記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Biochimica Et Biophysica Acta General Subjects  

Background: Hyaluronan (HA) is a major component of the extracellular matrix (ECM) and is involved in many cellular functions. In the adult brain, HA forms macromolecular aggregates around synapses and plays important roles in neural plasticity. In contrast to the well-characterized function of HA in the adult brain, its roles in the developing brain remain largely unknown. Methods: Biochemical and histochemical analyses were performed to analyze the amount, solubility, and localization of HA in the developing mouse brain. By combining in utero labeling, cell isolation, and in vitro cultures, we examined the expression of hyaluronan synthase (HAS) and morphological maturation of cortical neurons. Results: The amount of HA increased during perinatal development and decreased in the adult. HA existed as a soluble form in the early stages; however, its solubility markedly decreased during postnatal development. HA localized in cell-sparse regions in the embryonic stages, but was broadly distributed during the postnatal development of the cerebral cortex. Developing cortical neurons expressed both Has2 and Has3, but not Has1, suggesting the autonomous production of HA by neurons themselves. HA formed a pericellular matrix around the cell bodies and neurites of developing cortical neurons, and the inhibition of HA synthesis reduced neurite outgrowth. Conclusion: The formation of the pericellular HA matrix is essential for the proper morphological maturation of developing neurons. General significance: This study provides new insights into the roles of hyaluronan in the brain. development.

DOI： 10.1016/j.bbagen.2020.129679

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：DNA and Cell Biology  

Mammary gland involution is a regressive process for the gland to return to its prepregnancy state after lactation and comprises an initial reversible and second remodeling stage. Although many genes and the multiple expression profiles of their mRNAs have been found in this process, the mechanisms controlling the profiles are largely unknown. In this study, we identified and analyzed transcription factor Sox4 in mammary gland involution. Elevated expression of Sox4 gene in the first stage (48 h after weaning) was observed at the mRNA and protein levels in the mouse mammary gland. Immunohistochemistry of the involuting gland indicated that Sox4 was located in the nuclei of epithelial cells. Nuclear Sox4 was also detected in the second stage, but unlikely to be involved in cell death, one of the characteristic events of involution. To clarify the functional roles of Sox4 in involution, we introduced a model, including a normal mammary epithelial cell line, for finding candidate target genes of this transcription factor and examined its effect on tenascin C mRNA expression.

DOI： 10.1089/dna.2019.4700

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Biochemistry  

Casein (CN) is themajor milk protein to nourish infants but, in certain population, it causes cow'smilk allergy, indicating the uptake of antigenic CN and their peptides through the intestinal epithelium. Using human intestinal Caco-2 cell monolayers, the apical-to-basal transepithelial transport ofCNwas investigated.Confocal microscopy using component- specific antibodies showed that as1-CN antigens became detectable as punctate signals at the apical-side cytoplasm and reached to the cytoplasm at a tight-junction level within a few hours. Such intracellular CN signals were more remarkable than those of the other antigens, b-lactoglobulin and ovalbumin, colocalized in part with an early endosome marker protein (EEA1) and decreased in the presence of cytochalasin D or sodium azide and also at lowered temperature at 4°C. Liquid chromatography coupled with mass spectroscopy analysis of the protein fraction in the basal-sidemedium identified the αs1-CB fragment including the N-terminal region and the αs2-CN fragment containing the central part of polypeptide at 100°1,000 fmol per well levels. Moreover, ß-CN C-terminal overlapping peptides were identified in the peptide fraction below 10 kDa of the basal medium. These results suggest that CNs are partially degraded by cellular proteases and/or peptidases and immunologically active CN fragments are transported to basal side of the cell monolayers.

DOI： 10.1093/jb/mvy034

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Cell Biochemistry and Function  

