A Large Keratan Sulfate Proteoglycan Present in Human Cervical Mucous Appears to Be Involved in the Reorganization of the Cervical Extracellular Matrix at Term

Abstract

Objective: To elucidate the function of keratan sulfate proteoglycan (KS-PG) in the human uterine cervix, we analyzed its distribution with respect to physiologic conditions.

Methods: Immunohistochemistry was used to localize KS bearing proteoglycans (mAb 5D4) and decorin (mAb 6B6) in the lower uterine segment. Proteins present in cervical mucous were labeled with biotin, glycosaminoglycan chains were digested enzymatically, and the samples were analyzed by Western blot.

Results: Decorin was detected throughout the extracellular matrix, in tissues from menstruating nonpregnant women, in early pregnancy, from women who had cesarean at term, at postpartum hysterectomy, and from postmenopausal women. In menstruating nonpregnant women, in early pregnancy (first trimester), and in postmenopausal women, KS-PG was detectable only in epithelial, mucous-producing cells. Interestingly, in samples obtained either at the time of cesarean at term (lower uterine segment) or after postpartal hysterectomy, KS-PG was detectable throughout the extracellular matrix, indicating that the expression of KS-PG is associated with reorganization of the tissue. Biochemical analysis of the KS present in mucous revealed a core protein in the range of 220 kDa, suggesting an identity with the large KS-PG described previously.

Conclusion: At parturition, a large KS-PG, which is virtually exclusively present in the cervical mucous of either early or nonpregnant women, was detected in the extracellular matrix. This finding indicates that cervical ripening is accompanied not only by quantitative but also by qualitative changes in the composition of the extracellular matrix.

Keywords

Cervical mucous extracellular matrix human uterine cervix keratan sulfate proteoglycan parturition pregnancy

Get full access to this article

View all access options for this article.

References

1. Ruoslahti E . Structure and biology of proteoglycans. Annu Rev Cell Biol 1988;4:229-255.

2. Kjellen L , Lindahl U . Proteoglycans: Structure and interactions. Ann Rev Biochem 1991;60:443-475.

3. Massagué J . Receptors for the TGF-β family. Cell 1992;69:1067-1070.

4. Turnbull JE , Fernig DG , Youquiang K , Wilkinson MC , Gallagher JT . Identification of the basic fibroblast growth factor binding sequence in fibroblast heparan sulfate. J Biol Chem 1992;267:10337-10341.

5. Lin CQ , Bissell MJ . Multi-faceted regulation of cell differentiation by extracellular matrix. FASEB J 1993;7:737-743.

6. Lyon M , Deakin JA , Mizuno K , Nakamura T , Gallagher JT . Interaction of hepatocyte growth factor with heparan sulfate. J Biol Chem 1994;269:11216-11223.

7. Iozzo RV , Cohen IR , Grässel S , Murdoch AD . The biology of perlecan: The multifaceted heparan sulphate proteoglycan of basement membranes and pericellular matrices. Biochem J 1994;302:625-639.

8. Iozzo RV , Murdoch AD . Proteoglycans of the extracellular environ,ent: Clues from the gene and protein side offer novel perspectives in molecular diversity and function. FASEB J 1996;10:598-614.

9. Gallagher JT . Heparan sulphate proteoglycans: The control of cell growth. In: Comper WD , ed. Extracellular matrix. Vol. 2. Amsterdam: Harwood Academic Publishers. 1996:230-245.

10.

10. Salmivirta M , Lidholt K , Lindahl U . Heparan sulfate: A piece of information. FASEB J 1996;10:1270-1279.

11.

11. Hardingham TE , Fosang AJ . Proteoglycans: Many forms and many functions. FASEB J 1992;6:861-870.

12.

12. Lindblom A , Carlstedt I , Fransson LA . Identification of the core proteins in proteoglycans synthesized by vascular endothelial cells. Biochem J 1989;261:145-153.

13.

13. Hassel JM , Noonan DM , Ledbetter SR , Laurie GW . Biosynthesis and structure of the basement membrane proteoglycan containing heparan sulphate side-chains. In: Evered D , Whelan J , eds. Function of the proteoglycans. New York: Wiley, 1986:204-214.

