Ataxic Mutant Research Articles

Identification of a novel myosin-Va mutation in an ataxic mutant rat, dilute-opisthotonus.

Mutations of the myosin-Va gene (Myo5a) cause diluted coat color in mice and are occasionally associated with severe neurological disorders. Dilute-opisthotonus (dop) is a spontaneous gene mutation in the rat, and phenotypes of the homozygote (dop/dop) are similar to those of the Myo5a-deficient mouse, suggesting that the mutation resides in the rat Myo5a gene. To elucidate the molecular basis of the dop mutation, we cloned the rat Myo5a cDNA from the wild type and the dop/dop. The wild-type rat Myo5a cDNA contained a 5487-bp ORF and showed higher homology with Myo5a of the other species than Myr6 (Myo5b) in the rat. A 141-bp in-frame deletion was detected in the head region in the dop cDNA. An intragenic rearrangement consisting of a 306-bp inversion associated with 17-bp and 217-bp deletions were identified in the Myo5a gene of the dop genome. This rearrangement involved a 141-bp exon, which was skipped in the dop transcript. The MyoVA protein expression was severely impaired in the dop/dop brain. This is the first report to define the dop mutation as the Myo5a gene abnormality in the rat.

Ataxic mutant mice with defects in Ca2+channel α1Asubunit gene: morphological and functional abnormalities in cerebellar cortical neurons

This review summarizes recent studies in the morphological and functional abnormalities of cerebella in three ataxic mutant mice, i.e. tottering mouse, leaner mouse, and rolling mouse Nagoya (RMN). These mutants carry mutations in the Ca2+ channel α1A subunit gene, and become useful models for human neurological diseases such as episodic ataxia type-2, familial hemiplegic migraine, and spinocerebellar ataxia type-6. All three mutants exhibited altered morphology of the Purkinje cells, ectopic synaptic contacts between granule cell axons (parallel fibers) and Purkinje cell dendritic spines and abnormal expression of tyrosine hydroxylase in Purkinje cells. In leaner mice, Purkinje cell loss was observed in alternating sagittal compartments of the cerebellar cortex corresponding to the Zebrin II-negative zones. The mutated Ca2+ channel α1A subunit was highly expressed in granule and Purkinje cells, and the P-type Ca2+ currents in Purkinje cells were selectively reduced in the mutant mice. Therefore, we concluded that altered Ca2+ currents through the mutated Ca2+ channel α1A subunit might be involved in the functional and morphological abnormalities in granule and Purkinje cells, and might result in expressions of behavioral phenotypes including ataxia. Increased levels of corticotropin-releasing factor and cholecystokinin in some climbing and mossy fibers were observed in RMN. These neuropeptides modulated the excitability of granule and Purkinje cells, indicating the possible expression of ataxic symptoms.

Mutant mice with scrambled brains: understanding the signaling pathways that control cell positioning in the CNS.

The characterization of mouse neurological mutants has proven to be an extremely valuable approach to the analysis of central nervous system development (CNS). In particular, over the past four years, remarkable progress has been made toward the identification and characterization of genes that are required for correct cell positioning in the developing brains of mice and humans (D’Arcangelo and Curran 1998; Walsh 1999). This work has provided an impressive collection of genes (Table 1) that play key roles in both the radial and tangential migration of neurons (Pearlman et al. 1998; Hatten 1999). The classical ataxic mutant mouse reeler has served for many years as a prototype for the investigation of neurological mutations affecting neuronal migration and the organization of the CNS. The hallmark of this mutant is the disruption of neuronal cytoarchitecture in the cerebral cortex, cerebellum, and hippocampus (Caviness 1977; Rakic and Caviness 1995). The molecular basis of the reeler phenotype was uncovered by the fortuitous insertion of a fos transgene into the reeler locus, which led directly to the identification of the reelin (Reln) gene (D’Arcangelo et al. 1995). In short succession, the disabled-1 (Dab1) gene was found to be responsible for a reeler-like phenotype in both man-made and naturally occurring mouse mutants (Howell et al. 1997b; Sheldon et al. 1997; Ware et al. 1997), and related neuronal migration disorders were observed in mice deficient in either the cyclin dependent kinase 5 (Cdk5) gene or its neuronal specific activator p35 (Ohshima et al. 1996; Chae et al. 1997). The saga continued recently with reports that mice lacking both the very low density lipoprotein receptor (VLDLR) and the apolipoprotein E receptor 2 (ApoER2) exhibit anatomical trademarks of the reeler brain and that Reln binds directly to these receptors, activating a kinase signaling cascade that leads to tyrosine phosphorylation of Dab1 (D’Arcangelo et al. 1999; Hiesberger et al. 1999; Trommsdorff et al. 1999). These new findings, together with studies on the properties of the proteins encoded by these genes, are elaborating a molecular pathway that directs cell positioning in the developing CNS. The neurological mutant mouse reeler

