Biomedical subjects

Y Yoshimura

Publications and source records attributed to Y Yoshimura.

At least 73 records · Page 4Linked to original sources

PubMed2001

Role of leukocytes in uterine hypoperfusion and fetal growth retardation induced by ischemia-reperfusion.

We investigated leukocyte involvement in uterine hypoperfusion and intrauterine fetal growth retardation (IUGR) induced by ischemia-reperfusion (I/R) in Sprague-Dawley rats. On day 17 of gestation, leukocyte accumulation in the uterus and placenta subjected to 30 min of ischemia, followed by reperfusion, was assessed by measuring myeloperoxidase (MPO) activity. Uterine MPO activity was significantly higher after 1 h of reperfusion than it was before ischemia (P < 0.05), without any increase in placental MPO activity. Immunohistochemical staining showed leukocyte accumulation in the uterus subjected to I/R. The effects of treatment with monoclonal antibodies against CD11a (WT1) and CD18 (WT3) at a dose of 0.8 mg/kg on uterine blood flow and IUGR were investigated. Laser-Doppler flowmetry demonstrated that uterine hypoperfusion at 2 h after ischemia (blood flow, -51.7 +/- 1.2%; P < 0.01) was inhibited by WT1 and WT3 treatment. I/R-induced IUGR at full term (P < 0.05 vs. nonischemic horn) was prevented by WT1 and WT3 treatment on day 17. These results indicate that leukocyte accumulation may play an important role in the pathogenesis of uterine hypoperfusion and IUGR induced by I/R in pregnant rats.

Animals↗

PubMed2001

Promoting mechanism of menthol derivative, 1-O-ethyl-3-buthylcyclohexanol, on the percutaneous absorption of ketoprofen.

Menthol derivatives were synthesized and evaluated for their promoting activity on the percutaneous absorption of ketoprofen and skin irritation in vivo, choosing O-ethylmenthol (MET) as the mother compound. The compound having a C-3 positionned n-butyl group (1-O-ethyl-3-n-buthylcyclohexanol, OEBC) indicated the most promoting activity and caused relatively little skin irritation. In order to understand enhancement mechanism of OEBC an in vitro permeation study of ketoprofen was performed. The time course of the cumulative amounts of drug permeated through the rat skin exhibited a linear relation after an initial lag time. This was analyzed in membrane diffusion model and the diffusion and partition parameters of ketoprofen were estimated. Both parameters were remarkably enhanced when a hydrogel containing a small quantity of OEBC (0.5%) was applied. Furthermore, to clarify the site of action of OEBC, we also investigated in vitro permeation study of ketoprofen employing different skins of state, reversed skin and stratum corneum stripped skin. When OEBC was added to the hydrogels which were applied to the reversed and stripped skins, almost no changes of the flux were observed compared with the control (without OEBC). These results suggested that the site of action of OEBC was stratum corneum. Morphological changes of the stratum corneum surface were microscopically observed with 0-2% OEBC. The spaces between the stratum corneum cells treated with 0.5-2% OEBC became extended and the shape of each cell became clear. This may suggest that the site of action of OEBC was the intercellular of stratum corneum. Furthermore, an electron spin resonance study was performed to investigate the effect of OEBC on the intercellular lipid bilayer fluidity of the stratum corneum and the rotational correlation times were calculated. 2,2,6,6-Tetramethylpiperidine-1-oxyl (TEMPO) and 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL) were used as the spin label. In use of OEBC, the fluidity of TEMPO labeled the stratum corneum lipid increased as the addition of OEBC. The results suggested that OEBC promote the penetration of drugs by enhancing fluidity of the local lipid bilayers around TEMPO.

Algorithms↗

PubMed2001

Colocalization of noggin and bone morphogenetic protein-4 during fracture healing.

