Skip to main content
Log in

Partial characterization of a putative new growth factor present in pathological human vitreous

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

Abstract

• Background

Several growth factors have been implicated in the development of proliferative eye diseases, and some of those are present in human vitreous (HV). The effects of HV on cellular responses which modulate proliferative cell processes were studied. This study describes the partial characterization of a vitreous factor activity which does not correspond to any of the previously reported growth factors in pathological HV.

• Methods

Vitreous humour was obtained from medical vitrectomies, from patients with PDR and PVR. The biological activity of the vitreous factor was determined by its ability to increase cytosolic calcium concentration ([Ca2+ i]), increase production of inositol phosphates, and induce cell proliferation in the cell line EGFR T17. In some experiments other cell lines, such as NIH 3T3, 3T3-L1, FRTL5, A431, PC 12, Y79, and GH3, were also employed. Measurement of [Ca2+]i in cell suspensions was performed using the fluorescent Ca2+ indicator fura-2. The activity of the factor present in HV was compared with other growth factors by means of: (a) [Ca2+]i mobilization pattern, (b) sequential homologous and heterologous desensitization of receptors, (c) effects of phorbol esters on their action, and (d) inactivation after treatment with different proteolytic enzymes.

• Results

The HV-induced cell proliferation and increases in [Ca2+]i concentration were characterized by a peculiar time pattern. The different approaches used ruled out its identity with PDGF, bFGF, EGF, TGF-β, IGFs, TNF-α, NGF, and other compounds such as ATP, angiotensin I, and bradykinin. Vitreous factor actions are mediated by specific receptors apparently regulated by PKC. This factor is able to induce [Ca2+]i mobilization in most of the cell lines studied, indicating that its effects are not tissue specific.

• Conclusions

These results suggest the presence of a growth factor activity in pathological HV which may be due to the presence of an undescribed growth factor in the eye.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Berne RM, Rubio R, Curnish RR (1974) Release of adenosine from ischemic brain: effect on cerebral vascular resistance and incorporation into cerebral adenine nucleotides. Circ Res 35:262–271

    Google Scholar 

  2. Berridge MJ (1987) Inositol triphosphate and diacylglycerol: two interacting second messengers. Annu Rev Biochem 56:159–193

    Google Scholar 

  3. Block MR, Glick BS, Wilcox CA, Wieland FT, Rothman JE (1988) Purification of an N-ethylmaleimide-sensitive protein catalyzing vesicular transport. Proc Natl Acad Sci USA 85:7852–7856

    Google Scholar 

  4. Brooks SF, Hergert T, Broad S, Rozengurt E (1992) Expression of 80k/MARKS, a major substrate of protein kinase C (PKC), is down-regulated through both PKC-dependent and -independent pathways. J Biol Chem 267:14212–14218

    Google Scholar 

  5. Carpenter G, Wahl MI (1990) The epidermal growth factor family. In: Sporn MB, Roberts AB (eds) Peptide growth factors and their receptors. (Handbook of Experimental Pharmacology, vol 95, part 1) Springer, Berlin Heidelberg New York, pp 95:69–171

    Google Scholar 

  6. Casabiell X, Pandiella A, Casanueva FF (1991) Regulation of epidermal-growth-factor-receptor signal transduction by cis-unsaturated fatty acids. Biochem J 278:679–687

    Google Scholar 

  7. Casabiell X, Zugaza JL, Pombo CM, Pandiella A, Casanueva FF (1993) Oleic acid blocks epidermal growth factor-activated early intracellular signals without altering mitogenic response. Exp Cell Res 205:365–373

    Google Scholar 

  8. Danser AHJ, van der Dorpel MA, Deinum J, Derkx FHM, Franken AAM, Peperkamp E, de Jong, PTVM, Schalekamp MADH (1989) Renin, prorenin and immunoreactive renin in vitreous fluid from eye with and without diabetic retinopathy. J Clin Endocrinol Metab 68:160–169

    Google Scholar 

  9. Folkman J, Klagsburn M (1987) Angiogenic factors. Science 235:442–447

    Google Scholar 

  10. Grant M, Russell B, Fitzgerald C, Merimee TJ (1986) Insulin-like growth factor in vitreous. Studies in control and diabetic subjects with neovascularization. Diabetes 35:416–420

