Spontaneous resolution of isolated dissecting aneurysm on the posterior inferior cerebellar artery.

The authors report a rare example of an isolated dissecting posterior inferior cerebellar artery (PICA) aneurysm with spontaneous resolution. A 41 year-old male suffered sudden dizziness, nausea and vomiting. An angiogram and magnetic resonance imaging (MRI) detected an isolated PICA dissection. The patient was treated conservatively and recovered without any apparent neurological deficit. MRI detected the self-resolution of the dissecting aneurysm. Dissecting PICA aneurysms, especially non-haemorrhagic lesions, have the possibility of spontaneous resolution resulting in a favorable outcome. The treatment strategy for this vascular lesion may be decided based upon neuroradiological changes on careful follow-up.

Expression of neuron specific phosphatase, striatal enriched phosphatase (STEP) in reactive astrocytes after transient forebrain ischemia.

We studied the distribution and change of striatal enriched phosphatase (STEP) in the gerbil hippocampus after transient forebrain ischemia. STEP was expressed in the perikarya and in neuronal processes; it was not detected in non-neuronal cells of control animals. After 5-min forebrain ischemia, STEP immunoreactivity (STEP-IR) was preserved for 2 days; it disappeared 4 and more days after ischemia with completion of delayed neuronal death (DND) in the CA1 subfield. Furthermore, only in the CA1 after ischemia, STEP was expressed in reactive astrocytes for 4 to 28 days, showing different patterns of glial fibrillary acidic protein (GFAP)-positive reactive astrocytes. After non-or less-than lethal ischemia, STEP expression in reactive astrocytes corresponded with the degree of neuronal degeneration. Immunoblot analysis of the CA1 subfield revealed the expression of three isoforms, STEP45, -56 and -61; their expression patterns changed with time after ischemia. These data suggest that neuronal STEP is preserved until cell degeneration after ischemia and that STEP is expressed in reactive astrocytes only after lethal ischemia, with different expression patterns for its isoforms. Of STEP45, -56 and -61, STEP61 was the most strongly expressed in the reactive astrocytes; both STEP45 and -61 were expressed in neurons and the expression of STEP56 was weak. STEP may play an important role not only in neurons but also in reactive astrocytes after ischemia, depending on neuronal degeneration.

Activities of calcineurin and phosphatase 2A in the hippocampus after transient forebrain ischemia.

We investigated the changes in the enzyme activity and immunoreactivity of calcineurin in the rat hippocampus after transient forebrain ischemia. Immediately after 20-min transient forebrain ischemia, calcineurin activity decreased to about 40% of the control in the CA1 region and to about 55% in other regions. Protein phosphatase 2A activity showed no remarkable changes. By 12 h after ischemia, calcineurin activity recovered, more in the CA1 region than in other regions. At 24 h it decreased again, but only in the CA1 region. Immunohistochemical- and immunoblot analyses showed no remarkable change in calcineurin in any region of the hippocampus within 12 h after ischemia. Thus, the activity of calcineurin is dissociated from its immunoreactivity and quantity. Several studies have suggested that unknown inhibitory factor(s) and/or reversible changes in calcineurin act to modify enzyme activity after ischemia. In contrast, phosphatase 2A activity underwent no obvious changes during the post-ischemia period we examined. This unique time course of calcineurin activity may contribute to the mechanism of ischemic neuronal injury.

Cadherin-8 protein expression in gray matter structures and nerve fibers of the neonatal and adult mouse brain.

The topological distribution of mouse cadherin-8 protein in the neonatal and adult mouse brain was studied immunohistochemically using a rabbit antiserum. Cadherin-8 expression was restricted to several areas in neonatal brains constituting particular neural circuits, i.e. the limbic system, the basal ganglia-thalamocortical circuit, and the cerebellum and related nuclei. In addition, the nerve fibers linking some of the cadherin-8-positive areas, i.e. the habenulo-interpeduncular tract, decussation of the dorsal tegmentum, the medial longitudinal fasciculus, transverse pontine fibers, the brachium conjunctivum and the inferior cerebellar peduncle were cadherin-8 positive, as were the spinal tract of the trigeminal nerve, oculomotor nerve, facial nerve and trigeminal nerve. Cadherin-8 expression also showed a patch-like distribution in the intermediate gray layer of the superior colliculus, resembling acetylcholinesterase-rich patches in allocation. Segmentally organized cadherin-8-positive areas were found in the neonatal cerebellar Purkinje cell layer. Some nuclei and fibers in the brainstem and cerebellum, expressing cadherin-8 at neonatal stages, were also stained in the adult mouse brain. These findings suggest that cadherin-8 is involved in the formation of particular neural circuits by connecting areas expressing this molecule with positive nerve fibers, and indicate its possible implication in subdivisional organization in the superior colliculus and cerebellum.

