Expression of Syndecan-2,-4,and Fibroblast Growth Factor Receptor Type 1 in Human Periodontal Ligament Fibroblasts and Down-regulation of These Membrane Proteins during Maturation in Culture
Restricted accessResearch articleFirst published online December, 1999
Expression of Syndecan-2,-4,and Fibroblast Growth Factor Receptor Type 1 in Human Periodontal Ligament Fibroblasts and Down-regulation of These Membrane Proteins during Maturation in Culture
Syndecans are transmembrane heparan sulfate proteoglycans. They are known to interact with basic fibroblast growth factor (bFGF), and it has been suggested that they play important roles in the growth, morphology, and migration of a variety of cell types. We examined the expression of syndecans and fibroblast growth factor receptor type 1 (FGFR1) in periodontal ligament (PDL) cells, because these membrane proteins may play roles in the control of growth and differentiation during regeneration of PDL. Reverse-transcription/polymerase chain-reaction (RT-PCR) showed that PDL cells expressed syndecan-2 and -4 mRNAs. This was confirmed by sequence analysis of the PCR products. When PDL cells were maintained for 25 days, alkaline phosphatase (ALPase) activity gradually increased and reached a maximal level on day 20. Northern blotting analysis showed that PDL cells expressed 2.3-kb syndecan-2, 2.6-kb syndecan-4, and 2.8-kb FGFR1 mRNAs throughout the entire culture period, whereas no syndecan-1 mRNA was detectable by this method. Maximal levels of syndecan-2, -4, and FGFR1 mRNAs were observed on day 5. However, their levels were markedly decreased on days 20 and 25. Accordingly, the inhibitory effect of bFGF on ALPase activity was less on day 20 than on day 5. When PDL cells were pre-treated with heparitinase, a mitogenic response of PDL cells to bFGF was decreased. These observations indicate that PDL cells express syndecan-2, -4, and FGFR1 mRNAs, and that those levels are changed with the increase in ALPase activity in culture. The reductions in syndecan-2, -4, and FGFR1 levels may be involved in the control of growth and differentiation of PDL cells during development and regeneration.
Bai XM, Van der Schueren B., Cassiman JJ, Van den Berghe H., David G. (1994). Differential expression of multiple cell-surface heparan sulfate proteoglycans during embryonic tooth development. J Histochem Cytochem42:1043-1054.
2.
Bartold PM (1990). A biochemical and immunohistochemical study of the proteoglycans of alveolar bone. J Dent Res69:7-19. Basdra EK, Komposch G. (1997). Osteoblast-like properties of human periodontal ligament cells: an in vitro analysis. Eur J Orthod19:615-621.
3.
Bashkin P., Doctrow S., Klagsbrun M., Svahn CM, Folkman J., Vlodavsky I. (1989). Basic fibroblast growth factor binds to subendothelial extracellular matrix and is released by heparitinase and heparin-like molecules . Biochemistry28:1737-1743. Bemfield M., Kokenyesi R., Kato M., Hinkes MT, Spring J., Gallo RL, et al. (1992). Biology of the syndecans: a family of transmembrane heparan sulfate proteoglycans . Ann Rev Cell Biol8:365-393.
4.
Bessey O., Lowry O., Brock M. (1946). A method for the rapid determination of alkaline phosphatase with five cubic millimeters of serum. J Biol Chem164:321-329.
5.
Canalis E., Centrella M., McCarthy T. (1988). Effects of basic fibroblast growth factor on bone formation in vitro. J Clin Invest81:1572-1577.
6.
Chemousov MA, Carey DJ (1993). N-syndecan (syndecan 3) from neonatal rat brain binds basic fibroblast growth factor. J Biol Chem268:16810-16814.
7.
Chintala SK, Miller RR, McDevitt CA (1995). Role of heparan sulfate in the terminal differentiation of growth plate chondrocytes. Arch Biochem Biophys316:227-234.
8.
