Chang-Ling T, Neill A L, Hunt N H
Department of Anatomy, University of Sydney, Australia.
Am J Pathol. 1992 May;140(5):1121-30.
Changes in the cerebral microvasculature such as breakdown of the blood-brain barrier, petechial hemorrhages, congestion, and edema are observed in the later stages of murine cerebral malaria. These changes have been described from histologic sections of brain, but the need to section the material makes direct observation of the microvasculature in situ difficult. The retinal vasculature, in contrast, offers a unique opportunity to study rheologic, barrier, and functional properties of the microvasculature as a wholemount preparation with normal spatial relationship with other tissues and as an intact vascular plexus. A combination of techniques, including intravascular perfusion of Evan's Blue, Bisbenzimide and Monastral Blue, and fluorescence and transmitted light observation of retinal wholemounts, were developed to examine the progressive microvascular changes in murine cerebral malaria. These techniques allowed detection of phenomena such as monocyte adherence to endothelial cells, congestion, small hemorrhages, and breakdown of the blood-retinal barrier, with details of the location of this leakage, earlier than was possible by studying brain sections. Because the retina is intact, the phenomena were seen in greater detail and some, such as occlusion of vessel segments, were detectable only in retinal wholemounts. In addition, the covisualization of the blood elements, barrier properties, and vascular endothelial integrity that are possible with retinal wholemounts allowed detailed analysis of the interaction of different cellular elements in the pathogenesis of cerebral malaria. Except for detection of edema, the retinal wholemount technique offers a more powerful and less time-consuming technique for detecting early microvascular changes in murine cerebral malaria. This technique could find wider application in the study of other diseases that affect the microvasculature of the central nervous system, such as experimental allergic encephalitis and meningitis.
在鼠脑型疟疾后期可观察到脑微血管的变化，如血脑屏障破坏、瘀点出血、充血和水肿。这些变化已在脑的组织学切片中有所描述，但由于需要对材料进行切片，使得原位直接观察微血管变得困难。相比之下，视网膜血管系统提供了一个独特的机会，可作为一个与其他组织保持正常空间关系的整装标本以及一个完整的血管丛，来研究微血管的流变学、屏障和功能特性。我们开发了一系列技术，包括伊文思蓝、双苯甲酰亚胺和耐晒蓝的血管内灌注，以及视网膜整装标本的荧光和透射光观察，以研究鼠脑型疟疾中微血管的渐进性变化。这些技术能够检测到诸如单核细胞黏附于内皮细胞、充血、小出血以及血视网膜屏障破坏等现象，且能比研究脑切片更早地详细了解渗漏的位置。由于视网膜是完整的，这些现象能被更详细地观察到，有些现象，如血管段阻塞，仅在视网膜整装标本中可检测到。此外，视网膜整装标本能够同时观察血液成分、屏障特性和血管内皮完整性，从而可以详细分析脑型疟疾发病机制中不同细胞成分之间的相互作用。除了检测水肿外，视网膜整装技术为检测鼠脑型疟疾早期微血管变化提供了一种更强大且耗时更少的技术。该技术可能在研究其他影响中枢神经系统微血管的疾病，如实验性变应性脑脊髓炎和脑膜炎中得到更广泛的应用。