While transcription plays an obviously important role in gene expression, translation has recently been emerged as a key step that defines the composition and quality of the proteome in the cell of higher eukaryotes including mammals. Selective translation is supposed to be regulated by the structural heterogeneity of cytoplasmic ribosomes including differences in protein composition and chemical modifications. However, the current knowledge on the heterogeneity of mammalian ribosomes is limited. Here, we report mammalian Serbp1 as a ribosome-associated protein. The translated products of Serbp1 gene, including the longest isoform, were found to be localized in the nucleolus as well as in the cytoplasm. Subcellular fractionation indicated that most of cytoplasmic Serbp1 molecules were precipitated by ultracentrifugation. Proteomic analysis identified Serbp1 in the cytoplasmic ribosomes of the rodent testis. Polysome profiling suggested that Serbp1, as a component of the small 40S subunit, was included in translating ribosomes (polysomes). Cosedimentation of Serbp1 with the 40S subunit was observed after dissociation of the ribosomal subunits. Serbp1 was also included in the ribosomes of human cancer cells, which may lead to a mechanistic understanding of an emerging link between Serbp1 and tumour progression. Significance of the study: In mammalian cells, the final protein output of their genetic program is determined not only by controlling transcription but also by regulating the posttranscriptional events. Although mRNA-binding proteins and the cytoplasmic ribosome have long been recognized as central players in the posttranscriptional regulation, their physical and functional interactions are still far from a complete understanding. Here, we describe the intracellular localization of Serbp1, an mRNA-binding protein, and the inclusion of this protein in actively translating ribosomes in normal and cancer cells. These findings shed a new light into molecular mechanisms underlying Serbp1 action in translational gene regulation and tumour progression.

DOI： 10.1002/cbf.3350

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記述言語：日本語   掲載種別：研究論文（学術雑誌）  

記述言語：日本語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Genes to Cells  

Milk fat globule-EGF factor 8 (MFG-E8) is a divalent-binding secretory protein possessing an Arg-Gly-Asp (RGD) motif and a phosphatidylserine (PS)-binding motif. This protein has been shown to be involved in mammary gland development and morphogenesis. Integrin-binding activity is necessary for these MFG-E8-dependent cell processes. Although the target cells and molecules of MFG-E8 in the cellular microenvironment are important to understand its physiological function, its localization is largely unclear. Here, we found that mouse MFG-E8 localized to the basal lamina of the mammary gland during involution. In a model system of mammary COMMA-1D cells, exogenously and endogenously expressed MFG-E8 was deposited in the extracellular matrix (ECM) with membranous particles dependently on the PS-binding motifs in the discoidin domains that were essential for association ability to extracellular vesicles (EVs). These data showed the basal MFG-E8 localization mechanism in which EVs served as a scaffold. Such an immobilized MFG-E8 associating with cell substrata but not soluble one in the culture media promoted integrin-dependent suppression of β-casein expression. These results suggest that MFG-E8 requires EVs to transduce cellular signals from the basolateral side of the adhesion cells by accumulating in ECM.

DOI： 10.1111/gtc.12521

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：日本電気泳動学会  

<p>密度勾配超遠心分離とSDS-PAGEを用いて，マウス乳中に含まれる分泌膜小胞結合タンパク質，MFG-E8，の研究を行った．搾乳した新鮮なマウス乳に含まれる分泌膜小胞である乳脂肪球とエキソソームは密度勾配超遠心分離により，それぞれ低密度画分と高密度画分に分画され，SDS-PAGE免疫ブロット解析によりMFG-E8は主に乳脂肪球画分に検出された．このようなMFG-E8の分布は強制離乳により変化し，エキソソーム画分や可溶性タンパク質画分を含む幅広い密度範囲に分布した．乳脂肪球の蛍光免疫染色の結果などから，MFG-E8は分泌後間もない新鮮な乳脂肪球の表面には少なく時間経過とともに増加することが示された．MFG-E8の結合により乳脂肪球は貪食され易くなったことから，アポトーシス細胞と同様に，MFG-E8は古くなった乳脂肪球においてもeat-me signalとして機能することが示唆された．</p>

DOI： 10.2198/electroph.61.115

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1080/09168451.2015.1116927

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1074/jbc.M114.598821

Open Access

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1242/dev.121863

Open Access

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1095/biolreprod.114.119826

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1007/s11010-014-2128-x

記述言語：日本語  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1242/dev.102152

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1093/jb/mvt073

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1093/jb/mvt044

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.gene.2013.02.040

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： https://doi.org/10.1271/bbb.121011

