Jucker M, Bialobok P, Hagg T, Ingram D K
Laboratory of Cellular and Molecular Biology, National Institute on Aging, NIH, Baltimore, MD 21224.
Brain Res. 1992 Jul 17;586(1):166-70. doi: 10.1016/0006-8993(92)91390-z.
Normal adult and lesioned rat and mouse brains were fixed by formaldehyde perfusion by two methods that differ primarily in the length of the post-fixation period. Sections were subsequently immunostained using monoclonal and polyclonal antibodies to laminin. With relatively short post-fixation periods (up to 4 h), vascular basement membrane (BM)-laminin was immunostained, but intraneuronal laminin-like immunoreactivity was faint. With longer post-fixation periods (18-24 h), intraneuronal laminin-like immunoreactivity was distinct, while vascular BM-laminin immunoreactivity was reduced drastically. These findings are particularly relevant to studies examining laminin immunoreactive blood vessels in response to lesions, especially ischemic stroke. In fact, the present results suggest that the apparent neovascularization or up-regulation of vascular BM-laminin following CNS injury likely relates to differences in regional tissue fixation.
正常成年大鼠和小鼠的大脑以及损伤后的大脑，通过两种主要在固定后时间长度上有所不同的方法，用甲醛灌注固定。随后，使用抗层粘连蛋白的单克隆抗体和多克隆抗体对切片进行免疫染色。在相对较短的固定后时间（长达4小时）下，血管基底膜（BM）-层粘连蛋白被免疫染色，但神经元内的层粘连蛋白样免疫反应较弱。在较长的固定后时间（18-24小时）下，神经元内的层粘连蛋白样免疫反应明显，而血管BM-层粘连蛋白免疫反应则大幅降低。这些发现与研究层粘连蛋白免疫反应性血管对损伤（尤其是缺血性中风）的反应特别相关。事实上，目前的结果表明，中枢神经系统损伤后血管BM-层粘连蛋白明显的新生血管形成或上调可能与区域组织固定的差异有关。
