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Mutations in VMD2 have been implicated in Best vitelliform macular dystrophy and AMD, and bestrophin was originally identified as being localized to the basolateral plasma membrane of retinal pigment epithelium (RPE) cells [5].
Kvβ1 immunofluorescence was detected on the plasma membrane of retinal arteriolar myocytes.
Double immunostaining with GLUT1 (red, Figure 2e3), which is known to be expressed in both the abluminal and luminal membrane of retinal capillaries [ 17], showed that Oatp1c1 (green, Figure 2e2) overlapped on the abluminal (arrow head) and luminal membrane of retinal capillaries (double arrow head, yellow, Figure 2e1, overlap coefficient = 0.789).
We further demonstrated the immunoreactivities of Oatp1a4 and 1c1 on the inner and outer sides of the endothelium nuclei along with GLUT1 which is a marker for both the abluminal and luminal membrane of retinal capillaries [ 17] and showed that Oatp1a4 and 1c1 are located on both the abluminal and luminal membrane of retinal capillary endothelial cells.
Moreover, double immunostaining with GLUT1 (red, Figure 1e3), which is known to be expressed in both the abluminal and luminal membrane of retinal capillaries [ 17], showed that Oatp1a4 (green, Figure 1e2) overlapped on the abluminal (arrow head) and luminal membrane of retinal capillaries (double arrow head, yellow, Figure 1e1; overlap coefficient = 0.812).
At higher magnifications, Kvβ1 was also observed to be specifically localized to the plasma membrane of retinal arteriolar myocytes (Fig. 6 B, ii and iii), but was absent in the endothelial cell layer (Fig. 6 B, iv).
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The mechanism of BRB breakdown is multifactorial and secondary to changes in the tight junctions, pericyte loss, endothelial cell loss, retinal vessel leukostasis, upregulation of vesicular transport, and increased permeability of the surface membranes of retinal vessels and retinal pigment epithelium cells.
Recently, we observed that Ctgf+/− mice (lacking one functional allele for Ctgf) are protected from diabetes-induced basement membrane thickening of retinal and kidney glomerular capillaries ([ 19]; P. Roestenberg, F.A. Van Nieuwenhoven, R. Verheul et al., unpublished results).
The plasma membrane of the retinal cells in contact with PFCLs appeared irreversibly disrupted, and infiltration of the liquid within the retinal discs with cytoplasm degeneration was observed.
The relative fluorescence intensity for Oatp1a4 was strongly observed at the abluminal membrane of the retinal capillaries compared with the luminal membrane, indicating that Oatp1a4 is preferentially localized on the abluminal membrane of the retinal capillaries.
Oatp1a4 and 1c1 immunoreactivities were detected in the rat retinal capillaries and co-localized with GLUT1, suggesting that both proteins are located on the abluminal and luminal membrane of the retinal capillary endothelial cells.
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