Search PubMed⌕ Search

Biomedical subjects

A Privat

Publications and source records attributed to A Privat.

At least 55 records · Page 3Linked to original sources
PubMed1999

Neuroprotection of cultured foetal rat hippocampal cells against glucose deprivation: are GABAergic neurons less vulnerable or more sensitive to TCP protection?

In the rat brain, hippocampal neurons are particularly sensitive to secondary excitotoxic injury induced by ischaemia or hypoglycaemia. To determine some distinctive features of vulnerability among neuronal phenotypes in the hippocampus following a metabolic insult, we used an in vitro model of mild glucose deprivation. Primary cultures from the rat hippocampus (21 days in vitro) were deprived of glucose for 4 h and then were returned to the standard medium for 24 or 48 h. Survival of the GABAergic neuronal population was evaluated both by measuring [3H]GABA uptake and by counting GAD65-immunostained cells. This was compared with the survival of the total neuronal population evaluated by counting the neurofilament-200-immunostained cells. Glucose deprivation for 4 h followed by a recovery period of 48 h induced a decrease of 59% and 40% in the number of GAD65- and neurofilament-200-immunostained cells, respectively. Thus, GABAergic neurons were slightly more vulnerable to glucose deprivation than the other neurons in the hippocampal cell cultures. When the excitotoxic component of cellular death was blocked in the presence of TCP, an NMDA-antagonist, the survival of GABAergic neurons was almost complete after 48 h of recovery. In contrast, measurements of the release of lactate dehydrogenase in the medium indicated that TCP largely protected hippocampal cells after 24 h but was ineffective after 48 h. This observation was confirmed by immunostaining data which showed that after 48 h TCP did not significantly increase the survival of neurofilament-200-immunostained cells. These results indicate that after glucose deprivation and a recovery period of 48 h, GABAergic neurons in hippocampal cell cultures are not more resistant than other neurons but they are more sensitive to TCP protection.

Animals↗
PubMed1999

Cerebellar defect and impaired motor coordination in mice lacking vimentin.

Vimentin belongs to the family of intermediate filament (IF) proteins. During the nervous system development in mammals, it is transiently expressed in precursor cells of neuronal and glial lineages, and then it is progressively replaced by other types of IF proteins. Surprisingly, mice knock-out for vimentin develop and reproduce without any apparent defects (Colucci-Guyon et al. Cell 79:679-694, 1994). In adult rodents, Bergmann glia (BG) of the cerebellum continue to express vimentin together with glial fibrillary acidic protein (GFAP). A careful analysis of cerebellar morphology and ultrastructure in mutants showed poorly developed and highly abnormal BG, whereas the migration of granular neurons proceeded normally. Moreover, many Purkinje cells (PC) appeared stunted with a loss of spiny branchlets, and some of them were necrotic. Finally, impaired motor coordination was evidenced by behavioral tests. These observations demonstrate a role for vimentin in contributing to the normal development and morphology of BG and reveal a hitherto unreported functional relationship between BG and PC.

Animals↗
PubMed1998

Influence of hypergravity on the development of monoaminergic systems in the rat spinal cord.

We have investigated in this study the influence of a moderate hypergravity (1.8 G) on the development of monoaminergic projections to the spinal cord in the rat. Pregnant dams and their offspring were submitted to hypergravity from day 11 of gestation to postnatal day 15. Some animals were sacrificed at birth, other at postnatal day 15 and other after 8 months of normal gravity. In newborn animals, a substantial delay of the development of monoaminergic projections to the spinal cord was evidenced. In 15 days and 8 months animals, the pattern of innervation appeared anarchic, with numerous dystrophic profiles, mainly of serotonergic system. Ultrastructural examination of serotonergic projections revealed a paucity of synapses, and the frequent enveloping of serotonergic boutons by thin astrocytic profiles. We conclude that rats submitted to hypergravity during the critical period of onset of monoaminergic projections to the spinal cord are affected durably in the organization and the ultrastructure of these projections. Future studies are directed to the functional analysis of hypergravity animals, and to the influence of microgravity on the same system.

