Skip to main content
Download PDF

Early changes in retinal structure and BMP2 expression in the retina and crystalline lens of streptozotocin-induced diabetic pigs

Laboratory Animal Research volume 33, pages 216–222 (2017)Cite this article

Abstract

Purpose

This study aims to evaluate early changes in retinal structure and BMP2 expression in the retina and crystalline lens by comparing streptozotocin-induced diabetic pigs and normal control group pigs.

Methods

Five eye samples from five diabetic Micro-pigs (Medikinetics, Pyeongtaek, Korea) and five eye samples from five control pigs bred in a specific pathogen-free area were used. Diabetes was developed through intravenous injection of nicotinamide and streptozotocin, and the average fasting glucose level was maintained at 250 mg/dL or higher for 16 weeks. To evaluate BMP2 expression in the retina and crystalline lens, Western blotting was performed.

Results

In Hematoxylin and Eosin staining, most diabetic pigs showed structural abnormalities in the inner plexiform layer. The number of nuclei in the ganglion cell layer within the range of 104 μm2 was 3.78±0.60 for diabetic pigs and 5.57±1.07 for control group pigs, showing a statistically significant difference. In immunohistochemical staining, diabetic retinas showed an overall increase in BMP2 expression. In Western blotting, the average BMP2/actin level of diabetic retinas was 1.19±0.05, showing a significant increase compared to the 1.06±0.03 of the control group retinas (P=0.016). The BMP2/actin level of diabetic crystalline lenses was similar to the control group crystalline lenses (P=0.730).

Conclusions

Compared to control group pigs, the number of nuclei in the inner nuclear layer of retinas from streptozotocin-induced diabetic pigs decreased, while an increase in BMP2 expression was observed in the retina of diabetic pigs.

References

  1. Yau JW, Rogers SL, Kawasaki R, Lamoureux EL, Kowalski JW, Bek T, Chen SJ, Dekker JM, Fletcher A, Grauslund J, Haffner S, Hamman RF, Ikram MK, Kayama T, Klein BE, Klein R, Krishnaiah S, Mayurasakorn K, O’Hare JP, Orchard TJ, Porta M, Rema M, Roy MS, Sharma T, Shaw J, Taylor H, Tielsch JM, Varma R, Wang JJ, Wang N, West S, Xu L, Yasuda M, Zhang X, Mitchell P, Wong TY; Meta-Analysis for Eye Disease (META-EYE) Study Group. Global prevalence and major risk factors of diabetic retinopathy. Diabetes Care 2012; 35(3): 556–564.

    Article  Google Scholar 

  2. van Dijk HW, Verbraak FD, Kok PH, Garvin MK, Sonka M, Lee K, Devries JH, Michels RP, van Velthoven ME, Schlingemann RO, Abràmoff MD. Decreased retinal ganglion cell layer thickness in patients with type 1 diabetes. Invest Ophthalmol Vis Sci 2010; 51(7): 3660–3665.

    Article  Google Scholar 

  3. Sohn EH, van Dijk HW, Jiao C, Kok PH, Jeong W, Demirkaya N, Gannager A, Wit F, Kucukevcilioglu M, van Velthoven ME, DeVries JH, Mullins RF, Kuehn MH, Schlingemann RO, Sonka M, Verbraak FD, Abràmoff MD. Retinal neurodegeneration may precede microvascular changes characteristic of diabetic retinopathy in diabetes mellitus. Proc Natl Acad Sci USA 2016; 113(19): E2655–2664.

    Article  CAS  Google Scholar 

  4. Guo J, Wu G. The signaling and functions of heterodimeric bone morphogenetic proteins. Cytokine Growth Factor Rev 2012; 23(1-2): 61–573.

    Article  CAS  Google Scholar 

  5. Akeel S, El-Awady A, Hussein K, El-Refaey M, Elsalanty M, Sharawy M, Al-Shabrawey M. Recombinant bone morphogenetic protein-2 induces up-regulation of vascular endothelial growth factor and interleukin 6 in human pre-osteoblasts: role of reactive oxygen species. Arch Oral Biol 2012; 57(2): 445–452.

    Article  CAS  Google Scholar 

  6. Hussein KA, Choksi K, Akeel S, Ahmad S, Megyerdi S, El-Sherbiny M, Nawaz M, Abu El-Asrar A, Al-Shabrawey M. Bone morphogenetic protein 2: a potential new player in the pathogenesis of diabetic retinopathy. Exp Eye Res 2014; 125: 79–88.

