A new immunohistochemical method to evaluate the development of vestibular compensation after unilateral labyrinthectomy in rats.

These findings suggested that the number of MK801-induced Fos-LIR neurons in contra-MVe was decreased in concordance with the restoration of ipsi-MVe-activity during the late process of VC after UL and that thioperamide accelerated the late, but not the initial process of VC.


Native rotational knee kinematics are lost in bicruciate-retaining total knee arthroplasty when the tibial component is replaced.

The rotational kinematics of the native knee are not always preserved after BCR TKA. Native rotational kinematics are preserved after meniscectomy and femoral replacement, but are lost after tibial replacement in BCR TKA. Surgeons should pay close attention to maintain the anteroposterior stabilizing function of the ACL in BCR TKA, rather than to restore the native rotational kinematics.

Knee Surgery, Sports Traumatology, Arthroscopy


Location and direction of the nutrient artery to the first metatarsal at risk in osteotomy for hallux valgus.

Location and direction of the first metatarsal nutrient artery was established.


Pulsation and Collimation During Fluoroscopy to Decrease Radiation: A Cadaver Study.


A new concept of transforaminal ventral facetectomy including simultaneous decompression of foraminal and lateral recess stenosis: Technical considerations in a fresh cadaver model and a literature review.

Percutaneous endoscopic surgery for the lumbar spine, which was established in the last decade, requires only an 8-mm skin incision and causes minimal damage to the paravertebral muscles; thus, it is considered to be a minimally invasive technique for spinal surgery. It has been used to perform percutaneous endoscopic discectomy via two main approaches: the TF approach is a posterolateral one through the intervertebral foramen and can be done under local anesthesia; the IL approach is a more traditional one through the interlaminar space and is difficult to perform under local anesthesia. Recently, these techniques have been applied for lumbar spinal stenosis (LSS), the TF method for foraminal stenosis under local anesthesia, and the IL method for central and lateral recess stenosis under general anesthesia. In this study, using a fresh human cadaver model, we performed simultaneous decompression of the lateral recess and foraminal stenosis at L4-5 using the TF approach. Computed tomography confirmed enlargement of the lateral recess and intervertebral foramen. This technique, which can be performed under local anesthesia, should benefit elderly patients with LSS and poor general condition due to multiple comorbidities. Finally, we introduce the concept of percutaneous transforaminal ventral facetectomy using a spinal percutaneous endoscope. J. Med. Invest. 64: 1-6, February, 2017.

The Journal of Medical Investigation : JMI


Spinal nerve defects in mouse embryos prenatally exposed to valproic acid

To examine in detail spinal nerve defects induced by prenatal exposure to valproic acid in mice, pregnant ICR mice were subcutaneously injected with a single dose of 400 mg/kg valproic acid on gestational day 6, 7, 8, or 9, and their embryos were observed on gestational day 10. The whole-mount immunostaining using an anti-neurofilament antibody allowed us to identify spinal nerve defects, such as a loss of bundle, anastomosis among bundles arising from adjacent segment, and a disrupted segmental pattern of the dorsal root ganglia, in valproic acid-exposed embryos. The prevalence of spinal nerve defects was the highest in the embryos exposed to valproic acid on gestational day 8 among the experimental groups. Then, effects of the administration dose of valproic acid on the prevalence of spinal nerve defects were examined on gestational day 10 and found to be dose-dependently increased. It was noteworthy that all embryos exposed to 600 mg/kg of valproic acid on gestational day 8 suffered spinal nerve defects. Folic acid (3 mg/kg/day) supplementation during gestational day 6-10 suppressed the prevalence of valproic acid-induced neural tube defects, which are common malformations in offspring prenatally exposed to valproic acid, but not that of spinal nerve defects. Thus, the spinal nerve defects due to prenatal valproic acid exposure might be induced by mechanisms different from those of neural tube defects. Because spinal nerve defects were predicted to be caused by the disrupted segmental arrangement of the somites and/or that of neural crest cells, which was the origin of the dorsal root ganglia and/or abnormal polarity of the somite, this mouse model with spinal nerve defects at high incidence would be useful to examine the effects of valproic acid on the somitogenesis and morphogenesis of somite-associated structures.


Radiation Exposure to the Surgeon and Patient During a Fluoroscopic Procedure: How High is the Exposure Dose? A Cadaveric Study.

Seven fresh cadavers were irradiated for 1, 3, and 5 min with C-arm fluoroscopy. The X-ray source was positioned under the table, over the table, and laterally. Radiation exposure doses were measured at different simulated areas such as the center area, and the surgeon's hand or thyroid gland.


The Pullout Strength of Pedicle Screws following Re-direction after Lateral Wall Breach or End-plate Breach.


Influence of Medial collateral ligament release for internal rotation of tibia in posterior stabilized total knee arthroplasty: a cadaveric study

Previous studies suggested that changes in kinematics in total knee arthroplasty (TKA) affected satisfaction level. The aim of this cadaveric study was to evaluate the effect of medial collateral ligament (MCL) release by multiple needle puncture on knee rotational kinematics in posterior-stabilized TKA. Six fresh, frozen cadaveric knees were included in this study. All TKA procedures were performed with an image-free navigation system using a 10-mm polyethylene insert. Tibial internal rotation was assessed to evaluate intraoperative knee kinematics. Multiple needle puncturing was performed 5, 10, and 15 times for the hard portion of the MCL at 90° knee flexion. Kinematic analysis was performed after every 5 punctures. After performing 15 punctures, a 14-mm polyethylene insert was inserted, and kinematic analysis was performed. The tibial internal rotation angle at maximum knee flexion without multiple needle puncturing was significantly larger (9.42°) than that after 15 punctures (3°). Negative correlation (Pearson r = -0.715, P < .001) between tibial internal rotation angle at maximum knee flexion and frequency of puncture was observed. The tibial internal rotation angle with a 14-mm insert was significantly larger (7.25°) compared with the angle after 15 punctures. Tibial internal rotation during knee flexion was reduced by extensive MCL release using multiple needle puncturing and was recovered by increasing of medial tightness. From the point of view of knee kinematics, medial tightness should be allowed to maintain the internal rotation angle of the tibia during knee flexion which might lead to patient satisfaction.


Recurrence of type I Modic inflammatory changes in the lumbar spine: effectiveness of intradiscal therapy.

