Biomedical Research
Online ISSN : 1880-313X
Print ISSN : 0388-6107
ISSN-L : 0388-6107
36 巻 , 4 号
選択された号の論文の6件中1~6を表示しています
Full Papers
  • Ayuko KISHIMOTO, Toshina ISHIGURO-OONUMA, Ritei TAKAHASHI, Mamiko MAEK ...
    2015 年 36 巻 4 号 p. 225-234
    発行日: 2015/08/01
    公開日: 2015/08/22
    ジャーナル フリー
    Lactate represents a preferential energy substrate of germ cells rather than glucose. Testicular Sertoli cells are believed to produce lactate and pyruvate and to supply these to germ cells, particularly spermatocytes and spermatids. Monocarboxylate transporter (MCT), responsible for the transport of lactate and other monocarboxylates via the cell membrane, is abundant in the testes and sperm (MCT1, MCT2, and MCT4). For the uptake of glucose, germ cells within the seminiferous tubules and sperm have been known to intensely express GLUT3. The present study investigated expression profiles of MCTs and GLUTs and revealed their cellular and subcellular localization in the mouse and rat testis. An in situ hybridization analysis showed significant expressions of MCT1, MCT2, and GLUT3 mRNA in the testis. Immunohistochemically, spermatogonia, spermatocytes, and spermatids expressed MCT1 on their cell surfaces in a stage-dependent manner: in some seminiferous tubules, an intense expression of MCT1 was unique to the spermatogonia. MCT2 was restricted to the tails of elongated spermatids and sperm. An intense immunoreactivity for GLUT3 was shared by spermatocytes, spermatids, and sperm. Sertoli cells were devoid of any immunoreactivities for MCT1, MCT2, and GLUT3. The predominant energy source of germ cells may be lactate and other monocarboxylates—especially for spermatogonia, but glucose and other hexoses may be responsible for an energy supply to spermatocytes and spermatids.
  • Mohammad Khaja Mafij UDDIN, Wataru KIMURA, Tomoyuki ISHIKURA, Haruhik ...
    2015 年 36 巻 4 号 p. 235-245
    発行日: 2015/08/01
    公開日: 2015/08/22
    ジャーナル フリー
    The forkhead box C2 (Foxc2) protein is a member of the forkhead/winged helix transcription factor family and plays an essential role in cardiovascular development. Previous studies showed that Foxc2 null mouse embryos die during midgestation or just after birth with severe cardiovascular defects, including interruption, coarctation of the aortic arch and ventricular septal defects. These are also seen in human congenital heart disease. However, the tissue specific role of Foxc2 in aortic arch remodelling is not yet fully understood. Here we show that Foxc2 is expressed in a restricted pattern in several cell populations, including the mesenchyme and endothelium of pharyngeal arch arteries, which are important for cardiovascular development. In this study, we use a conditional knockout approach to examine the tissue specific role of Foxc2 in aortic arch remodelling. We demonstrate that mouse embryos lacking Foxc2 in Nkx2.5-expressing mesenchyme and endothelium of pharyngeal arch arteries display aortic arch interruption type B and ventricular septal defects. In contrast, conditional deletion of Foxc2 in Tie2-expressing endothelial cells does not result in aortic arch or ventricular septal defects, but does result in embryonic lethality due to peripheral oedema. Our data therefore provide for a detailed understanding of the role of mesenchymal Foxc2 in aortic arch remodelling and in the development of ventricular septum.
  • Shigenori ITO, Daisuke YAMAMOTO
    2015 年 36 巻 4 号 p. 247-252
    発行日: 2015/08/01
    公開日: 2015/08/22
    ジャーナル フリー
    The goal in this study was to clarify the color-change mechanisms of methyl orange (MO) bound to human serum albumin (HSA) and the structure of the binding site. The absorbance of the MOHSA complex was measured at 560 nm in solutions of varying pH (pH 2.4–6.6). The obtained pH-dependent experimental data were consistent with the data calculated using the Henderson-Hasselbalch equation and pKa values (3.8, MO; 1.4, carboxyl group). The extent of the binding of MO to an HSA molecule was determined to be 1–4 by performing surface plasmon resonance analysis. Furthermore, the binding of MO to HSA was inhibited by warfarin. A fitting model of MO to HSA was created to evaluate these results based on PDB data (warfarin-HSA complex: 2BXD) and protein-structure analysis. The color-change mechanism of the MO-HSA complex appears to be as follows: the dissociated sulfo group of MO binds to Arg218/Lys444 sidechains through electrostatic interaction in the warfarin-binding site, and, subsequently, the color change occurs through a proton exchange between the diazenyl group and the γ-carboxyl group of Glu292. The color-changed MO is fixed in the warfarin-binding site. These results could support the development of a reliable dye-binding method and of a new method for staining diverse tissues that is based on a validated mechanism.
