Abstract
Purpose:The aim of this project was to identify protein changes induced by short term diabetes in the rat retina.
Methods: Neuro–sensory retinae were pooled from Dark Agouti male rats after 10 weeks of streptozotocin–induced diabetes. Two–dimensional gel electrophoresis technology provided separation of solubilized proteins. After UV staining with Sypro–ruby (Bio–Rad)and Commassie Blue, PDQuest (Bio–Rad) software enabled comparison of protein spot profiles. Protein spots of interest were cut and peptide spectra generated using MALDI TOF–TOF (ABI 4700) and Q–TOF Ultima (Applied Biosystems) instruments. Spectra were compared to those of known proteins in the NCBI database for identification.
Results: Two–dimension gel profiles revealed a significant number of upregulated and down–regulated proteins. Diabetes was found to induce changes in retinal Crystallin subtype expression, Heat shock proteins, Retinaldehyde binding proteins, mitochondrial metabolic enzymes, chaperone proteins, adolase, tyrosine, and glycolytic metabolic enzymes .
Conclusions: Two–dimensional gel electrophoresis coupled with mass spectrometry technology has allowed identification of multiple changes at the protein level in an animal model of diabetic retinopathy.
Methods: Neuro–sensory retinae were pooled from Dark Agouti male rats after 10 weeks of streptozotocin–induced diabetes. Two–dimensional gel electrophoresis technology provided separation of solubilized proteins. After UV staining with Sypro–ruby (Bio–Rad)and Commassie Blue, PDQuest (Bio–Rad) software enabled comparison of protein spot profiles. Protein spots of interest were cut and peptide spectra generated using MALDI TOF–TOF (ABI 4700) and Q–TOF Ultima (Applied Biosystems) instruments. Spectra were compared to those of known proteins in the NCBI database for identification.
Results: Two–dimension gel profiles revealed a significant number of upregulated and down–regulated proteins. Diabetes was found to induce changes in retinal Crystallin subtype expression, Heat shock proteins, Retinaldehyde binding proteins, mitochondrial metabolic enzymes, chaperone proteins, adolase, tyrosine, and glycolytic metabolic enzymes .
Conclusions: Two–dimensional gel electrophoresis coupled with mass spectrometry technology has allowed identification of multiple changes at the protein level in an animal model of diabetic retinopathy.
| Original language | English |
|---|---|
| Pages (from-to) | 406-406 |
| Number of pages | 1 |
| Journal | Investigative Ophthalmology and Visual Science |
| Volume | 46 |
| Issue number | 13 |
| Publication status | Published - May 2005 |
| Externally published | Yes |
| Event | Meeting of the Association for Research in Vision and Ophthalmology (ARVO 2005) - Fort Lauderdale Duration: 1 May 2005 → 5 May 2005 |