Novel Diabetic Mouse Models as Tools for Investigating Diabetic Retinopathy

Peter F Kador, Peng Zhang, Jun Makita, Zifeng Zhang, Changmei Guo, James Randazzo, Hiroyoshi Kawada, Neena Haider, Karen Blessing

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Objective: Mouse models possessing green fluorescent protein (GFP) and/or human aldose reductase (hAR) in vascular tissues have been established and crossed with naturally diabetic Akita mice to produce new diabetic mouse models. Research Design and Methods: Colonies of transgenic C57BL mice expressing GFP (SMAA-GFP), hAR (SMAA-hAR) or both (SMAA-GFP-hAR) in vascular tissues expressing smooth muscle actin were established and crossbred with C57BL/6-Ins2Akita/J (AK) mice to produce naturally diabetic offspring AK-SMAA-GFP and AK-SMAA-GFP-hAR. Aldose reductase inhibitor AL1576 (ARI) was administered in chow. Retinal and lenticular sorbitol levels were determined by HPLC. Retinal functions were evaluated by electroretinography (ERGs). Growth factor and signaling changes were determined by Western Blots using commercially available antibodies. Retinal vasculatures were isolated from the neural retina by enzymatic digestion. Flat mounts were stained with PAS-hematoxylin and analyzed. Results: Akita transgenics developed DM by 8 weeks of age with blood glucose levels higher in males than females. Sorbitol levels were higher in neural retinas of AK-SMAA-GFP-hAR compared to AK-SMAA-GFP mice. AK-SMAA-GFP-hAR mice also had higher VEGF levels and reduced ERG scotopic b-wave function, both of which were normalized by AL1576. AK-SMAA-GFP-hAR mice showed induction of the retinal growth factors bFGF, IGF-1, and TGFβ, as well as signaling changes in P-Akt, P-SAPK/JNK and P-44/42 MAPK that were also reduced by ARI treatment. Quantitative analysis of flat mounts in 18 week AK-SMAA-GFP-hAR mice revealed increased loss of nuclei/capillary length and a significant increase in the percentage of acellular capillaries present which was not seen in AK-SMAA-GFP-hAR treated with ARI. Conclusions/Significance: These new mouse models of early onset diabetes may be valuable tools for assessing both the role of hyperglycemia and AR in the development of retinal lesions associated with diabetic retinopathy.

Original languageEnglish (US)
Article numbere49422
JournalPloS one
Volume7
Issue number12
DOIs
StatePublished - Dec 12 2012

Fingerprint

aldehyde reductase
diabetic retinopathy
Aldehyde Reductase
Diabetic Retinopathy
Green Fluorescent Proteins
green fluorescent protein
animal models
mice
Electroretinography
electroretinography
Sorbitol
vascular tissues
sorbitol
retina
growth factors
Blood Vessels
Retina
Intercellular Signaling Peptides and Proteins
genetically modified organisms
Tissue

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Kador, P. F., Zhang, P., Makita, J., Zhang, Z., Guo, C., Randazzo, J., ... Blessing, K. (2012). Novel Diabetic Mouse Models as Tools for Investigating Diabetic Retinopathy. PloS one, 7(12), [e49422]. https://doi.org/10.1371/journal.pone.0049422

Novel Diabetic Mouse Models as Tools for Investigating Diabetic Retinopathy. / Kador, Peter F; Zhang, Peng; Makita, Jun; Zhang, Zifeng; Guo, Changmei; Randazzo, James; Kawada, Hiroyoshi; Haider, Neena; Blessing, Karen.

In: PloS one, Vol. 7, No. 12, e49422, 12.12.2012.