Abnormal expression of tyrosine hydroxylase immunoreactivity in cerebellar cortex of ataxic mutant mice

Expression of tyrosine hydroxylase (TH) was examined immunohistochemically in the cerebellum of two ataxic mutants, Rolling mouse Nagoya (RMN) and dilute-lethal mice (DL). In littermate controls of both mutants, a few TH-positive Purkinje cells were distributed sparsely and their number was smaller than in the mutants at any ages examined. In RMN, TH-positive Purkinje cells were distributed in lobule IX and X, and were arranged into parasagittal bands at 2 weeks of age. TH-positive Purkinje cells increased in number and were widely distributed throughout the vermis at 3 weeks of age. In adult RMN, TH-positive Purkinje cells were found in all lobules of the cerebellum. Their parasagittal bands also became evident in the hemisphere. In DL, TH-positive Purkinje cells were mainly distributed in vermal lobules IX and X, and the flocculus at 3 weeks of age. They were also found as bands in lobules IX and X. The results suggest that abnormal expression of TH in Purkinje cells may not be specific to the allelic group. Since TH promoter is activated by Ca 2+, TH expression in the mutant Purkinje cells may predict neuronal dysfunction caused by alterations in cellular Ca 2+ currents.

Reeler: new tales on an old mutant mouse.

Neurological mouse mutants provide an opportunity to dissect the complex mechanisms that underlie vertebrate brain development. Advances in genetic technologies have permitted the identification of genes disrupted in many mutants, allowing a molecular interpretation of the phenotypes. For several decades, the spontaneous mutant mouse reeler has been used as a model for the analysis of the development of laminated brain structures. In this ataxic mutant, the migration of many neurons is aberrant, resulting in disrupted cellular organization. Recently, reelin, the gene disrupted in the reeler mouse, has been identified, reelin encodes a novel extracellular molecule that controls neural cell positioning through mechanisms that are not yet completely understood. Analysis of the expression pattern and the properties of the reelin gene product (Reelin) suggests models for its function during brain development. Furthermore, the recent identification of genes that may function in the Reelin signaling pathway advances our knowledge of the molecular basis of neuronal migration.

The stereo-specific effect of d-serine ethylester and the d-cycloserine in ataxic mutant mice

Spinocerebellar ataxia is one of the most common neurological disorders. However, few therapeutics are effective for the treatment of this disorder. In the present study, we investigated the efficacy of d-serine ethylester and a related substance, d-cycloserine, as therapeutic agents for ataxia in a murine model. Both compounds are known to stereospecific modulate N-methyl- d-aspartate type glutamate receptors, and impaired glutamate-mediated signaling has been implicated in spinocerebellar ataxia. Using a microdialysis method, we found that intraperitoneal administration of d-serine ethylester increases the extracellular content of endogenous d-serine in the mouse cerebellum for at least 3 h. Maximum elevation of the extracellular d-serine was observed at 40 min after injection. An open-field study was used to assay the effect of the d-serine derivatives on movement and ataxia. In mice exhibiting cytosine arabinoside-induced ataxia, d-serine ethylester reduced the falling index in a dose-dependent manner. The effect of d-serine ethylester was stereo-specific in that l-serine ethylester had no effect on the falling index at the maximum doses tested, and was partially inhibited by 5,7-dichlorokynurenate, an antagonist that binds to the glycine-binding site. Locomotor activity was not changed by the d-serine ethylester treatment. d-cycloserine also significantly reduced the falling index of the mice. Both d-serine ethylester and d-cycloserine had longer lasting effects than other potential therapeutic reagents for ataxia. Growing evidence suggests the essential involvement of endogenous d-serine in mammalian brain function, and our results suggest that d-serine derivatives may represent an effective new therapeutic for the treatment of spinocerebellar ataxia.