14.

14. Gallagher JT , Lyon M , Steward WP . Structure and function of the heparan sulphate proteoglycans. Biochem J 1986;236:313-325.

15.

15. Forman A , Ulmsten U , Banyi J , Wingerup L , Uldbjerg N . Evidence for a local effect of intracervical prostaglandin E2 gel. Am J Obstet Gynecol 1982;143:756-760.

16.

16. Aspden RM . Collagen organisation in the cervix and its relation to mechanical function. Collagen Relat Res 1988;8:103-112.

17.

17. Uldbjerg N , Malström A , Ekman G , Sheehan J , Ulmsten U , Wingerup L . Isolation and characterization of dermatan sulphate proteoglycan from human uterine cervix. Biochem J 1983;209:497-503.

18.

18. Uldbjerg N , Ekman G , Malmström A , Olsson K , Ulmsten U . Ripening of the human uterine cervix related to changes in collagen, glycoaminoglycans and collagenolytic activity. Am J Obstet Gynecol 1983;147:662-666.

19.

19. Norman M , Ekman G , Ulmsten U , Barchan K , Malmström A . Proteoglycan metabolism in the connective tissue of pregnant and non-regnant human cervix. Biochem J 1991;275:515-520.

20.

20. Norman M , Ekman G , Malmström A . Changed proteoglycan metabolism in human cervix immediately after spontaneous vaginal delivery. Obstet Gynecol 1993;81:217-223.

21.

21. Osmers R , Rath W , Pflanz MA , Kuhn W , Stuhlsatz HW , Szeverényi M . Glycosaminoglycans in cervical connective tissue during pregnancy and parturition. Obstet Gynecol 1993;81:88-92.

22.

22. Uldbjerg N , Danielsen CC . A study of the interaction in vitro between type I collagen and a small dermatan sulphate proteoglycan. Biochem J 1988;251:643-648.

23.

23. Fischer DC , Henning A , Winkler M , Rath W , Haubeck HD , Greiling H . Evidence for the presence of a large keratan sulphate proteoglycan in the human uterine cervix. Biochem J 1996;320:393-399.

24.

24. Rajabi MR , Dean DD , Beydoun SN , Woessner JF Jr . Elevated tissue levels of collagenase during dilatation of uterine cervix in human parturition. Am J Obstet Gynecol 1988;159:971-976.

25.

25. Winkler M , Fischer DC , Hlubek M , van de Leur E , Haubeck HD , Rath W . Interleukin-1β and interleukin-8 concentrations in the lower uterine segment during parturition at term. Obstet Gynecol 1998;91:945-948.

26.

26. Thomson AJ , Telfer JF , Young A , et al. Leukocytes infiltrate the myometrium during human parturition: Further evidence that labour is an inflammatory process. Hum Reprod 1999;14:229-236.

27.

27. Winkler M , Fischer DC , Ruck P , et al. Partutition at term: Parallel increases in interleukin-8 and proteinase concentrations and neutrophil count in the lower uterine segment. Hum Reprod 1999;14:1096-1100.

28.

28. Caterson B , Christner JE , Baker R . Identification of a monoclonal antibody that specifically recognizes corneal and skeletal keratan sulfate. J Biol Chem 1983;258:8848-8854.

29.

29. Sobue M , Nakashima N , Fukatsu TY , et al. Production and characterisation of monoclonal antibody to dermatan sulfate proteoglycan. J Histochem Cytochem 1988;36:479-485.

30.

30. Möller HJ , Heinegard D , Poulsen JH . Combined alcian blue and silver staining of subnanogram quantities of proteoglycans and glycosaminoglycans in sodium dodecyl sulfate-polyacrylamide gels. Anal Biochem 1993;209:169-175.

31.

31. Fischer DC , Haubeck HD , Eich K , et al. A novel keratan sulfate domain preferentially expressed on the large aggregating proteoglycan from human articular cartilage is recognized by the monoclonal antibody 3D12/H7. Biochem J 1996;318:1051-1056.

32.