The regulation of callus formation during fracture repair involves the coordinate expression of growth factors and their receptors. This article describes the temporal and spatial expression of noggin gene, an antagonist to bone morphogenetic protein (BMP), during the fracture repair process. Noggin expression was examined by means of Northern blotting and in situ hybridization and compared with the expression pattern of BMP-4 in a model of fracture repair in adult mice. Expression levels of noggin messenger RNA (mRNA) were enhanced in the early phase of fracture callus formation. The localization of the noggin mRNA was similar to that of BMP-4 mRNA. Distinct noggin mRNA signals were located predominantly in cells lining the periosteum and the cortical endosteum near the fracture site at 2 days after fracture. At 5, 10, and 21 days after fracture, noggin mRNA was detected in the chondrocytes and osteoblasts in the newly formed callus. The pattern of localization was indistinguishable from that of BMP-4. These results suggest that the noggin/BMP-4 balance could be an important factor in the regulation of callus formation during fracture healing.

Animals↗

PubMed2001

Autoreactive diabetogenic T-cells in NOD mice can efficiently expand from a greatly reduced precursor pool.

A broad repertoire of pancreatic beta-cell autoreactive T-cells normally contributes to the development of type 1 diabetes in NOD mice. However, it has been unknown if a large reduction in the precursor pool from which autoreactive T-cells are drawn would inhibit the development of type 1 diabetes. To address this issue, we reduced the precursor frequency of autoreactive T-cells in NOD mice through allelic exclusion induced by transgenic expression of an H2-Db class I-restricted T-cell receptor (TCR) specific for a pathologically irrelevant lymphocytic choriomeningitis virus (LCMV) peptide. TCR allelic exclusion greatly reduced the pool of T-cells from which diabetogenic effectors could be derived in these NODxLCMV TCR Tg mice. Surprisingly, this did not impair their type 1 diabetes susceptibility. Furthermore, a diabetogenic CD8 T-cell population that is prevalent in standard NOD mice was present at essentially equivalent levels in pancreatic islets of NODxLCMV TCR Tg mice. Other data indicated that the antigenic specificity of these CD8 T-cells is primarily the function of a shared TCR-alpha chain. Although the percentage of TCR transgenic T-cells decreased in NOD versus B6,D2 control mice, much higher total numbers of both the TCR transgenic and the nontransgenic T-cells accumulated in the NOD strain. This transgenic T-cell accumulation in the absence of the cognate peptide indicated that the NOD genetic background preferentially promotes a highly efficient antigen-independent T-cell expansion. This might allow diabetogenic T-cells in NOD mice to undergo an efficient expansion before encountering antigen, which would represent an important and previously unconsidered aspect of pathogenesis.

Alleles↗

PubMed2001

Elevation of N-(carboxymethyl)valine residue in hemoglobin of diabetic patients. Its role in the development of diabetic nephropathy.

OBJECTIVE: Advanced glycation end products (AGEs) are a risk factor for diabetic complications. We have developed an assay method for N-(carboxymethyl)valine (CMV) of the hemoglobin (CMV-Hb), which is an AGE generated from HbA1c. Herein we describe the clinical utility of CMV-Hb measurement for the diagnosis of diabetic nephropathy RESEARCH DESIGN AND METHODS: BALB/c mice were immunized with carboxy-methylated Hb and monoclonal antibody raised against CMV-Hb. This antibody was characterized by a surface plasmon resonance. We developed a latex immunoassay using the antibody and measured CMV-Hb from erythrocytes in type 2 diabetic patients and healthy control subjects (age 64.6 +/- 12.0 vs. 61.1 +/- 13.2 years, NS: HbA1c 69 +/- 1.5 vs. 5.2 +/- 0.4%, P < 0.0001). RESULTS: A monoclonal antibody against CMV-Hb beta-chain NH2-terminal and an assay method for measurement for CNMV-Hb were both developed in our laboratory. CMV-Hb levels were significantly greater in the diabetic patients than in the control subjects (18.2 +/- 6.9 vs. 12.7 +/- 0.9 pmol CMV/mg Hb, P < 0.0001). No correlation was found between CMV-Hb and HbA1c or CMV-Hb and glycated albumin. Levels of CMV-Hb increased as the diabetic nephropathy progressed. CONCLUSIONS: We established an assay method for CMV-Hb and confirmed the presence of CMV-Hb in circulating erythrocytes. CMV-Hb was more prevalent in diabetic patients than in healthy subjects. Furthermore, it was significantly higher in patients with diabetic nephropathy, suggesting that the presence of CMV-Hb may be a valuable marker for the progression of diabetic nephropathy.