Animals↗
PubMed1998

Recovery of locomotion following transplantation of monoaminergic neurons in the spinal cord of paraplegic rats.

Severe traumatic lesions of the spinal cord yield a permanent deficit of motricity in adult mammals and specifically a loss of locomotor activity of hindlimbs when the lesion is located at the lower thoracic level. To restore this function, we have developed a paradigm of transplantation in rats based on a transection model of the spinal cord and the subsequent injection at the sublesional level of a suspension of embryonic brainstem monoaminergic neurons which play a key role in the modulation of locomotion. A genuine locomotion was characterized in transplanted animals by electromyographic and electroneurographic recordings. This correlated with a specific reinnervation pattern of targets, where typical synapses were found, and with the normalization of biochemical parameters.

Animals↗
PubMed1998

The modulation by neurosteroids of the scopolamine-induced learning impairment in mice involves an interaction with sigma1 (sigma1) receptors.

Neurosteroids have been reported to modulate learning and memory processes in aged animals and in pharmacological models of amnesia. We report here the effects of dehydroepiandrosterone sulfate (DHEAS), pregnenolone sulfate (PREGS), and progesterone (PROG) on the learning impairment induced in mice by the muscarinic acetylcholine receptor antagonist, scopolamine. Spatial working memory was examined using the spontaneous alternation behavior in a Y-maze and long-term memory using place learning in a rectangular water-maze adapted for mice. Both DHEAS and PREGS (5-20 mg/kg, s.c.) prevented dose-dependently and significantly the scopolamine (2 mg/kg, s.c.)-induced alternation deficits. PROG (2-20 mg/kg, s.c.) failed to affect the scopolamine-induced deficits, but blocked, at 20 mg/kg, the beneficial effects induced by DHEAS or PREGS. In the water-maze, DHEAS (20 mg/kg) attenuated significantly the scopolamine-induced deficits, as observed during the acquisition sessions or the retention test. PROG (2, 20 mg/kg) did not affect the control or scopolamine-treated group performances, but blocked the ameliorating effect of DHEAS. Furthermore, in both tests, the selective sigma1 (sigma1) receptor antagonist NE-100 (1 mg/kg, i.p.) failed to affect the behaviors showed by the control or scopolamine-treated groups, but it blocked the ameliorating effects induced by DHEAS or PREGS. These results confirm the modulating role of neurosteroids in learning and memory processes and demonstrate that their modulation of the cholinergic systems involves an interaction with sigma1 receptors.

Animals↗
PubMed1998

Direct evidence for the link between monoaminergic descending pathways and motor activity: II. A study with microdialysis probes implanted in the ventral horn of the spinal cord.

In order to define precisely the relation between descending monoaminergic systems and the motor system, we measured in the ventral horn of spinal cord of adult rats the variations of extracellular concentrations of 5-HT, 5-HIAA, DA and MHPG. Measurements were performed during rest, endurance running on a treadmill, and a post-exercise period, with microdialysis probes implanted permanently for 45 days. We found a slight decrease in both 5-HT and 5-HIAA during locomotion with a more marked decrease during the post-exercise period compared to the mean of rest values. In contrast, the concentration of DA and MHPG increased slightly during the exercise and decreased thereafter. These results, when compared with those of a previous study, which measured monoamines in the spinal cord white matter [C. Gerin, D. Bécquet, A. Privat, Direct evidence for the link between monoaminergic descending pathways and motor activity: I. A study with microdialysis probes implanted in the ventral funiculus of the spinal cord, Brain Res. 704 (1995) 191-201], highlight the complex regulation of the release of monoamines that occurs in the ventral horn.

Animals↗
PubMed1998

Astrocytes and oligodendrocytes reactions after a total section of the rat spinal cord.

