Skip to main content
SpringerLink
Log in

Immunolocalization of prolyl 4-hydroxylase in fibroblasts cultured from Tenon's capsule of humans

  • Laboratory Investigation
  • Published:
Graefe's Archive for Clinical and Experimental Ophthalmology Aims and scope Submit manuscript

Abstract

• Background: The excessive accumulation of extracellular matrix (ECM) with the repopulation of fibroblasts may lead to an unsuccessful outcome of glaucoma filtering surgery. We examined the immunolocalization of ECM components and prolyl 4-hydroxylase, an enzyme involved in collagen biosynthesis, in cultured Tenon's capsule fibroblasts (TCFs) of humans to evaluate the production of ECM in the cells. • Methods: We used light microscopy to evaluate the immunolocalization of prolyl 4-hydroxylase and ECM components, collagen types I, III, and IV, cellular fibronectin, and laminin in TCFs. Ultrastructural localization of the enzyme was also evaluated by electron microscopy. • Results: Immunoreactivity with monoclonal antibodies against the α and β subunits of the enzyme or with the polyclonal antibody against it was detected in the cytoplasm of the cells in a fine granular pattern, indicating its localization in the indoplasmic reticulum (ER). Immunoreactivity for the enzyme was detected in the cisternae of the ER on electron microscopy. Types I and III collagen reactivities were also observed in the cytoplasm in a fine granular pattern. T reactivity was present diffusely on the cell surface. The distribution of laminin reactivity in the cytoplasm resembled that of types I and III collagen. Cellular fibronectin reactivity was observed in the ECM in a reticular pattern. • Conclusion: Prolyl 4-hydroxylase was located in the cisternae of the ER. TCFs produced a variety of ECM components in vitro. The results provide insight into the fibrotic process during scar formation at the site of a bleb following filtering surgery.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Al-Adnani MS, Patrick RS, McGee JO (1986) The subcellular distribution of prolyl hydroxylase determined immunohistochemically. J Cell Sci 16:639–650

    Google Scholar 

  2. Bai Y, Muragaki Y, Obata K, Iwata K, Ooshima A (1986) Immunological properties of monoclonal antibodies to human and rat prolyl 4-hydroxylase. J Biochem 99:1563–1570

    Google Scholar 

  3. Birk DE, Fitch JM, Linsenmayer TF (1986) Organization of collagen types I and V in the embryonic chicken cornea. Invest Ophthalmol Vis Sci 27:1470–1477

    Google Scholar 

  4. Birk DE, Fitch JM, Babiarz JP, Linsenmayer TF (1988) Collagen type I and type V are present in the same fibril in the avian corneal stroma. J Cell Biol 106:999–1008

    Google Scholar 

  5. Cheng SY, Gong QH, Parkinson C, Robinson EA, Appella E, Merlino GT, Pasten I (1987) The nucleotide sequence of a human cellular thyroid hormone binding protein present in endoplasmic reticulum. J Biol Chem 262:11221–11227

    Google Scholar 

  6. Costa VP, Spaeth GL, Eiferman RA, Orengo-Nania S (1993) Wound healing modulation in glaucoma filtration surgery. Ophthalmic Surg 24:152–170

    Google Scholar 

  7. Fuller GC, Langner RO (1970) Elevation of aortic proline hydroxylase: a biochemical defect in experimental arteriosclerosis. Science 168:987–988

    Google Scholar 

  8. Givens KT, Kitada S, Chen AK, Rothschiller J, Lee DA (1990) Proliferation of human ocular fibroblasts. An assessment of in vitro colorimetric assays. Invest Ophthalmol Vis Sci 31:1865–1862

    Google Scholar 

  9. Handa JT, Murad S, Jaffe GJ (1993) Minoxidil inhibits ocular cell proliferation and lysyl hydroxylase activity. Invest Ophthalmol Vis Sci 34:567–575

    Google Scholar 

  10. Hata R, Senoo H (1989) l-Ascorbic acid 2-phosphate stimulates collagen accumulation, cell proliferation, and formation of a three-dimensional tissuelike substance by skin fibroblasts. J Cell Physiol 138:8–16