Here we report a case of recurrence of Modic type I inflammatory changes in the lumbar spine. A 49-year-old man was referred to our department with a history of chronic low back pain of at least 20 years. At the first consultation, he complained of low back pain only and had no other symptoms such as leg pain, numbness, or weakness. Although his pain was typically mild, he experienced one or two episodes of severe and incapacitating low back pain a year. After two intradiscal steroid injections, his pain disappeared immediately and completely. After 6 months of conservative treatment, Modic type I change switched to Modic type II change. However, 12 months after the first treatment, he once again experienced severe low back pain. Follow-up magnetic resonance imaging (MRI) indicated recurrence of Modic type I change that was stronger than the first occurrence. Two intradiscal injections relieved the pain. Six months after the second episode, follow-up MRI showed another switch of Modic type I change to Modic type II change. Switches of Modic change have been controversial, with mixed findings on pain, natural history, and degenerative changes. The present case reinforces the notion that Modic type I change corresponds to reversible local inflammation.


Postnatal Change in Sulcal Length Asymmetry in Cerebrum of Cynomolgus Monkeys (Macaca fascicularis)

The purpose of this study was to determine the timing of the onset of adult-type sulcal length asymmetry during postnatal development of the male cynomolgus monkey cerebrum. The monkey brain has already reached adult size by 3 months of age, although the body weight only represents 1/8 of the adult body weight by that time. The fronto-occipital length and the cerebral width also reached adult levels by that postnatal age with no left/right bias. Consistently, lengths of the major primary sulci reached adult levels by 3 months of age, and then decreased slightly in sexually mature monkeys (4-6.5 years of age). Asymmetry quotient analysis showed that sulcal length asymmetry patterns gradually changed during postnatal development. The male adult pattern of sulcal length asymmetry was acquired after 24 months of age. In particular, age-dependent rightward lateralization of the arcuate sulcal length was revealed during cerebral maturation by three-way ANOVA. The results suggest that the regional difference in cerebral maturation from adolescence to young adulthood modifies the sulcal morphology with characteristic asymmetric patterns in male cynomolgus monkeys.


Fetal Gyrification in Cynomogus Monkeys: A Concept of Developmental Stages of Gyrificaiton

Our article summarizes a series of studies about fetal gyrification and its relation to cerebral growth in cynomolgus monkeys. Based on the cerebral growth (i.e., brain weight, cerebral volume, and frontooccipital length of the cerebral hemisphere) and the developmental pattern of gyrification in each sulcus of cynomolgus monkeys, we divided the gyrification process into four stages: Stage 1. Demarcation of cerebral lobes and limbic gyri; Stage 2. Demarcation of neocortical gyri; Stage 3. Emergence of secondary and tertiary sulci; and Stage 4. Growth of sulcal length and depth. Each stage of those gyrification processes was influenced by different developmental events, such as the emergence of corticocortical long-associative fiber tracts, cortical maturations, and subcortical white-matter development. This is the first report to systematically propose gyrification processes closely related to the order of phyologenetical development of the cerebral cortex in primates.


Neuroanatomic and magnetic resonance imaging references for normal development of cerebral sulci of laboratory primate, cynomolgus monkeys (Macaca fascicularis)

Cynomolgus monkey (Macaca fascicularis) is a popular laboratory primate belonging to Old World monkeys, which are the group most closely related to humans except for the apes. This paper summarizes a series of our studies regarding the development of cerebral sulci and gyri in this primate, and the stated possibility of evaluation of the sulcal development for assessing the developmental toxicity testing. The cerebrum of cynomolgus monkeys experienced a regular sequence of emergence of sulci and gyri on gross observation while such timetables corresponded to those obtained by magnetic resonance imaging (MRI) with a lag time of 10-30 days. When the timetables for the emergence of anatomically identical primary sulci and gyri were compared between cynomolgus monkeys and humans, their chronological sequences were comparable, while some sulci and gyri located on the phylogenetically newer cortical region in humans emerged earlier in monkeys. The present paper further indicates brief procedures for evaluating cerebral abnormalities and/or maturity using brain specimens without MRI measurements. The primary sulcal lengths measured by the 'cotton thread' method were a brief index of the degree of regional gyrification. As the development of a calcarine sulcus was closely correlated with morphological maturation of the lateral ventricle, which changed drastically during embryonic days (EDs) 90-100, the cerebral maturity on ED 100 could be evaluated by the infolding of that sulcus. Thus, the present paper provides gross anatomical and MRI references and brief procedures for investigating the normality of the development of cerebral sulci and gyri of laboratory primates, cynomolgus monkeys.


Laminar cytoarchitecture of cerebral cortex in CAv2.1 mutant, rolling mouse Nagoya


Sexual dimorphism of sulcal length asymmetry in the cerebrum of adult cynomolgus monkeys (Macaca fascicularis)

The present study aimed to quantitatively clarify the gross anatomical asymmetry and sexual dimorphism of the cerebral hemispheres of cynomolgus monkeys. While the fronto-occipital length of the right and left cerebral hemispheres was not different between sexes, a statistically significant rightward asymmetry was detected in the cerebral width at the perisylvian region in females, but not in males (narrower width of the left side in the females). An asymmetry quotient of the sulcal lengths revealed a rightward asymmetry in the inferior occipital sulcus and a leftward asymmetry in the central and intraparietal sulci in both sexes. However, the laterality of the lengths of other sulci was different for males and females. The arcuate sulcus was directed rightward in males but there was no rightward bias in females. Interestingly, the principle sulcus and lateral fissure were left-lateralized in the males, but right-lateralized in the females. The results suggest that lateralization patterns are regionally and sexually different in the cerebrum of cynomolgus monkeys. The present results provide a reference for quantitatively evaluating the normality of the cerebral cortical morphology in cynomolgus monkeys.


Development of Corticocortical Long Associative Fibers in Cynomolgus Monkey Fetal Cerebrum Analyzed Using DTI: its Relation to Sulcal Formation


Correlation between formation of the calcarine sulcus and morphological maturation of the lateral ventricle in cynomolgus monkey fetuses

In the present study developmental changes in the cerebral sulci and volumes of subcortical and archicortical structures of the cerebrum in cynomolgus monkey fetuses were examined with T(1)-weighted magnetic resonance (MR) images in 3D. On the embryonic day (ED) 90, the lateral ventricle had still an immature vesicular shape in the occipital region of the cerebrum, and it dramatically closed its lumen by ED 100. In that period the calcarine sulcus progressively infolded from the medial surface of the cerebral hemisphere narrowing the lumen of the lateral ventricle in the occipital region. Volume of the lateral ventricle decreased in the period ED 90-100, increasing afterwards in spite of increasing volumes of subcortical and archicortical structures such as the caudate nucleus, putamen, globus pallidus, amygdala and hippocampal formation. During the same time, the volume of the germinal matrix around lateral ventricles decreased to disappear completely by ED 120. These results suggest that the morphological maturation of lateral ventricle is linked to the development of calcarine sulcus in cynomolgus monkey fetuses. The degree of infolding of calcarine sulcus on ED 100 would be useful as a gross anatomical landmark for evaluating the cerebral maturation in cynomolgus monkey fetuses.


