Biomedical subjects

Kazuhiko Sawada

Publications and source records attributed to Kazuhiko Sawada.

15 recordsLinked to original sources

PubMed2006

Histological brain alterations following prenatal methamphetamine exposure in rats.

When pregnant women abuse methamphetamine, the foremost concern is the potential adverse effect of this substance on fetal development. Clinical studies in humans have found that exposure to methamphetamine during brain development can cause neurobehavioral abnormalities, such as aggressive behavior, learning problems, and poor social adaptation. In the present study, we examined the effects of prenatal methamphetamine exposure on brain development in rats. The first group of pregnant rats was administered methamphetamine at a dose of 5 mg/kg/day during gestational day (GD 10 to GD 20 [MA]. The second group of pregnant rats was injected with saline vehicle only [SAL]. On GD 21 their fetuses were removed and fetal brains were observed. We found various types of morphological damage in MA fetal brains, including microgyria, ectopia, and hemorrhage. In some cases, abnormal distribution of the leptomeninx, such as breach or accumulation, was observed in addition to these histological abnormalities. Therefore, we examined the expression of laminin, which is an important component of the pia mater, in the fetal brains. However, Western blot analysis revealed that there was no difference in expression amount of laminin in whole fetal brain between the MA and SAL groups. We concluded that methamphetamine use during pregnancy can cause histological brain alterations in fetuses. Morphological alterations of brain seen in the present study and previous human studies following prenatal exposure to methamphetamine might be related to the neurobehavioral abnormalities seen in patients who had been exposed to methamphetamine in utero.

Animals↗

PubMed2005

Regional difference in corticotropin-releasing factor immunoreactivity in mossy fiber terminals innervating calretinin-immunoreactive unipolar brush cells in vestibulocerebellum of rolling mouse Nagoya.

Unipolar brush cells (UBCs), a class of interneurons in the vestibulocerebellum, play roles in amplifying excitatory inputs from vestibulocerebellar mossy fibers. This study aimed to clarify whether corticotropin-releasing factor (CRF)-positive mossy fiber innervation of calretinin (CR)-positive UBCs was altered in rolling mouse Nagoya (RMN). The distribution and the number of CR-positive UBCs in the vestibulocerebellum were not different between RMN and control mice. Double immunofluorescence revealed that some CRF-positive mossy fiber terminals were in close apposition to CR-positive UBCs. In the lobule X of vermis, such mossy fiber terminals were about 5-fold greater in number in RMN than in controls. In contrast, the number of CRF-positive mossy fiber terminals adjoining CR-positive UBCs in the flocculus was not significantly different between RMN and controls. The results suggest increased number of CRF-positive mossy fiber terminals innervating CR-positive UBCs in the lobule X but not in the flocculus of RMN. CRF may alter CR-positive UBC-mediated excitatory pathways in the lobule X of RMN and may disturb functions of the lobule X such as cerebellar adaptation for linear motion of the head.

Animals↗

PubMed2004

Hydrocephalus following prenatal exposure to ethanol.

Pregnant rats were administered a liquid diet containing 5% (w/v) ethanol between gestational days 10 and 21, and the brains of nine offspring were examined at 8 weeks of age. Two ethanol-treated offspring showed obvious hydrocephalus characterized by markedly enlarged lateral ventricles. Most of the other ethanol-treated rats only showed a slight enlargement of the lateral ventricles. An ethanol-treated offspring showed no neuropathological signs of hydrocephalus. Histological observation of the hydrocephalic brains revealed dilation of the lateral ventricles, loosely bundled corpus callosum, hypoplasia of the septum and fimbria, and thinning of the cerebral cortices. There were no differences in the shape of the third ventricle and aqueduct between hydrocephalic and non-hydrocephalic rats. Ectopic cell clusters were found on the surface of the lateral ventricle and in the interventricular foramen in ethanol-treated rats with hydrocephalus. However, leptomeningeal heterotopias were found on the cortical surface in ethanol-exposed rats independently of whether or not they showed hydrocephalus. Thus, heterotopias within the ventricles may be related to the genesis of hydrocephalus following prenatal ethanol exposure. However, whether they could be a direct cause of the hydrocephalus is uncertain as they seem to be not enough large to block the current of the cerebrospinal fluid. We also examined the expression of L1, a cell adhesion molecule suspected of involvement in the genesis of hydrocephalus after prenatal ethanol exposure, in 1-day-old rats. Western blot analysis using specific antibody against L1 showed no significant change in L1 protein expression in ethanol-exposed rats. These data suggest that L1 protein expression is not affected by ethanol.

