Biomedical subjects

Jean Mariani

Publications and source records attributed to Jean Mariani.

33 records · Page 2Linked to original sources

PubMed2003

Lurcher GRID2-induced death and depolarization can be dissociated in cerebellar Purkinje cells.

The Lurcher mutation transforms the GRID2 receptor into a constitutively opened channel. In Lurcher heterozygous mice, cerebellar Purkinje cells are permanently depolarized, a characteristic that has been thought to be the primary cause of their death, which occurs from the second postnatal week onward. The more dramatic phenotype of Lurcher homozygotes is thought to be due to a simple gene dosage effect of the mutant allele. We have analyzed the phenotype of Lurcher/hotfoot heteroallelic mutants bearing only one copy of the Lurcher allele and no wild-type Grid2. Our results show that the absence of wild-type GRID2 receptors in these heteroallelic mutants induces an early and massive Purkinje cell death that is correlated with early signs of autophagy. This neuronal death is independent of depolarization and can be explained by the direct activation of autophagy by Lurcher GRID2 receptors through the recently discovered signaling pathway formed by GRID2, n-PIST, and Beclin1.

Animals↗

PubMed2003

p53 inactivation leads to impaired motor synchronization in mice.

We have combined genetic and pharmacological approaches to investigate the behavioural consequences of inactivation of the murine p53 protein. Our behavioural analysis revealed that p53-null mice (p53KO) exhibit a very specific and significant motor deficit in rapid walking synchronization. This deficit, observed using the rotarod test, was the only behavioural defect of p53KO mice. We demonstrated that it was not due to an increase in neuronal number or abnormal connectivity in the olivo-cerebellar system, thought to control motor synchronization. In order to test the role of p53 in the central nervous system, we injected a pharmacological inhibitor of p53 activation, pifithrin-alpha, into the cerebellum of wild-type mice. This treatment mimicked the walking synchronization deficit of p53KO mice, suggesting that presence of p53 protein in the cerebellum is necessary to execute this synchronization of walking. Our investigation reveals a functional role of cerebellar p53 protein in adult walking synchronization.

Animals↗

PubMed2003

Post-lesion transcommissural growth of olivary climbing fibres creates functional synaptic microzones.

In the adult mammalian central nervous system, reinnervation and recovery from trauma is limited. During development, however, postlesion plasticity may generate alternate paths, providing models to investigate reinnervating axon-target interactions. After unilateral transection of the neonatal rat olivocerebellar path, axons from the ipsilateral inferior olive grow into the denervated hemicerebellum and develop climbing fibre (CF)-like arbors on Purkinje cells (PCs). However, the synaptic function and extent of PC reinnervation remain unknown. In adult rats pedunculotomized on postnatal day 3 the morphological and electrophysiological properties of reinnervating olivocerebellar axons were studied, using axonal reconstruction and patch-clamp PC recording of CF-induced synaptic currents. Reinnervated PCs displayed normal CF currents, and the frequency of PC reinnervation decreased with increasing laterality. Reinnervating CF arbors were predominantly normal but 6% branched within the molecular layer forming smaller secondary arbors. CFs arose from transcommissural olivary axons, which branched extensively near their target PCs to produce on average 36 CFs, which is six times more than normal. Axons terminating in the hemisphere developed more CFs than those terminating in the vermis. However, the precise parasagittal microzone organization was preserved. Transcommissural axons also branched, although to a lesser extent, to the deep cerebellar nuclei and terminated in a distribution indicative of the olivo-cortico-nuclear circuit. These results show that reinnervating olivocerebellar axons are highly plastic in the cerebellum, compensating anatomically and functionally for early postnatal denervation, and that this reparation obeys precise topographic constraints although axonal plasticity is modified by target (PC or deep nuclear neurons) interactions.

Animals↗

PubMed2002

Abeta(25-35) and Abeta(1-40) act on different calcium channels in CA1 hippocampal neurons.

The acute effects of beta-amyloid (25-35) and (1-40) on high voltage activated calcium channels were compared in CA1 pyramidal cells of adult mouse hippocampal slices using the whole-cell patch-clamp recording. Bath application of oligomeric beta-amyloid (25-35) reversibly increased the barium current (I(Ba)) to 1.61 (normalized amplitude), while oligomeric beta-amyloid (1-40) reversibly enhanced the I(Ba) to 1.74. Reverse-sequence beta-amyloid [(35-25) and (40-1)] had no effect. The effect of beta-amyloid (25-35) was blocked by nifedipine, a selective antagonist of L-type calcium channels. In contrast, the effect of beta-amyloid (1-40) was not blocked by nifedipine and I(Ba) was enhanced to 4.96. It is concluded that these oligomeric peptides may act through different types of calcium channels and/or receptors. The toxicity of Abeta(25-35) implicates a potentiation of L-type calcium channels while the one of Abeta(1-40) is related to an increase of non-L-type calcium channels, which may involve an increase in transmitter release.