Cerebellar Brain-Derived Neurotrophic Factor–TrkB Defect Associated with Impairment of Eyeblink Conditioning inStargazerMutant Mice

In the spontaneous ataxic mutant mouse stargazer, there is a selective reduction of brain-derived neurotrophic factor (BDNF) mRNA expression in the cerebellum. BDNF protein levels in the cerebellum are reduced by 70%. Despite normal levels of full-length and truncated TrkB receptor, constitutive and neurotrophin-4/5-induced tyrosine phosphorylation was significantly reduced in several signal transduction molecules, including phospholipase-Cgamma1, erk1, and erk2. Morphological examination revealed an increased number of external granule cells at postnatal day 15 and the presence of abnormal neurons resembling immature granule cells in the adult. These abnormalities are associated with a severe impairment in the acquisition of classical eyeblink conditioning, indicating cerebellar malfunction. Our data suggest that normal BDNF expression and TrkB signal transduction in the cerebellum are necessary for learning and plasticity in this model.

Reeler: new tales on an old mutant mouse

Neurological mouse mutants provide an opportunity to dissect the complex mechanisms that underlie vertebrate brain development. Advances in genetic technologies have permitted the identification of genes disrupted in many mutants, allowing a molecular interpretation of the phenotypes. For several decades, the spontaneous mutant mouse reeler has been used as a model for the analysis of the development of laminated brain structures. In this ataxic mutant, the migration of many neurons is aberrant, resulting in disrupted cellular organization. Recently, reelin, the gene disrupted in the reeler mouse, has been identified. reelin encodes a novel extracellular molecule that controls neural cell positioning through mechanisms that are not yet completely understood. Analysis of the expression pattern and the properties of the reelin gene product (Reelin) suggests models for its function during brain development. Furthermore, the recent identification of genes that may function in the Reelin signaling pathway advances our knowledge of the molecular basis of neuronal migration. BioEssays 20:235–244, 1998.© 1998 John Wiley & Sons, Inc.

Brain D-serine and tyrosine levels in ataxic mutant mice

Since D-serine occurs at high concentrations in mammalian forebrains, the brain D-serine content was analyzed in hyperkinetic and ataxic mutant mice as well as normal control mice in a search for a physiological role. The concentrations of free D-serine (nmol/g wet weight) were 392 ± 114 (mean ± S.D.), 43 ± 17 and 18 ± 8.4 in the cerebrum, brain stem and cerebellum of the BUS mouse, respectively; and 336 ± 93,58 ± 11 and 18 ± 8.5 in the cerebrum, brain stem and cerebellum of the Rolling mouse, respectively. These values were not significantly different from those for each control animal. The present results suggest that brain D-serine may not be a cause of the abnormal movements of the mutant mice. On the contrary, among many amino acids examined, tyrosine level was found to be lower in the brain stem of BUS mouse compared to the normal control animal by amino acid analysis.

Chronic anti-ataxic actions of the novel thyrotropin-releasing hormone (TRH) analog, TA-0910, during and after repeated administration in Rolling mouse Nagoya: behavioral and pharmacokinetic studies.

Anti-ataxic effects of TA-0910, a novel thyrotropin-releasing hormone analog, in mice of the ataxic mutant mouse strain Rolling mouse Nagoya (RMN) are sustained beyond its 2-week oral administration period (Kinoshita et al., Eur. J. Pharmacol., 274, 65-72, 1995). We examined the concentration of TA-0910 in the central nervous system (CNS) of RMN after repeated administration in an attempt to clarify the mechanism of the sustained effect of the drug. Repeated administration of TA-0910 (3 mg/kg/d, i.p.) for 2 weeks produced a long-lasting ameliorating effect on ataxia in RMN, and this effect was maintained until 3 weeks after drug withdrawal. The concentrations of TA-0910 in the cerebrum and brain stem 24 h after the final administration were twice the concentration observed 24h after single administration. The cerebellum cencentration of TA-0910 was more than 4 times that observed 24 h after final administration. After repeated administrations, the drug concentrations in the brain tissues gradually decreased, but the drug was still detectable in the cerebrum and brain stem 3 weeks after withdrawal. However, these concentrations of TA-0910 3 weeks after withdrawal were as low those observed 24 h after single administration when there were no anti-ataxic effects. These observations suggest that the long-lasting ameliorating effect on the ataxia during and after repeated administration of TA-0910 is not ascribable to the drug remaining in the CNS of RMN.