    Google Scholar 

  11. Grynckiewickz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450

    Google Scholar 

  12. Kirchhof B, Kirchhof E, Ryan SJ, Sorgente N (1989) Vitreous modulation of migration and proliferation of retinal pigment epithelial cells in vitro. Invest Ophthalmol Vis Sci 30:1951–1957

    Google Scholar 

  13. Lambert TL, Kent RS, Whorton AR (1986) Bradykinin stimulation of inositol polyphosphate production in porcine aortic endothelial cells. J Biol Chem 261:15288–15293

    Google Scholar 

  14. Lutty GA, Mello RJ, Chandler C, Fait C, Bennett A, Patz A (1985) Regulation of cell growth by vitreous humour. J Cell Sci 76:53–65

    Google Scholar 

  15. Lutty GA, Chandler C, Bennet A, Fait C, Patz A (1986) Presence of endothelial cell growth factor activity in normal and diabetic eyes. Curr Eye Res 5:9–17

    Google Scholar 

  16. Morgan-Boyd R, Stewart JM, Vavrek RJ, Hassid A (1987) Effects of bradykinin and angiotensin II on intracellular Ca2+ dynamics in endothelial cells. Am J Physiol C588–C598

  17. Pandiella A, Massague J (1991) Cleavage of the membrane precursor for transforming growth factor a is a regulated process. Proc Natl Acad Sci USA 88:1276–1730

    Google Scholar 

  18. Pandiella A, Beguinot L, Velu TJ, Meldolesi J (1988) Transmembrane signalling at epidermal growth factor receptors overexpressed in NIH 3T3 cells. Biochem J 254:223–228

    Google Scholar 

  19. Pandiella A, Magni M, Meldolesi J (1989) Plasma membrane hyperpolarization and [Ca2+]i increase induced by fibroblast growth factor in NIH 3T3 fibroblasts: resemblance to early signals generated by platelet-derived growth factor. Biochem Biophys Res Commun 163:1325–1331

    Google Scholar 

  20. Penie M, Alderton J, Steindhart R, Tsien RY (1986) Calcium raises abruptly and briefly throughout the cell at onset of anaphase. Science 233:886–889

    Google Scholar 

  21. Robertson PL, Diane AR, Goldstein GW (1990) Phosphoinositide metabolism and prostacyclin formation in retinal microvascular endothelium: stimulation by adenine nucleotides. Exp Eye Res 50:37–44

    Google Scholar 

  22. Ryan SJ (1985) The pathophysiology of proliferative vitreoretinopathy and its managements. Am J Ophthalmol 100:188–193

    Google Scholar 

  23. Sivaligam A, Kenny J, Brown GC, Benson WE, Donoso L (1990) Basic fibroblast growth factor levels in vitreous of patients with proliferative diabetic retinopathy. Arch Ophthalmol 108:869–872

    Google Scholar 

  24. Sporn M, Roberts A, Wakefield L, de Comrugge B (1987) Some recent advances in the chemistry and biology of transforming growth factor-beta. J Cell Biol 105:1039–1045

    Google Scholar 

  25. Stauderman KA, Pruss RM (1989) Dissociation of Ca2+ entry and Ca2+ mobilization responses to angiotensin II in bovine adrenal chromaffin cells. J Biol Chem 264:18349–18355

    Google Scholar 

  26. Vilceck J, Lee TH (1991) Tumor necrosis factor. New insight into the molecular mechanismas of its multiple actions. J Biol Chem 266:7313–7316

    Google Scholar 

  27. Wahl SM, Wong H, McCartney-Francis N (1989) Role of growth factors in inflamation and tissue repair. J Cell Biochem 40:193–199

    Google Scholar 

  28. Wiedemann P (1992) Growth factors in retinal diseases: proliferative vitreoretinopathy, proliferative diabetic retinopathy, and retinal degeneration. Surv Ophthalmol 36:373–384

    Google Scholar 

  29. Wiedemann P, Ryan SJ, Novak P, Sorgente N (1985) Vitreous stimulates proliferation of fibroblasts and retinal pigment epithelial cells. Exp Eye Res 41:619–628

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pombo, C., Bokser, L., Casabiell, X. et al. Partial characterization of a putative new growth factor present in pathological human vitreous. Graefe's Arch Clin Exp Ophthalmol 234, 155–163 (1996). https://doi.org/10.1007/BF00462027

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00462027

Keywords

Navigation