Animals↗

PubMed2001

Blood pressure, serum cholesterol concentration and their related factors in urban and rural elderly of Ho Chi Minh City.

In Vietnam, information about blood pressure, serum lipids and their factors is limited. To obtain some of this information, a cross sectional nutrition survey was carried out in an urban and rural area of Ho Chi Minh City with 217 participants aged 60-69 y (148 females and 69 males). Anthropometry and blood pressure were measured. For three consecutive weekdays, 24 h dietary recalls were performed. Single 24 h urine was collected for sodium and potassium analysis. A fasting blood sample was taken and biochemical parameters were measured. Results indicate a high percentage of hypertension in urban (female: 35.5%, male: 43.8%) and rural areas (female: 22.2%, male: 35.1%). Blood pressure was correlated with body mass index (BMI) and 24 h urinary sodium-to-potassium (Na/K) ratio. A high prevalence of serum total cholesterol (TC) above 220 mg/dL (female: 55.3%, male: 31.3%) and overweight (female: 34.2%, male: 25.0%) were observed in urban residents. By contrast, 5.6% and 24.3% of rural females and males respectively had TC below 150 mg/dL and both genders had the same prevalence of underweight (32.4%). TC was positively correlated with body weight, BMI, dietary protein and dietary lipids. Overweight might be a major risk factor for hypertension in our urban elderly. A high Na/K intake ratio might be a risk factor for hypertension in both areas. The high prevalence of elevated TC in the urban area might to be related to the high lipid intake, and the high prevalence of low TC in the rural area might to be related to the low lipid intake.

Aged↗

PubMed2001

Ovarian autoimmunity in relation to egg production in laying hens.

The aim of this study was to determine whether anti-ovarian autoantibodies appear in the circulation of laying hens and whether the concentrations of these antibodies change with respect to ageing and egg laying rate. Autoantibodies to ovarian tissues in the circulation of aged (aged approximately 670 days) White Leghorn hens with low (< 50%) and high (> 90%) egg laying rates were examined by ELISA and western blotting. Young laying hens (aged 185 days) with > 95% egg production were used as controls. The results of the ELISA indicated that IgG, which bound to the ovary and small white follicles, was present in the circulation of old laying hens. More hens that laid few eggs had circulatory autoantibodies to the ovary and small white follicles, as determined by the cut-off value in ELISA (mean absorbance + 2 SD of young laying hens), than did hens that laid greater numbers of eggs, and the concentration of IgG was significantly higher in the hens that laid few eggs. In contrast, when the muscle proteins were used as antigens there were no significant differences in the absorbance values among low and high laying frequency old hens or young hens. Western blotting revealed many bands of immunoprecipitates formed by ovarian antigens and antibodies in the serum of old hens, indicating the presence of many binding sites for circulatory IgG in ovarian tissues. These results indicate that antibodies to ovarian tissues appear in the circulation of laying hens during ageing, and that the concentration of these autoantibodies is related inversely to the rate of egg laying by hens.

Aging↗

PubMed2001

[The voiding after the suburethral sling operation, obstructive or non-obstructive?].