Animals↗

PubMed2004

Abnormal expression of tyrosine hydroxylase not accompanied by phosphorylation at serine 40 in cerebellar Purkinje cells of ataxic mutant mice, rolling mouse Nagoya and dilute-lethal.

This study examined immunohistochemically the expression of an enzymatically active form of tyrosine hydroxylase (TH), phosphorylated TH at Ser40 (phospho-TH), in the cerebellum of ataxic mutant mice, rolling mouse Nagoya (RMN) and dilute-lethal (DL). TH immunostaining appeared in some Purkinje cells in RMN and DL, but in a few of the Purkinje cells of littermate controls for both mutants. In all groups of mice, there were no phospho-TH immunoreactive Purkinje cells in the cerebellum, although the subsets of TH immunoreactive Purkinje cells were found in the adjacent sections. The results suggest that TH expression in the Purkinje cells of ataxic mutants abnormally increases without activation of this enzyme by phosphorylation. This may mean that TH in Purkinje cells is not related to catecholamine synthesis.

Animals↗

PubMed2004

Purkinje cell loss in the cerebellum of ataxic mutant mouse, dilute-lethal: a fractionator study.

This study estimated total number of Purkinje cells in the cerebellum of an ataxic mutant mouse, dilute-lethal (DL), with reference to severe ataxic symptoms of this mutant. On postnatal day (PD) 21, the cerebellar weight is significantly lower in DL than in non-ataxic littermates (control mice). Total number of Purkinje cells is also significantly lower in DL than in the controls; approximately 25% less in DL than in the controls. Furthermore, we performed in situ nick end labeling (TUNEL) -staining in the cerebellum of DL during prenatal and postnatal periods in order to examine the cause of the reduced Purkinje cell number. For analyzes of the mutant fetuses, it is necessary to identify the homozygous mutant. We succeeded in identifying the homozygous DL fetuses from the control fetuses (wild-type or heterozygous fetuses) by the hair color of the grafted skin pieces on nude mice. The histological features of the cerebellar primordium did not differ between the DL and controls on embryonic and postnatal ages examined. In DL, a significantly greater number of TUNEL-positive Purkinje cells was detected on embryonic day (ED) 12, but not throughout ED 14 to PD 21. The results suggest that the Purkinje cell loss in the DL cerebellum is attributed to increased apoptotic cell death of the progenitors. This may be involved in the development of severe ataxic symptoms of DL.

Animals↗

PubMed2003

Prenatal exposure to alcohol alters the light response in postnatal circadian rhythm.

We studied the effect of prenatal exposure to alcohol on later circadian rhythm in the rat. In the normal light-dark cycle, an 8-h phase advance brought forward the deep body temperature rhythm in control rats, although it had a smaller effect in prenatally ethanol-exposed rats. In long constant darkness, the phase response of the deep body temperature rhythm to a light pulse at the early subjective night was less marked in ethanol-exposed rats in comparison to controls. These results indicate that prenatal exposure to alcohol has a long-lasting effect on the light responsiveness of the deep body temperature circadian rhythm.

Animals↗

PubMed2003

Over-expression of corticotropin-releasing factor mRNA in inferior olivary neurons of rolling mouse Nagoya.

Expression of corticotropin-releasing factor (CRF) mRNA was examined in the inferior olivary nucleus (ION) of an ataxic mutant, rolling mouse Nagoya (RMN) by semi-quantitative in situ hybridization. The most marked difference in the level of CRF mRNA signals between RMN and non-ataxic littermates (control mice) was observed in the beta-subnucleus and ventrolateral protrusion of the ION. The level of signals in these subnuclei was about twofold higher in RMN than in the controls. Signal levels in the dorsal nucleus, principal nucleus and subnucleus A were slightly but significantly higher in RMN than in the controls. In the other subnuclei, there were no differences in signal level between RMN and controls. These results suggest a region-related over-expression of CRF mRNA in the ION of RMN. This may be responsible for the increased sensitivity of some Purkinje cells to glutamate, resulting in ataxic symptoms of RMN.

Animals↗

PubMed2003

Expression of neural cell adhesion molecule L1 in the brain of rats exposed to X-irradiation in utero.