32. Stöcker G , Lückge J , Greiling H , Wagener C . Characterization of biotin-labeled proteoglycans by electrophoretic separation in minigels and blotting onto nylon membranes prior and after enzymatic digestion. Anal Biochem 1989;179:245-250.

33.

33. Brown GM , Huckerby TN , Morris HG , Abram BL , Nieduszynski IA . Oligosaccharides derived from bovine articular cartilage keratan sulfates after keratanase II digestion: Implications for keratan sulfate structural fingerprinting. Biochemistry 1994;33:4836-4846.

34.

34. Mehmet H , Scudder P , Tang PW , Hounsell EF , Caterson B , Feizi T . The antigenic determinants recognized by three monoclonal antibodies to keratan sulphate involved sulphated hepta- or larger oligosaccharides of the pol(N-acetyllactosamine) series. Eur J Biochem 1986;157:385-391.

35.

35. Tang PW , Scudder P , Mehmet H , Hounsell EF , Feizi T . Sulphate groups are involved in the antigenicity of keratan sulphate and mask antigen expression on their poly-N-lactosamine backbone. Eur J Biochem 1986;160:537-545.

36.

36. Liggins GC . Cervical ripening as an inflammatory reaction. In: Ellwood DA , Anderson ABM , eds. The cervix in pregnancy and labor: Clinical and biochemical investigations. Edinburgh: Churchill-Livingstone, 1981:1-9.

37.

37. Barclay CG , Brennand JE , Kelly RW , Calder AA . Interleukin-8 production by the human cervix. Am J Obstet Gynecol 1993;169:625-632.

38.

38. Osmers RGW , Bläser J , Kuhn W , Tschesche H . Interleukin-8 synthesis and the onset of labor. Obstet Gynecol 1995;86:223-229.

39.

39. Bokström H , Brännströ M , Alexandersson M , Norströ A . Leukocyte subpopulations in the human uterine cervical stroma at early and term pregnancy. Hum Reprod 1997;12:586-590.

40.

40. Winkler M , Fischer DC , Ruck P , Kemp B , Hlubek M , Ruth W . Cervical dilatation as an inflammatory reaction. Eur J Inflamm Immunol Dis Obstet Gynecol 1997;1:67-71.

41.

41. Osmers R , Rath W , Adelmann-Grill BC , et al. Origin of cervical collagenase during parturition. Am J Obstet Gynecol 1992;166:1455-1460.

42.

42. Rath W , Osmers R , Szeverényi M , Stuhlsatz HW , Kuhn W . Changes of glycosaminoglycans in cervical connective tissue during pregnancy and parturition. In: Leppert PC , Woessner FJ , eds. The extracellular matrix of the uterus, cervix, and fetal membranes. Ithaca, New York: Perinatalogy Press, 1991:105-112.

43.

43. Rath W , Osmers R , Stuhlsatz HW , Adelmann-Grill BC . Biochemische grundlagen der cervixreifung und der muttermundseröffnung. Z Geburtsh u Perinatol 1994;98:186-195.

44.

44. Aplin JD , Hoadley ME , Seif MW . Hormonally regulated secretion of keratan sulphate by human endometrial epithelium. Biochem Soc Trans 1988;17:136-137.

45.

45. Hoadley ME , Seif MW , Aplin JD . Menstrual-cycle-dependent expression of keratan sulphate in human endometrium. Biochem J 1990;266:757-763.

46.

46. Shiozawa T , Tsukahara Y , Nakayama J , Ishii K , Katsuyama T . Immunohistochemical reactivity of antikeratan sulphate monoclonal antibody 5D4 to various conditions of human endometrial tissues and its application as a useful marker for identifying endometrial epithelia. Gynecol Obstet Invest 1991;32:239-242.

47.

47. Graham RA , Li TC , Cooke ID , Aplin JD . Keratan sulphate as a secretory product of human endometrium: Cyclin expression in normal women. Hum Reprod 1994;9:926-930.

48.

48. Fischer DC , Kolbe-Busch S , Stöcker G , Hoffmann A , Haubeck HD . Development of enzyme immuno-assays specific for keratan sulphate- and core-protein epitopes of the large aggregating proteoglycan from human articular cartilage. Eur J Clin Chem Clin Biochem 1994;32:285-291.