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Biomedical subjects

Takahisa Taguchi

Publications and source records attributed to Takahisa Taguchi.

27 records · Page 2Linked to original sources

Nerve growth factor-induced glutamate release is via p75 receptor, ceramide, and Ca(2+) from ryanodine receptor in developing cerebellar neurons.

Very little is known about the contribution of a low affinity neurotrophin receptor, p75, to neurotransmitter release. Here we show that nerve growth factor (NGF) induced a rapid release of glutamate and an increase of Ca2+ in cerebellar neurons through a p75-dependent pathway. The NGF-induced release occurred even in the presence of the Trk inhibitor K252a. The release caused by NGF but not brain-derived neurotrophic factor was enhanced in neurons overexpressing p75. Further, after transfection of p75-small interfering RNA, which down-regulated the endogenous p75 expression, the NGF-induced release was inhibited, suggesting that the NGF-induced glutamate release was through p75. We found that the NGF-increased Ca2+ was derived from the ryanodine-sensitive Ca2+ receptor and that the NGF-increased Ca2+ was essential for the NGF-induced glutamate release. Furthermore, scyphostatin, a sphingomyelinase inhibitor, blocked the NGF-dependent Ca2+ increase and glutamate release, suggesting that a ceramide produced by sphingomyelinase was required for the NGF-stimulated Ca2+ increase and glutamate release. This action of NGF only occurred in developing neurons whereas the brain-derived neurotrophic factor-mediated Ca2+ increase and glutamate release was observed at the mature neuronal stage. Thus, we demonstrate that NGF-mediated neurotransmitter release via the p75-dependent pathway has an important role in developing neurons.

Adenoviridae↗

Estrogen enhances depolarization-induced glutamate release through activation of phosphatidylinositol 3-kinase and mitogen-activated protein kinase in cultured hippocampal neurons.

Changes in synaptic efficacy are considered necessary for learning and memory. Recently, it has been suggested that estrogen controls synaptic function in the central nervous system. However, it is unclear how estrogen regulates synaptic function in central nervous system neurons. We found that estrogen potentiated presynaptic function in cultured hippocampal neurons. Chronic treatment with estradiol (1 or 10 nm) for 24 h significantly increased a high potassium-induced glutamate release. The estrogen-potentiated glutamate release required the activation of both phosphatidylinositol 3-kinase and MAPK. The high potassium-evoked release with or without estradiol pretreatment was blocked by tetanus neurotoxin, which is an inhibitor of exocytosis. In addition, the reduction in intensity of FM1-43 fluorescence, which labeled presynaptic vesicles, was enhanced by estradiol, suggesting that estradiol potentiated the exocytotic mechanism. Furthermore, protein levels of synaptophysin, syntaxin, and synaptotagmin (synaptic proteins, respectively) were up-regulated by estradiol. We confirmed that the up-regulation of synaptophysin was blocked by the MAPK pathway inhibitor, U0126. These results suggested that estrogen enhanced presynaptic function through the up-regulated exocytotic system. In this study, we propose that estrogen reinforced excitatory synaptic transmission via potentiated-glutamate release from presynaptic sites.

Animals↗

Cloning and characterization of a novel synaptosome-enriched mRNA that encodes 31 kDa protein.

Although a subpopulation of mRNAs has been identified as translocated to the dendrites or the synaptic regions of neurons, the translocational mechanism has not been elucidated. To find mRNAs enriched in synapses, we compared the synaptosomal mRNAs with those from whole forebrain using differential display (DD). We cloned one of these mRNAs, which encoded a novel 31 kDa protein (PMES-2). PMES-2 mRNA was specifically transcribed in the brain and was present in the dendrites of the hippocampal neurons. PMES-2 protein was partly localized in the postsynaptic density. Although this protein is very similar to human NABC1 protein, its function is still unknown.

Amino Acid Sequence↗

Regulatory expression of MDP77 protein in the skeletal and cardiac muscles.

The mdp77 gene was first cloned from the cDNA library of denervated chick muscles, while its role(s) in vivo was unknown. In the present study, using specific polyclonal antibodies against MDP77, we show that MDP77 was expressed specifically in the skeletal and cardiac muscle, and confirm its presence in the cytoplasm of the extrafusal muscle fibers. In mature muscles, MDP77 immunoreactivity was observed in a repetitive manner along the sarcomere. The onset of MDP77 expression occurred just after myotube formation both in vivo and in vitro. Furthermore, MDP77 was enriched in the intrafusal muscle fibers. Our findings suggest that MDP77 plays an important role(s) in the differentiation, maturation and function of both the skeletal and cardiac muscles.

Animals↗

Basic fibroblast growth factor evokes a rapid glutamate release through activation of the MAPK pathway in cultured cortical neurons.