Zebrin II Expressing Purkinje Cell Phenotype-Related and Unrelated Cerebellar Abnormalities in Cav2.1 Mutant, Rolling Mouse Nagoya

Rolling mouse Nagoya is an ataxic mutant mouse that carries a mutation in a gene encoding for the alpha 1A subunit of the voltage-gated P/Q-type Ca2+ channel (Cav2.1). This report summarizes our studies and others concerning cerebellar abnormalities in rolling mice based on chemical neuroanatomy. While there are no obvious cerebellar deformations in this mutant mouse, the altered functions of Purkinje cells can be revealed as a reduced expression of type 1 ryanodine receptor (RyR1) in all Purkinje cells uniformly throughout the cerebellum, and as an ectopic expression of tyrosine hydroxylase (TH) in the Purkinje cell subsets with the zebrin II-immunopositive phenotype. As the mutated Cav2.1 channel is expressed at uniform levels in all Purkinje cells, its copresence with RyR1 staining suggests that a Cav2.1 channel dysfunction links with the expression of RyR1 in Purkinje cells of rolling mice. However, an ectopic expression of TH in the Purkinje cells is topologically related to the projection of corticotrophin-releasing factor-immunopositive climbing fibers rather than expression of the mutated Cav2.1 channel. On the other hand, increased levels of serotonin (5-HT) in 5-HTergic fibers were revealed immunohistochemically in Purkinje cells of the vermis of rolling cerebellum. Thus, to determine whether or not cerebellar abnormalities are related to Purkinje cell populations revealed by zebrin II expression is essential for enhancing our understanding of the pathogenesis of hereditary cerebellar ataxic mutants such as rolling mice.


Alternating array of tyrosine hydroxylase and heat shock protein 25 immunopositive purkinje cell stripes in zebrin II-defined transverse zone of the cerebellum of rolling mouse Nagoya

The present study examined the spatial organization of tyrosine hydroxylase (TH) immunopositive Purkinje cells in the cerebellum of rolling mouse Nagoya with reference to the distribution pattern of the cerebellar compartmentation antigen, heat shock protein 25 (HSP25). Whole-mount immunostaining revealed a striking pattern of parasagittal stripes of TH staining in the rolling mouse cerebellum but not in the control cerebellum. Although the TH stripes resembled the zebrin II stripes in the rolling cerebellum, these two distributions did not completely overlap. The TH stripes were present in the lobules VI and VII (central zone), the lobule X (nodular zone), and the paraflocculus, where zebrin II immunostaining was uniformly expressed. Double immunostaining revealed that TH stripes were aligned in an alternative fashion with HSP25 stripes within the caudal half of lobule VIb, lobules IXb and X, and paraflocculus. Some, but not all, TH stripes shared boundaries with HSP25 stripes. These results revealed an alternating array of TH immunopositive Purkinje cell subsets with HSP25 immunopositive Purkinje cells in the zebrin II-defined transverse zone of the rolling mouse cerebellum. The constitutive expression of HSP25 may prevent the ectopic expression of TH in zebrin II immunopositive Purkinje cell subsets.


Ontogenetic pattern of gyrification in fetuses of cynomolgus monkeys

The ontogenetic pattern of gyrification and its relationship with cerebral cortical volume were examined in cynomolgus monkey fetuses. T(1)-weighted coronal magnetic resonance (MR) images at 7 T were acquired from the fixed cerebra of three male fetuses, each at embryonic days (EDs) 70 to 150, and the gyrification index (GI) of each slice was estimated. The mean GI was low (1.1-1.2) during EDs 70 to 90, and then increased dramatically on ED 100. The developmental profiles of the rostrocaudal GI distribution revealed that cortical convolution was more frequent in the parietooccipital region than in other regions during EDs 100 to 150, forming an adult-like pattern by ED 150. The mean GI was closely correlated with the volume of cortical gray matter (r=0.9877), and also with the volume of white matter/intermediate zone (r=0.8961). These findings suggest that cortical convolution is correlated with either the maturation of cortical gray matter or the development of white matter bundles. The characteristic GI distribution pattern of catarrhines was formed by ED 150 in correlation with the progressive sulcal infolding in the parietooccipital region of the cerebrum.


Embryonic exposure to ethanol disturbs regulation of mitotic spindle orientation via GABA(A) receptors in neural progenitors in ventricular zone of developing neocortex.

Neural progenitors in the ventricular zone of the developing neocortex divide oriented either parallel or perpendicular to the ventricular surface based on their mitotic spindle orientation. It has been shown that the cleavage plane orientation is developmentally regulated and plays a crucial role in cell fate determination of neural progenitors or the maintenance of the proliferative ventricular zone during neocortical development. We tested if fetal exposure to ethanol, the most widely used psychoactive agent and a potent teratogen that may cause malformation in the central nervous system, alters mitotic cleavage orientation of the neural progenitors at the apical surface of the ventricular zone in the developing neocortex. Fetal exposure to ethanol on E10.5 and 11.5 increased the occurrence frequency of a horizontal cleavage plane that is parallel to the ventricular surface on E 12.5. Administration of picrotoxin, a GABA(A) receptor antagonist, prior to ethanol administration canceled the effect of ethanol with the frequency of horizontal division similar to the control level, although picrotoxin itself did not show any effect on cleavage plane orientation. Phenobarbital, a GABA(A) receptor agonist, induced horizontal cleavage to an extent similar to that induced by ethanol administration. (+)MK801, an antagonist of NMDA receptor that is another major target of ethanol in neural cells, did not affect the cleavage plane of dividing progenitors. These results suggest that fetal ethanol exposure induced alterations in the cleavage plane orientation of neural progenitors in the ventricular zone of the neocortex via the enhancement of the function of GABA(A) receptors.