To examine whether or not the suburethral sling operation produces obstruction during voiding, seven females who underwent the sling operation using synthetic material (Vesica Sling Kit) were studied postoperatively urodynamically. Uroflowmetry and residual urine measurement showed no overt voiding difficulties in any cases. However, in one case, a pressure flow study indicated equivocal and the position of the sling material was judged to be too proximal by fluoroscopic monitoring. In all other 6 cases pressure flow was shown to be non-obstructive. Fluoroscopic finding also demonstrated in these 6 cases an appropriate bladder neck opening at the time of voiding and the sling was positioned just from the bladder neck to mid-urethra. Thus, it is concluded that the suburethral sling operation produces no obstruction as long as the position of the sling material is carefully determined from bladder neck to mid-urethra and excessive tension is avoided.

Aged↗

PubMed2001

Changes in the localization of MHC class II positive cells in hen ovarian follicles during the processes of follicular growth, postovulatory regression and atresia.

The aim of this study was to determine the changes in the population of major histocompatibility complex class II positive (MHC-II(+)) cells in ovarian follicles during the processes of follicular growth, postovulatory regression and follicular atresia in hens. Cryostat sections of ovarian stroma containing cortical follicles, small white follicles, the largest (F(1)) and third largest (F(3)) preovulatory follicles, postovulatory and atretic follicles of laying hens were prepared. The sections were immunostained for MHC-II molecules using mouse anti-chicken MHC-II monoclonal antibody and positive cells were counted using a computer-assisted image analyser under a light microscope. MHC-II(+) cells were localized in the theca layer of normally growing follicles including cortical follicles, small white follicles and F(3) and F(1) preovulatory follicles, whereas they were found in both the theca and granulosa layers in postovulatory and atretic follicles. The frequency of MHC-II(+) cells in the theca layer was significantly increased during follicular growth from cortical follicles to F(3) preovulatory follicles. Although the population of MHC-II(+) cells did not differ between F(3) and F(1) preovulatory follicles, it increased significantly in postovulatory follicles (P < 0.01). The population of MHC-II(+) cells was significantly greater in the theca layer of atretic follicles than in non-atretic follicles (P < 0.01). These results indicate that the antigen-presenting function via MHC-II increases in association with follicular growth. A marked increase in MHC-II(+) cells indicates that these cells may be involved in regression of postovulatory and atretic follicular tissues.

Animals↗

PubMed2001

Immunochemical assay of hemoglobin with N(epsilon)-(carboxymethyl)lysine at lysine 66 of the beta chain.

BACKGROUND: N(epsilon)-(Carboxymethyl)lysine (CML), a well-characterized and major advanced glycation end product structure, is produced via a Maillard reaction by nonenzymatic glycation and/or oxidation. Although few of the carboxymethylation sites of lysine residues on proteins have been identified, it is known that the possible lysine glycation site in hemoglobin (Hb) is Lys-66 on the beta chain. We aimed to develop an assay for the Hb with a CML (CML-Hb) site specific to Lys-66 on the Hb beta chain and to determine whether the lysine residue at that site is carboxymethylated. METHODS: Ala-His-Gly-Lys-Lys(CM)-Val-Leu-Gly-Ala-Phe-Ser-Cys, the peptide derived from the beta chain of human Hb, was synthesized as an immunogen, and a monoclonal antibody against the peptide was prepared. A latex immunoassay method was established using the antibody on an automatic analyzer. In this study, 20 samples from healthy subjects and 80 samples from nondiabetic patients undergoing hemodialysis (HD) were analyzed. RESULTS: The latex immunoassay method using the antibody correlated significantly with the ELISA method using the antibody (r = 0.95; P <0.001). Between healthy subjects (n = 20) and nondiabetic HD patients (n = 80), a significant difference was seen in circulating CML-Hb (525 +/- 76 vs 778 +/- 137 pmol CML/mg of Hb; P <0.0001). CONCLUSION: The latex method for the CML-Hb site specific to Lys-66 on the beta chain can measure large numbers of samples on an automatic analyzer.

