April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Basement Membrane Alterations in Diabetic Retina of STZ-Treated Mice
Author Affiliations & Notes
  • E. Abari
    Department of Ophthalmology, Heinrich-Heine-University of Duesseldorf, Duesseldorf, Germany
  • M. Huemmeke
    Department of Ophthalmology, Heinrich-Heine-University of Duesseldorf, Duesseldorf, Germany
  • I. Semkova
    Department of Ophthalmology, Heinrich-Heine-University of Duesseldorf, Duesseldorf, Germany
  • U. Hartmann
    Center for Biochemistry and Molecular Medicine, University of Cologne, Cologne, Germany
  • A. Kunze
    Center for Biochemistry and Molecular Medicine, University of Cologne, Cologne, Germany
  • N. Kociok
    Department of Ophthalmology, Heinrich-Heine-University of Duesseldorf, Duesseldorf, Germany
  • M. Paulsson
    Center for Biochemistry and Molecular Medicine, University of Cologne, Cologne, Germany
  • A. M. Joussen
    Department of Ophthalmology, Heinrich-Heine-University of Duesseldorf, Duesseldorf, Germany
  • Footnotes
    Commercial Relationships  E. Abari, None; M. Huemmeke, None; I. Semkova, None; U. Hartmann, None; A. Kunze, None; N. Kociok, None; M. Paulsson, None; A.M. Joussen, None.
  • Footnotes
    Support  DFG Jo 324/4-1, DFG Jo 324-10-1
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 4991. doi:
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      E. Abari, M. Huemmeke, I. Semkova, U. Hartmann, A. Kunze, N. Kociok, M. Paulsson, A. M. Joussen; Basement Membrane Alterations in Diabetic Retina of STZ-Treated Mice. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4991.

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Abstract

Purpose: : Vascular and parenchyma basement membranes (BMs) are known to be thickened in diabetes, but alterations in individual BM components in diabetic retinopathy are obscure. The major basement membranes of the eye are Bruch’s membrane, the inner limiting membrane (ILM), and the capillary BMs forming a part of the blood-retina barrier. All BMs are formed by members of three protein families: laminins, nidogens, collagen IV, and by the proteoglycan perlecan. We aim to investigate the expression of individual basement membrane components in STZ induced diabetes in the eye.

Methods: : To identify abnormalities in the distribution of specific constituents, we analyzed eyes obtained from C57BL/6J mice that were rendered diabetic by five consecutive daily intraperitoneal injections of STZ (65 mg/kg). After 4 and 6 months of diabetes (defined by blood glucose >300 mg/dl) eyes were enucleated and examined by histological and immunohistochemical techniques. We used a panel of antibodies against the three BM protein families (laminin, collagen, nidogen) and perlecan to describe differences in the composition between normal and diabetic retinal BMs. Western blot was performed to quantify the changes in the BM protein expression in diabetic and non-diabetic retinae.

Results: : Compared to the controls, there was no difference in the perlecan expression in diabetic retina both 4 and 6 months of diabetes. The expression of collagen-IV was also not changed. In contrast, immunoreactivity for laminin-1, laminin alpha5 and gamma1 chain was enhanced in the capillaries and Bruch’s membrane of the diabetic retinae. Furthermore, immunoreactivity for nidogen-1 (normally moderate expressed in the Bruch’s membrane) appeared to increase in the retinae after 4 and 6 months of diabetes in comparison to the controls. Western blot analysis showed approximately 2-fold increase in the nidogen-1 expression after 4 months of diabetes. Nidogen-2 (normally high expressed) was not changed.

Conclusions: : These data give evidence the participation of laminins, nidogens, collagen IV in the BM composition for the capillaries in diabetic retinopathy. It appears that nidogen-1 was up-regulated while the levels of collagen IV and laminin are unchanged. These changes in the BMs composition may affect both the assembly and function and ultimately lead to pathological consequences such as macula edema and retinal fibrosis.

Keywords: diabetic retinopathy • microscopy: light/fluorescence/immunohistochemistry • pathology: experimental 
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