Ultrast ructural study of cytoplasmic structures underlying lipid raft

2007 Fiscal Year Final Research Report Summary

• Project/Area Number: 17590156
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General anatomy (including Histology/Embryology)
• Research Institution: Ibaraki Prefectural University of Health Science (2006-2007); University of Yamanashi (2005)
• Principal Investigator: BABA Takeshi Ibaraki Prefectural University of Health Science, Department of Health Sciences, Professor (90208710)
• Project Period (FY): 2005 – 2007
• Keywords: lipid raft / cholera toxin / lysenin / immunological synapse / ultrastructure / immunocvtochemistry

Research Abstract

Recent studies have revealed that lipid composition of cell membranes are not uniform but heterogeneous mixture of various membrane microdomains such as lipid rafts. However, ultrastructure of cytoplasmic counterparts of those membrane microdomains on the cell surface and corresponding signal transducing molecules are not well understood.
In the present research period, we studied the ultrastructural distribution of various lipid raft markers and obtained the following results.
We successfully induced immunological synapse formation by incubating Jurkat cells with coverslips coated with anti-D3 antibody. Next, we studied the distribution of lipid raft markers during immunological synapse formation. The cells were incubated with 5m-colloidal gold-labeled lysenin and 10nm-colloidal gold-labeled cholera toxin at 4℃. Labeled cells were incubated with anti-D3 antibody-coated coverslips at 4℃, and warmed up to 37℃ to induce immunological synapse formation. Fluorescence microscopic observations using HV161-lysenin and Alexa594-labeled cholera toxin revealed that randomly distributed lipid raft markers at 4℃ were accumulated surrounding attached area after incubation at 37℃. Ultrastructural study have revealed that both marker were accumulated around the immunological synapse, but lysenin was also distributed on the attached area. These results suggested that redistribution of lipid rafts during immunological synapse formation differs according to lipid type.

Research Products

• [Journal Article] Immunohistochemical study of serum albumin in normal and cadmium-treated mouse testis organs by in vivo cryotechnique (2006)
  • Author(s): Liao X, Terada N, Ohno N, Ii Z, Fujii Y, Baba T, Ohno S.
  • Journal Title: Histol Histopathol 21(1) Pages: 35-40
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Morphological and histochemical analyses of living mouse livers by new cryobionsy technique (2006)
  • Author(s): Fujii Y, Ohno N, Ii Z, Terada N, Baba T, Ohno S.
  • Journal Title: J Electron Microsc(Tokyo) 55 Pages: 113-122
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Dynamic study of intramembranous particles in human fresh erythrocytes using an "in vitro cryotechnique" (2006)
  • Author(s): Terada N, Ohno N, Fujii Y, Baba T, Ohno S.
  • Journal Title: Microsc Res Tech 69 Pages: 291-295
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Application of in vivo cryotechnique to the examination of cells and tissues in living animal organs (2006)
  • Author(s): Terada N, Ohno N, Ii Z, Fujii Y, Baba T, Ohno S.
  • Journal Title: Histol Histopathol 21 Pages: 265-272
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Morphological and histochemical analyses of living mouse livers by new 'cryobiopsy' technique (2006)
  • Author(s): Fujii, Y., Ohno, N., Li, Z., Terada, N., Baba, T., Ohno, S
  • Journal Title: Morphological and histochemical analyses of living mouse livers by new 'cryobiopsy' technique 55 Pages: 113-122
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Dynamic study of intramembranous particles in human fresh erythrocytes using an "in vitro cryotechnique" (2006)
  • Author(s): Terada, N., Ohno, N., Fujii, Y., Baba, T., Ohno, S
  • Journal Title: Microsc Res Tech 69 Pages: 291-295
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Application of in vivo cryotechnique to the examination of cells and tissues in living animal organs (2006)
  • Author(s): Terada, N., Ohno, N., Li, Z., Fujii, Y., Baba, T., Ohno, S
  • Journal Title: Histol Histopathol 21 Pages: 265-272
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Detection of injected fluorescence-conjugated IgG in living mouse organs using in vivo cryotechnique with freeze-substitution (2005)
  • Author(s): Terada N, Ohno N, Ii Z, Fujii Y, Baba T, Ohno S.
  • Journal Title: Microsc Res Tech 66(4) Pages: 173-178
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Spatial and functional heterogeneity of sphingolipid-rich membrane domains (2005)
  • Author(s): Kiyokawa E, Baba T, Otsuka N, Makino A, Ohno S, Kobayashi T.
  • Journal Title: J Biol Chem 280(25) Pages: 24072-24084
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Immunohistochemical study of a membrane skeletal molecule,protein 4.1G,in mouse seminiferous tubules (2005)
  • Author(s): Terada N, Ohno N, Yamakawa H, Ohara O, Iiao X, Baba T, Ohno S.
  • Journal Title: Histochem Cell Biol 124(3-4) Pages: 303-311
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Protein 4.1 G localizes in rodent microglia (2005)
  • Author(s): Ohno N, Terada N, Tanaka J, Yokoyama A, Yamakawa H, Fujii Y, Baba T, Ohara O, Ohno S.
  • Journal Title: Histochem Cell Biol 124(6) Pages: 477-486
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Detection of injected fluorescence-conjugated IgG in living mouse organs using "in vivo cryotechnique" with freeze-substitution (2005)
  • Author(s): Terada, N., Ohno, N., Li, Z., Fujii, Y., Baba, T., Ohno, S
  • Journal Title: Microsc Res Tech 66(4) Pages: 173-178
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Spatial and functional heterogeneity of sphingolipid-rich membrane domains (2005)
  • Author(s): Kiyokawa, E., Baba, T., Otsuka, N., Makino, A., Ohno, S., Kobayashi, T
  • Journal Title: J Biol Chem 280(25) Pages: 24072-24084
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Immunohistochemical study of a membrane skeletal molecule, protein 4.1G, in mouse seminiferous tubules (2005)
  • Author(s): Terada, N., Ohno, N., Yamakawa, H., Ohara, O., Liao, X., Baba, T., Ohno, S
  • Journal Title: Histochem Cell Biol 124(3-4) Pages: 303-311
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Protein 4.1 G localizes in rodent microglia (2005)
  • Author(s): Ohno, N., Terada, N., Tanaka, J., Yokoyama, A., Yamakawa, H., Fujii, Y., Baba, T., Ohara, O., Ohno, S
  • Journal Title: Histochem Cell Biol 124(6) Pages: 477-486
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Immunohistochemical study of serum albumin in normal and cadmium-treated mouse testis organs by "in vivo cryotechnique" (2005)
  • Author(s): Liao, X., Terada, N., Ohno, N., Li, Z., Fujii, Y., Baba, T., Ohno, S
  • Journal Title: Histol Histopathol 21(1) Pages: 35-40
  • Description: 「研究成果報告書概要(欧文)」より