Acetaldehyde↗

PubMed2001

An in situ hybridization study of the effects of artificial insemination on the localization of cells expressing MHC class II mRNA in the chicken oviduct.

The aim of this study was to determine the effects of artificial insemination on the localization of antigen-presenting cells expressing MHC class II mRNA in chicken oviducts. Laying hens (35 weeks old) were inseminated with fresh semen or sham-inseminated with saline daily for 3 days. In situ hybridization was performed to detect chicken MHC class II (B-LB21 major gene) mRNA on frozen sections of oviductal infundibulum, uterovaginal junction and vagina by using digoxigenin-labelled PCR probes. Cells expressing MHC class II were observed mainly in the oviductal mucosal stroma and occasionally in the mucosal epithelium. After 24 h, the population of cells expressing MHC class II in the infundibulum was significantly higher in laying hens inseminated with fresh semen than in the control hens sham-inseminated with saline (P < 0.05). However, there was no significant difference in the population of cells expressing MHC class II in the uterovaginal junction and vagina between the artificially inseminated and control hens. These results indicate that anti-sperm immune responses, including the influx of cells expressing MHC class II and enhanced MHC class II mRNA expression, probably occur in the infundibulum after artificial insemination.

Animals↗

PubMed2000

Ultra-hydrophobic fluorine polymer by Ar-ion bombardment.

A study has been made of the improvement of the hydrophobicity of a fluorine-polymer (polytetra-fluorinethylene, PTFE) by Ar-ion bombardment. Ar-ion bombardment to PTFE) by Ar-ion bombardment. Ar-ion bombardment to PTFE sheets was carried out at the fluences ranging from 3.1x10(20) to 18.3x10(20) ions/m(2). The beam current densities were ranged from 1.25 to 7.5 A/m(2). The hydrophobicity of modified materials has been investigated by means of a sessile drop method of water. The roughness, morphology and structure of bombarded specimens have been estimated as a function of beam current density and acceleration energy. It was found that Ar-bombardment causes the contact angle of water to increase from the original value 103 to 170 degrees. The results indicate that Ar-bombardment at an optional condition results in the formation of ultrahydrdophobic surfaces. The effects of Ar-bombardment to hydrophobicity of PTFE are discussed with the relationship among roughness, morphology and chemical bonding states of the surfaces.

Journal Article↗

PubMed2000

Determination of bisphenol A in human serum by high-performance liquid chromatography with multi-electrode electrochemical detection.

A simple and sensitive method using high-performance liquid chromatography with multi-electrode electrochemical detection (HPLC-ED) including a coulometric array of four electrochemical sensors has been developed for the determination of bisphenol A in water and human serum. For good separation and detection of bisphenol A, a CAPCELL PAK UG 120 C18 reversed-phase column and a mobile phase consisting of 0.3% phosphoric acid-acetonitrile (60:40) were used. The detection limit obtained by the HPLC-ED method was 0.01 ng/ml (0.5 pg), which was more than 3000-times higher than the detection limit obtained by the ultraviolet (UV) method, and more than 200-times higher than the detection limit obtained by the fluorescence (FL) method. Bisphenol A in water and serum samples was pretreated by solid-phase extraction (SPE) after removing possible contamination derived from a plastic SPE cartridges and water used for the pretreatment. A trace amount (ND approximately 0.013 ng/ml) of bisphenol A was detected from the parts of cartridges (filtration column, sorbent bed and frits) by extraction with methanol, and it was completely removed by washing with at least 15 ml of methanol in the operation process. The concentrations of bisphenol A in tap water and Milli-Q-purified water were found to be 0.01 and 0.02 ng/ml, respectively. For that reason, bisphenol A-free water was made to trap bisphenol A in water using an Empore disk. In every pretreatment, SPE methods using bisphenol A-free water and washing with 15 ml of methanol were done in water and serum samples. The yields obtained from the recovery tests using water to which 0.5 or 0.05 ng/ml of bisphenol A was added were 83.8 to 98.2%, and the RSDs were 3.4 to 6.1%, respectively. The yields obtained from the recovery tests by OASIS HLB using serum to which 1.0 ng/ml or 0.1 ng/ml of bisphenol A was added were 79.0% and 87.3%, and the RSDs were 5.1% and 13.5%, respectively. The limits of quantification in water and serum sample were 0.01 ng/ml and 0.05 ng/ml, respectively. The method was applied to the determination of bisphenol A in healthy human serum sample, and the obtained detection was 0.32 ng/ml. From these results, the HPLC-ED method should be the most useful in the determination of bisphenol A at low concentration levels in water and biological samples.

