Quantitative receptor autoradiographic analysis for angiotensin II receptors in bovine retinal microvessels: quantitation with radioluminography.
1. Specific 125I-Sar1, Ile8-Angiotensin II (125I-Sar1, Ile8-AII) binding sites in bovine retinal microvessels were investigated using the quantitative receptor autoradiographic method with pellet sections. 2. A quantitation was made with the computerized radioluminographic imaging plate system, a newly developed and highly sensitive method. Binding characteristics of the retinal microvessels were compared with those of the cerebral microvessels and the retinal macrovessels. 3. We isolated microvessels from the bovine retina and bovine cerebral cortex using the method composed of two-size sievings and high-speed homogenization with a Polytron. The isolated microvessels were composed of capillaries, and the retinal macrovessels contained vessels with smooth muscle. 4. There were specific binding sites for 125I-Sar1, Ile8-AII which were single and of a high affinity, in both the cerebral and the retinal microvessels and the retinal macrovessels. There were no differences in affinity between the vessels, but the retinal microvessels did have a higher density of binding sites than the cerebral microvessels. 5. The method we used is simple and sensitive for detecting and characterizing 125I-Sar1, Ile8-AII binding sites in retinal capillaries. Knowledge of the existence of large numbers of specific binding sites, candidates of physiologically active angiotensin II receptors, aids with understanding the regulatory roles of angiotensin II in the blood-retinal barrier.
Sato
T
T
Department of Ophthalmology, Nagasaki University School of Medicine, Japan.
Niwa
M
M
Himeno
A
A
Tsutsumi
K
K
Amemiya
T
T
D015059:1-Sarcosine-8-Isoleucine Angiotensin II / Q000378:metabolism; D000804:Angiotensin II / Q000378:metabolism; D000818:Animals; D001345:Autoradiography / Q000379:methods; D002196:Capillaries / Q000378:metabolism; D002417:Cattle; D002540:Cerebral Cortex / Q000098:blood supply / Q000378:metabolism; D003720:Densitometry / Q000379:methods; D005136:Eye Proteins / Q000378:metabolism*; D007091:Image Processing, Computer-Assisted; D008163:Luminescent Measurements; D008858:Microscopy, Phase-Contrast; D009124:Muscle Proteins / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D011945:Receptors, Angiotensin / Q000378:metabolism*; D012171:Retinal Vessels / Q000378:metabolism*
D005136:Eye Proteins; D009124:Muscle Proteins; D011945:Receptors, Angiotensin; D000804:Angiotensin II; D015059:1-Sarcosine-8-Isoleucine Angiotensin II