Versican and fibrillin-1 form a major hyaluronan-binding complex in the ciliary body

Akiko Ohno-Jinno, Zenzo Isogai, Masahiko Yoneda, Kenji Kasai, Osamu Miyaishi, Yoko Inoue, Takuya Kataoka, Jing Song Zhao, Huili Li, Masayuki Takeyama, Douglas R. Keene, Lynn Sakai, Koji Kimata, Masayoshi Iwaki, Masahiro Zako

Research output: Contribution to journalArticle

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Abstract

PURPOSE. In this study, biochemistry, molecular biology, immunohistochemistry, and electron microscopy techniques were used to examine whether versican, which is known to bind fibrillin-1, interacts with fibrillin-1 in the ciliary body and vitreous, and whether the versican in this complex binds to hyaluronan. METHODS. The new polyclonal antibodies against the amino and carboxyl termini of versican were raised and characterized. The mRNA expression levels of versican and fibrillin-1 were analyzed by RT-PCR and real-time PCR, and their protein levels were evaluated by Western blot analysis and immunohistochemistry. Isolation of versican bound to fibrillin-1-containing microfibrils from ciliary bodies was performed by extraction studies. Slot-blot analyses and rotary shadowing electron microscopy were applied to identify versican associated with fibrillin-1-containing microfibrils after gel filtration chromatography and density gradient centrifugation. RESULTS. The newly prepared polyclonal antibodies recognized amino and carboxyl termini of chicken versican. Versican, principally V0 and V1, was found to be securely bound to fibrillin-1-containing microfibrils, forming a major hyaluronan-binding structure in the ciliary nonpigmented epithelium. In addition, Western blot analysis revealed two cleaved complexes, the carboxyl-terminal end of versican bound to fibrillin microfibrils and the amino terminal end of versican bound to hyaluronan in the vitreous body. CONCLUSIONS. Fibrillin-1, versican, and hyaluronan form a unique complex in the ciliary nonpigmented epithelium, and two cleavage products of this complex were shown to exist in the vitreous body. This newly clarified fibrillin-versican-hyaluronan (FiVerHy) complex and its cleavage products may be indispensable for the physiological properties important to the ciliary body and vitreous.

Original languageEnglish (US)
Pages (from-to)2870-2877
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume49
Issue number7
DOIs
StatePublished - Jul 1 2008

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Versicans
Ciliary Body
Hyaluronic Acid
Microfibrils
Vitreous Body
Fibrillin-1
Electron Microscopy
Epithelium
Western Blotting
Immunohistochemistry
Density Gradient Centrifugation
Antibodies
Biochemistry
Gel Chromatography

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience
  • Medicine(all)

Cite this

Ohno-Jinno, A., Isogai, Z., Yoneda, M., Kasai, K., Miyaishi, O., Inoue, Y., ... Zako, M. (2008). Versican and fibrillin-1 form a major hyaluronan-binding complex in the ciliary body. Investigative Ophthalmology and Visual Science, 49(7), 2870-2877. https://doi.org/10.1167/iovs.07-1488

Versican and fibrillin-1 form a major hyaluronan-binding complex in the ciliary body. / Ohno-Jinno, Akiko; Isogai, Zenzo; Yoneda, Masahiko; Kasai, Kenji; Miyaishi, Osamu; Inoue, Yoko; Kataoka, Takuya; Zhao, Jing Song; Li, Huili; Takeyama, Masayuki; Keene, Douglas R.; Sakai, Lynn; Kimata, Koji; Iwaki, Masayoshi; Zako, Masahiro.

In: Investigative Ophthalmology and Visual Science, Vol. 49, No. 7, 01.07.2008, p. 2870-2877.

Research output: Contribution to journalArticle

Ohno-Jinno, A, Isogai, Z, Yoneda, M, Kasai, K, Miyaishi, O, Inoue, Y, Kataoka, T, Zhao, JS, Li, H, Takeyama, M, Keene, DR, Sakai, L, Kimata, K, Iwaki, M & Zako, M 2008, 'Versican and fibrillin-1 form a major hyaluronan-binding complex in the ciliary body', Investigative Ophthalmology and Visual Science, vol. 49, no. 7, pp. 2870-2877. https://doi.org/10.1167/iovs.07-1488
Ohno-Jinno, Akiko ; Isogai, Zenzo ; Yoneda, Masahiko ; Kasai, Kenji ; Miyaishi, Osamu ; Inoue, Yoko ; Kataoka, Takuya ; Zhao, Jing Song ; Li, Huili ; Takeyama, Masayuki ; Keene, Douglas R. ; Sakai, Lynn ; Kimata, Koji ; Iwaki, Masayoshi ; Zako, Masahiro. / Versican and fibrillin-1 form a major hyaluronan-binding complex in the ciliary body. In: Investigative Ophthalmology and Visual Science. 2008 ; Vol. 49, No. 7. pp. 2870-2877.
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abstract = "PURPOSE. In this study, biochemistry, molecular biology, immunohistochemistry, and electron microscopy techniques were used to examine whether versican, which is known to bind fibrillin-1, interacts with fibrillin-1 in the ciliary body and vitreous, and whether the versican in this complex binds to hyaluronan. METHODS. The new polyclonal antibodies against the amino and carboxyl termini of versican were raised and characterized. The mRNA expression levels of versican and fibrillin-1 were analyzed by RT-PCR and real-time PCR, and their protein levels were evaluated by Western blot analysis and immunohistochemistry. Isolation of versican bound to fibrillin-1-containing microfibrils from ciliary bodies was performed by extraction studies. Slot-blot analyses and rotary shadowing electron microscopy were applied to identify versican associated with fibrillin-1-containing microfibrils after gel filtration chromatography and density gradient centrifugation. RESULTS. The newly prepared polyclonal antibodies recognized amino and carboxyl termini of chicken versican. Versican, principally V0 and V1, was found to be securely bound to fibrillin-1-containing microfibrils, forming a major hyaluronan-binding structure in the ciliary nonpigmented epithelium. In addition, Western blot analysis revealed two cleaved complexes, the carboxyl-terminal end of versican bound to fibrillin microfibrils and the amino terminal end of versican bound to hyaluronan in the vitreous body. CONCLUSIONS. Fibrillin-1, versican, and hyaluronan form a unique complex in the ciliary nonpigmented epithelium, and two cleavage products of this complex were shown to exist in the vitreous body. This newly clarified fibrillin-versican-hyaluronan (FiVerHy) complex and its cleavage products may be indispensable for the physiological properties important to the ciliary body and vitreous.",
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T1 - Versican and fibrillin-1 form a major hyaluronan-binding complex in the ciliary body