Alteration in anxiety-related behaviors and reduction of serotonergic neurons in raphe nuclei in adult rats prenatally exposed to ethanol

It is known that the developing serotonergic system is one of the targets of ethanol teratogenicity. Because serotonin has multiple functions in both mature and immature brains, disturbance of the serotonergic system by ethanol exposure in utero can be cause of a wide range of psychiatric problems in adulthood. In the present study, we observed serotonergic neurons in the midbrain raphe nuclei and anxiety-like behaviors which would be affected by an altered serotonergic system in adult rats prenatally exposed to ethanol. Pregnant rats were fed a liquid diet containing 2.5-5.0% (w/v) ethanol on gestational days 10-21. Their offspring were examined at 60-70 days of age. A significant decrease in the number of serotonergic cells in the midbrain raphe nuclei was shown in prenatally ethanol-exposed offspring. In an open field test, they spent more time in a central area compared to controls. Also in an elevated plus maze test, prenatally ethanol-exposed offspring spent more time on the open arms than controls. These behavioral results suggested that prenatally ethanol-exposed rats were less sensitive to anxiety. However, 44% of prenatally ethanol-exposed offspring exhibited freezing behavior on the open arms of the elevated plus maze, causing strong anxiety, compared with 0% in intact control and 12.5% in isocaloric sucrose-fed control groups. These findings suggest that prenatal ethanol exposure decreases both susceptibility and resistance of anxiety. Insufficient serotonergic actions caused by reduced serotonergic neurons in the raphe nuclei might contribute to the alterations in anxiety-related behaviors observed in our prenatally ethanol-exposed rats.


Torpedoes of Purkinje cell axons in deep cerebellar nuclei of an ataxic mutant, rolling mouse Nagoya


Striking pattern of Purkinje cell loss in cerebellum of an ataxic mutant mouse, tottering

Tottering mouse is an ataxic mutant that carries a mutation in a gene encoding for the apha1A subunit of P/Q-type Ca2+ channel (Cav2.1). This study revisited to examine whether a Purkinje cell loss occurred in the cerebellum of tottering mice. In tottering mice, Calbindin D-28k negative gaps were apparent in the vermis but not in the hemisphere. Calbindin D-28k immunofluorescence with DAPI staining demonstrated the absence of Purkinje cells in the Calbindin D-28k negative gaps. The Purkinje cell loss seemed to be observed prominently in the zebrin II negative compartments of the anterior vermis, but in the zebrin II positive compartments of the posterior vermis. Quite consistent with the histopathological observations, quantitation of the density of Calbindin D-28k and zebrin II immunopositive Purkinje cells in the tottering cerebellum revealed that the Purkinje cells were selectively lost in the zebrin II immunonegative compartments of the lobules I and II but in the zebrin II immunopositive compartments in the lobule IX. Those results predict that the susceptibility to the Cav2.1 gene defect is different among Purkinje cell phenotypes of the tottering cerebellum rather than the expression pattern of mutated Cav2.1 channels. This may result in the reproducible parasagittal pattern of Purkinje cell loss.


Reactive nitrogen species-dependent DNA damage in EBV-associated nasopharyngeal carcinoma: The relation to STAT3 activation and EGFR expression

Nasopharyngeal carcinoma (NPC) is strongly associated with Epstein-Barr virus (EBV) infection. Recently, reactive nitrogen and oxygen species are considered to participate in inflammation-related carcinogenesis through DNA damage. In our study, we obtained biopsy and surgical specimens of nasopharyngeal tissues from NPC patients in southern China, and performed double immunofluorescent staining to examine the formation of 8-nitroguanine, a nitrative DNA lesion and 8-oxo-7,8-dihydro-2'-deoxyguanosine, an oxidative DNA lesion, in these specimens. Strong DNA lesions were observed in cancer cells and inflammatory cells in stroma of NPC patients. Intensive immunoreactivity of iNOS was detected in the cytoplasm of 8-nitroguanine-positive cancer cells. DNA lesions and iNOS expression were also observed in epithelial cells of EBV-positive patients with chronic nasopharyngitis, although their intensities were significantly weaker than those in NPC patients. In EBV-negative subjects, no or little DNA lesions and iNOS expression were observed. EGFR and phosphorylated STAT3 were strongly expressed in cancer cells of NPC patients, but NF-kappaB was not expressed, suggesting that STAT3-dependent mechanism is important for NPC carcinogenesis. IL-6 was expressed mainly in inflammatory cells of nasopharyngeal tissues of EBV-infected patients. EBV-encoded RNAs (EBERs) and latent membrane protein 1 (LMP1) were detected in cancer cells from all EBV-infected patients. In vitro cell system, nuclear accumulation of EGFR was observed in LMP1-expressing cells, and IL-6 induced phosphorylated STAT3 and iNOS. These data suggest that nuclear accumulation of EGFR and STAT3 activation by IL-6 play the key role in iNOS expression and resultant DNA damage, leading to EBV-mediated NPC.


Spatial distribution of corticotropin-releasing factor immunopositive climbing fibers in the mouse cerebellum: analysis by whole mount immunohistochemistry.

This study examined the spatial organization of corticotropin-releasing factor (CRF) immunopositive climbing fibers in the mouse cerebellum by whole mount immunohistochemistry. A striking pattern of parasagittal stripes of CRF staining was revealed. Cryosections of whole mount CRF stained cerebellum showed that anti-CRF immunostaining is restricted to climbing fibers in the molecular layer and does not penetrate deeper into the granular layer. The array of CRF stripes was reminiscent of zebrin II immunopositive Purkinje cell stripes in the anterior vermis and the hemispherical lobules. However, a direct comparison of the two distributions showed that the CRF-defined parasagittal stripes and transverse zones in the posterior vermis are different from those defined by the expression of zebrin II: in particular, CRF immunostaining revealed a transverse boundary between lobules VIb and VII and the presence of four CRF-immunopositive climbing fiber stripes in lobule VIII. Furthermore, an array of CRF stripes was seen in lobule X, the flocculus and the paraflocculus, despite uniform zebrin II expression in these areas. In these cases some, but not all, CRF-immunopositive stripes shared boundaries with Purkinje cell stripes that were visualized by the expression of heat shock protein 25. The results reveal a reproducible pattern of CRF-immunopositive climbing fiber innervation in the mouse cerebellum that bears a complex relationship to the stripes delineated by Purkinje cell compartmentation antigens.


Development of cerebral sulci and gyri in fetuses of cynomolgus monkeys (Macaca fascicularis). II. Gross observation of the medial surface.