  • Yoshinori TAOKA, Kazumasa MATSUMOTO, Kazuya OHASHI, Satoru MINAMID ...
    2015 年 36 巻 4 号 p. 253-261
    発行日: 2015/08/01
    公開日: 2015/08/22
    ジャーナル フリー
    We used a proteomic approach to compare the differentially regulated protein expression profiles of cisplatin-naïve and cisplatin-resistant bladder cancer cell lines to screen candidate molecules related to cisplatin resistance. The cisplatin-resistant cell line T24 was established by the stepwise exposure of T24 cells to up to 40 μM of cisplatin. We performed a comprehensive study of protein expression in bladder cancer cell lines that included cisplatin-naïve (T24) and cisplatin-resistant cells (T24CDDPR) by means of agarose two-dimensional gel electrophoresis followed by analysis of liquid chromatography tandem mass spectroscopy. We identified 25 obviously different spots for T24 and T24 CDDPR. Seven spots had increased expression and 18 spots had decreased expression in T24CDDPR compared to those in T24. Cytoskeletal proteins and enzyme modulators were prominent among differential proteins. Of the 25 proteins, we selected HNRNPA3, PCK2, PPL, PGK1, TKT, SERPINB2, GOT2, and EIF3A for further validation by Western blot. HNRNPA3, PGK1, TKT, and SERPINB2 had more than 1.5-times incremental expression in T24CDDPR compared to that in T24. PCK2 and PPL expressions were decreased less than 20% in T24CDDPR compared to that in T24. The results of 25 new proteins in this study could be valuable and could lead to the development of a new molecular marker.
  • Shunsuke KIMURA, Ayuko KISHIMOTO, Mami MUTOH, Hiromi TAKAHASHI-IWAN ...
    2015 年 36 巻 4 号 p. 263-272
    発行日: 2015/08/01
    公開日: 2015/08/22
    ジャーナル フリー
    GP2 is a membrane-associated secretory protein originally identified in zymogen granules of pancreatic acinar cells. Recently, this glycoprotein has attracted attention as a marker substance of M cells of Peyer’s patches and for its involvement in the selective uptake of pathological bacteria via M cells. When we stained the conjunctiva and tear ducts of mice using a GP2 antibody, all goblet cells in the squamous stratified epithelium of the conjunctiva were intensely immunolabeled, while goblet cells in the intestine and airway were devoid of the immunoreactivity, indicating that the conjunctiva contains a special type of goblet cell. Further immunostaining for GP-2 labeled dispersed cells of peculiar shapes within the stratified squamous epithelium in the lacrimal canaliculi, lacrimal sac, and nasolacrimal duct. The GP2-immunoreactive cells in the tear duct projected arched or branched processes toward the basement membrane. Electron-microscopically, immunogold particles for GP2 outlined the basolateral plasma membrane of both the conjuntival goblet cells and the peculiarly shaped cells in the tear duct. Intracellularly, GP2 products of the goblet cells were localized around secretory granules in the apical cytoplasm and those of the tear duct cells inside the vesicles. The luminal contents close to apical plasma membrane were heavily labeled with immunogold particles, suggesting an exocytosis-based targeting of GP2 to the plasma membrane and its release into the lumen. The possible function of GP2 in tear ducts is discussed in relation to a defense system against invasive microoranisms and antigens.
Communication
  • Ken YAMAGUCHI, Hirohisa KATAGIRI, Mitsuru TAKAHASHI, Yuji ISHIDA, Aki ...
    2015 年 36 巻 4 号 p. 273-277
    発行日: 2015/08/01
    公開日: 2015/08/22
    ジャーナル フリー
    We analyzed serum ProGRP levels in patients with Ewing sarcoma, and found that 5 out of 9 patients had elevated levels; the values range equally with those of patients with limited disease of small-cell lung carcinoma. Serum ProGRP levels in patients with bone and soft tissue malignancies other than Ewing sarcoma are not elevated. Immunohistochemical studies demonstrated that ProGRP-like immunoreactivities were detected in Ewing sarcoma tissues obtained from 2 patients with elevated serum ProGRP levels, suggesting that ProGRP is a product of tumor cells of Ewing sarcoma. These results indicate that serum ProGRP could serve as a specific tumor marker for Ewing sarcoma. Since ProGRP is a major hormonal product of tumor cells of small-cell lung carcinoma, a typical neuroendocrine carcinoma, it is reasonable to postulate that the present study provides an evidence for Ewing sarcoma to possess neuroendocrine differentiation.
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