Heterogeneity of cadherin-8 expression in the neonatal rat striatum: comparison with striatal compartments.

Mechanisms of organization of the striatal compartments are poorly understood, although involvement of cell adhesion molecules in the compartmentalization has been suggested. Cadherin-8 distribution in the neonatal rat striatum was immunohistochemically studied using a rabbit anti-cadherin-8 antiserum. Intensity of cadherin-8 immunolabeling in the striatum was heterogeneous from postnatal day 0 to postnatal day 7. At postnatal day 9, cadherin-8 immunoreactivity was so weak that heterogeneity was no longer clearly seen. Cadherin-8 immunoreactivity was not detectable at postnatal day 14. Cadherin-8-rich and cadherin-8-poor areas were identical to calbindin-rich areas and tyrosine hydroxylase-rich patches, respectively, in allocation, indicating that cadherin-8 was predominantly expressed in the striatal matrix. These results suggest that cadherin-8 is involved in formation of the striatal compartmentalized structures during brain development.

Serine/threonine phosphatase activity of calcineurin is inhibited by sodium orthovanadate and dithiothreitol reverses the inhibitory effect.

Orthovanadate is known to be an inhibitor of protein tyrosine phosphatases. However, we found that it inhibited calcineurin which has the activity of a serine/threonine protein phosphatase, using casein phosphorylated by cyclic AMP-dependent protein kinase as a substrate. Orthovanadate inhibits the Mn2+-activated activity of purified calcineurin to 20%; this is not the case without Mn2+. Furthermore, 10 mM dithiothreitol (DTT) reversed the inhibitory effects of orthovanadate. Orthovanadate showed the same inhibitory effect for calcineurin activity in homogenates as for the purified enzyme; the inhibitory effect was reversed by DTT. These results indicate that orthovanadate inhibits not only protein tyrosine phosphatases as reported, but also serine/threonine phosphatase activity of calcineurin.

Expression of cadherin-8 mRNA in the developing mouse central nervous system.

The expression of cadherin-8 was mapped by in situ hybridization in the embryonic and postnatal mouse central nervous system (CNS). From embryonic day 18 (E18) to postnatal day 6 (P6), cadherin-8 expression is restricted to a subset of developing brain nuclei and cortical areas in all major subdivisions of the CNS. The anlagen of some of the cadherin-8-positive structures also express this molecule at earlier developmental stages (E12.5-E16). The cadherin-8-positive neuroanatomical structures are parts of several functional systems in the brain. In the limbic system, cadherin-8-positive regions are found in the septal region, habenular nuclei, amygdala, interpeduncular nucleus, raphe nuclei, and hippocampus. Cerebral cortex shows expression in several limbic areas at P6. In the basal ganglia and related nuclei, cadherin-8 is expressed by parts of the striatum, globus pallidus, substantia nigra, entopeduncular nucleus, subthalamic nucleus, zona incerta, and pedunculopontine nuclei. A third group of cadherin-8-positive gray matter structures has functional connections with the cerebellum (superior colliculus, anterior pretectal nucleus, red nucleus, nucleus of posterior commissure, inferior olive, pontine, pontine reticular, and vestibular nuclei). The cerebellum itself shows parasagittal stripes of cadherin-8 expression in the Purkinje cell layer. In the hindbrain, cadherin-8 is expressed by several cranial nerve nuclei. Results from this study show that cadherin-8 expression in the embryonic and postnatal mouse brain is restricted to specific developing gray matter structures. These data support the idea that cadherins are a family of molecules whose expression provides a molecular code for the regionalization of the developing vertebrate brain.