This study aimed to clarify chronological sequences of the appearances of sulci and gyri on the medial cerebral surface and its relation to the regional development of the cerebrum in cynomolgus monkeys. The lengths of cingulate and calcarine sulci were measured, and the ratios of these lengths to fronto-occipital length were estimated as indices of the size of the "frontoparietal" and "occipital" regions, respectively. The relative length of cingulate sulcus showed a biphasic increase: a slow phase from EDs 100 to 110, and a rapid phase from EDs 110 to 130. The gyri in the "frontoparietal region" were convoluted in the limbic cortex during the initial slow phase and in the neocortical region during the rapid phase. The relative length of calcarine sulcus lineally increased between EDs 90 and 130, and the gyri in the "occipital region" generated in a dorso-ventral manner: the gyrus convolutions occurred first in the "phylogenetically older" striate and dorsal extrastriate cortices, and then in the "phylogenetically newer" ventral extrastriate cortex. The results suggest that the chronological order of appearance of sulci and gyri is closely associated with the order of phylogenetical development of the cerebral cortex. The present study provides a standard reference for the development of cerebral sulci and gyri of cynomolgus monkeys together with our previous study (Fukunishi et al. Anat Embryol 211:757-764, 2006).


Differential alterations in expressions of ryanodine receptor subtypes in cerebellar cortical neurons of an ataxic mutant, rolling mouse Nagoya

This study aimed to clarify changes in the spatial expressions of types 1, 2 and 3 ryanodine receptors (RyR1, RyR2 and RyR3) in the cerebellum of a Ca(2+) channel alpha(1A) subunit mutant, rolling mouse Nagoya. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) revealed that the mRNA signal levels of RyR1 and RyR3 were altered in the rolling cerebellum, which exhibited lower densities of RyR1 bands and higher densities of RyR3 bands than in the control cerebellum. Quite consistent with the RT-PCR results, the staining intensity of RyR1 and RyR3 was altered in the rolling cerebellum. RyR1 immunostaining appeared in somata and the proximal dendrites of Purkinje cells, and the staining intensity of both subcellular regions was equally lower in all cerebellar lobules of rolling mice than in those of controls. Although RyR3 immunostaining appeared in the dendrites of granule cells, more intense RyR3 staining in rolling mice than in controls was uniformly observed throughout all cerebellar lobules. The present study further examined co-localizations of ryanodine receptor subtypes and voltage-gated Ca(2+) channel alpha(1) subunits in the rolling cerebellum. Somatodendritic RyR1 immunostaining in Purkinje cells overlapped with either a mutated Ca(2+) channel alpha(1A) subunit (P/Q-type), or a Ca(2+) channel alpha(1C) subunit (L-type; dihydropyridine receptor) immunostaining. Immunostaining of these alpha(1) subunits also emerged in granule cells. Those results suggest non-region-related alterations in RyR1 and RyR3 expressions in the rolling mouse cerebellum. Such expressional changes in ryanodine receptor subtypes may be involved in Ca(2+) channel alpha(1A) subunit gene mutation, and may alter regulation of intracellular Ca(2+) concentrations in cerebellar cortical neurons.


Spatial learning and expression of neural cell adhesion molecule L1 in rats X-irradiated prenatally

The present study was designed to present evidence to clarify the relationships between learning ability, neuronal cell adhesion molecule L1 expression and hippocampal structural changes in the rat model received X-irradiation at an embryonic stage (E15). Water maze task indicated that all of the irradiated rats failed to learn the task in the whole training procedure. Their latency to the platform and swimming distance were significant differences from those sham-treated controls. Histological studies showed that the hippocampal ectopias induced by X-rays in the CA1 were involved in the spatial learning impairment, in which they hampered normal processes in learning development and transmission of information. Number, size and positions of the ectopias in the dorsal parts of the hippocampus were confirmed to be related to degrees of spatial learning impairment. On the other hand, L1 expression in the hippocampus was examined with Western blot analysis. The results indicated a lower content of L1 in the irradiated rats. A decrease in L1 might be one of reasons to cause disorganization of the septohippocampal pathways. These findings suggest some mechanisms of spatial learning impairment can be attributed to the formation of the hippocampal ectopias and redaction of L1 following prenatal exposure to X-irradiation.

The Journal of Medical Investigation : JMI


The effect of prenatal X-irradiation on the developing cerebral cortex of rats

The developing central nervous system is known to be highly vulnerable to X-irradiation. Although glial cells are involved in various brain functions, knowledge on the effects of X-irradiation on glial cells is limited. Therefore, the purpose of the present study was to evaluate the effects of prenatal X-irradiation on glial cells. Pregnant Wistar rats were exposed to X-irradiation at a dose of 1.0 Gy on day 15 of gestation. Their offspring were examined at 7 weeks of age. The forebrain weight of X-irradiated rats was significantly lower than that of the age-matched controls. Histological quantification with stereology of the somatosensory cortex (SC) revealed no significant difference in the numerical density of glial cells between the X-irradiated and control rats. However, the glial cells in the X-irradiated animals had significantly larger nuclear size. We had previously reported that a similar X-irradiation paradigm resulted in no significant change in the numerical density of neurons in the SC. According to the results of the present study, there were no significant differences in the glial cell-to-neuron ratios between the X-irradiated and control animals. Taken together, it is speculated that prenatal X-irradiation has an equal effects on the numerical densities of glial cells and neurons.

International Journal of Developmental Neuroscience


Increased serotonergic innervation of lumbosacral motoneurons of rolling mouse Nagoya in correlation with abnormal hindlimb extension

Rolling Mouse Nagoya (RMN) carries a mutation in a gene encoding for alpha(1A) subunit of P/Q-type Ca(2+) channel (Ca(v)2.1). In addition to ataxia, this mutant mouse exhibits abnormal hindlimb extension, which is characterized by a sustained excessive tone of hindlimb extensor muscles. This study aimed to clarify whether serotonergic (5-HTergic) innervation of the spinal motoneurons was altered in RMN in relation to the abnormal hindlimb extension. The density of 5-HT immunoreactive fibres in the ventral horn of lumbar and sacral regions of spinal cord was significantly greater in RMN than in controls. Retrograde wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) labelling combined with 5-HT immunostaining revealed that the number of 5-HT immunoreactive terminals adjoining femoris quadriceps motoneurons was about 2.5-fold greater in RMN than in controls. Furthermore, 5-HT immunostaining in the lumbar cord ventral horn was examined in three other Ca(v)2.1 mutant mice (tottering, leaner and pogo) as to whether or not they showed the abnormal hindlimb extension. Among these mutants, the increased density of 5-HT immunoreactive fibres was observed in correlation with the presence of the abnormal hindlimb extension. The results suggest an increased 5-HTergic innervation of the lumbosacral motoneurons in correlation with the abnormal hindlimb extension in RMN and other Ca(v)2.1 mutant mice. As 5-HT is known to induce the sustained membrane depolarizations without continuous excitatory synaptic inputs (plateau potentials) in spinal motoneurons, the increased 5-HTergic innervation may cause the sustained excitation of hindlimb extensor motoneurons, resulting in the abnormal hindlimb extension.


