ANGIOARCHITECTURE OF THE CNS, PITUITARY-GLAND, AND INTRACEREBRAL GRAFTS REVEALED WITH PEROXIDASE CYTOCHEMISTRY
ANGIOARCHITECTURE OF THE CNS, PITUITARY-GLAND, AND INTRACEREBRAL GRAFTS REVEALED WITH PEROXIDASE CYTOCHEMISTRY
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DOI：
10.1002/cne.902600105
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发表时间：
1987-06-01
影响因子：
2.5
通讯作者：
SALCMAN, M
中科院分区：
文献类型：
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作者：
BROADWELL, RD;CHARLTON, HM;SALCMAN, M
胎儿、新生儿和成年亚灵长类动物和灵长类动物CNS的血管，包括脑室周围器官（例如，正中隆起、脑下垂体等），以及固态中枢神经系统和非神经系统用过氧化物酶细胞化学方法观察了成年哺乳动物宿主脑内的同种异体（垂体前叶）移植物：（1）在灌注固定前全身注射天然辣根过氧化物酶（HRP）或与凝集素麦胚凝集素（WGA）结合的过氧化物酶的动物组织;（2）灌注固定后将天然HRP注入主动脉的动物组织;和（3）通过浸泡固定并孵育保留在血管内的红细胞中的内源性过氧化物酶活性的动物组织。在血管内接受天然HRP的新生和成年动物中，当使用四甲基联苯胺（TMB）而不是二氨基联苯胺（DAB）作为色原时，用HRP反应产物概述了有助于血脑屏障的无孔血管;由于血管外反应产物的密度，室周器官的有孔血管不可辨别。当在TMB和DAB溶液中孵育时，暴露于血源性WGA-HRP的有孔和无孔脑血管和脑外血管可见。将天然HRP注入固定动物的主动脉中，在暴露于TMB或DAB时，同样标记了整个脑中的无孔血管，但遮蔽了脑室周围器官的有孔血管。红细胞的内源性过氧化物酶活性，与TMB和DAB同样良好，勾勒出整个大脑灰质和白色物质以及胎仔、新生儿和成年动物的所有脑室周围器官的血管。应用三种过氧化物酶细胞化学方法研究脑内同种异体移植物中血脑屏障的形成或缺失，结果表明，16-19天胎儿/1天新生儿CNS同种异体移植物的血管化在脑内植入移植物后7天之前并不明确。从预期具有血脑屏障的供体部位固定的CNS同种异体移植物显示出对经静脉注射到宿主中的HRP不渗漏的血管。与垂体前叶同种异体移植物相关的有孔血管在移植后3天内在宿主脑内是明显的，并允许宿主中的血源性HRP进入移植物和周围的宿主脑实质。这三种过氧化物酶细胞化学的方法是有用的可视化中枢神经系统血管，有助于或不有助于血脑屏障在胎儿，新生儿和成年实验室动物;这三种方法有潜在的应用脑血管研究中风，创伤和肿瘤。
Blood vessels of the fetal, neonatal, and adult subprimate and primate CNS, including circumventricular organs (e.g., median eminence, pituitary gland, etc.), and of solid CNS and nonneural (anterior pituitary gland) allografts placed within brains of adult mammalian hosts were visualized with peroxidase cytochemistry applied in three ways: (1) to tissues from animals injected systemically with native horseradish peroxidase (HRP) or peroxidase conjugated to the lectin wheat germ agglutinin (WGA) prior to perfusion fixation; (2) to tissues from animals infused with native HRP into the aorta subsequent to perfusion fixation; and (3) to tissues from animals fixed by immersion and incubated for endogenous peroxidase activity in red cells retained within blood vessels. In neonatal and adult animals receiving native HRP intravascularly, non-fenestrated vessels contributing to a blood-brain barrier were outlined with HRP reaction product when tetramethylbenzidine (TMB) as opposed to diaminobenzidine (DAB) was used as the chromogen; fenestrated vessels of circumventricular organs were not discernible due to the density of extrasvascular reaction product. Fenestrated and nonfenestrated cerebral and extracerebral blood vessels exposed to blood-borne WGA-HRP were visible when incubated in TMB and DAB solutions. Native HRP infused into the aorta of fixed animals likewise labeled nonfenestrated vessels throughout the brain upon exposure to TMB or DAB but obscured fenestrated vessels of the circumventricular organs. Endogenous peroxidase activity of red cells, seen equally well with TMB and DAB, outlined blood vessels throughout the cerebral gray and white matter and all circumventricular organs in fetal, neonatal, and adult animals. Application of the three peroxidase cytochemical approaches to study the development or absence of a blood-brain barrier in intracerebral allografts demonstrated that the vascularization of day 16-19 fetal/1 day neonatal CNS allografts is not well defined prior to 7 days following intracerebral placement of the grafts. CNS allografts secured from donor sites expected to possess a blood-brain barrier exhibited blood vessels that were not leaky to HRP injected intravenously in the host. Fenestrated blood vessels associated with anterior pituitary allografts were evident prior to 3 days posttransplantation within the host brain and permitted blood-borne HRP in the host to enter the graft and surrounding host brain parenchyma. The three peroxidase cytochemical approaches are useful for visualizing CNS blood vessels that contribute or do not contribute to a blood-brain barrier in fetal, neonatal, and adult laboratory animals; the three approaches have potential application to cerebrovascular studies of stroke, trauma, and neoplasia.