Effectiveness of retrograde perfusion of the cerebral vein for attenuating neuronal injury after transient ischemia following reperfusion in the rat brain.

The effect of retrograde perfusion of the cerebral vein (RPCV) with antioxidant LY231617 on neuronal injury after transient ischemia in rat brain was examined. Transient ischemia was caused by left middle cerebral artery (MCA) occlusion and reperfusion. Rats were assigned to three groups: Group A (n = 6), MCA occlusion only; Group B (n = 8), RPCV with saline (flow rate 4.95 ml/hr) into the left inferior cerebral vein; and Group C (n = 6), RPCV with LY231617 (20 mg/kg/hr, flow rate 4.95 ml/hr). RPCV in Groups B and C was performed simultaneously with occlusion. Both occlusion and RPCV were performed for 30 minutes in awake animals. Seven days later, all rats were investigated for rotational behavior elicited by apomorphine (1.0 mg/kg), and then immunohistochemical analysis of brain specimens was carried out using calcineurin as a neuronal marker in the striatum to detect the ischemic damaged area. The number of turns to the left (lesioned side) in both Groups B (42 +/- 12) and C (46 +/- 14) was significantly lower (p < 0.01) than in Group A (222 +/- 45), but there was no significant difference between Groups B and C. The percentage ischemic damaged area in both Groups B (17.9 +/- 6.2%) and C (1.6 +/- 1.0%) was significantly less (p < 0.01) in Group A (51.1 +/- 2.1%). RPCV with and without LY231617 during occlusion was effective for attenuating reperfusion injury.

Restricted expression of cadherin-8 in segmental and functional subdivisions of the embryonic mouse brain.

We have cloned full-length cDNA of a novel mouse cadherin ("mCad8"). The deduced amino acid sequence of the mature form of mCad8 shows 98.2% identity with the sequence of human cadherin-8. The expression of mCad8 was studied by in situ hybridization in mouse embryos of 9.5-14 days gestation (E9.5-E14). Results show that mCad8 expression is restricted to particular subdivisions of the early central nervous system (CNS) and to the thymus. In the CNS, mCad8 expression was observed from E11.5. In the telencephalon, mCad8 is expressed by the ventricular layer of the ganglionic eminence, by cortical areas, and by cells at the caudato-pallial angle. In the diencephalon, the margins of one mCad8-positive area correspond to the borders of the ventral thalamic neuromere, as confirmed by mapping the expression of gene regulatory proteins (Dlx-2, Pax-6, and Gbx-2). In the rhombencephalon, two large groups of mCad8-expressing cells were seen in the pons and in an area of the lateral basal plate of the myelencephalon. These groups of cells extend from the intermediate zone to the mantle zone at E12.5 and later form the anlage of the pontine and the facial nuclei. In conclusion, the expression of mCad8 reflects, in part, the neuromeric organization of the early embryonic CNS. In the mantle layer, mCad8 is expressed by developing gray matter structures, such as brain nuclei, suggesting a role for mCad8 in brain morphogenesis.

Tortuosity of the Cervical Segment of the Internal Carotid Artery in AVM Patients.

SUMMARY: The correlation between the degree of tortuosity of the cervical segment of the internal carotid artery (CICA) and the size of the nidus in AVM patients was investigated on angiograms. Fifteen AVM patients underwent embolization of vessels that were fed primarily by the internal carotid artery. The patients were divided into two groups according to the size of nidus: in group 1 (n=5), the size of the nidus was rated as small (less than 6 cm), in group 2 (n=10) as large (greater than 6 cm). The tortuosity was rated as mild, moderate, or severe. The tortuosity of the ipsilateral CICA in group 1 was severe while in group 2 it was either mild or moderate. The tortuosity of the CICA seems to be an important factor to be considered when distal embolization is contemplated in AVM patients.

Neural BC1 RNA in mouse skeletal muscle is a denervation-induced RNA whose expression is developmentally regulated.

