Biomedical subjects

Yoshimi Kawasaki

Publications and source records attributed to Yoshimi Kawasaki.

4 recordsLinked to original sources

PubMed2004

AMPA receptor synaptic targeting regulated by stargazin interactions with the Golgi-resident PDZ protein nPIST.

Regulation of AMPA receptors (AMPARs) at synapses plays a critical role in alterations of synaptic strength in the brain. Stargazin, an AMPAR-interacting protein, is critical for clustering and regulation of synaptic AMPARs. Stargazin interacts with AMPARs via its extracellular domain and with PDZ [postsynaptic density-95 (PSD-95)/Discs large (Dlg)/zona occludens-1 (ZO-1)] proteins via its C-terminal PDZ-binding motif, and these interactions are necessary for stargazin and AMPAR synaptic targeting. By studying the expression of stargazin mutant constructs in cultured hippocampal neurons, we identified a novel domain corresponding to residues 243-283 within the cytoplasmic C terminus of stargazin that is also required for stargazin and AMPAR synaptic clustering. To identify proteins that interact with this stargazin synaptic clustering domain, we performed a yeast two-hybrid assay and found that this stargazin domain binds to nPIST (neuronal isoform of protein-interacting specifically with TC10), a Golgi-enriched protein implicated in trafficking of transmembrane proteins. Using in situ hybridization, immunohistochemistry, coimmunoprecipitation studies, and biochemical fractionation, we found that stargazin and nPIST colocalize and interact in the brain. Finally, by studying AMPAR clustering in transfected hippocampal neurons, we found that overexpression of nPIST enhances AMPAR synaptic clustering, whereas transfection of a dominant-negative nPIST construct attenuates AMPAR synaptic clustering. These studies identify a novel stargazin domain necessary for synaptic clustering of AMPARs and suggest that nPIST and stargazin interactions play a critical role in AMPAR trafficking to the synapse.

Animals↗

PubMed2003

Functional studies and distribution define a family of transmembrane AMPA receptor regulatory proteins.

Functional expression of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in cerebellar granule cells requires stargazin, a member of a large family of four-pass transmembrane proteins. Here, we define a family of transmembrane AMPA receptor regulatory proteins (TARPs), which comprise stargazin, gamma-3, gamma-4, and gamma-8, but not related proteins, that mediate surface expression of AMPA receptors. TARPs exhibit discrete and complementary patterns of expression in both neurons and glia in the developing and mature central nervous system. In brain regions that express multiple isoforms, such as cerebral cortex, TARP-AMPA receptor complexes are strictly segregated, suggesting distinct roles for TARP isoforms. TARPs interact with AMPA receptors at the postsynaptic density, and surface expression of mature AMPA receptors requires a TARP. These studies indicate a general role for TARPs in controlling synaptic AMPA receptors throughout the central nervous system.

Animals↗

PubMed2002

Postsynaptic targeting of alternative postsynaptic density-95 isoforms by distinct mechanisms.

Members of the postsynaptic density-95 (PSD95)/synapse-associated protein-90 (SAP90) family of scaffolding proteins contain a common set of modular protein interaction motifs including PDZ (postsynaptic density-95/Discs large/zona occludens-1), Src homology 3, and guanylate kinase domains, which regulate signaling and plasticity at excitatory synapses. We report that N-terminal alternative splicing of PSD95 generates an isoform, PSD95beta that contains an additional "L27" motif, which is also present in SAP97. Using yeast two hybrid and coimmunoprecipitation assays, we demonstrate that this N-terminal L27 domain of PSD-95beta, binds to an L27 domain in the membrane-associated guanylate kinase calcium/calmodulin-dependent serine kinase, and to Hrs, an endosomal ATPase that regulates vesicular trafficking. By transfecting heterologous cells and hippocampal neurons, we find that interactions with the L27 domain regulate synaptic clustering of PSD95beta. Disrupting Hrs-regulated early endosomal sorting in hippocampal neurons selectively blocks synaptic clustering of PSD95beta but does not interfere with trafficking of the palmitoylated isoform, PSD95alpha. These studies identify molecular and functional heterogeneity in synaptic PSD95 complexes and reveal critical roles for L27 domain interactions and Hrs regulated vesicular trafficking in postsynaptic protein clustering.

Adaptor Proteins, Signal Transducing↗

PubMedDate not supplied

Retinal glial cells stimulate microvascular pericyte proliferation via fibroblast growth factor and platelet-derived growth factor in vitro.

PURPOSE: To investigate whether retinal glial cells (RGCs), which are believed to play an important role in the development and maintenance of microvessels, stimulate the proliferation of retinal bovine microvascular pericytes, an essential component of the vessels. METHODS: Conditioned medium (CM) was collected from a primary culture of RGC obtained from chick embryonic retina. The cell number was assayed after stimulation by RGC-CM. Also, by neutralizing antibody and reverse transcription polymerase chain reaction (RT-PCR), we tried to identify which factor of the RGCs contributes to the pericyte stimulation. RESULTS: Pericyte proliferation was stimulated by RGC-CM in a dose-dependent manner. Platelet-derived growth factor-BB (PDGF-BB), acidic fibroblast growth factor (aFGF), and basic fibroblast growth factor (bFGF) stimulated pericyte proliferation; however, PDGF-AA, transforming growth factor-beta2 (TGF-beta2), and vascular endothelial growth factor (VEGF) did not. The RGC-CM-dependent stimulative effect was blocked, in part, by the neutralizing antibodies for aFGF, bFGF, and PDGF. A mixture of these three antibodies completely blocked the stimulation. RT-PCR revealed that RNA for aFGF, bFGF, and TGF-beta2 were expressed in RGCs. CONCLUSIONS: Pericyte growth is stimulated in vitro by RGC-CM through aFGF, bFGF, PDGF-BB, at least in part. This finding suggests that RGCs may modulate in vivo pericyte cell growth through these three growth factors.

Animals↗