Distribution and densitometry mapping of L1-CAM Immunoreactivity in the adult mouse brain – light microscopic observation

BACKGROUND: The importance of L1 expression in the matured brain is suggested by physiological and behavioral studies showing that L1 is related to hippocampal plasticity and fear conditioning. The distribution of L1 in mouse brain might provide a basis for understanding its role in the brain. RESULTS: We examined the overall distribution of L1 in the adult mouse brain by immunohistochemistry using two polyclonal antibodies against different epitopes for L1. Immunoreactive L1 was widely but unevenly distributed from the olfactory bulb to the upper cervical cord. The accumulation of immunoreactive L1 was greatest in a non-neuronal element of the major fibre bundles, i.e. the lateral olfactory tract, olfactory and temporal limb of the anterior commissure, corpus callosum, stria terminalis, globus pallidus, fornix, mammillothalamic tract, solitary tract, and spinal tract of the trigeminal nerve. High to highest levels of non-neuronal and neuronal L1 were found in the grey matter; i.e. the piriform and entorhinal cortices, hypothalamus, reticular part of the substantia nigra, periaqueductal grey, trigeminal spinal nucleus etc. High to moderate density of neuronal L1 was found in the olfactory bulb, layer V of the cerebral cortex, amygdala, pontine grey, superior colliculi, cerebellar cortex, solitary tract nucleus etc. Only low to lowest levels of neuronal L1 were found in the hippocampus, grey matter in the caudate-putamen, thalamus, cerebellar nuclei etc. CONCLUSION: L1 is widely and unevenly distributed in the matured mouse brain, where immunoreactivity was present not only in neuronal elements; axons, synapses and cell soma, but also in non-neuronal elements

alpha-lipoic acid suppresses 6-hydroxydoparnine-induced ROS generation and apoptosis through the stimulation of glutathione synthesis but not by the expression of heme oxygenase-1

We previously reported that the generation of reactive oxygen species (ROS) is the initial event in cell death induced by 6-hydroxydopamine (6-OHDA), an experimental model of Parkinsonism. Since recent studies suggested the important role of antioxidant activity of alpha-lipoic acid (LA) in the suppression of apoptosis of various types, we studied the effect on 6-OHDA-induced apoptosis of PC12 cells. Biochemical analysis revealed that LA suppressed the 6-OHDA-induced ROS generation, increase of caspase-like activity and chromatin condensation. The suppression of 6-OHDA-induced apoptosis by LA required pre-incubation of PC12 cells with LA for 12-24 h. LA increased the intracellular levels of heme oxygenase-1 (HO-1) and glutathione (GSH) and stimulated the expression of GSH synthesis-related genes such as cystine/glutamate antiporter and gamma-glutamylcysteine synthetase (gamma-GCS). However, Sn-mesoporphyrin IX, an inhibitor of HO-1, did not attenuate the LA-induced suppression of apoptosis. In contrast, buthionine sulfoximine, an inhibitor of gamma-GCS, attenuated the LA-induced suppression of ROS generation and chromatin condensation. in addition, a transcription factor Nrf2, which regulates the expression of antioxidant enzymes such as gamma-GCS, translocated to the nucleus by LA. These results suggested that LA suppressed the 6-OHDA induced-apoptosis by the increase in cellular glutathione through stimulation of the GSH synthesis system but not by the expression of HO-1.</p

Behavioural and biochemical effects of an ICV injection of streptozotocin in old Lewis rats

Intacerebroventricularly (ICV) injected streptozotocin (STREP) decreases central glucose metabolism and energy metabolism, which has also been observed in patients with dementia. In the present study we examined the behavioral (open-field behavior and two-way active avoidance learning) and biochemical (hippocampal ChAT activity) effects of STREP treatment in old Lewis rats. The results suggest that hippocampal function was affected by STREP. STREP-treated rats acquired the two-way active avoidance task faster than the control rats, which indicates that STREP treatment does not lead to a general learning deficit. Hippocampal ChAT activity was decreased in STREP-treated rats. The present results suggests also that susceptibility to STREP amy not be related to age in Lewis rats

Microwave Accelerated Transglycosylation of Rutin by Cyclodextrin Glucanotransferase from Bacillus sp. SK13.002