    Article  CAS  Google Scholar 

  7. Lee MS, Song KD, Yang HJ, Solis CD, Kim SH, Lee WK. Development of a type II diabetic mellitus animal model using Micropig®. Lab Anim Res 2012; 28(3): 205–208.

    Article  Google Scholar 

  8. van Dijk HW, Kok PH, Garvin M, Sonka M, Devries JH, Michels RP, van Velthoven ME, Schlingemann RO, Verbraak FD, Abràmoff MD. Selective loss of inner retinal layer thickness in type 1 diabetic patients with minimal diabetic retinopathy. Invest Ophthalmol Vis Sci 2009; 50(7): 3404–3409.

    Article  Google Scholar 

  9. Park HS, Park SJ, Park SH, Chun MH, Oh SJ. Shifting of parvalbumin expression in the rat retina in experimentally induced diabetes. Acta Neuropathol 2008; 115(2): 241–248.

    Article  CAS  Google Scholar 

  10. Kern TS, Barber AJ. Retinal ganglion cells in diabetes. J Physiol 2008; 586(18): 4401–4408.

    Article  CAS  Google Scholar 

  11. Zhang M, Zhou SH, Zhao S, Li XP, Liu LP, Shen XQ. Pioglitazone can downregulate bone morphogenetic protein-2 expression induced by high glucose in human umbilical vein endothelial cells. Pharmacology 2008; 81(4): 312–316.

    Article  CAS  Google Scholar 

  12. Belecky-Adams TL, Adler R, Beebe DC. Bone morphogenetic protein signaling and the initiation of lens fiber cell differentiation. Development 2002; 129(16): 3795–3802.

    CAS  PubMed  Google Scholar 

  13. Boswell BA, Overbeek PA, Musil LS. Essential role of BMPs in FGF-induced secondary lens fiber differentiation. Dev Biol 2008; 324(2): 202–212.

    Article  CAS  Google Scholar 

  14. Perry RE, Swamy MS, Abraham EC. Progressive changes in lens crystallin glycation and high-molecular-weight aggregate formation leading to cataract development in streptozotocin-diabetic rats. Exp Eye Res 1987; 44(2): 269–282.

    Article  CAS  Google Scholar 

  15. Chung SS, Ho EC, Lam KS, Chung SK. Contribution of polyol pathway to diabetes-induced oxidative stress. J Am Soc Nephrol 2003; 14: S233–236.

    Article  CAS  Google Scholar 

  16. Hussein KA, Zakhary IE, Elawady AR, Emam HA, Sharawy M, Baban B, Akeel S, Al-Shabrawey M, Elsalanty ME. Difference in soft tissue response between immediate and delayed delivery suggests a new mechanism for recombinant human bone morphogenetic protein 2 action in large segmental bone defects. Tissue Eng Part A 2012; 18(5-6): 665–675.

    Article  CAS  Google Scholar 

  17. Mathura JR Jr, Jafari N, Chang JT, Hackett SF, Wahlin KJ, Delia NG, Okamoto N, Zack DJ, Campochiaro PA. Bone morphogenetic proteins-2 and -4: negative growth regulators in adult retinal pigmented epithelium. Invest Ophthalmol Vis Sci 2000; 41(2): 592–600.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Ophthalmology and Inha Vision Science Laboratory, Inha University School of Medicine Hospital 27, Inhangro, Jung-gu, Incheon, 22332, Korea

    Jae Seung Jeong, Yeon Sung Moon & Na Rae Kim

  2. Laboratory of Developmental Genetics, Department of Biomedical Sciences, Inha University School of Medicine, Incheon, Korea

    Woon-Kyu Lee

Authors
  1. Jae Seung Jeong
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Woon-Kyu Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yeon Sung Moon
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Na Rae Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Yeon Sung Moon or Na Rae Kim.

Rights and permissions

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://doi.org/creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jeong, J.S., Lee, WK., Moon, Y.S. et al. Early changes in retinal structure and BMP2 expression in the retina and crystalline lens of streptozotocin-induced diabetic pigs. Lab Anim Res 33, 216–222 (2017). https://doi.org/10.5625/lar.2017.33.3.216

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.5625/lar.2017.33.3.216

Keywords

  • BMP2
  • diabetes mellitus
  • lens
  • pigs
  • retina
  • streptozotocin

Laboratory Animal Research

ISSN: 2233-7660