Abstract
Glycosaminoglycans (GAG) from the velvet antlers of Sika deer (Cervus nippon) at the different growing stages (Fukurozuno, Anshi, and Santajo) of bred and wild deer were isolated and their concentrations and sulfation patterns were analyzed. GAG were digested with chondroitinase ABC, ACI, heparinase-I and -III, and keratanase-II into the corresponding repeating disaccharides of chondroitin sulfate (CS), dermatan sulfate (DS), hyaluronan, heparan sulfate (HS), and keratan sulfate. Cartilaginous tissues contained CS-DS at high concentrations with an almost equal ratio of 4- and 6-sulfates, while 4-sulfate-type CS-DS predominantly occupied ossified tissues, but at low concentrations. High O- and N-sulfation degrees of HS correspond to high ossification. Dynamic quantitative changes in CS-DS and compositional changes in CS-DS and HS were closely associated with the mineralization of deer antlers.
Similar content being viewed by others
References
Dinsmore, C.E., Goss, R.J., Lenz, M.E., Thonar, E.J.-M.A.: Correlation between phases of deer antler regeneration and levels of serum keratin sulfate. Calcif. Tissue Int. 39, 244–247 (1986)
Zhao, Q.-C., Kiyohara, H., Nagai, T., Yamada, H.: Structure of the complement-activating proteoglycan from the pilose antler of Cervus nippon Temminck. Carbohydr. Res. 230, 361–372 (1992)
Sunwoo, H.H., Nakano, T., Hudson, R.J., Sim, J.S.: Chemical composition of antlers from wapiti (Cervus elaphus). J. Agric. Food Chem. 43, 2846–2849 (1995)
Sunwoo, H.H., Nakano, T., Hudson, R.J., Sim, J.S.: Isolation, characterization and localization of glycosaminoglycans in growing antlers of wapiti (Cervus elaphus). Comp. Biochem. Physiol. Part B. 120, 273–283 (1998)
Ha, Y.W., Jeon, B.T., Moon, S.H., Toyoda, H., Toida, T., Linhardt, R.J., Kim, Y.S.: Characterization of heparin sulfate from the unossified antler of Cervus elaphus. Carbohydr. Res. 340, 411–416 (2005)
Pothacharoen, P., Kodchakorn, K., Kongtawelert, P.: Characterization of chondroitin sulfate from deer tip antler and osteogenic properties. Glycoconj. J. 28, 473–480 (2011)
Arima, K., Fujita, H., Toita, R., Imazu-Okada, A., Tsutsumishita-Nakai, N., Takeda, N., Nakao, Y., Wang, H., Kawano, M., Matsushita, K., Tanaka, H., Morimoto, S., Nakamura, A., Kitagaki, M., Hieda, Y., Hatto, R., Watanabe, A., Yumura, T., Okuhara, T., Hayashi, H., Shimizu, K., Nakayama, K., Masuda, S., Ishihara, Y., Yoshioka, S., Yoshioka, S., Shirade, S., Tamura, J.I.: Amounts and compositional analysis of glycosaminoglycans in the tissue of fish. Carbohydr. Res. 366, 25–32 (2013)
Yoshida, K., Miyauchi, S., Kikuchi, H., Tawada, A., Tokuyasu, K.: Analysis of unsaturated disaccharides from glycosaminoglycuronan by high-performance liquid chromatography. Anal. Biochem. 177, 327–332 (1989)
Hjerpe, A., Antonopoulos, C.A., Engfeldt: Determination of hyaluronic acid using high-performance liquid chromatography chondroitinase digests. J. Chromatogr. 245, 365–368 (1982)
Mizumoto, S., Sugahara, K.: Glycosaminoglycan chain analysis and characterization (glycosylation/epimerization) “proteoglycans: methods and protocols”. Methods Mol. Biol. 836, 99–115 (2012)
Uchimura, K.: Keratran sulfate: biosynthesis, structures, and biological functions. Methods Mol. Biol. 1229, 389–400 (2015)
Zhang, Z., Ohtake-Niimi, S., Kadomatsu, K., Uchimura, K.: Reduced molecular size and altered disaccharide composition of cerebral chondroitin sulfate upon Alzheimer’s pathogenesis in mice. Nagoya J. Med. Sci. 78, 293–301 (2016)
Takeda, N., Horai, S., Tamura, J.: Facile analysis of contents and compositions of the chondroitin sulfate/dermatan sulfate hybrid chain in shark and ray tissues. Carbohydr. Res. 424, 54–58 (2016)
Harab, R.C., Mourão, P.A.S.: Increase of chondroitin 4-sulfate concentration in the endochondral ossification cartilage of normal dogs. Biochim. Biophys. Acta. 992, 237–240 (1989)
Robinson, H.C., Dorfman, A.: The sulfation of chondroitin sulfate in embryonic chick cartilage epiphyses. J. Biol. Chem. 244, 348–352 (1969)
Kitagawa, H., Tsutsumi, K., Tone, Y., Sugahara, K.: Developmental regulation of the sulfation profile of chondroitin sulfate chains in the chicken embryo brain. J. Biol. Chem. 272, 31377–31381 (1997)
Habuchi, H., Kimata, K., Suzuki, S.: Changes in proteoglycan composition during development of rat skin. The occurrence in fetal skin of a chondroitin sulfate proteoglycan with high turnover rate. J. Biol. Chem. 261, 1031–1040 (1986)
Roughly, P.J., White, R.J.: Age-related changes in the structure of the proteoglycan subunits from human articular cartilage. J. Biol. Chem. 255, 217–224 (1980)
Acknowledgements
We are grateful to Mr. Tsuyoshi Kawato for the kind gift of wild velvet antlers. We also thank Amano Enzyme Inc. (Nagoya, Japan) for kindly gifting PROTIN NY100. The authors thank Dr. Kenji Uchimura (UMR8576-CNRS, University of Lille), Ms. Kwon-Jung Yi (Konkuk University), Mr. Takamu Fujii, Ms. Yukino Ito (Meijo University), Mr. Yuga Inoue, Ms. Asumi Uemura, Mr. Daiki Sugita, Ms. Yuna Uemura, Ms. Yuka Omura, Ms. Akari Tani, and Ms. Marin Mitani (Tottori University) for their helpful assistance.
Author information
Authors and Affiliations
Contributions
N.T.-O. performed the preparation of glycans and analyzed CS/DS and HA. S.M. and S.Y. performed HS analyses. Z.Z. and K.K. performed KS analyses. S.-K.K., C.-H.L., and B.-T.J. collected deer antlers. Y.Z.H. performed histological examinations. J.T. designed the study and wrote the manuscript.
Corresponding author
Ethics declarations
Conflicts of interest
The authors declare that they have no conflicts of interests.
Ethical approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Takeda-Okuda, N., Mizumoto, S., Zhang, Z. et al. Compositional analysis of the glycosaminoglycan family in velvet antlers of Sika deer (Cervus nippon) at different growing stages. Glycoconj J 36, 127–139 (2019). https://doi.org/10.1007/s10719-019-09859-1
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10719-019-09859-1