Rutin was subjected to intermolecular transglycosylation assisted with microwave irradiation using cyclodextrin glucanotransferase (CGTase) produced from Bacillus sp. SK13.002. Compared with the conventional enzymatic method for rutin transglycosylation (without microwave irradiation), microwave-assisted reaction (MAR) was much faster and thus more efficient. While the conventional reaction took dozens of hours to reach the highest conversion rate of rutin and yield of transglycosylated rutin, MAR of rutin transglycosylation completed within only 6 min providing almost the same conversion rate of rutin and yield of products consisting of mono-, di-, tri-, tetra-, penta-glucosylated rutins. The optimum transglycosylation conditions for microwave irradiation were 40 °C and 60 W with the reaction system consisting mainly of the mixture of 0.3 g rutin (0.49 mmol) pre-dissolved in 15 mL methanol, 1.8 g maltodextrin in 15 mL of 0.2 M sodium acetate buffer (pH 5.5) and CGTase (900 U). Results from this study indicated that MAR could be a potentially useful and economical technique for a faster and more efficient transglycosylation of rutin

Distinct populations of presympathetic-premotor neurons express orexin or melanin-concentrating hormone in the rat lateral hypothalamus

Orexin and melanin-concentrating hormone (MCH) have been implicated in mediating a variety of different behaviors. These include sleep and wakefulness, locomotion, ingestive behaviors, and fight-or-flight response, as well as anxiety- and panic-like behaviors in rodents. Despite such diversity, all these processes require coordinated recruitment of the autonomic and somatomotor efferents. We have previously mapped the locations of presympathetic-premotor neurons (PSPMNs) in the rat brain. These putative dual-function neurons send trans-synaptic projections to somatomotor and sympathetic targets and likely participate in somatomotor-sympathetic integration. A significant portion of these neurons is found within the dorsomedial (DMH) and lateral hypothalamus (LH), areas of the brain that contain MCH- and orexin- synthesizing neurons in the central nervous system. Thus, we hypothesized that hypothalamic PSPMNs utilize MCH or orexin as their neurotransmitter. To test this hypothesis, we identified PSPMNs by using recombinant strains of the pseudorabies virus (PRV) for trans-synaptic tract tracing. PRV-152, a strain that expresses enhanced green fluorescent protein, was injected into sympathectomized gastrocnemius muscle, whereas PRV-BaBlu, which expresses Β-galactosidase, was injected into the adrenal gland in the same animals. By using immunofluorescent methods, we determined whether co-infected neurons express MCH or orexin. Our findings demonstrate that PSPMNs synthesizing either MCH or orexin are present within LH, where they form two separate populations. PSPMNs located around the fornix express orexin, whereas those located around the cerebral peduncle are more likely to express MCH. These two clusters of PSPMNs within LH likely play distinct functional roles in autonomic homeostasis and stress coping mechanisms. J. Comp. Neurol. 505:586–601, 2007. © 2007 Wiley-Liss, Inc.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/57335/1/21511_ftp.pd

血清アデノシンデアミナーゼ活性の測定法の改良とその臨床的応用

An improved method was devised for the estimation of serum adenosine deaminase activity. The principle of the method consists in measurement of a liberated ammonia by the action of the deaminase with a direct colorimetric determination of ammonia concentration [Okuda et al.]. The technique is relatively easy to estimate the serum enzyme activity and can be applied for routine clinical use. The serum adenosine deaminase activity assayed by this method was elevated in patients with hepatic disease and the others, such as auto-immune diseases, rheumatoid arthritis, malignant lymphoma, leukemia and multiple myeloma. The presented experiments were designed to clarify the origin of the elevated adenosine deaminase in serum. The heat stability and pH optimum were found to be essentially identical in sera from hepatic disease and the others. However, Michael is constant (Km) for adenosine and V max were higher in the sera from patients with hepatic diseases than those of the others. The isoelectrofocusing pattern of the serum enzyme was shown quite different between hepatic disease and the others. The results in the present investigation indicated that the enhanced serum adenosine deaminase activity and its isoenzyme pattern were very useful for clinical diagnosis

Expression of neuropsin in oligodendrocytes after injury to the CNS

Elsevier, Xiao-Ping He, Sadao Shiosaka and Shigetaka Yoshida, Neuroscience Research, 39(4), 2001, 455-462. authorProteases are involved in a variety of processes including demyelination after injury to the central nervous systyem. Neuropsin is a serine protease, which is constitutively expressed in the neurons of the limbic system. In the present study, intrahippocampal kainate injection and enucleation were performed on adult mice. Neuropsin mRNA and protein expression was detected by in situ hybridization and immunohistochemistry. Double in situ hybridization confirmed that the mRNA expression was induced in oligodendrocytes. One day after kainate injection to the hippocampus, neuropsin mRNA was expressed, peaking 4–8 days postoperatively and disappearing at 14 days. Immunohistochemistry and immunoelectron microscopy revealed that neuropsin was expressed in the cell body of oligodendrocytes and myelin. To see if neuropsin degrades myelin protein, purified myelin was incubated with recombinant neuropsin. A decrease in the intensity of the bands of myelin basic protein was observed. These results indicate that neuropsin is involved in demyelination