Regeneration after an injury in the Central Nervous System is dependent on intrinsic and extrinsic factors. Among the latter are the reactions of glial cells. Using the model of total section of adult rat spinal cord, we have studied the spatial and temporal responses of astrocytes and oligodendrocytes to the lesion of spinal cord axons. We studied at molecular and cellular levels the specific markers GFAP (glial fibrillary acidic protein) for the astrocytes, CNP (2'-3' cyclic 3' nucleotide phosphodiesterase) which is principally expressed by immature oligodendrocytes, and MBP (myelin basic protein) implicated later in the myelin compaction, and which is more specific of mature oligodendrocytes. After injury, all astrocytes, but more markedly those of the grey matter, reacted by an increase of GFAP messenger and protein. This increase was very rapid for messenger, and peaked at 3 days. This increase was more protracted for the protein and persisted after 3 weeks. Messenger increase is more marked and more protracted below than above the lesion. Oligodendrocytes also reacted quickly by an increase of CNP and MBP messengers. For CNP, both messenger and protein increased rapidly and returned to control level after 1 week. MBP showed the same time course of changes, with lower and slower decrease above the lesion. Counts of oligodendrocytes showed that the percentage of the less mature form (light oligodendrocytes) increased dramatically above and below the lesion. After 1 week, above the lesion, this percentage was well below that of the control, whereas below the lesion, it reverted to control value. These results indicate that, following a lesion, astrocytes react quickly and intensely, but more so below the lesion; oligodendrocytes resume a sequence of maturation which is eventually completed above the lesion where remyelinisation can occur and which is prematurely interrupted below the lesion. However, intact oligodendrocytes persist below the lesion, where they constitute a potential for remyelinisation of regenerated and/or transplanted axons.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗
PubMed1998

Sigma1 (sigma 1) receptor agonists and neurosteroids attenuate B25-35-amyloid peptide-induced amnesia in mice through a common mechanism.

The sigma1 (sigma 1) receptor agonists exert potent anti-amnesic effects, as they apparently block the learning impairments either induced by the muscarinic receptor antagonist scopolamine, the N-methyl-D-aspartate receptor antagonist dizocilpine or inherently due to the age-related deficits in senescence-accelerated mice. We recently described the amnesia induced by the beta-amyloid-related peptide beta 25-35, administered centrally in an aggregated form, in mice. The deficits were sensitive to cholinomimetics or to N-methyl-D-aspartate/glycine modulatory site agonists. Herein, we examined the effects of sigma 1 receptor ligands on the beta 25-35 peptide-induced amnesia. The effects of neuro(active) steroids, which interact in vitro and in vivo with sigma 1 receptors were examined in parallel. Mnesic capacity was evaluated seven days after administration of aggregated beta 25-35 peptide (3 nmol), using spontaneous alternation in the Y-maze for spatial short-term memory, or after 14 days, using the step-down type passive avoidance test for long-term memory. The sigma 1 receptor agonists (+)-pentazocine, PRE-084, or SA4503 attenuated, in a dose-dependent and bell-shaped manner, the beta 25-35 peptide-induced deficits on both tests. These effects were antagonized by haloperidol or BMY-14802, confirming the sigma 1 receptor pharmacology. Pregnenolone, dehydroepiandrosterone, and their sulphate esters, but not progesterone, also dose-dependently attenuated the beta 25-35 peptide-induced deficits. Progesterone blocked the beneficial effects of each other neurosteroid, behaving as an antagonist. Furthermore, haloperidol blocked the effects induced by neurosteroids, whereas progesterone antagonized the effects of the non-steroidal sigma 1 receptor agonists, showing a clear crossed pharmacology of different drug classes. These results demonstrate that: (i) the anti-amnesic effect of sigma 1 receptor agonists may be of therapeutic relevance in pathological states affecting the cholinergic and/or glutamatergic systems, such as in pathological aging; (ii) neurosteroids play an important role in learning processes and may collectively constitute a therapeutic target; (iii) the interaction between sigma 1 systems and neurosteroids appears indeed of behavioural relevance.

Amnesia↗
PubMed1998

Adenoviral-mediated transfection of the Lac-Z gene into rat dissociated embryonic central nervous system cells before and after seeding.