We detected neural BC1 RNA in mouse skeletal muscle. The level of BC1 RNA was high in the fetus, but it declined progressively to the adult level as development proceeded. These observations suggest that this RNA is involved in the prenatal development and differentiation of muscles. Although its developmental expression correlates with the fetal period of polyneuronal innervation, BC1 RNA does not seem to play a direct role(s) in synaptogenesis, since its expression was not restricted to the neuromuscular junction. We also demonstrated that the BC1 RNA level in adult muscle was elevated after denervation, suggesting that changes in the activity of muscles or neural factors caused by axotomy, or both may result in BC1 RNA upregulation.

Cytochrome oxidase activity during acute focal ischaemia in rat brain. A pathophysiology of acute focal ischaemia: Part 2.

An enzyme-histochemical technique was used to examine the changes in cytochrome oxidase activity during acute focal ischaemia in the rat. In the somatosensory cortex, the enzyme activity began to increase significantly (p < 0.01) 1 hour after middle cerebral artery occlusion (MCAO) and continued to increase up to 3 hours, during which ischaemic cell damage was not detected. In the striatum, the enzyme activity increased significantly (p < 0.01) 1 hour after MCAO in the absence of morphological evidence of ischaemic cell damage; a peak activity was reached at 2 hours, and began to decline 3 hours after MCAO when moderate ischaemic change was detected. In both cortical and subcortical areas, the enzyme activity tended to decrease from 4 hours after MCAO, and was reduced to a level similar to or below that of the non-ischaemic hemisphere 5 hours after MCAO, when severe ischaemic damage was demonstrated. The relation of this transient increase of cytochrome oxidase activity in the early stage of acute ischaemia and the hypermetabolism of neuronal cells during ischaemic insult was discussed.

Change of phosphotyrosine immunoreactivity on microglia in the rat substantia nigra following striatal ischemic injury.

Using immunohistochemistry, we investigated changes in phosphotyrosine (P-Tyr) immunoreactivity on the microglia of the rat substantia nigra (SN) following striatal ischemic injury produced by transient middle cerebral artery (MCA) occlusion. Anterograde axonal degeneration in the SN due to striatal ischemic injury was detected by depletion of calcineurin immunoreactivity in that region from 1 day after operation. From 3 days to 1 month (the longest period examined in this study) after MCA occlusion, there was a significant increase in P-Tyr immunoreactivity in the SN ipsilateral to the MCA occlusion. Also, light microscopic observation showed that the microglia exhibited an increased immunoreactivity for P-Tyr and characteristic morphological changes in the ipsilateral SN. The present results indicate that a signal transducing cascade(s) associated with tyrosine phosphorylation may be involved in the activation of the microglia in the SN responding to anterograde degeneration of the striatonigral pathway.

Cellular localization of type II Ca2+/calmodulin-dependent protein kinase in the rat basal ganglia and intrastriatal grafts derived from fetal striatal primordia, in comparison with that of Ca2+/calmodulin-regulated protein phosphatase, calcineurin.

We investigated immunohistochemically the cellular localization of multifunctional type II Ca2+/calmodulin-dependent protein kinase in the rat basal ganglia and intrastriatal grafts derived from fetal striatal primordia, in comparison with that of calcineurin, a reliable marker for striatal medium-sized spinous neurons. The type II Ca2+/calmodulin-dependent protein kinase-positive neurons were of medium size, with a mean diameter of 16.1 +/- microns (average +/- S.D., n = 72, range 13.6-18.3 microns) and comprised approximately 70% of the total neuronal population in the striatum. Light microscopy showed that the type II Ca2+/calmodulin-dependent protein kinase-positive cells had round, triangular or polygonal cell bodies with relatively little cytoplasm. Analysis of serial sections showed that type II Ca2+/calmodulin-dependent protein kinase and calcineurin immunoreactivities were co-localized in the striatal neurons examined with a similar distribution pattern. Type II Ca2+/calmodulin-dependent protein kinase-positive cells were always immunoreactive for calcineurin and cells negative for type II Ca2+/calmodulin-dependent protein kinase showed no apparent calcineurin immunoreactivity. Type II Ca2+/calmodulin-dependent protein kinase-positive nerve fibers in the globus pallidus and substantia nigra almost disappeared following striatal ischemic injury produced by transient middle cerebral artery occlusion and cerebral hemitransection, respectively, suggesting that these immunopositive fibers were striatal projections. Thus, most type II Ca2+/calmodulin-dependent protein kinase-positive neurons in the rat striatum are considered to be of the medium-sized spinous type. Type II Ca2+/calmodulin-dependent protein kinase or calcineurin immunoreactivity was also observed in a large number of neurons in transplants derived from fetal striatal primordia grafted into striatal ischemic lesions. In addition, type II Ca2+/calmodulin-dependent protein kinase- or calcineurin-immunoreactive nerve fibers appeared in the deafferented globus pallidus of the host rats, suggesting that the striatopallidal pathway was reformed by striatal projection neurons of the transplants. This finding may also indicate that Ca2+/calmodulin-regulated enzymes are useful for tracing striatal projection fibers as endogenous marker proteins.

