Long J M, Kalehua A N, Muth N J, Hengemihle J M, Jucker M, Calhoun M E, Ingram D K, Mouton P R
Gerontology Research Center, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA.
J Neurosci Methods. 1998 Oct 1;84(1-2):101-8. doi: 10.1016/s0165-0270(98)00100-9.
Microglia are brain cells of considerable interest because of their role in CNS inflammatory responses and strong association with neuritic plaques in Alzheimer's disease (AD). In the present study, immunocytochemistry was combined with unbiased stereology to estimate the mean total number of microglia in dentate gyrus and CA1 regions of the mouse hippocampus. Systematic-uniform-random (SUR) sections were cut through the hippocampal formation of male C57BL/6J mice (n = 7, 4-5 months) and immunostained with Mac-1, an antibody to the complement subunit 3 receptor (CR3). The total number of Mac-1 immunopositive cells was determined using the optical fractionator method. The mean total number of microglia in the mouse dentate gyrus was estimated to be 20,300 (CV = 0.21) with a mean coefficient of error (CE) = 0.09. The mean total number of microglia in the mouse CA1 was estimated to be 43,200 (CV = 0.24) with a CE = 0.11. Comparison of total number estimates, derived from fraction- or volume-based methods, supported stereological theory regarding the equivalence of the two techniques. The time required to determine total microglia number in both hippocampal sub-regions was approximately 6 h per mouse from stained sections. The combination of immunocytochemistry and stereology provides a reliable means to assess microglia number that can easily be adopted for studies of transgenic and lesion-based models of aging and neurodegenerative diseases.
小胶质细胞是备受关注的脑细胞，因为它们在中枢神经系统炎症反应中发挥作用，且与阿尔茨海默病（AD）中的神经炎性斑块密切相关。在本研究中，免疫细胞化学与无偏倚立体学相结合，以估计小鼠海马齿状回和CA1区域中小胶质细胞的平均总数。对雄性C57BL/6J小鼠（n = 7，4 - 5个月）的海马结构进行系统均匀随机（SUR）切片，并使用针对补体亚基3受体（CR3）的抗体Mac-1进行免疫染色。使用光学分割器方法确定Mac-1免疫阳性细胞的总数。小鼠齿状回中小胶质细胞的平均总数估计为20300（CV = 0.21），平均误差系数（CE）= 0.09。小鼠CA1中小胶质细胞的平均总数估计为43200（CV = 0.24），CE = 0.11。基于分数或体积的方法得出的总数估计值的比较，支持了关于这两种技术等效性的立体学理论。从染色切片确定两个海马亚区域中小胶质细胞总数所需的时间约为每只小鼠6小时。免疫细胞化学和立体学的结合提供了一种可靠的方法来评估小胶质细胞数量，这种方法可轻松应用于衰老和神经退行性疾病的转基因和基于损伤模型的研究。