Chomczynski P., Sacchi N. (1987). Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem162:156-159.
9.
Coutts JC, Gallagher JT (1995). Receptors for fibroblast growth factors. Immunol Cell Biol73:584-589.
10.
David G., van der Schueren B., Marynen P., Cassiman JJ, van den Berghe H. (1992). Molecular cloning of amphiglycan, a novel integral membrane heparan sulfate proteoglycan expressed by epithelial and fibroblastic cells. J Cell Biol118:961-969. Elenius K., Jalkanen M. (1994). Function of the syndecans-a family of cell surface proteoglycans. J Cell Sci107:2975-2982.
11.
Gao J., Jordan TW, Cutress Tvv (1996). Immunolocalization of basic fibroblast growth factor (bFGF) in human periodontal ligament (PDL) tissue. J Periodont Res31:260-264.
12.
Grover J., Roughley PJ (1995). Expression of cell-surface proteoglycan mRNA by human articular chondrocytes. Biochem J309:963-968.
13.
Hakkinen L., Oksala O., Salo T., Rahemtulla F., Larjava H. (1993). Immunohistochemical localization of proteoglycans in human periodontium. J Histochem Cytochem41:1689-1699.
14.
Iwamoto M., Shimazu A., Nakashima K., Suzuki F., Kato Y. ( 1991). Reduction of basic fibroblasts growth factor receptor is coupled with terminal differentiation of chondrocytes. J Biol Chem266:461-467.
15.
Iwamoto M., Jikko A., Murakami H., Shimazu A., Nakashima K.,
16.
Iwamoto M., et al. (1994). Changes in parathyroid hormone receptors during chondrocyte cytodifferentiation. J Biol Chem269:17245-17251.
17.
Jalkanen M., Elenius K., Salmivirta M. (1992). Syndecan-a cell surface proteoglycan that selectively binds extracellular effector molecules. Adv Exp Med Biol313:79-85.
18.
Jikko A., Aoba T., Murakami H., Takano Y., Iwamoto M., Kato Y. ( 1993). Characterization of the mineralization process in cultures of rabbit growth plate chondrocytes. Dev Biol156:372-380.
19.
Kato Y., Iwamoto M. (1990). Fibroblast growth factor is an inhibitor of chondrocyte terminal differentiation. J Biol Chem265:5903-5909.
20.
Kojima T., Katsumi A., Yamazaki T., Muramatsu T., Nagasaka T., Ohsumi K., et al. (1996). Human ryudocan from endothelium-like cells binds basic fibroblast growth factor, midkine, and tissue factor pathway inhibitor . J Biol Chem271:5914-5920. Labarca C., Paigen K. (1980). A simple, rapid, and sensitive DNA assay procedure . Anal Biochem102:344-352.
21.
Last KS, Stanbury JB, Embery G. (1985). Glycosaminoglycans in human gingival crevicular fluid as indicators of active periodontal disease. Arch Oral Biol30:275-281.
22.
Lories V., Cassiman JJ, Van den Berghe H., David G. (1992). Differential expression of cell surface heparan sulfate proteoglycans in human mammary epithelial cells and lung fibroblasts . J Biol Chem267:1116-1122.
23.
Mali M., Jaakkola P., Arvilommi AM, Jalkanen M. (1990). Sequence of human syndecan indicates a novel gene family of integral membrane proteoglycans. J Biol Chem265:6884-6889.
24.
Mali M, Elenius K, Miettinen HM, Jalkanen M (1993). Inhibition of basic fibroblast growth factor-induced growth promotion by overexpression of syndecan-1. J Biol Chem 268:24215-24222. Marynen P, Zhang J, Cassiman JJ, Van den Berghe H, David G (1989). Partial primary structure of the 48- and 90-kilodalton core proteins of cell surface-associated heparan sulfate proteoglycans of lung fibroblasts. Prediction of an integral membrane domain and evidence for multiple distinct core proteins at the cell surface of human lung fibroblasts. J Biol Chem 264:7017-7024.