Open Access

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1093/jb/mvs154

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Milk fat globules (MFGs) secreted by lactating mammary gland are unique lipid surrounded by a phospholipid bi-layer. We report here post-weaning changes in milk fat globule EGF factor VIII (MFG-E8) and annexin V-accessible phosphatidylserine on the surface of MFGs. The MFG content in milk markedly decreased to about half within 2 days after forced weaning, despite a slight increase in milk protein content. Immunofluorescence-staining of MFGs using anti-MFG-E8 and annexin V indicated that MFG-E8 was present on some, but not all, MFGs before weaning, whereas most of MFGs were MFG-E8-positive and annexin V-negative after weaning. Free MFG-E8 with binding activity to phosphatidylserine was present abundantly in the post-weaning milk, and indeed exhibited binding to MFGs in pre-weaning milk. MFGs were taken up by HC11 mouse mammary epithelial cells in vitro, and those from post-weaning milk were remarkable for such cellular uptake. Moreover, the uptake of MFGs by the cells was inhibited by an anti-MFG-E8 antibody. Taken together, these findings suggest that MFG-E8 plays a critical role in regulation of MFG dynamics after weaning or during the suckling interval through the control of MFG-epithelial cell interaction in lactating mammary glands.

DOI： 10.1093/jb/mvs116

記述言語：英語   掲載種別：研究論文（学術雑誌）  

In lactating mammary glands, milk fat is secreted as fat globules surrounded by a cell plasma membrane containing characteristic membrane-associated proteins. Among these, butyrophilin has been shown to be specific and intrinsic to the fat globule membrane, whereas milk-fat globule EGF-factor VIII (MFG-E8) is uncertain. We characterized in the present study MFG-E8 in milk fat globules and in the culture medium of HC11 mammary epithelial cells. MFG-E8 was immunologically detected in the mammary tissues of both pregnant and lactating mice. Double-immunofluorescence staining for MFG-E8 and butyrophilin showed diversity in the MFG-E8-staining intensity among different fat globules in milk. HC11 cells secreted monomeric MFG-E8 with phosphatidylserine-binding activity, despite no fat globules being detected in the cells. This secretion was upregulated by not only prolactin but also by insulin or EGF. These results suggest that milk MFG-E8 was not equally present among fat globules and not necessarily intrinsic to the fat globules.

DOI： 10.1271/bbb.120412

Open Access

記述言語：英語   掲載種別：研究論文（学術雑誌）  

... These results suggest that degraded but Dectin - 1 - reactive β- glucan is discharged from macrophage cells ... ROS -production inhibitors reduced soluble β- glucan discharge from the ... Soluble β- glucan produced by in vitro ROS - driven degradation activated macrophages . ...

DOI： 10.1016/j.bbrc.2012.04.009

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Microtubules form flexible fibers, which are utilized in cell proliferation and differentiation. Although the flexibility of microtubules was shown to be regulated by various microtubule-associated proteins, this regulation is still far from complete understanding. Here, we report a new potential regulator of microtubules in mammals. Gcap14 colocalizes with microtubules in mammalian cells transfected with Gcap14 expression vector. Association of Gcap14 with microtubules was confirmed by biochemical subcellular fractionation. Recombinant Gcap14 protein cosedimented with pure microtubules, indicating a direct binding between the two. Furthermore, recombinant Gcap14 was shown to have the ability of inducing microtubule bundling in vitro.

DOI： 10.1016/j.febslet.2012.04.008

担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1242/jcs.087700

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.devcel.2011.02.001

担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1242/dev.052605

担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.cub.2006.06.032

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1042/BJ20051459

記述言語：英語   掲載種別：研究論文（学術雑誌）  

MFG-E8 (milk fat globule-EGF factor 8) is a peripheral membrane glycoprotein, which is expressed abundantly in lactating mammary glands and is secreted in association with fat globules. This protein consists of two-repeated EGF-like domains, a mucin-like domain and two-repeated discoidin-like domains (C-domains), and contains an integrin-binding motif (RGD sequence) in the EGF-like domain. To clarify the role of each domain on the peripheral association with the cell membrane, several domain-deletion mutants of MFG-E8 were expressed in COS-7 cells. The immunofluorescent staining of intracellular and cell-surface proteins and biochemical analyses of cell-surface-biotinylated and secreted proteins demonstrated that both of the two C-domains were required for the membrane association. During the course of these studies for domain functions, MFG-E8, but not C-domain deletion mutants, was shown to be secreted as membrane vesicle complexes. By size-exclusion chromatography and ultracentrifugation analyses, the complexes were characterized to have a high-molecular mass, low density and higher sedimentation velocity and to be detergent-sensitive. Not only such a exogenously expressed MFG-E8 but also that endogenously expressed in a mammary epithelial cell line, COMMA-1D, was secreted as the membrane vesicle-like complex. Scanning electron microscopic analyses revealed that MFG-E8 was secreted into the culture medium in association with small membrane vesicles with a size from 100 to 200 nm in diameter. Furthermore, the expression of MFG-E8 increased the number of these membrane vesicle secreted into the culture medium. These results suggest a possible role of MFG-E8 in the membrane vesicle secretion, such as budding or shedding of plasma membrane (microvesicles) and exocytosis of endocytic multivesicular bodies (exosomes).