Benzhydryl Compounds↗

PubMed2000

Ca(2+)-independent activity of Ca(2+)/calmodulin-dependent protein kinase II involved in stimulation of neurite outgrowth in neuroblastoma cells.

We investigated the involvement of Ca(2+)-independent activity of Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) in stimulation of neurite outgrowth. When neuroblastoma Neruo2a (Nb2a) cells expressing the alpha isoform of CaM kinase II (Nb2a/alpha cells) were stimulated by plating, they changed shape from round to flattened, and began to form neurites within 15 min. Numbers of cells bearing neurites increased from 15 min to about 2 h. Neurite length increased markedly from 30 min to 2 h after stimulation. Ca(2+)-independent activity of CaM kinase II increased immediately after stimulation, peaked at about 30 min, and then gradually decreased. Autophosphorylation of Thr-286 followed the same time course as the increase in Ca(2+)-independent activity. The autophosphorylation and appearance of Ca(2+)-independent activity preceded the formation of neurites. The effect of mutation of the autophosphorylation site in the kinase whose Thr-286 was replaced with Ala (alphaT286A kinase) or Asp (alphaT286D kinase) was examined. alphaT286A kinase was not converted to a Ca(2+)-independent form, and alphaT286D kinase had Ca(2+)-independent activity significantly as an autophosphorylated kinase. Cells expressing alphaT286A kinase did not form neurites, and were indistinguishable from control Nb2a cells. Cells expressing alphaT286D kinase had much longer neurites than Nb2a/alpha cells expressing the wild type kinase, although the initiation of neurite outgrowth was very late. These results indicated that Ca(2+)-independent activity of the kinase autophosphorylated at Thr-286 involves for neurite outgrowth.

Animals↗

PubMed2000

Immunocytochemical study of cell proliferation in the cystic ovarian follicles in cows.

We examined the frequency of proliferating cells in cystic, atretic and healthy antral follicles to determine whether a disorder of cell proliferation was responsible for the occurrence of bovine cystic follicles. Paraffin sections of healthy follicles and various stages of atretic and cystic follicles were immunostained with mouse monoclonal antibody to proliferating cell nuclear antigen (PCNA). The PCNA-positive cells were counted in 4 different regions of a follicle from the apical to the basal side. In the granulosa layer, a significantly higher frequency of PCNA-positive cells was observed in the healthy follicle in the basal region as compared with the apical region. A similar pattern of PCNA-positive cells population was observed in the granulosa layer of atretic follicles, although the frequency in the basal region was significantly lower in the atretic than the healthy follicle. The rate of cell proliferation in the granulosa layer of cystic follicles was markedly lower at the basal region than that of atretic follicles. In the theca interna, the frequency of PCNA-positive cells in atretic follicles at the early stages was higher than that in cystic follicles at the early stages. These results suggest that in the healthy follicle the proliferative activity of granulosa cells is higher in the basal than the apical region, and that the cell proliferation activity in the granulosa and theca interna may decrease in association with the induction of a follicular cyst.

Animals↗

PubMed2000

Activity-dependent maintenance of long-term potentiation at visual cortical inhibitory synapses.

