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1.
Figure 1

Figure 1. Acellular capillaries. From: Bone marrow-CNS connections: Implications in the pathogenesis of diabetic retinopathy.

Trypsin digested image of diabetic rat retina showing acellular capillary (red arrow) which is the result of loss of both pericytes and endothelial cells. Acellular capillaries are the pathologic hallmark of diabetic retinopathy and their quantification is a useful way to assess the extent of retinopathy.

Yellowlees Douglas, et al. Prog Retin Eye Res. 2012 September;31(5):481-494.
2.
Figure 5

Figure 5. Diabetic retina contains higher numbers of CD45+cells. From: Bone marrow-CNS connections: Implications in the pathogenesis of diabetic retinopathy.

Immunohistochemical analysis of CD45+ cells in retinal cross sections from control (A) and mice with one month of STZ-induced diabetes was performed. (C) Quantitation of the number of CD45+ cells/section is shown and demonstrates increased number in diabetic compared to controls.

Yellowlees Douglas, et al. Prog Retin Eye Res. 2012 September;31(5):481-494.
3.
Figure 4

Figure 4. VASP redistribution is impaired in diabetic EPCs. From: Bone marrow-CNS connections: Implications in the pathogenesis of diabetic retinopathy.

Immunofluorescence images of EPCs from normal (A, B) and diabetic (C, D) donors left untreated (A, C) or treated with 100nM SDF-1 for 4 hours (B, D). Normal EPCs show a dramatic VASP redistribution following treatment (A, B) while EPCs from a diabetic donor do not show any response to SDF-1 (C, D).

Yellowlees Douglas, et al. Prog Retin Eye Res. 2012 September;31(5):481-494.
4.
Figure 2

Figure 2. EPC staining in rat retina. From: Bone marrow-CNS connections: Implications in the pathogenesis of diabetic retinopathy.

(A) Representative trypsin digested rat retina stained with CD133 i) CD14 (ii) and lectin (iii) showing presence of EPCs (white arrow) in the retinal vasculature of a diabetic rat. These cells appeared at the bifurcations of the retinal capillaries (B) with occasional clusters (white arrow) (C) or a single cell (D).

Yellowlees Douglas, et al. Prog Retin Eye Res. 2012 September;31(5):481-494.
5.
Figure 6

Figure 6. Immature progenitors are present in the diabetic retina. From: Bone marrow-CNS connections: Implications in the pathogenesis of diabetic retinopathy.

(A) Trypsin digested diabetic retinal vasculature (i) shows presence of CD133+ cells in diabetic vasculature (ii) with positive staining for CD14 marker (iii). Surprizingly, vehicle control rat retina (iv) did not show CD133+ cells (v) in 60% of the trypsin-digests although monocytic CD14+ expression were detected (vi). N=6 each vehicle control or diabetic treatment. (B) Individual count of number of CD133+ and CD14+ cells showing two fold increase in CD133+ cells (p<0.001).

Yellowlees Douglas, et al. Prog Retin Eye Res. 2012 September;31(5):481-494.
6.
Figure 7

Figure 7. Bone marrow-CNS connection in diabetes. From: Bone marrow-CNS connections: Implications in the pathogenesis of diabetic retinopathy.

Our hypothesis suggests a strong connection between the bone marrow and CNS and retina. An imbalance in the bone marrow leads to too many monocytes being produced and too few reparative endothelial progenitors. The inflammatory cells extravasate and promote local activation of resident microglia in the retina and brain contributing to both CNS and retinal pathology. Specific CNS centers that regulate the sympathetic nervous system (SNS) make direct connections to the bone marrow while the bone marrow, via the cells it releases, communicates back to these specific SNS centers. CNS inflammation may play a central role, not only in disease process such as neurogenic hypertension and metabolic syndrome, but also in perpetuating the inflammation that plays a critical role in the pathogenesis of diabetic retinopathy. (BN = basal amygdala, GIC = granular insular cortex, MC = middle commissure, PVN = paraventricular nucleus of hypothalamus)

Yellowlees Douglas, et al. Prog Retin Eye Res. 2012 September;31(5):481-494.
7.
Figure 3

Figure 3. Human peripheral blood CD34+ cells from diabetic patients are dysfunctional in vascular homing and association and cannot repair damaged retinal vessels. From: Bone marrow-CNS connections: Implications in the pathogenesis of diabetic retinopathy.

Mouse eyes subjected to ischemia/reperfusion insult underwent injection (into the vitreous) with CD34+ cells isolated from long-term diabetic patients or age- and gender-matched non-diabetic controls. Two days after injection of the cells, the mouse eyes were harvested and examined by immunofluorescent laser scanning confocal microscopy. (A) CD34+ cells (identified by green fluorescence) from diabetic patients are found near vessels (visualized by red fluorescence), albeit in a random distribution (Zheng et al., 2007). (B) Most cells from non-diabetic control patients are associated with vessels. The yellow color is the result of co-localized fluorescence and indicates the CD34+ cells are associating and possibly some of the cells actually incorporating into vessel walls (Caballero et al., 2007). Images are compressed z-series (approximately 30μm total thickness). Scale bar=50μm.

Yellowlees Douglas, et al. Prog Retin Eye Res. 2012 September;31(5):481-494.

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