    Google Scholar 

  11. Heuer DK, Parrish RK II, Gressel MG, Hodapp E, Palmberg PF, Anderson DR (1984) 5-Fluorouracil and glaucoma filtering surgery. III. Intermediate follow-up of a pilot study. Ophthalmology 91:384–394

    Google Scholar 

  12. Jampel HD (1992) Effect of brief exposure to mitomycin C on viability and proliferation of human Tenon's capsule fibroblasts. Ophthalmology 99:1471–1476

    Google Scholar 

  13. Kitazawa Y, Kawase K, Matsushita H, Minobe M (1991) Trabeculectomy with mitomycin. A comparative study with fluorouracil. Arch Ophthalmol 109:1693–1698

    Google Scholar 

  14. Koivu J, Myllyla R, Helaakoski T, Pihlajaniemi T, Tasanen K, Kivirikko KI (1987) A single polypeptide acts both as the β-subunit of prolyl 4-hydroxylase and as a protein disulfideisomerase. J Biol Chem 262:6447–6449

    Google Scholar 

  15. Lee DA, Shapourifar-Tehrani S, Kitada S (1990) The effect of 5-fluorouracil and cytarabine on human fibroblasts from Tenon's capsule. Invest Ophthalmol Vis Sci 31:1848–1855

    Google Scholar 

  16. Linsenmayer TF, Fitch JM, Mayne R (1984) Exracellular matrices in the developing avian eye: type V collagen in corneal and noncorneal tissues. Invest Ophthalmol Vis Sci 25:41–47

    Google Scholar 

  17. Linsenmayer TF, Gibney E, Igoe F, Gordon MK, Fitch JM, Fessler LI, Birk DE (1993) Type V collagen: molecular structure and fibrillar organization of the chicken al(V) NH2-terminal domain, a putative regulator of corneal fibrillogenesis. J Cell Biol 121:1181–1189

    Google Scholar 

  18. Matsumoto E, Muragaki Y, Ooshima A (1989) Increased amount of serum type IV collagen peptide in human liver fibrosis as determined by enzyme-immunoassay with monoclonal antibodies. Acta Pathol Jpn 39:217–223

    Google Scholar 

  19. McGuigan LJB, Mason RP, Sanchez R, Quigley HA (1987) d-Penicillamine and beta-aminopropionitrile effects on experimental filtering surgery. Invest Ophthalmol Vis Sci 28:1625–1629

    Google Scholar 

  20. McLean IW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixative. A new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083

    Google Scholar 

  21. Murad S, Pinnell SR (1987) Suppression of fibroblast proliferation and lysyl hydroxylase activity by minoxidil. J Biol Chem 262:11973–11978

    Google Scholar 

  22. Mussini E, Hutton JJ Jr, Udenfriend S (1974) Collagen proline hydroxylase in wound healing, granuloma formation, scarring, and growth. Science 157:927–929

    Google Scholar 

  23. Olsen BR, Berg RA, Kishida Y, Prockop DJ (1973) Collagen synthesis: localization of prolyl hydroxylase in tendon cells detached with ferritinlabeled antibodies. Science 182:825–827

    Google Scholar 

  24. Ooshima A, Fuller GC, Cardinale GJ, Spector S, Udenfriend S (1974) Increased collagen synthesis in blood vessels of hypertensive rats and its reversal by antihypertensive agents. Proc Natl Acad Sci USA 71:3019–3023

    Google Scholar 

  25. Ooshima A, Fuller GC, Cardinale GJ, Spector S, Udenfriend S (1975) Collagen biosynthesis in blood vessels of brain and other tissues in the hypertensive rat. Science 190:898–900

    Google Scholar 

  26. Pasqale LR, Thibault D, Dorman-Pease ML, Quigley HA, Jampel HD (1992) Effect of topical mitomycin C on glaucoma filtration surgery in monkeys. Ophthalmology 99:14–18

    Google Scholar 

  27. Pihlajaniemi T, Helaakoski T, Tasanen K, Myllyla R, Huhtala ML, Koivu J, Kivirikko KI (1987) Molecular cloning of the β-subunit of human 4prolyl hydroxylase. This subunit and protein disulfide isomerase are products of the same gene. EMBO J 6:643–649