We examined the possibility that basic fibroblast growth factor (bFGF) is involved in synaptic transmissions. We found that bFGF rapidly induced the release of glutamate and an increase in the intracellular Ca2+ concentration through voltage-dependent Ca2+ channels in cultured cerebral cortical neurons. bFGF also evoked a significant influx of Na+. Tetanustoxin inhibited the bFGF-induced glutamate release, revealing that bFGF triggered exocytosis. The mitogen-activated protein kinase (MAPK) pathway was required for these acute effects of bFGF. We also found that pretreatment with bFGF significantly enhanced high K+-elicited glutamate release also in a MAPK activation-dependent manner. Therefore, we propose that bFGF exerts promoting effects on excitatory neuronal transmission via activation of the MAPK pathway.

Amino Acids↗

A synaptic potentiation by a protein factor distinct from those induced by neurotrophins.

We report a protein factor(s) contained in the conditioned medium (CM) of the Mg(2+)-free treatment induced the synaptic potentiation. This type of potentiation shared a different pathway from those induced by neurotrophins. Neurotrophins were confirmed to induce a synaptic potentiation in the dissociated chick neurons. Furthermore, K252a, an inhibitor of tyrosine kinase, abolished this potentiation. Nevertheless, the potentiation induced by the CM was not blocked by K252a. In addition, the CM prepared from the chick neurons induced a similar potentiation in rat and mouse neurons. These results suggest that the protein factor is a novel protein molecule for inducing the potentiation and it plays a critical role in the common mechanism for the potentiation between avian and mammal.

Animals↗

A simple exploratory algorithm for the accurate and fast detection of spontaneous synaptic events.

We have developed a program for the fast and accurate detection of spontaneous synaptic events. The algorithm identifies each event of which the slope and amplitude which meet criteria. The significant feature of this algorithm is its stepwise and exploratory search for the onset and the peak points. During the first step, the program employing the algorithm makes a rough estimate of the candidate for a synaptic event, and determines a 'temporary' onset data point. The next step is the detection of the true onset data point and 'temporary' peak data point, which probably exist several points after the temporary onset data point. The third step is a backward search to detect the true peak data point. The final step is to check whether the amplitude of the detected event exceeds the threshold. This stepwise and shuttlewise search allows for the accurate detection of the peak points. Using this program, we succeeded in detecting an increased frequency and amplitude of spontaneous excitatory postsynaptic currents in chick cerebral neurons following the application of 12-O-tetradecanoyl-phorbol-13-acetate (TPA). In addition, we demonstrated that the program employing the algorithm was able to be used for the detection of extracellular action potentials.

Action Potentials↗

Development of two transmitter release components during the critical period for imprinting in the chick IMHV.

Transmitter release at an excitatory synapse has two components, fast synchronous and slow asynchronous transmitter release. Using the whole cell recording technique, we investigated the developmental properties of neurotransmitter release, which is composed of the two components in the intermediate and medial part of the hyperstriatum ventral (IMHV) of chicks during the critical period for imprinting. Analysis of the paired-pulse responses revealed that the depression of the excitatory postsynaptic currents (EPSCs), driven mainly by fast synchronous release, was frequently observed in P0-1 chicks but not in those at P5-8. The spontaneous excitatory postsynaptic currents (sEPSCs) after the paired-pulse stimulation, which were thought to be driven by asynchronous transmitter releases, were observed more frequently in P0-1 chicks than P5-8 chicks. Furthermore, examination of Ca2+ dependency in the evoked EPSCs showed that the amplitudes in P5-8 chicks were more sensitive to reduction of the extracellular Ca2+ concentration than younger chicks. Considering that the Ca2+ dependency of EPSCs is defined by both Ca2+ sensitivity and the proportion of each type of release machineries at the release site, these results indicate that the ratio of fast synchronous to slow asynchronous transmitter release machinery changed during the critical period. These changes may play critical roles in the capacity of the avian brain to consolidate novel experience in the immediate period after hatching.

Aging↗

Pseudoalteromonas ruthenica sp. nov., isolated from marine invertebrates.

On the basis of phenotypic and genotypic characteristics and analysis of 165 rRNA sequences, a novel species belonging to the genus Pseudoalteromonas is described. Two pale-orange-pigmented strains, KMM 300T and KMM 290, isolated respectively from a mussel, Crenomytilus grayanus, and a scallop, Patinopecten yessoensis, are marine, gram-negative, aerobic, rod-shaped bacteria that produce a number of antimicrobial compounds. The strains are able to degrade gelatin, elastin, starch, DNA and Tween 80. Chitin and agar are not degraded. The isolates from marine invertebrates grew at NaCl concentrations of 1-9% and a temperature range of 10-35 degrees C and did not utilize most of the wide range of carbohydrates tested, with the exception of D-glucose, cellobiose and sucrose. The DNA G+C content was 48.4-48.9 mol%. The level of DNA homology of the two strains was 98%. DNA from the strains isolated from marine invertebrates showed 5-15% genetic relatedness to the DNA of other type strains of the genus Pseudoalteromonas. 16S rRNA analysis indicated a clear affiliation of the novel bacteria to other species of the genus. The strains are assigned to a novel species, Pseudomonas ruthenica sp. nov., with the type strain KMM 300T (= LMG 19699T = CIP 106857T).

Animals↗