an effect which was far more marked in the HZ genotype. Brain levels of serotonin (5-HT) were markedly increased, striatal dopamine (DA) was increased, whereas metabolites and turnover were decreased, in SEP mice of both genotypes. The spontaneous home-cage activity was generally lower in HZ than WT mice, and OLZ treatment contrasted this hypoactivity profile. Maternal separation also decreased the interest toward an unknown mouse proposed as a social stimulus, but only in WT mice. CONCLUSION We investigated the interplay between genetic vulnerability (reelin haploinsufficiency), the outcome of early stressful experiences, and the efficacy of the antipsychotic drug therapy. The reeler HZ genotype exhibited a slightly lower sensitivity to the environmental insult as well as an enhanced response to the atypical antipsychotic treatment.

Age-related changes of the mossy fibre system in rat hippocampus: effect of long term acetyl-l-carnitine treatment

Age-related changes of the hippocampal mossy fibre system were studied in rats of different ages (4, 12, and 24 months) using the Timms histochemical technique for the detection of tissue stores of zinc and heavy transitional metals. Quantitative image analysis and microdensitometry techniques were applied for the evaluation of the changes themselves. The area occupied by mossy fibres and the intensity of Timms staining in field CA3 of hippocampus were greater in adult rats than in young and old animals. These findings are consistent with previous behavioural data indicating that young and old rats exhibited a reduced passive avoidance learning with respect to adult animals. The above techniques were also used in a second session of this study designed to investigate the effect of a 6-months treatment with acetyl-L-carnitine (ALC) on the hippocampal mossy fibre system. ALC was found to increase significantly the area occupied by mossy fibres and the intensity of Timms staining in the hippocampus of old rats. These findings are in agreement with and support from an anatomical point of view, the data indicating that long-term treatment with ALC improves passive avoidance performance.

Spatial learning and memory in the radial maze: A longitudinal study in rats from 4 to 25 months of age

This longitudinal study was designed to investigate whether previous experience may influence performances and strategies of rats tested in the radial maze without external cues when aged 4, 13, and 25 months. Their performances and strategies were compared with those of another group of rats tested only when aged 25 months. Expert old animals showed a good retention of previous experiences, whereas age-matched nonexpert animals exhibited some acquisition deficits. On the contrary, in the course of aging, the animals kept modifying their strategies independently of experience. In summary, we can conclude that previous experience is likely to influence performances of the aged rat but not the strategies adopted which are strictly age-dependent and independent of acquired experience.

Age-dependent nerve cell loss in the brain of Sprague-Dawley rats: effect of long term acetyl-L-carnitine treatment.

The age dependent loss of nerve cells was investigated in 22 brain areas from young (3 month), adult (13 month) and old (25 month) Sprague-Dawley rats. As in previous studies, we observed an age-related neuronal loss primarily in the archicortex and in the hippocampus and in other subcortical structures (amigdaloid nucleus, pontine nuclei, cerebellar cortex). In sensory areas of cerebral cortex the neuronal loss was less marked. The effect of a 6 month treatment with acetyl-L-carnitine (ALCAR) on the number of nerve cells in the same brain areas was also investigated. ALCAR treatment began when the rats were aged 16 months. ALCAR treatment was able to counteract the age-dependent decrease in nerve cell number primarily in the temporal and occipital cortical areas, in the archicortex and hippocampus. The above findings suggest that long term ALCAR treatment may be effective in slowing down the age-related nerve cell loss in some rat brain areas.

A positron emission tomography microdosing study with a potential antiamyloid drug in healthy volunteers and patients with Alzheimer's disease

This work describes a microdosing study with an investigational, carbon 11‐labeled antiamyloid drug, 1,1′‐methylene‐di‐(2‐naphthol) (ST1859), and positron emission tomography (PET) in healthy volunteers (n = 3) and patients with Alzheimers disease (n = 6). The study aimed to assess the distribution and local tissue pharmacokinetics of the study drug in its target organ, the human brain. Before PET studies were performed in humans, the toxicologic characteristics of ST1859 were investigated by an extended single‐dose toxicity study according to guidelines of the Food and Drug Administration and European Medicines Agency, which are relevant for clinical trials with a single microdose. After intravenous bolus injection of 341 ± 21 MBq [11C]ST1859 (containing <11.4 nmol of unlabeled ST1859), peripheral metabolism was rapid, with less than 20% of total plasma radioactivity being in the form of unchanged parent drug at 10 minutes after administration. In both the control and patient groups, uptake of radioactivity into the brain was relatively fast (time to reach maximum concentration, 9–17 minutes) and pronounced (maximum concentration [standardized uptake value], 1.3–2.2). In both healthy volunteers and patients, there was a rather uniform distribution of radioactivity in the brain, including both amyloid‐beta‐rich and ‐poor regions, with slow washout of radioactivity (half‐life, 82–185 minutes). In conclusion, these data provide important information on the blood‐brain barrier penetration and metabolism of an investigational antiamyloid drug and suggest that the PET microdosing approach is a useful method to describe the target‐organ pharmacokinetics of radiolabeled drugs in humans.