Amyloid beta-Peptides↗

PubMed2002

Identification of genes involved in ceramide-dependent neuronal apoptosis using cDNA arrays.

BACKGROUND: Ceramide is important in many cell responses, such as proliferation, differentiation, growth arrest and apoptosis. Elevated ceramide levels have been shown to induce apoptosis in primary neuronal cultures and neuronally differentiated PC 12 cells. RESULTS: To investigate gene expression during ceramide-dependent apoptosis, we carried out a global study of gene expression in neuronally differentiated PC 12 cells treated with C2-ceramide using an array of 9,120 cDNA clones. Although the criteria adopted for differential hybridization were stringent, modulation of expression of 239 genes was identified during the effector phase of C2-ceramide-induced cell death. We have made an attempt at classifying these genes on the basis of their putative functions, first with respect to known effects of ceramide or ceramide-mediated transduction systems, and then with respect to regulation of cell growth and apoptosis. CONCLUSIONS: Our cell-culture model has enabled us to establish a profile of gene expression during the effector phase of ceramide-mediated cell death. Of the 239 genes that met the criteria for differential hybridization, 10 correspond to genes previously involved in C2-ceramide or TNF-alpha signaling pathways and 20 in neuronal disorders, oncogenesis or more broadly in the regulation of proliferation. The remaining 209 genes, with or without known functions, constitute a pool of genes potentially implicated in the regulation of neuronal cell death.

Animals↗

PubMed2002

HRP injection in lobule VI-VII of the cerebellar cortex reveals a bilateral inferior olive projection in granuloprival rats.

In immature rats, Purkinje cells receive synapses from multiple climbing fibers. During development, this multi-innervation regresses and only one climbing fiber innervates each Purkinje cell in the adult. The multi-innervation of immature rats is maintained in the adult if the precursors of the cerebellar granule cells are destroyed by early postnatal X-irradiation. The present study was undertaken to determine the origin of climbing fibers projecting to lobule VI-VII of the cerebellum in X-irradiated granuloprival rats. Olivary neurons were labelled by retrograde transport of wheat germ agglutinin conjugated to horseradish peroxidase, which was injected by iontophoresis in the right vermis of lobule VI-VII. Three-dimensional reconstructions of the inferior olive were made for granuloprival and control rats. No significant variation in the shape and dimension of the olive was observed between the two groups. Labeled cells were found in the middle part of the median accessory olive (MAO). In control rats, stained cells were found only in the contralateral MAO, whereas in the granuloprival rats they were located in both the contralateral and the ipsilateral MAO. Homologous zones were marked in control and granuloprival rats in the middle part of MAO. In granuloprival rats, there was a symmetry in the distribution of the stained cells in the ipsi- and contralateral MAO along the three axes. Therefore, polyinnervation involves homologous regions of both inferior olivary nuclei.

Animals↗

PubMed2002

Smooth muscle dysfunction in resistance arteries of the staggerer mouse, a mutant of the nuclear receptor RORalpha.

Retinoic acid receptor-related orphan receptor alpha (RORalpha) is a member of the nuclear receptor superfamily. The mouse mutant staggerer (sg/sg) carries a deletion within the RORalpha gene. RORalpha plays a major role in cellular differentiation during development and growth. In the present study, we found a lower mean arterial blood pressure in sg/sg than in +/+ mice (80.1+/-1.2 and 87.0+/-0.9 mm Hg, respectively; P<0.0002) and a smaller increase in blood pressure after in vivo injections of phenylephrine. To elucidate the mechanisms responsible for this phenotype, we investigated the vascular reactivity of large vessels (aorta and carotid arteries) and small resistance mesenteric arteries in response to mechanical forces or vasoactive agents. Arteries from sg/sg and +/+ mice were studied in vitro in arteriographs. Vascular responses of large vessels to all stimuli were similar in both groups. However, we found a markedly altered vascular function in mesenteric arteries from sg/sg mice. Flow-induced dilation, pressure-induced myogenic tone, responses to endothelium-dependent or -independent vasodilators, and responses to vasoconstrictors were significantly reduced in sg/sg compared with +/+ mice. We also determined by Western blot analysis the expression of smooth muscle (SM)-myosin, calponin, and heavy (h)-caldesmon, in large and small arteries of sg/sg and +/+ mice, and found a marked decrease in the expression of these contractile proteins in mesenteric arteries of sg/sg mice. Our findings provide the first evidence that functional RORalpha is required for normal contractile phenotype of smooth muscle cells (SMCs) in small resistance arteries and suggest that RORalpha might be involved in the differentiation of SMCs in mesenteric arteries.