The adenovirus carrying a reporter gene--the Lac Z gene--is known to infect central nervous system (CNS) cells in primary cell cultures. The percentage of infected neurons with respect to the total number of neurons was studied in primary dissociated cultures as a function of the day of inoculation and the age of three rat CNS cultures: spinal cord, mesencephalon and cortex. Two methods of viral inoculation were compared: the first inoculation was performed on the cultured cell at 2, 3 or 6 days in vitro (DIV) whereas the second inoculation was performed on the cell suspensions before seeding. All the infected CNS cells has the same aspect as the control cultures. In the spinal cord and the mesencephalic cultures, the glial cells were preferentially infected, especially when the cells were inoculated at 6 DIV. In the cortical cultures, there were more infected neurons than infected glial cells. The number of CNS cells was lower when inoculation was performed at 6 DIV as compared with 3 DIV. Very few infected GABA cells were found in the cultures. A high percentage of infected neuronal cells relative to the total number of neuronal cells was found when infection of the three types of cultures was performed on the dissociated embryonic cell suspension before seeding.

Adenoviridae↗
PubMed1998

Measurement by digital autoradiography with a beta-imager of [35S]methionine incorporated by rat central nervous system cells in primary cultures: a marker of in vitro development.

Mixed glial neuronal cultures prepared from rat embryonic cortical cells were either treated with aracytosine or infected with an adenovirus encoding the Lac-Z gene according to two protocols of infection. In each experiment, 24 h before the end of the incubation period, [35S]methionine was added to one set of cultures which were performed in plastic chamber slides. At 10-13 days in vitro, control and treated cultures were processed either for immunocytochemical detection of neuron-specific enolase (NSE)-stained cells or for measurement of [35S]methionine incorporation. For the latter, cultures grown in the chamber slides were fixed with 4% paraformaldehyde, dehydrated, and air-dried. After removal of the upper structures of the chambers, the slides were directly transferred to a 1200 beta-imager, a gaseous detector which displays a digital image of the cultured cells and permits the quantitative measurement of incorporated [35S]methionine within a few hours. In aracytosine-treated cultures, we observed that the numbers of NSE(+) cells as well as [35S]methionine incorporation were decreased compared with control cultures. After viral infection, the number of NSE(+) neurons and the amount of radioactivity incorporated were either the same in control and infected cultures or decreased for the cultures treated according to the different protocols. In all cases, the amount of [35S]methionine incorporated varied in the same direction as the number of NSE(+) neurons in cultures. The digital imaging of the cultures permitted observation of the layer of cultured cells. It appears that such a rapid and direct measurement of incorporation of a radiolabeled indicator of protein synthesis may be considered as a quick and reliable marker of cell survival and/or proliferation.

Animals↗
PubMed1998

Biological interventions for spinal cord injury.

Spinal cord injury is frequently followed by the loss of supraspinal control of sensory, autonomus and motor functions at sublesional level. To enhance recovery in patients with spinal cord injuries, three fundamental strategies have been developed in experimental models. These strategies involve three different time points for postlesional intervention in the spinal cord. Neuroprotection soon after injury uses pharmacological tools to reduce the progressive secondary injury processes that follow during the first week after the initial lesion occurs, in order to limit tissue damage. A second strategy, which is initiated shortly after the lesion occurs, aims at promoting axonal regeneration by acting pharmacologically on inhibitors or barriers of regeneration, or by the application of cell or gene therapy as a source of neurotrophic factors or as a bridge or support to enhance the regeneration of lesioned axons. Finally, a mid-term substitutive strategy is the management of the sublesional spinal cord by sensorimotor stimulation or the supply of missing key afferents, such as monoaminergic systems. These three strategies are reviewed. Only a combination of these different approaches can provide an optimal basis for potential therapeutic interventions aimed at functional recovery after spinal cord injury.

Animals↗
PubMed1998

Partial deafferentation of the developing rat spinal cord delays the spontaneous repression of GAD67 mRNAs in spinal cells.