Developmental alteration of local circuits in the somatosensory cortex of the ataxic mutant rat

Developmental alteration of local circuits in the somatosensory cortex of the ataxic mutant rat

Endoplasmic reticulum is missing in dendritic spines of Purkinje cells of the ataxic mutant rat

Dilute-opisthotonus ( dop) is a spontaneous ataxic mutation in the rat, regulated by an autosomal recessive gene. Immunohistochemical staining with anti-inositol 1,4,5-trisphosphate receptor antibody and electron microscopic examinations revealed that the endoplasmic reticulum in dendritic spines of Purkinje cell was missing in the ataxic rat. This could impair the intracellular signal transduction in the parallel fiber-Purkinje cell synapse, and be a cause of the severe ataxic movement.

Selective failure of brain-derived neurotrophic factor mRNA expression in the cerebellum of stargazer, a mutant mouse with ataxia

In search of the possible involvement of neurotrophic factors in inherited neurological disease, we examined brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), and neurotrophin-3 (NT-3) mRNA expression patterns in the ataxic mutant mouse stargazer (stg). Using in situ hybridization, we found a selective and near total reduction in BDNF mRNA in the cerebellar granule cell layer. NT-3 or NGF mRNA expression in the cerebellum was normal. Northern blot analysis demonstrated a 70% reduction in BDNF mRNA in the whole cerebellum. BDNF mRNA levels in other mutant brain regions were unchanged. Absence of BDNF mRNA in granule cells was observed at postnatal age (P15), coincident with the onset of ataxia, and expression levels failed to follow the developmental increase found in the wild type at later ages (P20 and P30). Despite the severe BDNF reduction, in situ hybridization patterns for both the full-length and the truncated BDNF TrkB receptor mRNA were unaltered. No major cytoarchitectural abnormalities were apparent in the stg/stg cerebellum. BDNF expression in a related ataxic mutant, tottering, was unaltered. These data show that BDNF can be regulated selectively in distinct brain regions, possibly by differential activation of its multiple promoters. Absence of cerebellar granule cell BDNF mRNA in stg/stg mice demonstrates that sustained expression of this neurotrophin is not required for cell survival in the developing cerebellar cortex. Our data, in contrast, suggest a role of BDNF in maturation of specific cerebellar neurons and pathways. Early failure of cerebellar BDNF expression may be related to the ataxic phenotype in stg mice.

Alterations in Local Cerebral Glucose Metabolism and Endogenous Thyrotropin-Releasing Hormone Levels in Rolling Mouse Nagoya and Effect of Thyrotropin-Releasing Hormone Tartrate

To identify the brain region(s) responsible for the expression of ataxic gaits in an ataxic mutant mouse model, Rolling mouse Nagoya (RMN), changes in local cerebral glucose metabolism in various brain regions and the effect of thyrotropin-releasing hormone tartrate (TRH-T), together with alterations in endogenous thyrotropin-releasing hormone (TRH) levels in the brains of RMN, were investi gated. Ataxic mice [RMN (rol/rol)] showed significant decreases in glucose metabolism in regions of the diencephalon: thalamic dorsomedial nucleus, lateral geniculate body and superior colliculus; brain stem: substantia nigra, raphe nucleus and vestibular nucleus; and cerebellar nucleus as compared with normal controls [RMN (+/+)]. When RMN (rol/rol) was treated with TRH-T (10 mg/kg, equivalent to 7 mg/kg free TRH), glucose metabolism was significantly increased in these regions. These results suggest that these regions may be responsible for ataxia. We also found that TRH levels in the cerebellum and brain stem of RMN (rol/rol) were significantly higher than those of RMN (+/+). These results suggest that ataxic symptoms in RMN (rol/rol) may relate to the abnormal metabolism of TRH and energy metabolism in the cerebellum and/or brain stem and that exogenously given TRH normalizes them.

Mapping of the dilute-opisthotonus (dop) gene on chromosome 8 of the rat.