Corticotropin-Releasing Factor Immunoreactivity Increases in the Cerebellar Climbing Fibers in the Novel Ataxic Mutant Mouse,pogo

The ataxic pogo mouse (pogo/pogo) is a novel neurological mutant, which was derived as an inbred strain (KJR/MsKist) from a Korean wild mouse. The pathological manifestations include a difficulty in maintaining a normal posture, the failure of inter-limb coordination and an inability to walk straight. In this study, we examined the distribution of corticotropin-releasing factor (CRF) immunoreactive cerebellar climbing fibres and their projections to tyrosine hydroxylase (TH) immunoreactive Purkinje cells in the cerebellum of the pogo mutant mouse using immunohistochemistry. In the pogo/pogo mouse, a subset of climbing fibres was stained more intensely for CRF than in the control. Moreover, ataxic pogo mouse, neurons of the inferior olivary nucleus projecting climbing fibres were also more intensely stained for CRF than in the control. In the pogo/pogo mouse, TH immunoreactivity was located in the Purkinje cells, whereas no TH expression was found in the control. Double immunostaining for CRF and TH in the pogo/pogo cerebellum revealed that the distribution of TH-immunoreactive Purkinje cells corresponded to terminal fields of CRF-immunoreactive climbing fibres but not to the CRF-immunoreactive mossy fibres. Therefore, we suggest that an increase of CRF level may alter the function of targeted Purkinje cells and that it is related to the ataxic phenotype in the pogo mutant mouse.


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.


オーストラリア クイーンズランド大学における解剖学教育

オーストラリア クイーンズランド大学における解剖学教育


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


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.


Comments from the Behavioral Teratology Committee of the Japanese Teratology Society on OECD Gudeline for the Testing of Chemicals, Proposal for a New Guideline 426, Developmental Neurotoxicity Study, Draft Document (September 2003)

In September 2003, a new revision of the draft guideline (Organization for Economic Co-operation and Development [OECD] Guideline for the Testing of Chemicals, Proposal for a New Guideline 426, Developmental Neurotoxicity Study) was distributed. The draft guideline consists of 51 paragraphs and an appendix. The National Coordinators were requested to arrange national expert reviews of the guideline proposal in their member countries. The member of the Behavioral Teratology (BT) Committee of the Japanese Teratology Society (JTS) reviewed, discussed and commented on the draft Test Guideline proposal. The BT Committee of the JTS also commented that the International Collaborative Study to validate this protocol should be definitely performed. These comments were sent to the OECD Secretariat. The BT Committee of the JTS expects that the comments are useful for further discussion.


Postnatal Changes of p39nck5ai Expression in the Developing Rat Cerebellum


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


Cyclin-dependent kinase 5 (Cdk5) is primarily associated with brain development but it is also implicated in lens and muscle differentiation. We found that Cdk5 is also expressed in mouse ovary, and explored the possibility that it plays a role in that tissue. We show by Western blotting and immunohistochemistry that the known Cdk5 activator, p35, is also present in the mouse ovary. Cdk5 and p35 were detected in oocytes at all stages of the follicle. While Cdk5 was present in the cytoplasm and nucleus of the oocyte, p35 was observed only in the cytoplasm. Both proteins were detected in the cytoplasm of luteinized cells in the corpus luteum. Immunoprecipitation and histone H1 kinase assays revealed that they form an ovarian complex with considerable kinase activity. Phosphorylation assays showed that several ovarian proteins are substrates for Cdk5/p35 in vitro. Together our findings suggest that p35-associated Cdk5 activity plays an important role in the ovary, where it may regulate cell differentiation and apoptosis as it does in the brain.


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.


Prenatal exposure to alchol 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.


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.

Brain Research. Molecular Brain Research


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.

The Journal of Medical Investigation : JMI


A genetic mouse model carrying the nonfunctional xeroderma pigmentosum group G gene

A genetic mouse model with a disrupted XPG allele was generated by insertion of neo cassette sequences into exon 3 of the XPG gene by using embryonic stem (ES) cell techniques. The xpg-deficient mice showed distinct developmental characteristics. Their body was marked smaller than that in wild-type littermates since the postnatal day 6, and this postnatal growth failure became more severe with developmental proceeding. Their life span was very short, all of the mutants died by postnatal day 23 after showing great weakness and emaciation. In addition, the mutant homozygous mice also showed some progressive neurological signs, like the lower level of activity and a progressive ataxia. Further examination indicated there was developmental retardation of the brain in the mutant mice. Their brain weight, and thickness of cerebral cortex and cerebellar cortex were significant different from the controls. These characteristics, like small size brain, brain developmental retardation and progressive neurological dysfunctions in the homozygotes were similar to the typical clinical phenotype of the XPG patients and Cockayne syndrome, we believe that the xpgdeficient mice will be an animal model for studying the function of the XP-G protein in nucleotide-excision repair and mechanisms related to the clinic symptoms of XP-G and Cockayne syndrome in humans.


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.


ローリングマウス小脳におけるカルレチニン陽性unipolar brush cellとCRF陽性苔状線


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.


Differential expression of two distinct vesicular glutamate transporters in the rat retina.

Expression and cellular localization of vesicular glutamate transporters (BNPI and DNPI) were studied in the rat retina. RT-PCR showed expression of both transporter mRNAs. hybridization demonstrated BNPI mRNA signals in the inner segments of photoreceptors and the inner nuclear layer, whereas DNPI mRNA signals were confined to the ganglion cell layer. Punctate BNPI immunoreactivity was localized in the inner and outer plexiform layers, and weak DNPI immunoreactivity was detectable only in some cells and fibers of the ganglion cell layer. The present study suggests that BNPI exists in photoreceptors and bipolar cells, while DNPI is present in ganglion cells, as specific systems in distinct glutamatergic neurons of the retina.


Immunohistochemistry of Voltage-Gated Calcium Channel α1B 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.


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.


p35, Cdk5 Activator, is Expressed in Mouse Epididymis


Cdk5 and Its Activator, p35, are Expressed in Mouse Ovary


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

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.