To gain insight to the cellular and molecular mechanisms involved abnormal neuronal migration induced by irradiation, we investigated expression of neuronal cell adhesion molecule L1 and neuronal migration in the brains through comparison between rats prenatally exposed to X-ray and controls. To observe the pattern of neuronal migration, bromodeoxyuridine (BrdU) was chosen as a marker to label migrating cells. The results showed some of the labeled cells remained in the lower of the cortical plate in the irradiated rats, suggesting that neuronal migration was disrupted by X-ray. To study change of expressing neural cell molecule L1, rat brains were analyzed by SDS-PAGE after isolation of L1 by immunoaffinity chromatography. In the all brain membrane fraction, immunoaffinity purified L1 had bands at 200, 180, 140 and 80 kDa. However, the bands in the irradiated group were very weak when compared with the control. Taking these results into account, abnormal neuronal migration and reduction of expression L1 found in the irradiated brain indicated that migration of neural cells may be largely dependent on radial glial fiber as well as neural cell molecules like L1. A decrease in L1 expression may be one of reasons of abnormal neuronal migration.

Animals↗

PubMed2003

Glutamate and GABA concentrations in the cerebellum of novel ataxic mutant Pogo mice.

The Pogo mouse is an autosomal recessive ataxic mutant that arose spontaneously in the inbred KJR/MsKist strain derived originally from Korean wild mice. The ataxic phenotype is characterized by difficulty in maintaining posture and side to side stability, faulty coordination between limbs and trunk, and the consequent inability to walk straight. In the present study, the cerebellar concentrations of glutamate and GABA were analyzed, since glutamate is a most prevalent excitatory neurotransmitter whereas gamma-aminobutyric acid (GABA) is one of the most abundant inhibitory neurotransmitters, which may be the main neurotransmitters related with the ataxia and epilepsy. The concentration of glutamate of cerebellum decreased significantly in ataxic mutant Pogo mouse compared to those of control mouse. However, GABA concentration was not decrease. These results suggested that the decrease in glutamate concentration may contribute to ataxia in mutant Pogo mouse.

Animals↗

PubMed2002

Expression of inorganic phosphate/vesicular glutamate transporters (BNPI/VGLUT1 and DNPI/VGLUT2) in the cerebellum and precerebellar nuclei of the rat.

Expression of inorganic phosphate/vesicular glutamate transporters (BNPI/VGLUT1 and DNPI/VGLUT2) was studied in the cerebellum and precerebellar nuclei of rats using immunohistochemistry and in situ hybridization. DNPI/VGLUT2-stained mossy fibers were principally seen in the vermis (lobules I and VIII-X) and flocculus, whereas BNPI/VGLUT1-stained mossy fibers were localized throughout the cortex. Some vermal and floccular mossy fibers were stained for both transporters. High levels of DNPI/VGLUT2 mRNA hybridization signals were demonstrated in many neurons throughout the vestibular nuclear complex as well as the lateral reticular, external cuneate, inferior olivary and deep cerebellar nuclei. Significant BNPI/VGLUT1 mRNA signals were demonstrated in the lateral reticular nucleus and vestibular nuclear complex but not in the inferior olivary nucleus, indicating that climbing fibers have DNPI/VGLUT2 only. These results show that DNPI/VGLUT2 is expressed preferentially to vestibulo-, reticulo- and cuneocerebellar neurons, some of which also possess BNPI/VGLUT1, suggesting some differential and co-operative functions between DNPI/VGLUT2 and BNPI/VGLUT1 in the cerebellum.

Animals↗

PubMed2002

Adverse effects of maternal ethanol consumption on development of dorsal hippocampus in rat offspring.

We examined the laminar structure and distribution of mossy fiber terminal fields in the dorsal hippocampus, an important area for spatial learning, in rats exposed to ethanol during gestational days 10-21. Pyramidal cells in the CA3a subfield were loosely packed compared to control rats. Aberrant infra- and intrapyramidal mossy fibers were found in the CA3 region, especially in the CA3a subfield, throughout the dorsal hippocampus of ethanol-exposed rats. Aberrant mossy fiber terminals were observed more frequently in the rostral than the caudal level of the dorsal hippocampus. At the most caudal level of the dorsal hippocampus, disarrangement of pyramidal cells was seen in the CA3c subfield along with disturbed mossy fiber terminals. Immunohistochemical studies revealed that neural cell adhesion molecule (NCAM) was not related to aberrant distribution of mossy fiber terminals after prenatal exposure to ethanol. Parvalbumin immunoreactivity was increased in the dorsal hippocampus of ethanol-exposed rats compared with control rats. Abnormal development of the dorsal hippocampus induced by prenatal ethanol exposure may be associated with the defect of spatial memory seen in fetal alcohol syndrome children and their animal models.