Animals↗

PubMed2002

The role of climbing and parallel fibers inputs to cerebellar cortex in navigation.

DA-HAN rats with partial or total lesion of climbing (CF) and parallel fibers (PF) inputs of the cerebellum were tested in a water task. Two different protocols were used, requiring to find either a non-visible or a visible platform. These two protocols were, respectively, designed to evaluate visuo-motor guidance (visible platform) and navigation (non-visible platform). Both groups of lesioned rats presented a deficit in the non-visible platform task but not in the visible platform one. The protocol of navigation we used was a fixed start-fixed arrival procedure. Totally lesioned animals were unable to learn to orient their body toward the non-visible platform and adopted instead a circling behavior. Our results suggest a role of cerebellar inputs (climbing (CF) and PF) in navigation.

Animals↗

PubMed2002

Expression and regulation of the nuclear receptor RORalpha in human vascular cells.

Retinoic acid receptor-related orphan receptor alpha (RORalpha) is a member of the nuclear receptor superfamily. Using RT-PCR, RORalpha mRNA was identified in human aortic smooth muscle cells (hASMC), endothelial cells (EC), as well as in human mammary arteries and atherosclerotic plaques. We found a predominant expression of RORalpha1 in hASMC, and RORalpha4 in EC. RORalpha2 and RORalpha3 were not detected. In arteries, RORalpha4 was predominant compared with RORalpha1. In atherosclerotic plaques, RORalpha expression was significantly decreased. In hASMC stimulated with cytokines, RORalpha expression was increased by 2.5-fold. RORalpha mRNA was also significantly increased (approximately 2-fold) in hASMC and EC cultured under hypoxia.

Aorta↗

PubMed2002

Age-related phenotypes in the staggerer mouse expand the RORalpha nuclear receptor's role beyond the cerebellum.

The homozygous mutant mouse staggerer (RORa(sg)/RORa(sg)), was initially described as ataxic, due to the presence of massive neurodegeneration in the cerebellum [Science 136 (1962) 610]. The identification of the widely expressed Retinoic acid receptor-related Orphan Receptor, NR1F1 (RORalpha) gene as the site of mutation in the staggerer mouse has led to great progress in understanding the molecular basis of its phenotype in recent years [Nature 379 (1996) 736]. RORalpha is a transcription factor, belonging to the nuclear receptor superfamily, for which no natural ligand has yet been identified. Mice engineered for the disruption of the gene encoding RORalpha display the same cerebellar atrophic phenotype as the staggerer mouse [Proc. Natl. Acad. Sci. USA 95 (1998) 3960]. More recently, it has been shown that the mutation is semi-dominant, as heterozygous animals display an increased loss of Purkinje cells with age. Furthermore, a number of additional phenotypes outside the nervous system have recently been identified. These include a greater susceptibility to atherosclerosis [Circulation 15 (1998) 2738], immunodeficiencies linked to the overexpression of inflammatory cytokines [J. Neurochem. 58 (1992) 192], abnormalities in the formation and maintenance of bone tissue [Proc. Natl. Acad. Sci. USA 97 (2000) 9197] and changes in muscle differentiation [Nucleic Acids Res. 27 (1999) 411]. Thus, RORalpha has been directly linked to a number of age-related pathologies of great medical interest.

Age Factors↗

PubMed2002

To die or not to die, does it change the function? Behavior of transgenic mice reveals a role for developmental cell death.

In humans, perturbations in the developmental neuronal death leading to an excess of neurons could be associated with developmental neuropsychiatric disorders. Hu-bcl-2 transgenic mice appear to be a valuable tool to study the functional role of developmental programmed cell death. Indeed, the over-expression of the anti-apoptotic gene bcl-2 decreases developmental neuronal death and Hu-bcl-2 mice present supernumerary neurons in several brain regions. A detailed behavioral analysis of these mice revealed selective deficits. Hu-bcl-2 mice have normal vision, general activity and motor skills. Only the most complex behavior like anxiety and learning abilities are impaired in these mice.

Animals↗

PubMedDate not supplied

Diminishing Purkinje cell populations in the cerebella of aging heterozygous Purkinje cell degeneration but not heterozygous nervous mice.