前額部の皮下腫瘍摘出術における頭髪生え際切除法 --- 局所解剖所見からの工夫

前額部の皮下腫瘍摘出術における頭髪生え際切除法 --- 局所解剖所見からの工夫

Journal of the Japan Society of Cranio-Maxillo-Facial Surgery

日本頭蓋顎顔面外科学会誌


A quantitative study of the optic nerve in diabetic mutant, Otsuka Long-Evans Tokushima Fatty (OLETF) rats.


An increased expression of Ca2+ channel alpha1A subunit immunoreactivity in deep cerebellar neurons of rolling mouse Nagoya.

Rolling mouse Nagoya (RMN) is an ataxic mutant and carries a mutation in the gene coding for the alpha(1A) subunit of the P/Q-type Ca(2+) channel. We examined the immunohistochemical expression of the alpha(1A) subunit in deep cerebellar nuclei of RMN. The antibody used recognized residues 865-883 of the mouse alpha(1A) subunit not overlapping the altered sequences in RMN. In RMN, many neurons exhibited definite alpha(1A) subunit-staining in the medial nucleus, interposed nucleus, and lateral nucleus of deep cerebellar nuclei. The number of positive neurons in these nuclei was significantly higher in RMN than in controls. Increased expression of the alpha(1A) subunit in deep cerebellar neurons might compensate for the altered function of the P/Q-type Ca(2+) channel of RMN.


Ethnomethodology of anatomy practice --- Medical education as social achievement

解剖実習のエスノメソドロジー --- 社会的達成としての医学教育


Abnormalities of cerebellar foliation in rats prenatally exposed to ethanol

Pregnant rats were given a liquid diet containing 5% (w/v) ethanol between gestational days 10 and 21. Cerebella of their offspring were examined at 7 weeks of age. Rats exposed prenatally to ethanol showed a fusion of folia V and VI in the cerebellar vermis. Around the fusion, the cortical laminar structure was disrupted: Purkinje cell dendrites derived from each adjacent folium were tangled, and solitary or clustered ectopic granule cells were in the molecular layer. Some ectopic granule cells surrounded basophilic rosette-like structures. Glial fibrillary acidic protein immunostaining revealed defects in the glia limitans, which is formed by Bergmann glial endfeet on the cerebellar surface. Absence of the glia limitans was observed corresponding to the fusion area. These findings suggested that prenatal exposure to ethanol might impair the formation of the glia limitans in the cerebellum, resulting in the fusion of folia and the disruption of the cortical structure. These malformations may be involved in the delayed motor development and ataxia seen in human fetal alcohol syndrome.


First Cutaneous Branch of the Internal Pudendal Artery --- An Anatomical Basis for the So-called Gluteal Fold Flap

We investigated the cutaneous blood supply in the gluteal and perineal regions of 35 donated cadavers to provide an anatomical basis for reliable vulvo-vaginal reconstruction using a skin flap such as the so-called gluteal fold flap. The cutaneous areas along the gluteal cleft and sulcus were likely to be supplied by 3 routes: 1) the internal pudendal artery (IPA), especially its first cutaneous branch; 2) perforators running through the gluteus maximus muscle and arising from the inferior gluteal artery (IGA); and 3) a non-perforator running around and inferior to the ischial tuberosity and originating from the IGA. Route 1 supplied the skin along the gluteal cleft, route 2 the gluteal fold (i.e., a bulky skin fold along the upper edge of the gluteal sulcus), and route 3, just along the gluteal sulcus. In those 3 routes, we noted the consistent morphology of the thick and long, first cutaneous branch of the IPA. The first arterial branch, 1.5 mm in diameter at its origin on average (ranging from 0.7-2.6 mm), usually originated from the IPA under the cover of or at the inferomedial or distal side of the sacrotuberous ligament (almost always less than 20 mm from the inferomedial margin of the ligament). The branch ran superomedially toward the coccyx or ran medially in the ischiorectal fat. It accompanied the vein and nerve at its distal (peripheral) course although the nerve often ran independently at its proxomal course near the ligament. Therefore, the first branch of the IPA seems to provide a reliable pedicle using the skin along the gluteal cleft whether the incision for approach is conducted along the gluteal sulcus or not. However, if the gluteus maximus muscle extended much inferomedially, the pedicle would be very short. In this case, preparation of the pedicle seems to be necessary along the arterial course under the cover of the muscle.


Differential localization and colocalization of two neuron-types of sodium-dependent inorganic phosphate cotransporters in rat forebrain

We studied by immunohistochemistry the distribution of differentiation-associated sodium-dependent inorganic phosphate (Pi) cotransporter (DNPI) in the rat forebrain, in comparison with brain-specific cotransporter (BNPI). DNPI-staining was principally seen in axonal synaptic terminals which showed a widespread but discrete pattern of distribution different from that of the BNPI-staining. In the diencephalon, marked DNPI-staining was seen in the dorsal lateral geniculate, medial geniculate, ventral posterolateral, ventral posteromedial, anterior, and reticular thalamic nuclei without the colocalization with BNPI-staining. DNPI-staining showed a strong mosaical pattern and overlapped well the BNPI-staining in the medial habenular nucleus. DNPI-staining was moderate over the hypothalamus and notably localized in neurosecretory terminals containing corticotropin-releasing hormone in the median eminence. In contrast, the BNPI-staining was region-related and strong in the ventromedial and mammillary nuclei. In the telencephalon, laminar DNPI-staining was seen over the neocortex, corresponding to the thalamocortical termination, and also found in the retrosplenial cortex and the striatum, with the highest intensity in the accumbens nucleus shell. The present results suggest that DNPI serves as a dominant Pi transport system in synaptic terminals of diencephalic neurons including thalamocortical and thalamostriatal pathways as well as the hypothalamic neuroendocrine system in the rat forebrain.


