It is estimated that 17-34 million have a varying stage of diabetic retinopathy (DR)1. In the US, it is the leading cause of blindness in working age adults and remains a public health problem throughout the world. The earliest manifestations of DR are believed to originate in capillary occlusion 2 resulting in both hypo- and hyperperfusion of regional capillaries. Additionally, vascular associated cells called pericytes which ensheathe the capillary endothelium, are characteristically lost with DR progression3,4. And while these manifestations have been identified as hallmarks of the disease, the pathogenic chronology of these events remain unclear. A major obstacle in identifying the primary and associated cause of capillary infarction, regression and proliferation has been lack of sufficient resolution to identify these microscopic events in the living eye. Traditional fundoscopy approaches lack necessary resolution to resolve sub-cellular structure because the eye's optics blur the retinal image. Therefore, we will use an adaptive optics scanning laser ophthalmoscope (AOSLO) which corrects for the eyes aberrations to achieve sub-cellular resolution needed for this research. Because AOSLO is non-invasive, a benefit of this approach is that the same subjects can be imaged over longitudinal progression of the disease;not requiring post-mortem analysis to acquire sufficient resolution. We will image both pericytes and capillary blood flow in a transgenic mouse model expressing fluorescent pericytes. In this model, we will induce a condition similar to human type-1 diabetes mellitus by injecting streptozotocin, an agent that selectively destroys insulin producing ?-cells in the pancreas. By tracking the progressive changes in capillary flow and pericyte density in the same animals over the course of weeks, we seek to better understand the chronology of the earliest events related to the human form of DR!
The mechanisms causing the earliest events of diabetic retinopathy have not been characterized in the living eye because they often exceed the resolution of conventional imaging. In this project we use a novel retinal camera that provides sub-cellular resolution to characterize the progression of two cellular patterns implicated with earliest stages of disease: the velocity of capillary flow and the morphology of pericytes, a cell type that is lost in diabetic retinopathy. Understanding the key events linked with early, preclinical stages of diabetic retinopathy is a central focus of this research and will be criticalfor developing therapies that target the primary cause of pathology. ! ! !
| Alarcon-Martinez, Luis; Yilmaz-Ozcan, Sinem; Yemisci, Muge et al. (2018) Capillary pericytes express ?-smooth muscle actin, which requires prevention of filamentous-actin depolymerization for detection. Elife 7: |
| Marcos, Susana; Werner, John S; Burns, Stephen A et al. (2017) Vision science and adaptive optics, the state of the field. Vision Res 132:3-33 |
| Guevara-Torres, A; Joseph, A; Schallek, J B (2016) Label free measurement of retinal blood cell flux, velocity, hematocrit and capillary width in the living mouse eye. Biomed Opt Express 7:4228-4249 |
| Guevara-Torres, A; Williams, D R; Schallek, J B (2015) Imaging translucent cell bodies in the living mouse retina without contrast agents. Biomed Opt Express 6:2106-19 |
| Schallek, Jesse; Geng, Ying; Nguyen, HoanVu et al. (2013) Morphology and topography of retinal pericytes in the living mouse retina using in vivo adaptive optics imaging and ex vivo characterization. Invest Ophthalmol Vis Sci 54:8237-50 |
