Biomedical subjects

Kazuyuki Kiyosue

Publications and source records attributed to Kazuyuki Kiyosue.

7 recordsLinked to original sources

PubMed2006

Development of new screening system for Alzheimer disease, in vitro Abeta sink assay, to identify the dissociation of soluble Abeta from fibrils.

Abeta is one of the primary therapeutic targets for Alzheimer disease (AD). Abeta vaccination induces the disappearance of Abeta deposits. Since few reports have focused on the reverse phase of Abeta aggregation, we established a new screening system, the in vitro Abeta sink assay, to clarify the process of dissociation of soluble forms from fibrils. Abeta42 was more resistant to dissociation from fibrils to monomers and/or low molecular weight (LMW) soluble oligomers than Abeta40. We applied this system to find a potential therapy for AD. Ultrasound irradiation significantly enhanced the dissociation of soluble Abeta from fibrils, while ultrasound experiments also confirmed the difference between Abeta40 and Abeta42. We found that some compounds enhanced the dissociation of Abeta from fibrils. Here, we proposed that Abeta42 was more resistant to dissociation from fibrils to monomers and/or LMW soluble oligomers than Abeta40, and this system might be useful to identify dissociation of soluble Abeta from fibrils.

Alzheimer Disease↗

PubMed2005

Diminished neuronal activity increases neuron-neuron connectivity underlying silent synapse formation and the rapid conversion of silent to functional synapses.

Neuronal activity regulates the synaptic strength of neuronal networks. However, it is still unclear how diminished activity changes connection patterns in neuronal circuits. To address this issue, we analyzed neuronal connectivity and relevant mechanisms using hippocampal cultures in which developmental synaptogenesis had occurred. We show that diminution of network activity in mature neuronal circuit promotes reorganization of neuronal circuits via NR2B subunit-containing NMDA-type glutamate receptors (NR2B-NMDARs), which mediate silent synapse formation. Simultaneous double whole-cell recordings revealed that diminishing neuronal circuit activity for 48 h increased the number of synaptically connected neuron pairs with both silent and functional synapses. This increase was accompanied by the specific expression of NR2B-NMDARs at synaptic sites. Analysis of miniature EPSCs (mEPSCs) showed that the frequency of NMDAR-mediated, but not AMPAR-mediated, mEPSCs increased, indicating that diminished neuronal activity promotes silent synapse formation via the surface delivering NR2B-NMDARs in mature neurons. After activation of neuronal circuit by releasing from TTX blockade (referred as circuit reactivation), the frequency of AMPAR-mediated mEPSCs increased instead, and this increase was prevented by ifenprodil. The circuit reactivation also caused an increased colocalization of glutamate receptor 1-specfic and synaptic NR2B-specific puncta. These results indicate that the circuit reactivation converts rapidly silent synapses formed during activity suppression to functional synapses. These data may provide a new example of homeostatic circuit plasticity that entails the modulation of neuron-neuron connectivity by synaptic activity.

2-Amino-5-phosphonovalerate↗

PubMed2004

Micropatterned composite membranes of polymerized and fluid lipid bilayers.

Micropatterned composite membranes of polymerized and fluid lipid bilayers were constructed on solid substrates. Lithographic photopolymerization of a diacetylene-containing phospholipid, 1,2-bis(10,12-tricosadiynoyl)-sn-glycero-3-phosphocholine (DiynePC), and subsequent removal of nonreacted monomers by a detergent solution (0.1 M sodium dodecyl sulfate (SDS)) yielded a patterned polymeric bilayer matrix on the substrate. Fluid lipid bilayers of phosphatidylcholine from egg yolk (egg-PC) were incorporated into the lipid-free wells surrounded by the polymeric bilayers through the process of fusion and reorganization of suspended small unilamellar vesicles. Spatial distribution of the fluid bilayers in the patterned bilayer depended on the degree of photopolymerization that in turn could be modulated by varying the applied UV irradiation dose. The polymeric bilayer domains blocked lateral diffusion of the fluid lipid bilayers and confined them in the defined areas (corrals), if the polymerization was conducted with a sufficiently large UV dose. On the other hand, lipid molecules of the fluid bilayers penetrated into the polymeric bilayer domains, if the UV dose was relatively small. A direct correlation was observed between the applied UV dose and the lateral diffusion coefficient of fluorescent marker molecules in the fluid bilayers embedded within the polymeric bilayer domains. Artificial control of lateral diffusion by polymeric bilayers may lead to the creation of complex and versatile biomimetic model membrane arrays.

Diffusion↗

PubMed2004

Re-expression of NR2B-containing NMDA receptors in vitro by suppression of neuronal activity.

N-methyl-D-aspartate receptors (NMDARs) are known to play critical roles in the development of the nervous system, and their expression is regulated in an activity-dependent fashion during development. However, the regulation of NMDAR expression after circuit formation is less well understood. To examine this, we performed patch-clamp recordings from chick cerebral neurons in an activity-controlled culture. Analysis of NMDAR channels from neurons before synapse formation showed that there are two components in channel open kinetics. The major slow component is clearly blocked by ifenprodil, a specific inhibitor of NR2B-containing NMDARs. In contrast, slow component of NMDAR channel opening from neurons after synapse formation became minor and ifenprodil had little effect on the NMDAR channel openings. Furthermore, this change is reversibly regulated by neuronal activity, in that suppression induces the re-expression of NR2B-containing NMDARs, even after circuit formation.

Action Potentials↗

PubMed2002

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↗

PubMed2002

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↗

PubMed2002

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↗