Alcohol Drinking↗

PubMed2002

Administration schedule for an ethanol-containing diet in pregnancy affects types of offspring brain malformations.

In the present study, we administered liquid diets containing ethanol to pregnant rats on different schedules, and examined the cerebral cortex of their pups on gestational day (GD) 21 by immunohistochemistry. The first group of pregnant rats was fed a liquid diet containing 5% (w/v) ethanol during GDs 10-21(5% Et). The second group was fed a liquid diet containing 2.5% (w/v) ethanol on GDs 10-12, a diet containing 4% (w/v) ethanol on GDs 13-15, and a diet containing 5% (w/v) ethanol on GDs 16-21 (2.5-5% Et). Pups of 5% Et dams had leptomeningeal heterotopias mainly in the parietal cortex. In 2.5-5% Et pups, other types of malformations such as grooves, microgyri, stacked-up cortices, and defects of layer I were found. The diet intake and body weight gain of 2.5-5% Et dams were significantly higher than those of 5% Et dams during GDs 11-16. There was no difference in total ethanol consumption during GDs 10-21 between the two groups. However, ethanol consumption on GD 15 in 2.5% Et was higher than in 5% Et. A different schedule for administration of an ethanol-containing diet in pregnancy might induce different types of cerebral malformations in rat fetuses.

Alcohol-Induced Disorders, Nervous System↗

PubMed2002

A selective loss of small-diameter myelinated optic nerve axons in rats prenatally exposed to ethanol.

Pregnant rats were fed an ethanol-containing liquid diet between gestational days 10 and 21. The optic nerves of their litters at 49 days of age were examined using quantitative stereological procedures. Cross-sectional areas of the optic nerve in ethanol-exposed rats were significantly smaller than those in controls. This was reflected in the reduced number of myelinated fibers, but not of non-myelinated fibers. The size distribution histogram indicated a decreased number of small axonal-diameter myelinated fibers in ethanol-exposed rats. The results suggested optic nerve hypoplasia in ethanol-exposed rats characterized by a selective loss of small-diameter myelinated fibers.

Animals↗

PubMed2002

Normal and abnormal neuronal migration in the developing cerebral cortex.

Neuronal migration is the critical cellular process which initiates histogenesis of cerebral cortex. Migration involves a series of complex cell interactions and transformation. After completing their final mitosis, neurons migrate from the ventricular zone into the cortical plate, and then establish neuronal lamina and settle onto the outermost layer, forming an "inside-out" gradient of maturation. This process is guided by radial glial fibers, requires proper receptors, ligands, other unknown extracellular factors, and local signaling to stop neuronal migration. This process is also highly sensitive to various physical, chemical and biological agents as well as to genetic mutations. Any disturbance of the normal process may result in neuronal migration disorder. Such neuronal migration disorder is believed as major cause of both gross brain malformation and more special cerebral structural and functional abnormalities in experimental animals and in humans. An increasing number of instructive studies on experimental models and several genetic model systems of neuronal migration disorder have established the foundation of cortex formation and provided deeper insights into the genetic and molecular mechanisms underlying normal and abnormal neuronal migration.

1-Alkyl-2-acetylglycerophosphocholine Esterase↗

PubMed2002

Immunohistochemistry of voltage-gated calcium channel alpha1B subunit in mouse cerebellum.

Secretion of neurotransmitters is initiated by voltage-gated calcium influx through presynaptic, voltage- gated N-type calcium channels. However, little is known about their cellular distribution in the mouse cerebellum. In the cerebellum, alpha1B immunoreactivity is found mainly on the cell bodies of all Purkinje cells. In addition, the immunoreactivity was detected on a subset of Purkinje cell dendrites, clustered to form a parasagittal array of bands. In the anterior lobe vermis, immunoreactive Purkinje cell dendrites form narrow stripes separated by broad bands of unstained dendrites. Moving caudally through the vermis, these stripes become thicker as a larger fraction of the Purkinje cell dendrites become immunoreactive. This localization study of the alpha1B pore-forming subunits in mouse cerebellum may guide future investigations of the role of calcium channels in neurological pathways.

Animals↗