Immunohistochemical analyses on serum proteins in nephrons of protein-overload mice by "in vivo cryotechnique".
Immunohistochemical analyses on serum proteins in nephrons of protein-overload mice by "in vivo cryotechnique".
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DOI：
10.14670/hh-22.137
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发表时间：
2007-02
影响因子：
2
通讯作者：
Daoyuan Zhou;N. Ohno;N. Terada;Z. Li;Hiroyuki Morita;K. Inui;A. Yoshimura;S. Ohno
中科院分区：
文献类型：
--
作者：
Daoyuan Zhou;N. Ohno;N. Terada;Z. Li;Hiroyuki Morita;K. Inui;A. Yoshimura;S. Ohno
使用传统的制备方法通常很难对活体小鼠肾脏中血清蛋白的局部分布进行免疫组织化学分析。通过使用我们的“体内冷冻技术”与冷冻替代相结合，我们检查了牛血清白蛋白（BSA）超载小鼠肾单位中血清蛋白的免疫定位，并将其与常规制备方法获得的结果进行了比较。在两天内每天注射 BSA 的小鼠中，可以在鲍曼间隙和尿管中观察到 BSA 的免疫定位，并伴有明显的蛋白尿，而另一种内源性小鼠白蛋白也有类似的免疫定位。在 55% 的肾单位中检测到 BSA 和小鼠白蛋白在鲍曼间隙中的渗漏以及它们被重吸收到近端小管中，但没有检测到免疫球蛋白 G1 (IgG1) 的渗漏。然而，除了 BSA 和小鼠白蛋白之外，在其他肾单位中也检测到了 IgG1 的渗漏。通过仔细检查BSA和IgG1的免疫定位，它们与没有正常血液循环进入肾脏的常规制备方法获得的结果明显不同。 BSA 和小鼠血清蛋白的免疫定位可以通过“体内冷冻技术”直接分析，这表明在 BSA 超载小鼠模型的早期阶段，肾小球滤过屏障的功能损伤是不同的，具体取决于活体小鼠的每个肾单位。
Immunohistochemical analyses on local distributions of serum proteins in living mouse kidneys are usually difficult to examine with conventional preparation methods. By using our "in vivo cryotechnique" combined with freeze-substitution, we have checked immunolocalizations of the serum proteins in nephrons of bovine serum albumin (BSA)-overload mice, and compared them with those obtained by the conventional preparation methods. In two days of daily BSA-injected mice, the immunolocalization of BSA could be observed in Bowman's space and urinary tubules with their overt proteinuria, where another endogenous mouse albumin was similarly immunolocalized. The leakage of BSA and mouse albumin in Bowman's space and their reabsorption into proximal tubules were detected in 55% of nephrons, where no leakage of immunoglobulin G1 (IgG1) was detected. However, the leakage of IgG1, in addition to BSA and mouse albumin, was detected in the other nephrons. By carefully examining immunolocalizations of BSA and IgG1, they were obviously different from those obtained by the conventional preparation methods without normal blood circulation into the kidneys. The immunolocalizations of both BSA and mouse serum proteins could be directly analyzed with the "in vivo cryotechnique", suggesting that functional damage to glomerular filtration barriers are different at early stages of the BSA-overload mouse model, depending on each nephron of living mice.