Early and ubiquitous detection of GABA in the rat spinal cord before the occurrence of synaptogenesis has led to the concept of a neurotrophic role of GABA, in addition to a promoting effect on neurite extension and neurodevelopment. The aim of this study was to further establish, in vivo, evidence for a link between the maturation of spinal cord innervation and the regulation of several isoforms of the synthetic enzymes of GABA, the glutamic acid decarboxylases GAD65, GAD67, and EP10, the embryonic truncated form of GAD67. Neonatal capsaicin treatment was used to induce a specific loss of afferent fibers (unmyelinated C fibers, thin myelinated fibers A delta) to the dorsal horn. The regulation of various GAD mRNAs was investigated using sensitive techniques such as RT-PCR and in situ hybridization. The sensitivity of the methods was further enhanced by the use of a gaseous detector (beta-imager) to quantitate the mRNAs species. After neonatal capsaicin treatment, higher levels of GAD67 mRNA were detected transiently during the postnatal development of the rat spinal cord. A maximum two-fold increase of GAD67 mRNA was found on the day following the capsaicin injection and reached control values within 3 weeks. In contrast, GAD65 mRNA levels remained low and were unaffected by the treatment, and EP10 was not detected. In addition, we have found a similar upregulation, with the same time course, of the cytoskeletal protein beta-actin. The capsaicin-induction of mRNA synthesis was, however, two-fold greater for beta-actin than for GAD67. Moreover, since this upregulation of GAD67 mRNA coincides with the sprouting of unaffected afferent fibers and of 5HT axons, one can hypothesize that GAD67 participates in the structural plasticity occurring in reaction to the capsaicin-induced partial deafferentation.

Actins↗
PubMed1997

SA4503, a novel cognitive enhancer with sigma1 receptor agonist properties, facilitates NMDA receptor-dependent learning in mice.

The selective sigma1 receptor agonist 1-(3,4-dimethoxyphenethyl)-4-(3-phenyl propyl)piperazine dihydrochloride (SA4503) was reported to reverse the amnesia induced by the muscarinic receptor antagonist scopolamine at sub-mg/kg doses. We examined its effect on the learning impairment induced in mice by the non-competitive NMDA receptor antagonist dizocilpine. Learning capacities were evaluated using spontaneous alternation in the Y-maze for spatial working memory, and step-down type passive avoidance. SA4503 (0.03-1 mg/kg s.c.) attenuated the dizocilpine (0.15 mg/kg i.p.)-induced memory deficits following a bell-shaped curve in both tests. These effects of SA4503 were blocked by haloperidol (0.05 mg/kg i.p.), implicating sigma1 receptors. SA4503 also reversed the alternation deficit induced by N(omega)-nitro-L-arginine methyl ester (L-NAME, 100 mg/kg i.p.) at the same dosage, indicating that it acted on working memory through the nitric oxide (NO)-mediated signalling pathway. Furthermore, progesterone (2 mg/kg s.c.) blocked the SA4503 effects in the dizocilpine- and L-NAME-amnesia models, in accordance with the purported neurosteroids/sigma1 receptors interaction. These results demonstrate a promising neurobehavioural profile of SA4503, a ligand equally efficient to reverse the deficit in the glutamatergic as well as in the cholinergic amnesia model. Pertinent informations on the potential mechanism of the anti-amnesic effects of sigma1 receptor ligands were also obtained.

Amnesia↗
PubMed1997

Prevention of motoneuron death by adenovirus-mediated neurotrophic factors.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive loss of motoneurons, and has no effective treatment. Experimental studies in rodents have shown that motoneurons respond to a variety of molecules including brain-derived neurotrophic factor (BDNF). and the glial-cell line-derived neurotrophic factor (GDNF). Here we investigated the neuroprotective effect of these growth factors, encoded by an adenovirus, on the death of axotomized facial motoneurons in newborn rats. We used a new gene therapy strategy that involves gene transfer to motoneurons by intramuscular injection of an adenoviral vector, which is retrogradely transported from injected target muscle (Finiels et al.,: NeuroReport 7:373-378, 1995). A significant increased survival of motoneurons was observed in animals pretreated with adenovirus encoding BDNF (34.5%, P < 0.05) ou GDNF (41.9%, P < 0.05) 1 week after axotomy. These results indicate that pretreatment with BDNF or GDNF, using this therapeutic strategy, is able to prevent the massive death of motoneurons that normally follows axotomy in the neonatal period, opening new perspectives to limit neuronal death in degenerative disorders.