Research output: Contribution to journalArticle

Kador, PF, Zhang, P, Makita, J, Zhang, Z, Guo, C, Randazzo, J, Kawada, H, Haider, N & Blessing, K 2012, 'Novel Diabetic Mouse Models as Tools for Investigating Diabetic Retinopathy', PloS one, vol. 7, no. 12, e49422. https://doi.org/10.1371/journal.pone.0049422
Kador, Peter F ; Zhang, Peng ; Makita, Jun ; Zhang, Zifeng ; Guo, Changmei ; Randazzo, James ; Kawada, Hiroyoshi ; Haider, Neena ; Blessing, Karen. / Novel Diabetic Mouse Models as Tools for Investigating Diabetic Retinopathy. In: PloS one. 2012 ; Vol. 7, No. 12.
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abstract = "Objective: Mouse models possessing green fluorescent protein (GFP) and/or human aldose reductase (hAR) in vascular tissues have been established and crossed with naturally diabetic Akita mice to produce new diabetic mouse models. Research Design and Methods: Colonies of transgenic C57BL mice expressing GFP (SMAA-GFP), hAR (SMAA-hAR) or both (SMAA-GFP-hAR) in vascular tissues expressing smooth muscle actin were established and crossbred with C57BL/6-Ins2Akita/J (AK) mice to produce naturally diabetic offspring AK-SMAA-GFP and AK-SMAA-GFP-hAR. Aldose reductase inhibitor AL1576 (ARI) was administered in chow. Retinal and lenticular sorbitol levels were determined by HPLC. Retinal functions were evaluated by electroretinography (ERGs). Growth factor and signaling changes were determined by Western Blots using commercially available antibodies. Retinal vasculatures were isolated from the neural retina by enzymatic digestion. Flat mounts were stained with PAS-hematoxylin and analyzed. Results: Akita transgenics developed DM by 8 weeks of age with blood glucose levels higher in males than females. Sorbitol levels were higher in neural retinas of AK-SMAA-GFP-hAR compared to AK-SMAA-GFP mice. AK-SMAA-GFP-hAR mice also had higher VEGF levels and reduced ERG scotopic b-wave function, both of which were normalized by AL1576. AK-SMAA-GFP-hAR mice showed induction of the retinal growth factors bFGF, IGF-1, and TGFβ, as well as signaling changes in P-Akt, P-SAPK/JNK and P-44/42 MAPK that were also reduced by ARI treatment. Quantitative analysis of flat mounts in 18 week AK-SMAA-GFP-hAR mice revealed increased loss of nuclei/capillary length and a significant increase in the percentage of acellular capillaries present which was not seen in AK-SMAA-GFP-hAR treated with ARI. Conclusions/Significance: These new mouse models of early onset diabetes may be valuable tools for assessing both the role of hyperglycemia and AR in the development of retinal lesions associated with diabetic retinopathy.",
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AU - Randazzo, James

AU - Kawada, Hiroyoshi

AU - Haider, Neena

AU - Blessing, Karen

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N2 - Objective: Mouse models possessing green fluorescent protein (GFP) and/or human aldose reductase (hAR) in vascular tissues have been established and crossed with naturally diabetic Akita mice to produce new diabetic mouse models. Research Design and Methods: Colonies of transgenic C57BL mice expressing GFP (SMAA-GFP), hAR (SMAA-hAR) or both (SMAA-GFP-hAR) in vascular tissues expressing smooth muscle actin were established and crossbred with C57BL/6-Ins2Akita/J (AK) mice to produce naturally diabetic offspring AK-SMAA-GFP and AK-SMAA-GFP-hAR. Aldose reductase inhibitor AL1576 (ARI) was administered in chow. Retinal and lenticular sorbitol levels were determined by HPLC. Retinal functions were evaluated by electroretinography (ERGs). Growth factor and signaling changes were determined by Western Blots using commercially available antibodies. Retinal vasculatures were isolated from the neural retina by enzymatic digestion. Flat mounts were stained with PAS-hematoxylin and analyzed. Results: Akita transgenics developed DM by 8 weeks of age with blood glucose levels higher in males than females. Sorbitol levels were higher in neural retinas of AK-SMAA-GFP-hAR compared to AK-SMAA-GFP mice. AK-SMAA-GFP-hAR mice also had higher VEGF levels and reduced ERG scotopic b-wave function, both of which were normalized by AL1576. AK-SMAA-GFP-hAR mice showed induction of the retinal growth factors bFGF, IGF-1, and TGFβ, as well as signaling changes in P-Akt, P-SAPK/JNK and P-44/42 MAPK that were also reduced by ARI treatment. Quantitative analysis of flat mounts in 18 week AK-SMAA-GFP-hAR mice revealed increased loss of nuclei/capillary length and a significant increase in the percentage of acellular capillaries present which was not seen in AK-SMAA-GFP-hAR treated with ARI. Conclusions/Significance: These new mouse models of early onset diabetes may be valuable tools for assessing both the role of hyperglycemia and AR in the development of retinal lesions associated with diabetic retinopathy.

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