The rat dilute-opisthotonus (dop) autosomal recessive gene, causing ataxia and coat color dilution, was mapped on chromosome 8 by PCR-amplified microsatellite markers. To facilitate the linkage analysis, an intersubspecific cross with a Japanese wild rat strain was used. The recombination frequencies were 12.8% between Apoc3 and dop, and 32.1% between dop and Mylc1v. The following order of three genes is proposed; Apoc3-dop-Mylc1v. This mutation appears to be homologous to dilute-lethal (d1) of the mouse in terms of clinical symptoms, coat color effect and chromosomal location of the gene loci. Key words: ataxic mutant rat, dilute-opisthotonus (dop), gene mapping.

CNS monoamine levels and motoric behaviors in the hotfoot ataxic mutant

Mice that were homozygous recessive for the single-gene mutation, hotfoot, showed profound and progressive motor disturbances in an open field after approximately the 4th postnatal week. Studies were undertaken to examine the role of the monoaminergic system in the behavioral and developmental expression of this neurological mutation. Relative to controls, 10- and 30-day-old hotfoot mice demonstrated a significantly attenuated response to the stimulating locomotor effects of amphetamine while adult hotfoot mice were motorically unaffected by amphetamine administration. 30-day-old and adult hotfoot mice also were hypothermic relative to phenotypically normal mice after amphetamine administration. Examination of monoamine levels and turnover revealed that hotfoot mice had significantly greater concentrations of norepinephrine associated with lower turnover in cerebellum and greater levels of serotonin in cerebellum and striatum, relative to phenotypic controls. In addition, mice born and raised by hotfoot dams demonstrated neurochemical alterations regardless of genotype. Both the neurochemical data and the developmental response to the general catecholamine agonist, amphetamine, suggest that the monaminergic neurotransmitter system may be altered as a consequence of the hotfoot mutation.

Ataxia-ameliorating effects of YM-14673, a potent analog of thyrotropin releasing hormone, in ataxic mutant mice

The effects of YM-14673 ( N α-[[( S)-4-oxo-2-azetidinyl]carbonyl]- L-histidyl- L-prolinamide dihydrate), a potent analog of thyroptropin releasing hormone (TRH), on the behavior of ataxic mutant mice (staggerer, realer, Purkinje cell degeneration and weaver mice) were investigated in comparison with those of TRH in an open-field appartuss. The fall index, the ratio of the number of falls to the distance moved, was defined as an index for ataxia. YM-14673 reduced the fall index in all types of ataxic mice examined and was more potent than TRH in reeler, staggerer and Purkinje cell degereration mice. YM-14673 may have a beneficial effect on patients with spinocerebellar degeneration.

Levels of somatostatin and cholecystokinin in the brain of ataxic mutant mice

Changes in immunoreactive somatostatin (SOM) and cholecystokinin (CCK-8) levels in the cerebellum and cerebrum were investigated in three types of genetically - determined ataxic mutant mice: rolling mouse Nagoya (RMN), weaver, and Purkinje cell degeneration (PCD) mice. The cerebellar pathology in each of these types differes. The concentration of both SOM and CCK-8 (ng/mg weight) was significantly higher in the cerebellum and the cerebrum of the three types of ataxic mutant mice than in these regions in the respective controls. SOM and CCK-8 content (ng/organ) was significantly higher in PCD and RMN than in controls but this was not in the weaver mice. The possible involvement of both peptides in manifestations of ataxia is discussed.

<b>Substance P-like immunoreactivity in the central nervous system of Wriggle mouse Sagami, an ataxic mutant </b><b>mouse </b>

<b>Substance P-like immunoreactivity in the central nervous system of Wriggle mouse Sagami, an ataxic mutant </b><b>mouse </b>

Concentrations of catecholamines and indoleamines in the central nervous system of wriggle mouse sagami

1. 1. The concentrations of norepinephrine (NE), 3-methoxy-4-hydroxyphenylglycol (MHPG), 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) in the central nervous system of Wriggle mouse Sagami (WMS), which is a new ataxic mutant mouse, were studied. 2. 2. NE and MHPG levels were increased most remarkably in the cerebellum. 3. 3. 5-HT and 5-HIAA levels were increased most remarkably in the brain stem and spinal cord. 4. 4. The present results suggest enhancement of catecholamine and indoleamine metabolism in the cerebellum and bulbospinal cord, respectively, of the WMS, and these changes seem relevant to the specific motor dysfunction of the WMS.