Nuclear Factor of Activated T Cells Is Activated in the Endothelium of Retinal Microvessels in Diabetic Mice

Figure 1

Expression of NFATc transcription factor genes in whole retina and in retinal microvessels. (a) RT-PCR analysis of NFAT isoform (c1–c4) expression in whole retinas and isolated retinal vessels from nondiabetic FVBN mice (). ((b)-(c)) All NFAT family members are expressed in mouse aorta (b) and in cultured human retinal microvascular endothelial cells (HRMVECs, (c)). Markers (M) for 300, 500, and 700 bp are indicated. (d)(i) Dark field/side illumination of a vascular network isolated from whole retina as described in material and methods (Section 2.2). (d)(ii) Confocal image showing smooth muscle α-actin positive staining (red) and nuclei (green) of an isolated retinal vessel preparation. Note the lack of surrounding nonvascular tissue (black background). (d)(iii) Confocal image of a smaller caliber vessel network stained as in (d)(ii) showing the absence of smooth muscle α-actin positive cells. Scale bars = 500 μm for (d)(i); 20 μm for (d)(ii)-(d)(iii). (e)-(f) Quantitative RT-PCR data showing expression levels of neuronal maker Tau (e) and glial marker GFAP (f) in whole retina and in isolated retinal vessels, both normalized to 18S. retinas/group.