Assessment of retinal and choroidal blood flow changes using laser Doppler flowmetry in rats.

PURPOSE: A new noninvasive laser Doppler flowmetry (LDF) probe (one emitting fiber surrounded by a ring of eight collecting fibers, 1-mm interaxis distance) was tested for its sensitivity to assess the retinal/choroidal blood flow variations in response to hypercapnia, hyperoxia, diverse vasoactive agents and following retinal arteries photocoagulation in the rat. MATERIALS AND METHODS: After pupil dilation, a LDF probe was placed in contact to the cornea of anesthetized rats in the optic axis. Hypercapnia and hyperoxia were induced by inhalation of CO(2) (8% in medical air) and O(2) (100%) while pharmacological agents were injected intravitreously. The relative contribution of the choroidal circulation to the LDF signal was estimated after retinal artery occlusion by photocoagulation. RESULTS: Blood flow was significantly increased by hypercapnia (18%), adenosine (14%) and sodium nitroprusside (16%) as compared to baseline values while it was decreased by hyperoxia (-8%) and endothelin-1 (-11%). Photocoagulation of retinal arteries significantly decreased blood flow level (-45%). CONCLUSIONS: Although choroidal circulation most likely contributes to the LDF signal in this setting, the results demonstrate that LDF represents a suitable in vivo noninvasive technique to monitor online relative reactivity of retinal perfusion to metabolic or pharmacological challenge. This technique could be used for repeatedly assessing blood flow reactivity in rodent models of ocular diseases.

Modulation of retinal blood flow by kinin B1 receptor in Streptozotocin-diabetic rats.

The vasoactive kinin B1 receptor (B1R) is overexpressed in the retina of diabetic rats in response to hyperglycemia and oxidative stress. The aim of the present study was to determine whether B1R could contribute to the early retinal blood flow changes occurring in diabetes. Male Wistar rats were rendered diabetic with a single i.p. injection of Streptozotocin (STZ) and studied 4 days or 6 weeks after diabetes induction. The presence of B1R in the retina was confirmed by Western blot. The impact of oral administration of the B1R selective antagonist SSR240612 (10mg/kg) was measured on alteration of retinal perfusion in awake diabetic rats by quantitative autoradiography. Data showed that B1R was upregulated in the STZ-diabetic retina at 4 days and 6 weeks. Retinal blood flow was not altered in 4-day diabetic rats compared with age-matched controls but was significantly decreased following SSR240612 treatment. In 6-week diabetic rats, retinal blood flow was markedly reduced compared to control rats and SSR240612 did not further decrease the blood flow. These results suggest that B1R is upregulated in STZ-diabetic retina and has a protective compensatory role on retinal microcirculation at 4 days but not at 6 weeks following diabetes induction.

Quantitative and regional measurement of retinal blood flow in rats using N-isopropyl-p-[14C]-iodoamphetamine ([14C]-IMP).

Quantitative and regional measurement of retinal blood flow in rodents is of prime interest for the investigation of regulatory mechanisms of ocular circulation in physiological and pathological conditions. In this study, a quantitative autoradiographic method using N-isopropyl-p-(14)C-iodoamphetamine ([(14)C]-IMP), a diffusible radioactive tracer, was evaluated for its ability to detect changes in retinal blood perfusion during hypercapnia. Findings were compared to cerebral blood flow values measured simultaneously. Hypercapnia was induced in awaken Wistar rats by inhalation of 5% or 8% CO(2) in medical air for 5 min. [(14)C]-IMP (100 microCi/kg) was injected in the femoral vein over a 30 s period and the rats were sacrificed 2 min later. Blood flow was calculated from whole-mount retinae and 20 microm thick brain sections in discrete regions of interest by quantitative autoradiography or from digested samples of retina and brain by liquid scintillation counting. Retinal blood flow values measured with quantitative and regional autoradiography were higher in the central (108 +/- 20 ml/100 g/min) than in peripheral (84 +/- 15 ml/100 g/min) retina. These values were within the same range as cortical blood flow values (97 +/- 4 ml/100 g/min). The retinal blood flow values obtained on whole-mount retinae were validated by the sampling method. Hypercapnia significantly increased overall blood flow in the retina (24-53%) with a maximal augmentation in the peripheral region and in the brain (22-142%). The changes were stronger in the brain compared to retina (p = 0.016). These results demonstrate that retinal blood flow can be quantified using [(14)C]-IMP and compared with cerebral blood flow. This technique is a powerful tool to study how retinal blood flow is regulated in different regions of the rat retina.