Microglial response to transient focal cerebral ischemia: an immunocytochemical study on the rat cerebral cortex using anti-phosphotyrosine antibody.

Microglial response to transient focal ischemia was examined using an immunohistochemical method with a monoclonal antibody to phosphotyrosine (P-Tyr). For this purpose, a rat model of reversible middle cerebral artery occlusion for 1 h was used. Compared with results in the noninsulted hemisphere, there was a significant increase in P-Tyr immunolabeling of the microglia in the insulted cerebral cortex 3 h postreperfusion. This microglial reaction progressed up to 24 h after ischemic insult. In the affected cerebral cortex, morphological changes of the microglial positive for P-Tyr were also observed, with shortened and thickened processes, enlarged cell bodies, and ameboid features. Cell density analysis did not show any apparent change in number of P-Tyr-positive microglia in the insulted cortex at 6, 12, and 24 h after reperfusion, suggesting that the cells with increased P-Tyr immunoreactivity were resident microglia. The present findings suggest that signal transduction mediated by tyrosine phosphorylation is involved in the microglial response to ischemic injury in the rat cerebral cortex.

Subdivisional ischemic injury of the unilateral striatum causes apomorphine-induced rotational behavior in rats.

Behavioral and histological studies were performed on a reversible ischemia model in rats. At 60 days after unilateral transient middle cerebral artery occlusion for 30 min, the operated rats exhibited the ipsiversive rotational behavior elicited by systemic administration of dopamine receptor agonist apomorphine in a dose-dependent manner. Histologically, the ipsilateral striatum of the rats showed a subdivisional ischemic injury, while the nigral dopaminergic neurons appeared intact. The striatal lesions having a cell type-specific injury were located in the dorsolateral portion of the rostral striatum and in the lateral portion of the caudal part of the nucleus. Thus, the transient cerebral ischemia could successfully produce selective damage of a striatal subdivision, which causes an abnormality in motor controls in response to dopamine receptor stimulation. The present data may provide a part of functional and anatomical basis for understanding the movement disorders associated with basal ganglia dysfunction (e.g., parkinsonism), which may occur in patients with cerebrovascular disorders.

Disruption of the blood-cerebrospinal fluid barrier by transient cerebral ischemia.

The influence of transient cerebral ischemia on blood-brain and blood-cerebrospinal fluid (CSF) barrier permeability was studied sequentially by magnetic resonance imaging (MRI) contrast enhancement using gadolinium-diethylene triamine pentaacetic acid (Gd-DTPA) in rats. The unilateral internal carotid and middle cerebral arteries were transiently occluded by inserting a nylon thread into the carotid artery and removing it following a variable interval of 5 to 60 min. Contrast enhancement of the lateral ventricle on the affected side was seen in the enhanced T1-weighted image at the early stage of reperfusion 6 h after the start of ischemia in most of the rats subjected to 30- and 60-min ischemia, and in 3 of 6 rats in the 15-min ischemia group. Autoradiograms of Gd-[14C]DTPA in rats subjected to 60-min ischemia demonstrated that the tracer strongly accumulated in the choroid plexus, the wall of the lateral ventricle and its surrounding brain tissue. On the other hand, parenchymal enhancement of the striatum was seen only in the 60-min ischemia group and appeared later on Day 1 or Day 7. These results indicate that ventricular enhancement on MRI in this model is caused by disruption of the blood-CSF barrier at the choroid plexus of the lateral ventricle. This is the first reported study to demonstrate blood-CSF barrier disruption by transient ischemia.