Adenoviridae↗
PubMed1997

Morphological and morphometric analysis of serotonin-containing neurons in primary dissociated cultures of human rhombencephalon: a study of development.

Primary dissociated cultures of rhombencephalon were prepared from 5-9-week-old human fetuses. Half of some cultures were treated by two non-competitive N-methyl-D-aspartate antagonists, namely 1-(2-thienyl)cyclohexylpiperidine (TCP) and cis-Pip/Mel-[1-(2-thienyl)-2-methyl-cyclohexyl]piperidine (GK11) in negative enantiomeric form, which enhance the survival of human fetal central nervous system cells in culture. At different days in vitro, the treated and the control cultures were processed for immunocytochemical detection of serotonin-containing neurons which were studied by morphological and morphometric analysis. Statistical analysis showed that the surface of the stained neurons increased as a function of two parameters of time, the gestational age of the cells and the duration of the cultures. The complexity of the shape of the serotonin neurons characterized by the shape factor, the number of bifurcations and the morphological feature (bipolar or multipolar) was found to increase with the gestational age. It appears that the in vitro development of the embryonic cells which represents stages of maturation and differentiation can be specifically evaluated. Such an analysis of fetal central nervous system cells improves the knowledge of factors important in grafting experiments. We verified that the two drugs do not appreciably alter the in vitro development of the treated cells; thus they may be considered as promising drugs for human neuroprotection.

Cell Differentiation↗
PubMed1997

Long-term monitoring of extracellular dopamine concentration in the rat striatum by a repeated microdialysis procedure.

This study examined a protocol for repeated measurement of the extracellular dopamine (DA) concentration in the rat striatum by microdialysis. Rats were implanted with a guide cannula in the striatum and the probe was inserted on each dialysing day, i.e. ten times over a 23 day period. During this period the animals were submitted to a control saline treatment. DA concentration was measured using the no-net-flux method. In these conditions, DA concentration remained remarkably constant over the 23 day period. The histological analysis using glial fibrillary acidic protein (GFAP), dopamine (DA) and tyrosine hydroxylase (TH) immunocytochemistry showed a moderate gliosis and a discrete increase of immunoreactivity of catecholaminergic fibres around the probe implantation site. This increase is probably related to a plasticity of the dopaminergic system in response to the lesion due to the probe implantations. This study shows that such a paradigm makes possible to measure the whole time course of the DA concentration in the rat striatum during chronic treatments with psychoactive drugs such cocaine or other compounds acting in the nigrostriatal system.

Animals↗
PubMed1997

In vitro aggregation facilities beta-amyloid peptide-(25-35)-induced amnesia in the rat.

The beta-amyloid peptide-(25-35) fragment, but not beta-amyloid peptide-(1-28), shares with beta-amyloid protein-(1-42) the ability to self-aggregate and to induce neurotoxicity in vitro. This study examined the induction of amnesia in rats given intracerebroventricularly soluble or aggregated beta-amyloid peptide-(25-35) (5-45 nmol), or beta-amyloid peptide-(1-28) (15 nmol). Memory deficit in the water-maze test, examined 14 days after aggregated beta-amyloid peptide-(25-35) injection, was more pronounced than with soluble beta-amyloid peptide-(25-35). beta-Amyloid peptide-(1-28) only affected retention. These results confirm the direct amnesic properties of beta-amyloid peptides in the rat brain and showed that prior peptide aggregation markedly facilitates the appearance of amnesia.

Amnesia↗