25.
Morishita M, Yamamura T, Bachchu MAH, Shimazu A, Iwamoto Y (1998). The effect of estrogen on osteocalcin production by human periodontal ligament cells. Arch Oral Biol 43:329-333. Narisawa S, Frohlander N, Millan JL (1997). Inactivation of two mouse alkaline phosphatase genes and establishment of a model of infantile hypophosphatasia. Dev Dynam 208:432-446. Nohutcu RM, McCauley LK, Koh AJ, Somerman MJ (1997). Expression of extracellular matrix proteins in human periodontal ligament cells during mineralization in vitro. J Periodontol 68:320-327.
26.
Ogata Y., Niisato N., Sakurai T., Furuyama S., Sugiya H. (1995). Comparison of the characteristics of human gingival fibroblasts and periodontal ligament cells. J Periodontol66 :1025-1031.
27.
Ohgi S., Johnson PW (1996). Glucose modulates growth of gingival fibroblasts and periodontal ligament cells: correlation with expression of basic fibroblast growth factor. J Periodont Res31:579-588.
28.
Okamoto T., Yatsuzuka N., Tanaka Y., Kan M., Yamanaka T., Sakamoto A., et al. (1997). Growth and differentiation of periodontal ligament-derived cells in serum-free defined culture.
29.
In VitroCell Dev Biol Anim33:302-309.
30.
Oksala O., Haapasalmi K., Hakkinen L., Uitto VJ, Larjava H. (1997). Expression of heparan sulphate and small dermatan/chondroitin sulphate proteoglycans in chronically inflamed human periodontium. J Dent Res76:1250-1259.
31.
Shimazu A., Nah HD, Kirsch T., Koyama E., Leatherman JL, Golden EB, et al. (1996). Syndecan-3 and the control of chondrocyte proliferation during endochondral ossification. Exp Cell Res229:126-136.
32.
Somerman MJ, Foster RA, Imm GM, Sauk JJ, Archer SY (1989). Periodontal ligament cells and gingival fibroblasts respond differently to attachment factors in vitro. J Periodontol60:73-77. Somerman MJ, Young MF, Foster RA, Moehring Jim, Imm G., Sauk JJ (1990). Characteristics of human periodontal ligament cells in vitro. Arch Oral Biol35:241-247.
33.
Takano-Yamamoto T., Takemura T., Kitamura Y., Nomura S. (1994). Site-specific expression of mRNAs for osteonectin, osteocalcin, and osteopontin revealed by in situ hybridization in rat periodontal ligament during physiological tooth movement. J Histochem Cytochem42:885-896.
A, et al. (1997). Effects of basic fibroblast growth factor on human periodontal ligament cells. J Periodont Res8:667-675. Terranova VP, Odziemiec C., Tweden KS, Spadone DP (1989). Repopulation of dentin surfaces by periodontal ligament cells and endothelial cells. Effect of basic fibroblast growth factor. J Periodontol60:293-301.
36.
Thesleff I., Vainio S., Salmivirta M., Jalkanen M. (1990). Syndecan and tenascin: induction during early tooth morphogenesis and possible interactions. Cell Differ Dev32:383-389.
37.
Turnbull JE, Fernig DG, Ke Y., Wilkinson MC, Gallagher JT (1992). Identification of the basic fibroblast growth factor binding sequence in fibroblast heparan sulfate. J Biol Chem267:10337-10341.
38.
Vainio S., Thesleff I. (1992). Sequential induction of syndecan, tenascin and cell proliferation associated with mesenchymal cell condensation during early tooth development. Differentiation50:97-105.
39.
Yayon A., Klagsbrun M., Esko JD, Leder P., Ornitz DM (1991). Cell surface, heparin-like molecules are required for binding of basic fibroblast growth factor to its high affinity receptor . Cell64:841-848.