Neuronal induction of 72-kDa heat shock protein following methamphetamine-induced hyperthermia in the mouse hippocampus.

By means of an immunohistochemical technique, we examined the neuronal induction of 72-kDa heat shock protein (HSP72) in response to methamphetamine-induced hyperthermia in the mouse hippocampus. Strong HSP72 immunoreactivity (ir) was found in the neurons of hippocampus proper, particularly in the CA1/2 and medical CA3 subfields, at 10 h after drug injection. By 18 h, those neurons still revealed HSP72-ir, while neurons of the dentate gyrus also appeared positive for HSP72. At this stage, intense HSP72-ir was first detected in non-neuronal cells, i.e. glial and vascular endothelial cells. At 24 h, no apparent HSP72-ir was found in the hippocampal neurons, while only non-neuronal cells still revealed immunoreactivity for HSP72. In addition, no morphological evidence of cell degeneration or loss was noted in the CA1 sector or other hippocampal regions at 5 days after hyperthermic insult. In conclusion, (1) methamphetamine-induced hyperthermia per se is a stressful stimulant causing neuronal induction of HSP72 in the hippocampus neurons, particularly of CA1/2 and medial CA3 sectors, but does not prove fatal to the cells; (2) there is a cell type-specific difference in response to hyperthermic insult by inducing HSP72 and the timing of the induction response in the hippocampal formation; and (3) the animals that underwent drug-induced hyperthermia may be useful as an experimental model for the study of the protective mechanism of heat shock proteins against subsequent harmful stimuli.

Therapeutic effects of moxibustion on delayed uterine involution in postpartum dairy cows.

Moxibustion on 12 specific points (Keiketsu in Japanese) was applied for treatment of delayed uterine involution in 16 cows that were diagnosed on the basis of rectal palpation and vaginoscopic examination 21 to 35 days after parturition. The treatment was continued for three consecutive days. Other 32 cows with the delayed uterine involution were either injected intramuscularly with 25 mg PGF2 alpha (17 cows) or infused in utero with 500 mg ampicillin (15 cows). The uterine involution following the treatment was monitored by rectal palpation and vaginoscopic examination. Milk samples were collected three times weekly and used for milk progesterone assay to monitor the ovarian function. No significant difference was observed in the uterine involution among the groups treated with moxibustion, PGF2 alpha or ampicillin. Percentages of cows with abnormal cervical mucus and bacterial isolation from cervical swab decreased remarkably in all groups during 4 weeks after treatment. Forty-six percent of cows with delayed uterine involution was diagnosed as having inactive ovaries. Percentage of cows that responded with ovulation and corpus luteum formation after moxibustion was 67 percent, slightly higher than those in cows treated with PGF2 alpha or ampicillin. Reproductive performance after the moxibustion was well-comparable to those after PGF2 alpha or ampicillin treatment. Result indicates that the moxibustion could be used as the alternative to PGF2 alpha and antibiotics for treating delayed uterine involution in cows.

Changes of immunoreactivity for synaptophysin ('protein p38') following a transient cerebral ischemia in the rat striatum.

We assessed the chronological change of the expression of synaptophysin, an integral glycoprotein on the presynaptic vesicles, after a transient cerebral ischemic insult in the rat. The ischemic lesion was consistently localized in the dorsolateral part of the striatum, which was clearly visualized by a depletion of calcineurin immunostaining or increases of immunoreactivities for glial fibrillary acidic protein and tyrosine hydroxylase. Immunoreactivity for synaptophysin was transiently increased in the ischemic lesions from 3 to 7 days after cerebral ischemia. Thereafter, synaptophysin immunostaining in the damaged areas gradually decreased and finally almost disappeared one month after surgery. Because synaptophysin is located in the presynaptic vesicle, and thought to be involved in presynaptic functions such as vesicle-membrane fusion and release of neurotransmitters, present findings suggest that loss of the postsynaptic site after ischemic insult induces a transient increase of the presynaptic functions, followed by a decrease of functional presynaptic activity or trans-synaptic retrograde degeneration of axon terminals.