Measurement of region-specific nitrate levels of the posterior chamber of the rat eye using low-flow push-pull perfusion.
Measurement of region-specific nitrate levels of the posterior chamber of the rat eye using low-flow push-pull perfusion.
复制标题
使用低流量推拉灌注测量大鼠眼后房区域特异性硝酸盐水平。
DOI：
10.1021/ac800238d
复制
发表时间：
2008
影响因子：
7.4
通讯作者：
Shippy,ScottA
中科院分区：
文献类型：
--
作者：
Pritchett,JeanitaS;Pulido,JoseS;Shippy,ScottA
以区域选择性的方式确定大鼠玻璃体腔中是否存在一氧化氮代谢物是由眼睛的大小和形状以及分子及其代谢物的扩散性决定的。在这项工作中，活体低流量推式−拉式灌流采样和快速毛细管电泳法监测了正常和老年模型大鼠玻璃体视网膜界面的主要NO代谢物硝酸盐的水平。通过29号导针将取样探头尖端放置在后房内的三个不同位置。在视神经头和视神经头周围区域的VRI上进行采样。此外，从玻璃体中部区域采集样本，以与VRI样本进行比较。灌流液中硝酸盐浓度在每个部位都有显著差异(P<；0.05)，这可能是由于NO产生的来源或分子从玻璃体腔中清除的机制所致。注射L-NAME和生理盐水后，观察到的硝酸盐水平显著下降(35%)。然后使用LFPPP观察平均老化4.5个月后的硝酸盐水平。与年轻的对照动物相比，成年动物视神经头上方的硝酸盐平均水平增加了3倍。准确测量VRI沿线的NO代谢物可能有助于深入了解NO在维持体内平衡状态中的作用以及病变视网膜的分子变化。
The determination of the presence of nitric oxide metabolites in the rat vitreous cavity in a regioselective manner is complicated by the size and shape of the eye as well as the diffusivity of the molecule and its metabolites. In this work, in vivo low-flow push−pull perfusion sampling was utilized with a rapid capillary electrophoretic assay to monitor levels of the major NO metabolite, nitrate, at the vitreoretinal interface (VRI) of normal and aged rat models. The sampling probe tips were placed in three different positions in the posterior chamber through a 29-gauge guide needle. Sampling was performed along the VRI over the optic nerve head and regions peripheral to the optic nerve head. Additionally, samples were collected from the middle vitreous region to compare to VRI sampling. A significant (P< 0.05) difference in the perfusate nitrate concentration was observed in each location, which may be due to the source of NO production or the clearance mechanism of the molecule from the vitreous cavity. Infusion of L-NAME with physiological saline led to a significant decrease (35%) in the observed nitrate level. LFPPP was then utilized to observe nitrate levels after an average of 4.5 months of aging. A 3-fold increase in the mean level of nitrate over the optic nerve head was observed in mature animals compared to younger control animals. Precise measurement of NO metabolites along the VRI may provide insights into the function of NO in maintaining homeostatic conditions and the molecular changes at the diseased retina.