Topological relationship between corticotropin-releasing factor-immunoreactive cerebellar afferents and tyrosine hydroxylase-immunoreactive Purkinje cells in a hereditary ataxic mutant, rolling mouse Nagoya

Using immunohistochemistry we examined the distribution of corticotropin-releasing factor-positive cerebellar afferents and the topological relationship between their projections and the distribution of tyrosine hydroxylase-positive Purkinje cells in an ataxic mutant, rolling mouse Nagoya. In the mutants, some climbing fibers were more intensely stained for corticotropin-releasing factor, but their zonal distribution remained the same as in non-ataxic littermates (control mice). These climbing fibers arose from the dorsal accessory nucleus, the ventral lamella of principal nucleus, the dorsomedial cell group, the subnucleus A, the beta subnucleus and the ventrolateral protrusion of the inferior olive, since perikarya in these olivary subdivisions were more intensely stained for corticotropin-releasing factor than in controls. Some mossy fiber rosettes in the vermal lobules, the simple lobule, the crus I of ansiform lobule, the copula pyramidis and the flocculus also exhibited corticotropin-releasing factor immunoreactivity and were more densely stained in the mutants than in controls. Double immunostaining for corticotropin-releasing factor and tyrosine hydroxylase in the mutant cerebellum revealed that the distribution of tyrosine hydroxylase-positive Purkinje cells corresponded to terminal fields of corticotropin-releasing factor-positive climbing fibers but not corticotropin-releasing factor-positive mossy fibers. This study indicated an increased corticotropin-releasing factor immunoreactivity in some climbing or mossy fibers in the cerebellum of rolling mouse Nagoya. We also found that the distribution of tyrosine hydroxylase-positive Purkinje cells corresponded to terminal fields of corticotropin-releasing factor-positive climbing fibers in the mutant cerebellum. As the transcription of the tyrosine hydroxylase gene is facilitated by Ca2+, abnormal tyrosine hydroxylase expression in the mutant Purkinje cells may indicate functional abnormality by alterations in intracellular Ca2+ concentrations. Therefore, we suggest that an increased level of corticotropin-releasing factor in a specific population of climbing fibers may alter the function of their target Purkinje cells.


Prenatal exposure to ethanol induces leptomeningeal heterotopia in the cerebral cortex of the rat fetus.

Pregnant rats were fed an ethanol-containing liquid diet between gestational day (GD) 10 and GD 21. Leptomeningeal heterotopias were observed in the cerebral cortex of ethanol-exposed fetuses. They appeared on the brain surface of the lateral cortical region near the rhinal fissure, and were found more numerously in the rostral than the caudal region. These abnormalities contained certain neuronal perikarya, microtubule-associated protein (MAP) 1b-positive neuronal processes, and Rat-401-positive radial glial fibers. Immunostaining for Rat-401 revealed that the heterotopias protruded through breaches in the glia limitans. In adult rats exposed to ethanol prenatally, the heterotopias persisted in the lateral cortical region. We conclude that prenatal exposure to ethanol might induce defects in the glia limitans, resulting in the genesis of leptomeningeal heterotopias. These abnormalities may be related to mental retardation or the cognitive deficits associated with human fetal alcohol syndrome (FAS).


Abnormal expression of tyrosine hydroxylase immunoreactivity in Purkinje cells precedes the onset of ataxia in dilute-lethal mice

Expression of tyrosine hydroxylase (TH) immunostaining in the cerebellum was examined in dilute-lethal mice (DL) prior to and following the onset of ataxia. DL walked normally on postnatal days 7 and 8. Falling over when walking was exhibited by about 20% of DL on day 9 and by all DL by day 10. TH-positive Purkinje cells in lobules IX and X of the vermis of either ataxic or non-ataxic DL were clearly observed on day 9 when compared to control mice, and had drastically increased by day 10. These results revealed that abnormal TH expression occurred in some Purkinje cells of DL cerebella, preceding the onset of ataxia.


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 Ca2+, TH expression in the mutant Purkinje cells may predict neuronal dysfunction caused by alterations in cellular Ca2+ currents.


Abnormal distribution of hippocampal mossy fibers in rats exposed to X-irradiation in utero


Acute regulation by dietary phosphate of the sodium-dependent phosphate transporter (NaP(i)-2) in rat kidney.


Observations of brain morphogenesis and its abnormalities using MRI

MRIを用いた脳発達の研究


外耳道に生じたリンパ腫様丘疹症例

外耳道に生じたリンパ腫様丘疹症例


ローリングマウス小脳におけるカルレチニン陽性 unipolar brush cell と CRF 陽性苔状線維終末


ラット視神経の morphometryー糖尿病，胎児性アルコール症候群ラットの解析ー


A 5-HT2A/2C receptor agonist, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane, mitigates developmental neurotoxicity of ethanol to serotonergic neurons.

Prenatal ethanol exposure causes the reduction of serotonergic (5-HTergic) neurons in the midbrain raphe nuclei. In the present study, we examined whether an activation of signaling via 5-HT2A and 5-HT2C receptors during the fetal period is able to prevent the reduction of 5-HTergic neurons induced by prenatal ethanol exposure. Pregnant Sprague-Dawley rats were given a liquid diet containing 2.5 to 5.0% (w/v) ethanol on gestational days (GDs) 10 to 20 (Et). As a pair-fed control, other pregnant rats were fed the same liquid diet except that the ethanol was replaced by isocaloric sucrose (Pf). Each Et and Pf group was subdivided into two groups; one of the groups was treated with 1 mg/kg (i.p.) of 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), an agonist for 5-HT2A/2C receptors, during GDs 13 to 19 (Et-DOI or Pf-DOI), and another was injected with saline vehicle only (Et-Sal or Pf-Sal). Their fetuses were removed by cesarean section on GD 19 or 20, and fetal brains were collected. An immunohistological examination of 5-HTergic neurons in the fetuses on embryonic day 20 using an antibody against tryptophan hydroxylase revealed that the number of 5-HTergic neurons in the midbrain raphe nuclei was significantly reduced in the Et-Sal fetuses compared to that of the Pf-Sal and Pf-DOI fetuses, whereas there were no significant differences between Et-DOI and each Pf control. Thus, we concluded that the reduction of 5-HTergic neurons that resulted in prenatal ethanol exposure could be alleviated by the enhancement of signaling via 5-HT2A/2C receptors during the fetal period.


Prenatal ethanol exposure impairs passive avoidance acquisition and enhances unconditioned freezing in rat offspring

Previous studies have suggested that ethanol exposure during brain development affects responses to fear and anxiety after maturity. To clarify in detail the impaired behavior related to fear and anxiety seen in rat offspring prenatally exposed to ethanol, their behaviors were observed using an elevated T-maze (ETM) test, which allows assessment of passive avoidance acquisition and one-way escape separately, and an elevated open platform (EOP) test for the assessment of unconditioned freezing against innate fear. The ETM test revealed that acquisition of passive avoidance was significantly inhibited in prenatally ethanol-exposed rats, while their escape behavior was not altered. In the EOP test, the duration of the freezing behavior was significantly elongated in prenatally ethanol-exposed offspring. Thus, we concluded that prenatal ethanol exposure could impair acquisition of passive avoidance, while it could facilitate a response related to unconditioned fears in rat offspring.