AU - Ohno-Jinno, Akiko

AU - Isogai, Zenzo

AU - Yoneda, Masahiko

AU - Kasai, Kenji

AU - Miyaishi, Osamu

AU - Inoue, Yoko

AU - Kataoka, Takuya

AU - Zhao, Jing Song

AU - Li, Huili

AU - Takeyama, Masayuki

AU - Keene, Douglas R.

AU - Sakai, Lynn

AU - Kimata, Koji

AU - Iwaki, Masayoshi

AU - Zako, Masahiro

PY - 2008/7/1

Y1 - 2008/7/1

N2 - PURPOSE. In this study, biochemistry, molecular biology, immunohistochemistry, and electron microscopy techniques were used to examine whether versican, which is known to bind fibrillin-1, interacts with fibrillin-1 in the ciliary body and vitreous, and whether the versican in this complex binds to hyaluronan. METHODS. The new polyclonal antibodies against the amino and carboxyl termini of versican were raised and characterized. The mRNA expression levels of versican and fibrillin-1 were analyzed by RT-PCR and real-time PCR, and their protein levels were evaluated by Western blot analysis and immunohistochemistry. Isolation of versican bound to fibrillin-1-containing microfibrils from ciliary bodies was performed by extraction studies. Slot-blot analyses and rotary shadowing electron microscopy were applied to identify versican associated with fibrillin-1-containing microfibrils after gel filtration chromatography and density gradient centrifugation. RESULTS. The newly prepared polyclonal antibodies recognized amino and carboxyl termini of chicken versican. Versican, principally V0 and V1, was found to be securely bound to fibrillin-1-containing microfibrils, forming a major hyaluronan-binding structure in the ciliary nonpigmented epithelium. In addition, Western blot analysis revealed two cleaved complexes, the carboxyl-terminal end of versican bound to fibrillin microfibrils and the amino terminal end of versican bound to hyaluronan in the vitreous body. CONCLUSIONS. Fibrillin-1, versican, and hyaluronan form a unique complex in the ciliary nonpigmented epithelium, and two cleavage products of this complex were shown to exist in the vitreous body. This newly clarified fibrillin-versican-hyaluronan (FiVerHy) complex and its cleavage products may be indispensable for the physiological properties important to the ciliary body and vitreous.

AB - PURPOSE. In this study, biochemistry, molecular biology, immunohistochemistry, and electron microscopy techniques were used to examine whether versican, which is known to bind fibrillin-1, interacts with fibrillin-1 in the ciliary body and vitreous, and whether the versican in this complex binds to hyaluronan. METHODS. The new polyclonal antibodies against the amino and carboxyl termini of versican were raised and characterized. The mRNA expression levels of versican and fibrillin-1 were analyzed by RT-PCR and real-time PCR, and their protein levels were evaluated by Western blot analysis and immunohistochemistry. Isolation of versican bound to fibrillin-1-containing microfibrils from ciliary bodies was performed by extraction studies. Slot-blot analyses and rotary shadowing electron microscopy were applied to identify versican associated with fibrillin-1-containing microfibrils after gel filtration chromatography and density gradient centrifugation. RESULTS. The newly prepared polyclonal antibodies recognized amino and carboxyl termini of chicken versican. Versican, principally V0 and V1, was found to be securely bound to fibrillin-1-containing microfibrils, forming a major hyaluronan-binding structure in the ciliary nonpigmented epithelium. In addition, Western blot analysis revealed two cleaved complexes, the carboxyl-terminal end of versican bound to fibrillin microfibrils and the amino terminal end of versican bound to hyaluronan in the vitreous body. CONCLUSIONS. Fibrillin-1, versican, and hyaluronan form a unique complex in the ciliary nonpigmented epithelium, and two cleavage products of this complex were shown to exist in the vitreous body. This newly clarified fibrillin-versican-hyaluronan (FiVerHy) complex and its cleavage products may be indispensable for the physiological properties important to the ciliary body and vitreous.

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