記述言語：英語   掲載種別：研究論文（学術雑誌）  

... Biochem., 64 (4), 785-792, 2000 M An ELISA - Based Assay for Detergent - solubilized Cellular {\ss

DOI： 10.1271/bbb.64.785

Open Access

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Expression of mRNA encoding MFG-E8, a milk fat-associated glycoprotein was investigated in mouse mammary gland. Two forms of mRNA, long and short variants, were shown to be expressed in the mammary tissue by RT-PCR analysis. Sequence analyses of these two variants and an isolated MFG-E8 gene segment indicated that the long and short mRNA variants resulted from an alternative splicing of a single pre-mRNA through in-flame inclusion and skipping of one exon, which encodes a proline/threonine (Pro/Thr)-rich domain. The long variant was expressed predominantly in mammary gland and the expression level was remarkably increased at late gestation and kept high during lactation. On the contrary, the short variant was detected ubiquitously in various tissues and its expression in the mammary gland was rather decreased in a lactation dependent manner. Expression of the long variant was also detected in a mouse mammary epithelial cell line, COMMA-1D, and enhanced by incubation with lactogenic hormones. Glycosylation inhibition analyses using tunicamycin and alpha-benzyl-GalNAc were conducted with COS7 cells transfected with plasmids expressing each mRNA variant, demonstrating that a fully glycosylated product of the long mRNA variant was not only N-glycosylated but also multiply O-glycosylated, whereas a product of the short one had only N-glycan(s). These results suggest that the alternative splicing plays a critical role for the mammary-specific and lactation-dependent expression of the MFG-E8 isoform and that the multiply O-glycosylated Pro/Thr-rich domain of this isoform is functionally important for formation of milk fat globules in mammary epithelial cells.

DOI： 10.1006/bbrc.1998.0107

記述言語：日本語   掲載種別：記事・総説・解説・論説等（学術雑誌）   出版者・発行元：日本医学館  

開催年月日： 2023年9月 - 2023年10月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：三重大学生物資源学部   国名：日本国  

開催年月日： 2023年5月

記述言語：日本語   会議種別：ポスター発表  

開催地：札幌コンベンションセンター   国名：日本国  

開催年月日： 2022年10月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京工科大学八王子キャンパス  

開催年月日： 2020年3月

記述言語：日本語  

開催年月日： 2019年

記述言語：英語   会議種別：口頭発表（一般）  

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開催年月日： 2018年10月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：広島大学広仁会館  

開催年月日： 2018年9月

記述言語：日本語   会議種別：ポスター発表  

開催地：名古屋大学豊田講堂  

開催年月日： 2016年11月 - 2016年12月

記述言語：日本語   会議種別：ポスター発表  

開催地：パシフィコ横浜   国名：日本国  

開催年月日： 2016年11月

記述言語：日本語   会議種別：公開講演，セミナー，チュートリアル，講習，講義等  

国名：日本国  

開催年月日： 2016年10月

記述言語：日本語   会議種別：ポスター発表  

開催地：昭和大学旗の台キャンパス   国名：日本国  

開催年月日： 2016年9月

記述言語：日本語   会議種別：口頭発表（一般）  

国名：日本国  

開催年月日： 2015年12月

記述言語：日本語   会議種別：ポスター発表  

国名：日本国  

開催年月日： 2015年12月

記述言語：日本語   会議種別：ポスター発表  

国名：日本国  

開催年月日： 2015年11月

記述言語：英語   会議種別：口頭発表（一般）  

国名：日本国  

開催年月日： 2013年12月

記述言語：日本語   会議種別：ポスター発表  

国名：日本国  

開催年月日： 2013年12月

記述言語：日本語   会議種別：ポスター発表  

国名：日本国  

開催年月日： 2011年9月

記述言語：英語   会議種別：口頭発表（招待・特別）  

国名：日本国  

開催年月日： 2009年9月

記述言語：日本語   会議種別：口頭発表（招待・特別）  

開催地：筑波大学   国名：日本国