PURPOSE: Hyaluronic acid (HA) is the predominant glycosaminoglycan (GAG) of the human vitreous. Interaction of this HA with vitreous collagen is important for maintaining gel structure. The mechanism of HA homeostasis in the vitreous is incompletely understood. The aim of this study was to determine whether hyaluronidase, an endoglycosidase that degrades HA, was present in human vitreous. METHODS: Vitreous samples were collected from post-mortem eye bank specimens and from non-hemorrhagic, non-inflamed biopsy specimens. Vitreous hyaluronidase was purified by a series of column chromatographic steps, and its activity was measured by an ELISA-like assay and by substrate gel electrophoresis through and HA-impregnated gel. The purified hyaluronidase was also analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and by Western blotting. RESULTS: Hyaluronidase activity was detected in vitreous samples from both post-mortem and biopsy specimens. The enzyme was most active at acid pH, but demonstrated significant activity at neutral pH. The partially purified enzyme migrated as a 59 kDa protein on SDS-PAGE, and a single band on Western blots. CONCLUSIONS: Hyaluronidase is present in the human vitreous. Thus, hyaluronidase may be involved in HA catabolism in the vitreous and may play a role in determining its gel structure.
PURPOSE:Hyaluronic acid (HA) is the predominant glycosaminoglycan (GAG) of the human vitreous. Interaction of this HA with vitreous collagen is important for maintaining gel structure. The mechanism of HA homeostasis in the vitreous is incompletely understood. The aim of this study was to determine whether hyaluronidase, an endoglycosidase that degrades HA, was present in human vitreous. METHODS: Vitreous samples were collected from post-mortem eye bank specimens and from non-hemorrhagic, non-inflamed biopsy specimens. Vitreous hyaluronidase was purified by a series of column chromatographic steps, and its activity was measured by an ELISA-like assay and by substrate gel electrophoresis through and HA-impregnated gel. The purified hyaluronidase was also analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and by Western blotting. RESULTS: Hyaluronidase activity was detected in vitreous samples from both post-mortem and biopsy specimens. The enzyme was most active at acid pH, but demonstrated significant activity at neutral pH. The partially purified enzyme migrated as a 59 kDa protein on SDS-PAGE, and a single band on Western blots. CONCLUSIONS: Hyaluronidase is present in the human vitreous. Thus, hyaluronidase may be involved in HA catabolism in the vitreous and may play a role in determining its gel structure.
Authors: Gregor Cornelius Weber; Bettina Alexandra Buhren; Holger Schrumpf; Johannes Wohlrab; Peter Arne Gerber Journal: Adv Exp Med Biol Date: 2019 Impact factor: 2.622
Authors: Pierre C Dromel; Deepti Singh; Eliot Andres; Molly Likes; Motoichi Kurisawa; Alfredo Alexander-Katz; Myron Spector; Michael Young Journal: NPJ Regen Med Date: 2021-12-20
Authors: Soohyun Kim; Jennifer J Kang-Mieler; Wenqiang Liu; Zhe Wang; Glenn Yiu; Leandro B C Teixeira; William F Mieler; Sara M Thomasy Journal: Transl Vis Sci Technol Date: 2020-02-27 Impact factor: 3.283
Authors: Stefaniya Konstantinova Boneva; Julian Wolf; Dennis-Dominik Rosmus; Anja Schlecht; Gabriele Prinz; Yannik Laich; Myriam Boeck; Peipei Zhang; Ingo Hilgendorf; Andreas Stahl; Thomas Reinhard; James Bainbridge; Günther Schlunck; Hansjürgen Agostini; Peter Wieghofer; Clemens A K Lange Journal: Front Immunol Date: 2020-09-11 Impact factor: 7.561