    Google Scholar 

  28. Pihlajaniemi T, Myllyla R, Kivirikko KI (1991) Prolyl 4-hydroxylase and its role in collagen synthesis. J Hepatol 13 [Suppl]:S2-S7

    Google Scholar 

  29. Saika S (1992) Ultrastructural effect of L-ascorbic acid 2-phosphate on cultured keratocytes. Cornea 11:439–445

    Google Scholar 

  30. Saika S (1993) Ascorbic acid and proliferation of cultured rabbit keratocytes. Cornea 12:191–198

    Google Scholar 

  31. Saiko S, Kanagawa R, Uenoyama K, Hiroi K, Hiraoka J (1991) l-Ascorbic acid 2-phosphate, a phosphate derivative of l-ascorbic acid, enhances the growth of cultured rabbit keratocytes. Graefes Arch Clin Exp Ophthalmol 229:79–83

    Google Scholar 

  32. Saika S, Uenoyama K, Hiroi K, Ooshima A (1992) l-Ascorbic acid 2phosphate enhances the production of type I and type III collagen peptides in cultured rabbit keratocytes. Ophthalmic Res 24:68–72

    Google Scholar 

  33. Saika S, Hashizume N, Okada Y, Kobata S, Yamanaka O, Uenoyama K, Ooshima A (1994) Prolyl hydroxylase inhibitor and lysyl hydroxylase inhibitor inhibit spreading of corneal epithelium. Graefes Arch Clin Exp Ophthalmol 232:499–502

    Google Scholar 

  34. Smith LT, Holbrock KA, Madri JA (1986) Collagen types I, III, and V in human embryonic and fetal skin. Am J Anat 175:507–521

    Google Scholar 

  35. Tonoe O, Yamanaka O, Okada Y, Kobata S, Saika S (1994) Collagen biosynthesis and cellular proliferation after filtering surgery in rabbits (in Japanese). Folia Ophthalmol Jpn 46:242–245

    Google Scholar 

  36. Vartio T, Laitinen L, Narvanen O, Cutolo M, Thornell L-E, Zardi L, Virtanen I (1987) Differential expression of the ED sequence-containing form of cellular fibronectin in embryonic and adult human tissues. J Cell Sci 88:419–439

    Google Scholar 

  37. Woodley DT, Scheidt VJ, Reese MJ, Paller AS, Manning TO, Yoshiike T, Briggaman RA (1987) Localization of the alpha 3(V) chain of type V collagen in human skin. J Invest Dermatol 88:246–252

    Google Scholar 

  38. Yamamoto T, Varani J, Soong HK, Lichter PR (1990) Effects of 5-fluorouracil and mitomycin C on cultured rabbit subconjunctival fibroblasts. Ophthalmology 97:1204–1210

    Google Scholar 

  39. Yamanaka O, Saika S, Uenoyama K, Ooshima A (1994) Alpha- and betasubunits of prolyl 4-hydroxylase in rat limbal tissue and cultured subconjunctival fibroblasts. Invest Ophthalmol Vis Sci 35 [Suppl]:1429

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Ophthalmology, Wakayama Medical College, 7-Bancho 27, 640, Wakayama, Japan

    Shizuya Saika, Osamu Yamanaka, Osamu Tonoe, Yuka Okada & Yoshitaka Ohnishi

  2. Department of Pathology, Wakayama Medical College, 7-Bancho 27, 640, Wakayama, Japan

    Akira Ooshima

  3. Department of Anatomy, Kansai College of Acupuncture Medicine, 990, Ogaito, Kumatori, Sennan, 590-04, Osaka, Japan

    Michiro Kimura

Authors
  1. Shizuya Saika
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Akira Ooshima
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Osamu Yamanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Osamu Tonoe
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yuka Okada
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yoshitaka Ohnishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Michiro Kimura
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Saika, S., Ooshima, A., Yamanaka, O. et al. Immunolocalization of prolyl 4-hydroxylase in fibroblasts cultured from Tenon's capsule of humans. Graefe's Arch Clin Exp Ophthalmol 234, 251–257 (1996). https://doi.org/10.1007/BF00430418

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00430418

Keywords

Search

Navigation