Neural activity producing a transient increase in intracellular Ca(2+) concentration can induce long-term potentiation (LTP) at visual cortical inhibitory synapses similar to those seen at various excitatory synapses. Here we report that low-frequency neural activity is required to maintain LTP at these inhibitory synapses. Inhibitory responses of layer 5 cells evoked by layer 4 stimulation were studied in developing rat visual cortical slices under a pharmacological blockade of excitatory synaptic transmission using intracellular and whole-cell recording methods. Although LTP induced by high-frequency stimulation (HFS) persisted while test stimulation was applied at 0.1 Hz, it was not maintained in approximately two-thirds of cells after test stimulation was stopped for 30 min. In the rest of the cells, LTP seemed to be maintained by spontaneous presynaptic spikes, because presynaptic inhibitory cells discharged spontaneously in our experimental condition and because LTP was totally abolished by a temporary application of Na(+) channel blockers. Experiments applying various Ca(2+) channel blockers and Ca(2+) chelators after HFS demonstrated that LTP maintenance was mediated by presynaptic Ca(2+) entries through multiple types of high-threshold Ca(2+) channels, which activated Ca(2+)-dependent reactions different from those triggering transmitter release. The Ca(2+) entries associated with action potentials seemed to be regulated by presynaptic K(+) channels, presumably large-conductance Ca(2+)-activated K(+) channels, because the application of blockers for these channels facilitated LTP maintenance. In addition, noradrenaline facilitated the maintenance of LTP. These findings demonstrate a new mechanism by which neural activity regulates the continuation and termination of LTP at visual cortical inhibitory synapses.

Action Potentials↗

PubMed2000

Investigation of protein substrates of Ca(2+)/calmodulin-dependent protein kinase II translocated to the postsynaptic density.

To elucidate the physiological significance of the translocation of Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II), we investigated substrates of CaM kinase II in the postsynaptic density (PSD). PSD proteins were phosphorylated by CaM kinase II of its PSD complex, and separated by two-dimensional gel electrophoresis. More than 28 proteins were phosphorylated under experimental conditions. Proteins corresponding to CaM kinase II substrates were excised from the gels, eluted electrophoretically, and then sequenced. Several substrates were identified, including PSD95, SAP90, alpha-internexin, neurofilament L chain, cAMP phosphodiesterase, and alpha- and beta-tubulin. Some substrates were also identified by immunoblotting, including N-methyl-D-aspartic acid (NMDA) receptor 2B subunit, 1-alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptor 1 (GluR1), neurofilament H chain and dynamin. PSD95, SAP90, dynamin, and alpha-internexin were demonstrated for the first time to be substrates of CaM kinase II. NMDA receptor 2B subunit and GluR1 existed as major substrates in the PSD. Moreover, translocation of CaM kinase II was inhibited by phosphorylation of PSD proteins. These results suggest that CaM kinase II plays important roles in the regulation of synaptic functions through phosphorylation of PSD proteins.

Amino Acid Sequence↗

PubMed2000

Low calcium environment effects osteoprotegerin ligand/osteoclast differentiation factor.

In coculture with osteoblastic cell line MC3T3-E1 (E1) and mouse bone marrow cells, we reported that numbers of osteoclasts rose significantly on exposure to a low-calcium environment. Here we examined how osteoblasts influence osteoclastogenesis under a low-calcium environment. Comparing low extracellular calcium with a regular calcium environment, osteoprotegerin ligand (OPGL)/osteoclast differentiation factor (ODF) mRNA expression show more increase in the culture of low-calcium environment than in that of a regular calcium environment. Calcium-sensing receptor (CaSR), which was supposed as one of the mechanisms of recognizing extracellular calcium, existedon the surface of E1 cells. When E1 cells stimulated with agonists of CaSR, gadolinium, and neomycin, OPGL/ODF mRNA expression decreased. Moreover, these agonists reduced osteoclast formation in coculture. Taken together, it is possible that osteoblasts may recognize extracellular calcium via CaSR and regulate osteoclastogenesis.

3T3 Cells↗