Mice homozygous for the recessive, cerebellar affected mutations nervous and Purkinje cell degeneration display an almost complete loss of Purkinje cells during their first two postnatal months. We have recently shown a progressive and age-related loss of Purkinje cells in mutant mice heterozygous for mutations apparently recessive, such as staggerer and reeler, and have thus chosen to investigate whether a similar Purkinje cell loss occurred with aging in +/nr and +/pcd heterozygous mice. Purkinje cells were counted on serial sections stained with thionin obtained from 17-month-old male and female heterozygous mice and their respective wild-type controls. In the case of the +/nr and wild-type mice, no difference in cell counts was observed. However, +/pcd mice had significantly fewer (-18%) Purkinje cells (143.700+/-5.400) than control wild-types (175.100+/-2.300); p<0.0001) at 17 months. These results extend our previous findings and further support the idea that apparently recessive neurological mutations may exert, at the heterozygous state, a deleterious effect upon Purkinje cells during the aging process.

Aging↗

PubMedDate not supplied

Cell number in the inferior olive of nervous and leaner mutant mice.

Naturally occurring cell death is an important feature of neuronal network development: the absence of adequate postsynaptic target neurons during a critical period may result in the death of presynaptic neurons, the degree of death varying inversely with the size of the target population. Studies of mouse mutants with abnormal cerebellar development provide support for this neuron/target relationship in circuits within the CNS. In the present study we analysed the inferior olivary cell population in two cerebellar mutant mice, nervous (nr/nr) and leaner (Cacna1ala/la). In these mice Purkinje cell degeneration begins near the end of the first postnatal month. In nervous mice the loss starts at postnatal day 20 (P20) and by the end of second month almost 90% of the Purkinje cells in the hemisphere and 50% in the vermis have disappeared. In leaner mice Purkinje cell loss starts after P30 and by P60 almost 50% of these cells are lost. We report here a loss of one third of inferior olivary neurons in the nervous mutation while the entire population appears intact in the leaner mouse. These results allow better definition of the end of the period of target dependency of inferior olive neurons. Their implications for the cell-cell interactions in the developing olivo-cerebellar system are discussed.

Animals↗

PubMedDate not supplied

Quantitative analysis of microglial cells in the degenerating cerebellum of the staggerer (RORA(sg/sg)) mutant mouse.

Elevated levels of pro-inflammatory cytokines, such as IL-1ss and IL-6, have been detected in the cerebellum of Rora(sg/sg) mice during the first postnatal month of neurodegenerative process. This suggests the existence of a microglial reaction in the context of an inflammatory process that would be triggered by the massive neuronal loss. To test this hypothesis, we qualitatively and quantitatively studied the microglial cell population using lectin and nucleosidic diphosphatase labeling of the cerebellum of 30-day-old mice. The massive neuronal loss induces a 11.7-fold smaller size of the Rora(sg/sg) cerebellum compared to wild-types. We showed that the Rora(sg/sg) microglia population is exclusively composed of cells displaying the characteristic morphology of activated cells, with enlarged, heavily stained cell bodies and few thick processes, in contrast to microglial cells in the wild-type. The density of microglia is 2.7-fold higher in Rora(sg/sg) than wild-type mice (22444+/-5011 cells/mm(3) versus 8158+/-1584 cells/mm(3)), although the absolute number is 4-fold smaller. These results show that neurodegeneration in the Rora(sg/sg) cerebellum leads to persistance of microglial activation while in wild-type it disappears around P10.

Acid Anhydride Hydrolases↗

PubMedDate not supplied

Cerebellar purkinje cell loss in heterozygous rora+/- mice: a longitudinal study.

The staggerer (sg) mutation is a spontaneous deletion in the Rora gene that prevents the translation of the ligand-binding domain (LBD), leading to the loss of RORalpha activity. The homozygous Rorasg/sg mutant mouse, whose most obvious phenotype is ataxia associated with cerebellar degeneration, also displays a variety of other phenotypes. The heterozygous Rora+/sg is able to develop a cerebellum that is qualitatively normal but which suffers a significant loss of cerebellar neuronal cells with advancing age. A truncated protein synthesized by the mutated allele may play a role both in Rorasg/sg and Rora+/sg. To determine the effects during life span of true haplo-insufficiency of the RORalpha protein, derived from the invalidation of the gene, we compared the evolution of Purkinje cell numbers in heterozygous Rora knock-out males (Rora+/-) and in their wild-type counterparts from 1 to 24 months of age. We also compared the evolution of Purkinje cell (PC) numbers in Rora+/- and Rora+/sg males from 1 to 9 months. The main finding is that in Rora+/- mice, for which only one-half the normal amount of protein is produced, the deficit was established as early as 1 month and did not change during the animals' adult lifespans. Thus, the effects of aging on PC number were apparent much earlier in Rora+/- than in Rora+/sg, although at 24 months of age the degrees of deficit were similar.

Aging↗