Suitability of muscarinic acetylcholine receptor antibodies for immunohistochemistry evaluated on tissue sections of receptor gene-deficient mice.
Suitability of muscarinic acetylcholine receptor antibodies for immunohistochemistry evaluated on tissue sections of receptor gene-deficient mice.
复制标题
DOI：
10.1007/s00210-008-0365-9
复制
发表时间：
2009-04
影响因子：
3.6
通讯作者：
Kummer, Wolfgang
中科院分区：
文献类型：
--
作者：
Jositsch, Gitte;Papadakis, Tamara;Haberberger, Rainer V.;Wolff, Miriam;Wess, Juergen;Kummer, Wolfgang
乙酰胆碱（Acetylcholine，ACh）是内脏功能的主要调节剂，通过五种类型的毒蕈碱受体（M1 R-M5 R）对平滑肌张力和上皮功能发挥与内脏功能相关的作用。在这里，我们通过分析野生型和各自基因缺陷小鼠的标本，评估了MR抗体在组织切片免疫组织化学标记中的特异性。在本研究中评价的24种抗体中，16种抗体在18种不同条件下进行了检测，其中8种在21种不同方案中进行了检测，得到总共456种抗体/方案组合。每种方法至少在4种抗体稀释度下进行检测，因此最终至少评价了1，824种条件。对每一种动物的背根神经节、膀胱和所有胸部脏器的横截面进行了研究。在抗原可用的所有情况下，确定了至少一种孵育条件，其中仅选定细胞类型在阳性对照中进行免疫标记，但在吸收前对照中保持未标记。然而，除了两个例外（M2 R抗体），所有抗体在相应基因缺陷小鼠的组织中产生相同的免疫组织化学标记模式，即使在野生型小鼠中的预吸收对照表明特异性。因此，目前的数据表明，令人不快的事实，可靠的免疫组化定位MR亚型的抗体是例外，而不是规则。外周器官组织切片中MR亚型定位的免疫组织化学检测仅限于使用最常用的方法学途径的M2 R亚型。
Acetylcholine (ACh) is a major regulator of visceral function exerting pharmacologically relevant effects upon smooth muscle tone and epithelial function via five types of muscarinic receptors (M1R-M5R). Here, we assessed the specificity of MR antibodies in immunohistochemical labelling on tissue sections by analysing specimens from wild-type and respective gene-deficient mice. Of 24 antibodies evaluated in this study, 16 were tested at 18 different conditions each, and 8 of them in 21 different protocols, resulting in a total number of 456 antibody/protocol combinations. Each of them was tested at 4 antibody dilutions at minimum, so that finally at least 1,824 conditions were evaluated. For each of them, dorsal root ganglia, urinary bladder and cross sections through all thoracic viscera were investigated. In all cases where the antigen was available, at least one incubation condition was identified in which only select cell types were immunolabelled in the positive control but remained unlabeled in the preabsorption control. With two exceptions (M2R antibodies), however, all antibodies produced identical immunohistochemical labelling patterns in tissues taken from corresponding gene-deficient mice, even when the preabsorption control in wild-type mice suggested specificity. Hence, the present data demonstrate the unpleasant fact that reliable immunohistochemical localization of MR subtypes with antibodies is the exception rather than the rule. Immunohistochemical detection of MR subtype localization in tissue sections of peripheral organs is limited to the M2R subtype utilizing the most commonly used methodological approaches.
登录
查看更多内容
影响因子：
2.5
作者：
Haberberger, R;Scholz, A;Kress, M
通讯作者：
Kress, M
影响因子：
3.3
作者：
Fayon, M;De La Roque, ED;Marthan, R
通讯作者：
Marthan, R
影响因子：
158.5
作者：
GROSS, NJ;SKORODIN, MS
通讯作者：
SKORODIN, MS
DOI：
10.1007/bf00494642
发表时间：
1980-01-01
期刊：
HISTOCHEMISTRY
影响因子：
--
作者：
GRUBE, D
通讯作者：
GRUBE, D
影响因子：
2.5
作者：
Haberberger, R;Henrich, M;Kummer, W
通讯作者：
Kummer, W